A MODEL TO IMPLEMENT A STEM CURRICULUM IN PLANT BIOTECHNOLOGY AT THE SECONDARY SCHOOL LEVEL By WENDY VIDOR A DISSERTATION PRESENTED TO THE GRADUATE SCHOOL OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT OF THE REQUIR EMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY UNIVERSITY OF FLORIDA 2017
2017 Wendy Vidor
To my supportive and loving husband and family for always encouraging me to pursue my dreams and persevere
4 ACK NOWLEDGMENTS I would like to express the dee pest appreciation to my mentor and committee chair Dr. Michael Kane, in the Environmental Horticulture Department at the University of Florida, Gainesville, FL. His dedication in helping students succeed and his immense support and guidance has been mainly responsible for helping me to complete my work. His scholarly advice and support of teaching has helped me accomplish this task. He is an expert in micropropagation of plants and his knowledge and expertise has given me the knowledge and confidence to be successful in my future endeavors in this field. As a high school educator, I appreciate his unending support of teachers and willingness to help educators obtain the background knowledge, training and support that is so important for success in the classroom. I would like to thank my committee members at the University of Florida, Dr. Mary Jo Koroly, Dr. Grady Roberts, Dr. Charles Guy, and Dr. Carrie Adams, whose work and dedication to graduate students have g uided me to understand the academic knowledge need e d to teach biotechnology and the importance of educating students in pursuit of STEM careers in agriculture, horticulture, and biotechnology. Their guidance and support helped me develop the academic knowl edge and the skills needed to teach and support biotechnology education. Finally, my deepest thanks to my husband Robert my daughters Samantha, Matti e and Abbey for enduring my role as a full time teacher and doctoral student. Without their love, under standing, and sacrifice this dissertation would not be possible.
5 TABLE OF CONTENTS page ACKNOWLEDGMENTS ................................ ................................ ................................ .. 4 LIST OF TABLES ................................ ................................ ................................ ............ 8 LIST OF FIGURES ................................ ................................ ................................ .......... 9 LIST OF OBJECTS ................................ ................................ ................................ ....... 11 LIST OF ABBREVIATIONS ................................ ................................ ........................... 12 ABSTRACT ................................ ................................ ................................ ................... 13 CHAPTER 1 OVERVIEW OF PLANT BIOTECHNOLOGY IN THE CLASSROOM ..................... 15 Introduction to Biotechnology ................................ ................................ .................. 15 History and Developm ent of Biotechnology ................................ ...................... 18 STEM Education and Integration of Biotechnology ................................ .......... 20 Biotechnology Curriculum and Integration ................................ ........................ 22 Constructivism Theory and Sociocultural Theory ................................ ............. 25 Agriculture Biotechnology Curriculum Development ................................ ........ 27 Teacher Attitudes and Concerns About Teaching Biotechnology ..................... 30 Biotechnology Teacher Training ................................ ................................ ....... 31 Flipping the Classroom ................................ ................................ ..................... 35 Development of Multimedia Tools for Flipping ................................ .................. 42 Factors Affecting Implementation of Biotechnology ................................ .......... 47 Purpose of the Study ................................ ................................ .............................. 50 Statement of Objectives ................................ ................................ .......................... 50 Statement of the Hypotheses ................................ ................................ .................. 51 Definition of Terms ................................ ................................ ................................ .. 51 Potential Limitations of the Study ................................ ................................ ............ 52 2 T EACHER BELIEFS ABOUT TEACHING BIOTECHNOLOGY IN THE CLASSROOM ................................ ................................ ................................ ......... 54 Introduction ................................ ................................ ................................ ............. 54 Statement of the Problem ................................ ................................ ....................... 55 Purpose and Objectives ................................ ................................ .......................... 57 Methods ................................ ................................ ................................ .................. 57 General Procedures ................................ ................................ ......................... 57 Participants ................................ ................................ ................................ ....... 58 Survey Instrumentation ................................ ................................ .................... 58 Discussion ................................ ................................ ................................ .............. 65
6 Conclusions ................................ ................................ ................................ ............ 71 3 ASSESSMENT OF PLANT BIOTECHNOLOGY TEACHING MODULES ............... 79 Introduction ................................ ................................ ................................ ............. 79 Purpose and Objectives ................................ ................................ .......................... 81 Materials and Methods ................................ ................................ ............................ 82 Participants ................................ ................................ ................................ ....... 82 Workshop Date and Location ................................ ................................ ........... 82 Workshop Design ................................ ................................ ............................. 82 Development and Design of Teaching Less on Modules ................................ .. 84 Research Design ................................ ................................ .............................. 86 Results ................................ ................................ ................................ .................... 87 Discussion ................................ ................................ ................................ .............. 88 Conclusion and Recommendations ................................ ................................ ........ 95 Teaching Lesson Module #1 ................................ ................................ ............ 96 PowerPoint Ov erview of Carnation Module ................................ .................... 115 Teaching Lesson Module #2 ................................ ................................ .......... 119 Teaching Module Lesson #3 ................................ ................................ .......... 128 Teacher Plant Tissue Culture Workshop Pretest/Posttest .............................. 141 4 EFFECTS OF FLIPPING ON INCREASE IN TEACHER CONTENT KNOWLEDGE ................................ ................................ ................................ ...... 143 Introduction ................................ ................................ ................................ ........... 143 Purpose and Objectives ................................ ................................ ........................ 148 Methods ................................ ................................ ................................ ................ 148 Workshop Setting and Participant Selection ................................ ................... 149 Research Integrity and Compliance ................................ ............................... 149 Experimental Procedures ................................ ................................ ............... 150 Pretest Posttest Instrument ................................ ................................ ............ 152 Workshop Questionnaire ................................ ................................ ................ 153 Caladium Laborat ory Module ................................ ................................ ......... 153 Research Design ................................ ................................ ............................ 154 Rationale for Research Design and Statistical Analysis ................................ 155 Results ................................ ................................ ................................ .................. 156 Discussion and Summary ................................ ................................ ..................... 159 Limitations and Implications ................................ ................................ .................. 164 Conclusion ................................ ................................ ................................ ............ 166 5 CONCLUSIONS AND RECOMMENDATIONS ................................ ..................... 189 Recommendations ................................ ................................ ................................ 191 Recommendations for Teacher Education and Curriculum ................................ ... 192 Recommendations for Further Research ................................ .............................. 192
7 APPENDIX A IRB APPROVAL FOR SURVEY ................................ ................................ ........... 194 B INFORMED CONSENT SURVEY LETTER ................................ .......................... 195 C TEACHER PLANT BIOTECHNOLGY WORKSHOP AGENDA 2 015 ................... 197 D IRB APPROVAL TEACHER PLANT BIOTECHNOLOGY WORKSHOP ............... 200 E INFORMED CONSENT LETTER FOR THE EFFECT OF FLIPPING INSTRUCTION O N TEACHER PLANT BIOTECHNOLOGY CONTENT KNOWLEDGE ................................ ................................ ................................ ...... 202 F THE EFFECT OF FLIPPING WORKSHOP EVALUATION QUESTIONNAIRE .... 204 G TEACHER PLANT BIOTECHNOLOGY WORKSHOP AGENDA 2016 ................. 205 H PRETEST AND POSTTEST SHOOT CULTURE OF CALADIUM ........................ 209 I MICROPROPAGATION METHODS OF SHOOT C ULTURE, ORGANOGENENSIS AND NON ZYGOTIC EMBRYOGENENSIS POWERPOINT ................................ ................................ ................................ ..... 213 J SHOOT CULTURE OF CALADIUM OVERVIEW POWERPOINT LECTURE ...... 235 L IST OF REFERENCES ................................ ................................ ............................. 250 BIOGRAPHICAL SKETCH ................................ ................................ .......................... 261
8 LIST OF TABLES Table page 2 1 Survey res ults of teacher beliefs about teaching biotechnology ......................... 73 2 2 Teachers attitudes and concerns about teaching biotechnology subgroup percentages. ................................ ................................ ................................ ....... 74 2 3 Mean response of agriculture attitudes of teachers teaching biotechnology ...... 76 2 4 Difference in attitude mean scores of agriculture science teachers that teach biotechnology and teachers that do not teach biotechnology. ............................ 77 2 5 Participant pre service and in service training experiences in teaching ............. 77 3 1 Teacher perceived barriers in teaching biotechnology in the classroom expressed by the November 2015 workshop participants. ................................ 92 4 1 Randomized subjects, pretest posttest control group design .......................... 156 4 2 Pre test and posttest response rate totals ................................ ......................... 157 4 3 ................................ ... 157 4 4 Adjusted and unadjusted intervention mean scores and variability for post intervention of Caladium module exam. ................................ ........................... 158 4 5 ANCOVA test of C aladium module exam ................................ ......................... 158
9 LIST OF FIGURES Figure page 1 1 ................................ ................................ .............. 46 1 2 Chronological order of research tasks. ................................ ............................... 53 3 1 Mean change in test scores on teacher conten t knowledge ............................. 88 3 2 The shoot tip is comprised of the apica l meristem ................................ ............. 99 3 3 Materials used to complet e exercise ................................ ................................ 106 3 4 Tissue culture t ools ................................ ................................ ........................... 106 3 5 Carnation shoot node explants with leaves r emoved ................................ ....... 107 3 6 Carnation shoot explant rinsing procedure ................................ ....................... 107 3 7 Sequential steps to surface sterilize the Car nation shoot explants ................... 108 3 8 Rinsing procedure for Carnation sh oot node explants ................................ ...... 109 3 9 Cuts required to obtain stem i nternode explants ................................ .............. 121 3 10 Formation of shoots from in ternode explants ................................ ................... 121 3 11 Four day old Arabidopsis seedlings germinat ed on filter paper ........................ 138 3 12 Polypropylene funnel with alginic acid on ring stand ................................ ........ 139 3 13 Sprouted se edling placed into alginate bead ................................ .................... 139 4 1 Scatterplot comparison of teacher p retest and posttest performance ............... 159 4 2 The shoot tip is comprised of the apical me ristem ................................ ........... 175 4 3 Rinsin g off potting media from roots ................................ ................................ 177 4 4 Tools used to complete Caladium bicolor shoot culture experimen t ................. 178 4 5 Step by step pro cedure for Caladium shoot ex plant sterilization ...................... 179 4 6 Use stereomicroscope (left) to trim shoot expl ants ................................ ........... 179 4 7 Inoculat ion o f shoot explants ................................ ................................ ............ 180 4 8 Acclimatization of rooted Caladium bicolor shoots ................................ ........... 181
10 4 9 Micropropagation stages. ................................ ................................ ................. 182
11 LIST OF OBJECTS Object page 4 1 Interactive Lecture Overview ................................ ................................ ........... 158
12 LIST OF ABBREVIATIONS CPET Center for Precollegiate Education and Tra ining IRB International Review Board S&E Science and Engineering Related Careers SPSS Statistical Package for the Social Sciences TBT TB Teachers Belief About Teaching Biotechnology UbD Understanding by Design Framework UF University of Florida
13 Abstract of Dissertation Presented to the Graduate School of the University of Florida Requirements for the Degree of Doctor of Philosophy A MODEL TO IMPLEMENT A STEM CURRICULUM IN PLANT BIOTECHNOLOGY AT THE SECONDARY SCHOOL LEVEL By Wend y Vidor December 2017 Chair: Michael E. Kane Major: Horticultural Sciences Agricultural biotechnology encompasses diverse scientific technologies and tools to improve plants, animals and microorganisms and includes tec hniques to make or develop organism s for specific agricultural uses. B iotechnology training and curricula are underrepresented in the classroom. The creation of new career paths incorporating STEM and biotechnology in secondary classrooms will require teachers to be trained and curricula to be written. programs is to prepare students for STEM based occupations that are important to i n plant biotechnology. The purpose of this descriptive research study was to determine and alleviate barriers in teaching and integrating biotechnology in Florida A survey was completed to determine teacher current barri ers in Florida agriculture biotechnology teachers and subsequently develop effective teaching modules A maj or finding in the study was the agriculture teachers had concerns about teaching biotechnology including pre service and in service training, equipment, lack of instructional materi als, and time constraints A series of three comprehensive plant biotechnol ogy teaching modules meant to overcome these barriers were evaluated by
14 agriculture teachers in a wo rkshop experience. T eacher training modules were develop ed and assessed for effec tiveness in the classroom. An evaluation revealed a significant content k nowledge gain in posttest score compared to the pre test for teachers after completing the teaching modules during the training In a follow up workshop a plant biotechnology caladium shoot culture teaching module was used to determine the effectiveness o f classroom flipping instruction on content knowledge gain of teachers using a pretest and posttest. The results indicated there was no significant effect of flipping on teacher content knowledge gain but the teachers had positive views of the interactive flipped model for individualizing learning and increasing engagement in biotechnology content These results were limited to the extent that instruction was limited to a small group of teachers due to size of the teaching facility. of biotechnology curricula and instructional approaches is dependent on teacher knowledge and self efficacy of the subject T he r esults presented in this study are important for designing future biotechnology training workshops and effective plant biotechn ology curricula
15 CHAPTER 1 OVERVIEW OF PLANT BIOTECHNOLOGY IN THE CLASSROOM Introduction to Biotechnology Agricultural biotec hnology is a collection of scientific technologies and tools to improve plants, animals and microorganisms and includes tradition al breeding tec hniques to make or develop organisms for specific agricultural uses. One aspect of biotechnology includes crop biotechnology. A desired trait is transferred fro m a particular species to an entirely different species creating transgene crops that possess desirable characteristics for pest and disease resistance, flower color, flavor or to increase growth rate ( USAID 2004) Continual biotechnological advances in many areas including production of food and medicine to treat d isease, the creation of new energy sources, and manipulation of genomes have a significant impact and are transforming sci ence and society (Borgerding et al. 2013). Despite the new advances, biotechnology training and curricula are underrepresented in the classroom. The creation of new career paths in biotechnology in secondary classrooms will require teachers to be trained and curriculum to be written (Borgerding et al. 2013; Hanegan & Bigler, 2009) The AFNR, Agriculture Food and Natural Resources are the driving force s in the creation of educ ational programs to meet the needs of the workforce including agriculture biotechnology (Florida Department of Education, 2017)
16 STEM is an acronym for science, technology, engineering and mathematics. The definition of STEM education is the in tentional integration of science, technology, engineering and mathematics, and their associated practices to create a student centered learning environment in which students investigate and engineer solutions to problems, and construct evidence based expla nations of real world phenomena. This through shared contributions of schools, families, and community partners (Florida Department of Education, 2017) STEM education is critical to the ongoing economic success of the state of Florida. STEM careers outpace any other occupation with respect to salaries and career earning potent ial (Florida Department of Education, 2017) One of the major issues educators are facing is how to integrate agriculture biotechnology into STEM based h igh school curricula Being familiar with biotechnology concept s is important today because this content is essential for students to understand advancements in technology and science and how this relates to training for careers in the science and technical fields. The fields of mole cular breeding, bioinformatics and technology are in high demand in the global marke tplace. Thoron (2010) described the need for agriculture education to develop a working foundation in science and technology as well as formal reasoning skills crucial in t he scientific process to direct students towards the science and biological fields. There are also many challenges to address in how to train teachers to effectively teach curriculum in agriculture biotechnology. To address these challenges, information
17 is needed to determine the barriers faced by agriculture biotechnology teachers to implement a STEM based biotechnology curriculum. Mowen et. al (2007 a ) investigated teacher barriers, roles and information sources for teaching biotechnology. The major ba rriers reported were equipment availability, laboratory space, and time as the three major barriers related to teaching biotechnology. In a second study Mowen et al. (2007 b ) reported significant relationships biotechnology topics and the likelihood that the topic was taught in the classroom. Mowen et al. (2007 a ) also reported a weak their atti tudes toward biotechnology. Attendance of tea chers at biotechnology workshops was reported to be only 36%. This raises the question are teachers not getting the information the y need to teach biotechnology through in service workshops, then where are they getting instruction? In s everal studies th e need for professional development in biotechnology at both the pre service and in service level has been emphasized (Brown et al. 1998; Dunham et al. 2002; Scott et al ., 2006; Kwon, 2009) Brown et al (2008) indic a ted that several outside factors that are barriers to implementation of biotechnology include school setting, availabilit idence in the subject material. Wells and Kwon (2008) suggested that the lack of teacher confidence in the material resulted from insufficient professional development of the type necessary for effec tively teaching biotechnology content. T his review addresses 1) the history and development of biotechnology, 2) the influence of STEM education on the integration of biotechnology, and 3) the subsequent
18 development of agriculture biotechnology curricul a and their integration in the secon dary classroom In addition, the theoretical models of constructivism driving the study as well as development of a griculture biotechnology curricul a to incr ease teacher content knowledge will be probed. Importantly, facto rs that influence attitudes and concerns about the challenges faced by agriculture educa tion teachers and the current biotechnology training available will be described. T he benefits of incorporating blended learning methods including flipped instruction and the use of multi media tools to integrate biotechnological principles and skills in the secondary classroom will be reviewed. Finally, the factors affecting implementation of biotechnology curricula in the classroom will be discussed. History and Development of Biotechnology Biotechnology is technology based on biology and harnesses cellular and biomolecular processes to develop technologies and products to improve our lives and the planet (Biotechnology Innovation Organization, 2017) Plant biotechnology is widely used in agriculture, medicine and industrial biotechnology today. Vasil ( 2008) stated that totipotency and genetic Per Vasil (2008), this technology can be traced back to the Cell Theory proposed by Ma tthias Schleiden, Theodor Schwann and genetic transformation of bacteria, by Frederick Griffith, respectively. Vasil (2008) stated that cellular totipotency in plants is the ability of a single cell to divide and thr ough growth and differentiation ultimate ly produce a whole plant. Genetic transformation is an alteration in a plant s genetic makeup following uptake of foreign DNA which is then integrated and expressed in the DNA. Totipotency was first demonstrated by Herman Vochting (1878) in his dissections of small plant tissue fragments He discovered that plants could be
19 regenerated from very small fragments of tissue under the proper external growing conditions. Plant genetic transformation has been accomplished using agrobacterium mediated DNA transfer and by the gene gun developed by Sanford (1987) Arabidopsis has also been used widely in plant research to study genetics including gene expression, reg ulation and function (Vasil, 2008). Many economically important food crops including fruits, vegetables and trees species that h ave been genetically transformed for imp rovement. Biotech crops can be used to produce may vaccines and pharmaceuticals. Underst anding this technology is important to plant breeders and researchers to develop new and improved crops to overcome an ever changing food supply. Expansion of biotechnology will require better informed citizens equipped with the skills and education in bi otechnology and advanced science concepts to achieve technical skills, literacy and acceptance of biotechnology (Wells, 1994). To improve acceptance of biotechnology, both scientists and industry have recognized that biotechnology education is necessary an d needs to begin at the high school level (Kwon, 2009; National Scien ce Foundation, 2014) Biotechnology training is being implemented in many agriculture education classrooms nationally. The demands for trained workers in these highly technical fields are driving the change for more science, technology, engineering and ma th in the national curriculum. The National Agriculture Food and Natural Resources ( AFNR) Content Standards outlined rigorous standards designed to develop well planned curriculum (The National Council for Agricultural Education, 2014)
20 STEM Education and Integration of Biotechnology Through federal legislation, the Department of Education and Workforce Development require s career and technical ed ucation programs at the high school level (Kuenz i, 2008) Students need specialized job skills and industry relevant knowledge to compete in the global marketplace and be successful in science careers. Agriculture education is changing from curricula that are primarily focused on production agricultu re to those containing more highly rigorous science content. There is a need for secondary schools to integrate more science into the agriculture curriculum, improve academic content and help students to adequately prepare for science related careers (Des pain, 2016; Mowen et al ., 2007 a ). Agricultural biotechnology courses are now being developed an d implemented in the secondary career and t echnical education classroom as a method to enhance STEM Education and to add rigor t o agriculture education courses. Specifically, it may be important for students to have knowledge about cellular biology and biochemistry of cells to understand biotechnology. Mowen et al. (2007 b ) suggests that science integration is needed to prepare students for diverse careers in both science and technology including biotechnology. Career and technical education classrooms pr epare students in high school to be college and career ready. These programs prepare students to integrate academics with employability skills and technical, job sp ecific skills (A ssociation for Career and Technical Education, 2017) A ccording to the Federal STEM Education Strategic Plan
21 the United States is to continue its remarkable record of ( National Science and Technology Council, 2013) Mandated by Federal l egislation, the Department of Education, and Workforce Development requires career and technical education programs at the high school level to develo p and integrate High Skill, High work force (Florida Department of Education, 2016) In 2012, there were more than 14 million unemployed people in the United States while cur rently unemployment is 7.1 million (Bureau of Labor Statistics, 2017) Over the last decade there has been a considerable shortage of STEM worker s to meet labor market demands. skilled enough in math and technology to fill an estimated 3 million permanent job (Price, 2012) Many students in the U.S. are graduating with inadequate work solution is to motivate student interest i n STEM education (Price, 2012). Stubbs and Myers (2015) proposed that Agricultural Education should help create a 21 st Century workforce that will address social, economic and environmental challenges through STEM. Myers & Dyer (2004) reported that best methods in implement ing and integrating STEM curriculum need to be identified for teacher preparation. Connecting content knowledge to problem solving through real world experiences is the objective of STEM (Ejiwale, 2012) Agricultu re education is a component of career and technical programs in high schools and is also one of the primary vehicles for integration of STEM programs at the high school level Agriculture
22 education methods differ from traditional science courses by offerin g hands on experiences in agriculture farms or greenhouses and applies them to biological concepts. Biology courses are the other vehicle through which biotechnology concepts are introduced to high school students. Biology courses introduce the concepts of biotechnology in a limited capacity using laboratory experiences when they can be integrated into a unit of study (Borge rding et al., 2013) Because agriculture brings meaning and understanding of through application, the integration of biotechnology skills should offer students the opportunity to gain academic knowledge and skills. Career and technical courses can be used to support integrated coursework between science, math, engineering and technology by designing curriculum and projects that are student centered to meet these new academic and career and technical standards (Florida Department of Education, 2016) The STEM education programs help youth acquire key employability skills and provides students opportunities to pursue college and careers (National Science Foundation, 2014) Biotechnology Curri cul um and Integration As previously stated, b iotechnology is a field that combines current modern techniques including genetics and cell technologies (Garrett, 2009) and somewhat older techniques such as selective breeding of yeasts and fermentation processes. These are current l aboratory techniques taught in agriculture education. Modern biotechnology methods are being used in food products, medicines, and environmental processes to create energy and help in environmental restoration but, these technologies are currently under re presented in classrooms. Biotechnology has been a content area in educational technology for almost thirty years but still is not widely taught in the career and technology curriculum in many
23 high schools due to misconceptions regarding the field and cons equent lack of confidence in the teaching and delivery of material resulting from insufficient professional development to teach biotechnology content (Wells & Kwon, 2008). The potential of any curriculum to be adopted and accepted by a teacher is depende nt upon their pedagogical content knowledge and the ability to teach the content (Shulman, 1986) Teachers must also integrate their content knowledge of the subject matter while making connections between subject are a concepts and different instructional approaches (Davis & Krajcik, 2005). For example, science teachers need to help students understand how to make observation, collect data, analyze and interpret how scientific inquiry leads to generating evidence throu gh experimentation. It can be difficult for teachers to connect content to theory with practice Fenstermacher, ( 1994) and to apply that knowledge across multiple contexts, or a group of conditions and disciplines without the content knowledge and training. Both theory and practice needs to be integrated across disciplines from science to agriculture to biotechnology. Davis and Krajcik (2005) know how to help students understand the authentic acti vities of a discipline, the way knowledge is developed in a particular field, and the beliefs that represent a sophisticated ontent knowledge of biotechnology will have an impact on student learning. The effective design of curriculum materials should be focused on the intention of first promoting teacher learning (Davis & Krajcik, 2005) A teache r's efficacy is a measure of his or her capabilities to bring about desired outcomes of student engagement and the learning environment (Tschannen
24 Moran & Hoy, 2001) Teachers that have a higher efficacy in learning are open to new ideas and will invest both more time and in new methods to engage their students in learn ing These individuals are also more characteristically resilient and have a greater commitment to teaching (Tschannen Moran & Hoy, 2001) The attitudes and perceptions of agriculture teachers towards biotechnology are dependent upon their ability to deliver content with confidence to achieve student engagement. If teache r learning is the focus for adoption and implementa tion of biotechnology curricula we need to also examine teacher motivation and beliefs. Self efficacy theory is focused on expectancies for success. Bandura (1994) supports two types of expectancy : outcom e and efficacy expectations Outcome expectations are beliefs that certain behaviors encompass question certain outcomes while efficacy expectations encompass the question whether or not one can effectively perform the behaviors necessary to produce the ou tcome (Eccles & Wigfiel d, 2002) To understand teacher motivation, teacher behavior and attitudes toward the content areas in biotechnology need to be examined When teacher attitudes toward STEM fields are examined, Ejiwale ( 2012) cont ends that attitudes toward self efficacy are important for establishing a stable learning environment The instructional materials and methods are the essential tools that provide learners guidance as to what they are expected to do in a lesson or unit (Rothwell, 1992) The instructional materials and methods given to teachers will need to be designed to promote self efficacy and confidence in teaching the content of a biotechnology unit or course.
25 Recommendations provid e d by Borgerding et al. (2012); Bryce and Gray (2004) ; and Zeller (1994) document teacher concerns regarding the lack of biotechnology subject matter content and topics, the basic science underlying biotechnology, the applications of biotechnology, and the expectation for student participation on the social discourse about biotechnology. Borgerding (2012) also reported the need for additional curricula focused training, new innovative teaching methods, and activities for teaching bi otechnology. As mentioned in the studies above the opportunities for teachers to receive professional development opportunities in pre service and in service programs is essential for integration of the curriculum. Constructivism Theory and Sociocultural T heory The Constructivist Theory, developed by Bruner (1966) is based on the belief that we view knowledge as the result of connecting previous experiences together with the importance of societal and conte xtual parameters (Pelech & Pieper 2010). Construc Cognitive Development which is a description of cognitive development in children and how they interpret the world around them using a four stage model describing how the mind p ) Sociocultural Theory was a guiding principle and were used in the development of both the blend ed learning methods and plant biotechnology curriculum modules developed in th is study The objective of the constructivist class is to shift the focus from the teacher and to the student with the use of pr oblem solving and inquiry based learning methods employing a variety of resources used to find solutions and answe rs. Doolittle (1999) and Save ry and D interaction and navigating the environment. These learners experience problems and
26 create knowledge by interpreting the world around them and discussing them with oculture Theory facilitates this aspect of learning by altering a child environment through interpersonal skills and develops cognitive growth (Schunk, student to student inter actions, teacher to student interactions, and student to teacher Vygotsky 's theory di ffered from Piaget 's who believed that learnin place in stages or universally acr oss cultures as Piaget propose d ( in McLeod, 2015). Vygotsky further believed the environment in which children grow up will influence how they think and what they think about, and therefore he concentrated on the role of language in cognitive developmen t. Vygotsky (1978) advocated that cognitive functions are affec ted by beliefs, values and tools of intellectual adaptation of the culture and therefore are socio culturally developed ( in McLeod, 2015) Jean Piaget (1896 1980) is credited with having played a significant role in forming the constructivis m philosophy. Piaget emphasized that children think and learn through participation and construct knowledge of the world through interaction with objects. The learner is active, and readiness is based on maturation that takes place with time to develop prerequisites for the acquisition of knowledge. Intellectual growth includes three processes; assimilation, accomm odation, and equilibration. Assimilation uses an existing knowledge to deal with a new object or situation. Accommodation object or situation. Equilib ration involves the use of force of metacognitive processes to
27 move development along, not steadily but in leaps and bounds. It is the force that drives the learning process (McLeod, 2015). What are the implications of learning theory for educational curricula? The student centere d philosophy utilizes teaching methods that are not just programmed instruction but utilize assimilation and accommodation to acquire knowledge. This involves individualized learning and opportunities for teacher to student and student to peer discussion. Experimentation, discovery and creation are essential components of this philosophy of learning. Laboratories, workshops and technologies should be encouraged including interaction with media and virtual reality appl ications. Bruner (1985) believed that pe er to peer methods improve problem solving which build on important critical thinking skills and use them as tools to increase meta cognitive functioning (Gokhale,1995). C lassroom applications such as recipr ocal teaching, scaffolding, apprenticeship, and c ollaborative learning utilize peer to peer and teacher to peer interaction for arranging tasks for novice learners, so they will be successful (McLeod, 2015). Consequently constructivism is the philosophy that promotes learning through social interaction, provides context, creates active learning, and promotes discovery and engagement in the learning process It is the pedagogical foundation for agriculture education curriculum and learning. Agriculture Biotechnology Curriculum Development What are the cu rrent designs and content of a biotechnology curriculum? There have been several reports of the integration of biotechnology through agriculture and industrial technologies in the high school curriculum (Mowen, et al. 2007; Savage & Sterry, 1995; Wells, 1994)
28 Because of the accelerated advances in biotechnology, curricula must b e frequently revised and updated to incorporate needed content changes The significant growth in genetically modified crops and industrial applications for food and agriculture products are the driving for ce for the global economy which, in turn, requires continual revisions of education materials and training for individuals to prepare for careers in biotechnology The impact of biotechnology on society and the impact of this technology in solving problems has become increasingly pervasive. Because of co ntinually changing legislation and advancements in technology, high school students must learn and demonstrate technical skills that increase their mastery of advanced science concepts for jobs in the biotechnology industry. The importance of secondary lev el instruction in biotechnology education drives integration in the science and technology curriculum Sae z et al. ( 2008) T he National Council for Agriculture Educati on has developed standards and guidelines for teaching biotechnology. The National Council for Agriculture, Food and Natural Resources (AFNR) Career Cluster Standards (2015) outline the current content for biotechnology curriculum and the course standards in the agriculture education classroom. BS.01 NCAE Standard : Assess factors that have influenced the evolution of biotechnology in agriculture (e.g., historical events, societal trends, ethical and legal applications, etc.). BS.01.01 Investigate and ex plain the relationship between past, current, and emerging applications of biotechnology in agriculture. BS.01.02 Evaluate the scope and implications of regulatory agencies on applications of biotechnology in agriculture and protection of public interest s. BS.01.03 Analyze the relationship and implications of bioethics, laws and public perceptions on applications of biotechnology in agriculture.
29 BS.02 NCAE Standard: Demonstrate proficiency by safely applying appropriate laboratory skills to complete task s in biotechnology research and development. BS.02.01 Read, document, evaluate and secure accurate laboratory records of experimental protocols, observations and results. BS.02.02 Implement standard operating procedures for the proper maintenance, use and sterilization of equipment in a laboratory. BS.02.03 Apply standard operating procedures for the use of safe handling of biological and chemical materials in a laboratory. BS.02.04 Safely manage and dispose of biological materials, chemicals an d wastes according to standard operating procedures. BS.02.05 Examine and perform scientific procedures using microbes, DNA, RNA, and proteins in a laboratory. BS.03 NCAE Standard: Demonstrate the application of biotechnology to solve problems in Agric ulture, Food, and Natural Resources (AFNR) systems BS.03.01 Apply biotechnology principles, techniques and processes to create transgenic species through genetic engineering. BS.03.02 Apply biotechnology principles, techniques and processes to enhance the production of food using microorganisms and enzymes. BS.03.03 Apply biotechnology principles, techniques and processes to protect the environment and maximize use of natural resources. BS.03.04 Apply biotechnology principles, techniques and process es to enhance plant and animal care and production. To align with the national standards there are two distinct pathways in Florida to teach animal and plant biotechnology. These career pathways lead to industry certifications for high school students (Florida Department of Education, 2017) The national standards are a guideline for most of the states to follow in developing of their career and technical education programs and are interpreted by state legislation and work force demand! This can pr esent challenges in implementation at the local school level. Several studies have indicated that teaching biotechnology is not being broadly
30 implemented in classrooms (Brown, Kemp, & Hall, 1998, Sanders, 2001, Russell, 2003, Kwon, 2009). To address this n eed, the implementation and instructional methods of teaching plant biotechnology curriculum and the factors that affect implementation at the local and state level must b e determin ed. Teacher Attitudes and Concerns About Teaching Biotechnology In s everal studies teacher concerns about biotechnology and the subject matter have been documented These concerns include the need to understand the subject matter, the desire for additional training, curricula, new innovative teaching methods, and strategies and activities for teaching biotechnology (Bryce & Gray, 2004; Kwon & Change, 200 9; Reicks, Zeller, 1994) Additionally, teachers are concerned with time and resources required for adopting the se new programs and the lack of funding for biotechnology teaching mater ials and equipment (Steele & Aubusson, 2004; Zeller, 1994) Teachers expressed concern about not having sufficient instructional time for preparation for biotechnology demonstrati ons and exercises or biotechnology instruction (Borgerding et al., 2013; Michael, 2006; Steele & Aubusson, 2004; Zeller, 1994) In addition, teachers requested support from stakeholders in the biotechnolog y industry, including tools, technology, equipment, and resources on the global impact of biotechnology on the environment and society. Hord et al. (1987) used the Stages of Concern Framework to develop theoretical models to investigate teacher concerns a bout biotechnology instruction and their practices in an attempt to understand how teachers approach biotechnology education and the concerns they have teaching biotechnology. Moreland et al. (2006) outlined pedagogical content knowledge needed to teach bi otechnology including conceptual, procedural, societal and technical aspects of biotechnology, knowledge of curriculum in
31 relation to biotechnology, teaching assessments pertaining to biotechnology, and classroom environment and management To promote le arning, Mowen et al. (2007 b ) likewise studied barriers, roles, and information source preferences for teaching agr iculture biotechnology topics. The information from these studies and an investigation on current biotechnology curricula could be useful in d eveloping instructional materials focused on teacher professional development. Two of the most pressing obstacles in teaching biotechnology are the availability of a current and relevant curriculum and instructional methods and laboratory exercises that address the science and current technologies used in biotechnology research and industries. There is also an absence of significant research regarding the integration of technology and engineering in the design of biotechnology curricula in the STEM field. Biotechnology Teacher Training The opportunities for professional development training for teachers in biotechnology are limited to science teachers specializing in biology and chemistry. These trainings are offered through industry and universities as professional development workshops in the summer including The Biotility Applied Biotechnology Training and The Center for Precollegiate Education and Training Workshops both offered at the University of F lorida. Currently, there are few in state resources and training opportunities for current Florida agriculture education teachers. Teacher professional training programs are available through the universities and supporting agencies of agriculture, yet it has not been broadly implemented in technology prog rams (Kwon, 2009).
32 As mentioned, there are two well designed programs for content teacher training in other are as of biotechnology in Florida. The first program is Biotility Applied Biotech Training for industrial biotechnology education and training with the emphasis on industrial products utilizing biotechnology methods. Teachers are immersed in a two week, laboratory based workshop in current biotechnology procedures and skills (University of Florida Center of Regenerative Health Biotechnology 2017) Secondly, workshops developed through The Center for Precollegiate Education and Training (CPET) are available to science educa tors (Brown et al., 2014 ). These programs are offered to all educators, but they are structured primarily for science content area instructors with biology teaching certificat ions. These programs offer summer training opportunities for other teachers in the sciences incl uding agriculture educators to obtain certification credentials. The programs are developed through the National Science Foundation and the National Research Council (2012) to achieve creation of science curriculum based in emerging and new knowledge in sc ience that actively engage students in sciences and engineering practices and to apply these concepts to deepen their understanding of the core ideas in these fields (National Research Council, 2012) The focus of these professional development workshops is to facilitate l earning of the basic principles of DNA, RNA, and protein based technologies used in me dical and industrial research. Science teachers and students are involved in several different CPET workshops that emphasize STEM laboratory and skill development in biot echnology. Inquiry based methods of learning including problem based learning (Barrows,1986) active research and wet lab activities are offered during trainings to promote trans fer learning to participants. There are several biotechnology workshops
33 that a re offered including programs such as Bench to Bedside, a translational medical works hop for teachers (CPET, 2017). The summer research experience available from CPET integrate teachers into laboratory research at the University of Florida and other two we ek trainings offered through Biotility emphasizing protein technologies and industrial applications. The research studies by Borgerding et al. (2013) on biotechnology address t integration of biotechnology and are the basis for the design of these workshops. The workshops discussed address some of the challenges that teachers face integrating biotechnology curriculum including biological content training and laboratory applications for the classroom. A strength of these professional development trainings for teachers include working with professors and researchers at the university level in the Teacher Summer Immersion Experience (CPET, 2017). Training also includes bench research using current technologies and applications of resear ch that are occurring in the workforce. There are hands on wet lab opportunities utilizing up to date equipment used in biotechnology laboratories and presentations by research scientists on the current research in t he focus area of the workshop. CPET offe rs equipment lockers containing lab equipment that teachers usually cannot afford to purchase for their classroom. The workshops are mainly focused towards basic science teachers. Agriculture teachers are utilizing some of the same skill sets but the focu s is on food, pathogen control and environmental remediation. Consequently, this can be a motivational concern for agriscience teachers attending the workshop series. Pedagogical content knowledge as outlined by Moreland et al. (2006) describes the content knowledge
34 challenges teachers face to address content shortfalls. Each of the workshop series provides pedagogical content ba sed either in biomedical, environmental or industrial biotechnology and could be adapted to fit an agriculture biotechnology course. The Curriculum for Agricultural Science Education (CASE) is a national training program offered to agriculture education te achers. The Case Institute was developed as a training program in all the areas of AFNR. Teachers travel to different states during the summer for professional development in the career subject content of choice. This program is developed on the founda tion s of two curriculum designs: How People Learn by The National Research Council (2000) and Understanding by Design Framework by Wiggins & McTighe (2005). This training is offere d to agriculture educators across the nation on several different career themed content areas. The agriculture biotechnology 2 week summer training opportunities are offered to develop content knowledge and laboratory knowledge through inquiry based learni ng methods. Using these strategies within the agriculture and natural resources context, the goal is to provide academic rigor and opportunities to learn and prepare students for future careers in the agriculture industry utilizing biotechnology (CASE, 201 7). There are both strengths and weaknesses to be address ed in this workshop training series. The CASE curriculum major focus is on agriculture education teachers and the series is open to all agriculture education teachers around the nation. The cost of the workshop is expensive, but there are scholarships available through supporting donors for teachers to attend. The workshop mainly consists of wet laboratory experiences progressing from fermentation, DNA extraction, protein purification, immunoassays a nd basic research of emerging fields. There were many hands on
35 laboratory experiences using equipment, but the series lacked pedagogical content knowledge and scientific theory. The instruction was very focused on laboratory activities for a yearlong labor atory course. Curriculum and lesson plans were available to the teacher after successful completion of the course. The equipment for the laboratory activities is not included. The expectation to teach the curriculum is for teachers to purchase the biotechn ology equipment and have laboratory space to complete the exercises. Flipping the Classroom The flipped classroom has the potential to be an effective m ethod to transform learning Flipped Learning is a pedagogical approach in which direct instruction mov es from the group learning space to the individual learning space, and the resulting group space is transformed into a dynamic interactive learning environment where the educator guides students as they apply concepts and engage creatively with the subject matter (F lipped Learning Network, 2014). Removing the direct instructional method (a scripted version of the task) to an online format outside of class frees up class time for practice and activities that promote active learning (Bergman et al., 2012). Fl ipping the classroom employs easy to use technology to free class time from traditional lecture allowing for more active learning Active learning includes teacher to student mentoring, peer to peer collaboration and cross disciplinary engagement (Roehl, et al., 2013) According to Shimamato (2012) the constructivist model presents learning as an active, social process in which learners use existing knowledge and prior experiences to build understanding of new material. Teachers act as facilitators to guide learning and allow students to construct their own understanding of the material. Learning takes place when students are exposed to differentiated instruction (providing students with different
36 avenues of learning) and they subsequen tly gain knowledge through these personalized experiences. A flipped classroom offers the opportunity to accommodate student learning styles for more differentiated instruction i.e., providing students different avenues to learning. The additional time i n the classroom provided by flipped classroom develops active learners that take control of their learning, rather than passive learners that receive information from the instructor and internalize it, while receiving no feedback (Minhas, 2017; Sams & Bergmann, 2013) Collaborative learning, involves groups of students working together to solve a problem, complete a task or create a product. Flipping the classroom can lead to higher levels of collaboration betwe en students (Roehl et al. 2013) Collaboration can motivate students who are more interested in their learning to better interact with their peers and bec ome better at problem solving. Flipped classrooms basically allo w more classroom ti me for collaborative projects. During the collaboration process students learn from one another and partake in their learning which in turn builds confidence. The flipped classroom requires that teachers transform their instructional pr actices from teacher centered to student centered. The teachers are required to ts at their own pace. (Bergmann et al.,2012). T he flipped classroom teacher is required to develop effective instructional methods to promote student engagement and improve learning outcomes. Researchers have studied the flipped classroom in multiple disciplines and the benefits on learning. Flipped l earning studies have been completed in engineering
37 education Mason, et. al. ( 2013) technology education Blair et al. ( 2016) physics Deslauriers et. al. ( 2011) agriculture education and communication ( Conner et al. 2014; Gardner, ( 2012 ); McCubbins et al. ( 2016) and other areas su c h as mathematics and business. McCubbins et al. (2016) examined student perceptions of flipping using a teaching method called team based learning in a capstone agriculture class O ther studies have offered insight into student perceptions, engagement, and content learning gains of the flipped classroom and its effectiveness to motivate and engage learners. Flipped classrooms have an influence on student learning and impact stud ent engagement and perception. More recently, the flipped classroom has been a common subject of study with research conducted at the college level (McCallum et al ., 2015; Blair et al., 2015; Conner et al., 2014; Jamuldin et al., 2014; Abeysekera and Dawson, 2014). Empirical research has been conducted on the student centered method of motivation. Davies et al. (2013) re ported how flipping in an introductory level college course classroom on spreadsheets affected student achievement and motivation. I n this study a simulated instruction course on spreadsheets was compared to a flipped classroom approach. Their results did indicate some learning gains of the students utilizing the simulation based instructional model, but students expressed frustration with the focus motivation to learn. Results of this study support a position that technology enhanced flipped classroom was effective and better facilitated than the simul ati on based learning
38 training. This approach was ob served to motivate students to utilize different learning methods to assimilate instruction and content learning gains (Davies, et al., 2013) With the goal to increase student motivation and engagement and learning gains through flipping instruction, Deslauriers et al. (2011) flipped a large enrollment physics cla ss. One course was taught using a traditional lecture presented by an experienced instructor versus a course that was taught using flipped research based methods by an inexperienced instructor. It was concluded that the flipped method increased attendance, higher engagement and more than twice the learning gains in flipped experimental group. A factor that Deslauriers discovered during his study was the that the embedded assessments used during the content lecture delivery of the flipped group increased stu dent motivation and engagement. The formative assessments, completed during the intervention allowed direct measurement of learning gains because they were assessed at the time of learning. This may improve teacher effectiveness in delivering content with active learning (Pollock & Finkelstein, 2008, Turpan & Finkelstein, 2009). In contrast to Desla uriers et al. (2011), Andrews et al. (2011) reported that active learning could not be correlated with student learning gains in college introductory biology cou rses. Andrews et al. (2011) specifically concluded th at there was a serious limitation of the effectiveness of active learning by typical biology instructors. A major concern expressed was that results documenting impressive learning gains may not be representative of what a typical instructor achieves And r ews et al. (2011) further pointed out that previous studies used science educators that had science education research backgrounds and therefore were prepared to use active learning more
39 effectively (Pollock et a l., 2008; Turpan et al., 2009). Without this expertise, the instructor may lack the constructivist elements necessary for improving learning. Andrew et al. (2011) comments that constructivist theory argues what students know and believe at the beginning of the course is scientifically inaccurate and if instruction is not specifically designed to address prior knowledge active lea rning instruction may fail. Instructors should not assume they are teaching effectively just because they are using active learning techniques without assessing the effectiv eness of their instruction. Andrews et al. (2011) recommends instructors develop active learning exercises for a broad population of instructors and identify training and ongoing support for science faculty to ef fectively use active learning. Inst ructors w ill need to assess the effectiveness of their instruction using pre/posttest designs to assess the effectiveness of instruction as well as using more formative assessments throughout the course to monitor student learning to d ocument student learning gains Preparing and supporting instructors and effectively using active learning techniques including flipped learning (Bergmann et al,. 2012) can help improve student learning i n science courses. There have been other concerns expressed in studies by Conner & Stripling (2014); Gardner (2012); Shimamoto (2012); Strayer (2012); Zhang et al. (2006) that learning gains may not represent wha t typical instructors are likely to obtain without further training opportunities as an instructor as reported by Strayer (2012). In an inverted (flipped) classroom Strayer observed that students were less satisfied with how the structure of classroom was set up but found that they accepted cooperative learning as an innovation. Innovation is defined as the introduction of something new or a new
40 common goal. Strayer (2012) also noted that students cooperated more effectively in a flipped classroom than students in a traditional lecture classroom. Strayer recommended that a flipped classroom format used in an introductory course should provide step by step instructions to create m ore structure therefore reducing the dissatisfaction f rom the This new structure should inclu de scaffolding, breaking instruction into segments and then provide a tool for structure when using these instructional techniques to advan ce students towards understanding and indepe ndence in the learning process. Eventually the support is removed as the activity progresses to help students become more aware of their learning proc esses. Strayer (2012) emphasized the need for students to refl ect on their activities which will help them connect with their course and improve overall perception. Student engagement is defined as the degree of attention, curiosity, passion or interest that students show when learning and is another area impacted b y active learning. Day and Foley (2006) have concluded that learning gains could occur if student perceptions which include achievement, motivation, happiness, satisfaction and success occur with student engagement by using flipped instruction web lecture s. The experimental method used in this study consisted of a quasi experimental design because random sampling was not possible. The purpose of the study was to increase student engagement by using integrated web lectures online, instead of the traditional class lecture during a semester long course. In a research study by student engagement and motivation during a web lecture (a combination of audio video and
41 PowerPoint materials), were compared to a traditional lecture of a human computer interaction cour se (Day & Foley, 2006) The study inv olved 46 participants, meeting two days a week for 15 weeks. The motivation provided to the students was to observe the lectures with embedded assessment s in the homework assignments. The assignments consisted of two to four synthesis type questions of the material covered in the lecture plus an additional question related back to pr evious class work or projects. The class time was utilized for various hand s on learning activities and used the web based lecture as a stimulus for discussion. The two sections were matched on the factors of instructor teaching the course, topics, lecture slides were identical, assigned readings, time on task and assessments were the same. A The control group consisted of a traditional course with lecture consisting of the same homework questions as the experimental group. Results of the study revealed that the Web lectures enhanced classroom learning with students in the experimental section scoring higher grades than students in the control section. The experimental group cours e section average final grade was 88.2 + 6.1, while the control section was 79.9 + 4.7 with a significant (p < 0.01) eight point difference. in addition, 6 students reported more self reported learning in the experimental group thus validating the hypothesis that the web lecture format was more educationally effective and enjoyable than a traditional lecture based course. Student motivation is defined as something that promotes a person to participate in a learning process and influences the reasons underlyi ng their involvement in academic activities (Abeysekera & Dawson, 2015) To increase student motivation as suggested by Day & Foley (2006) students need to first perceive the mat erial as useful and engaging before they will express more satisfaction with the course (Abeysekera &
42 Dawson, 2015). There are two ty pes of motivation, intrinsic motivation, behavior that is driven by internal rewards, and extrinsic motivation that con tribute to student engagement. Cole et al. (2004) define motivation as the will ingness to learn new material. The level of motivation inf luences a learners focus and level of effort expended on a given learning activity (Abeysekera & Dawson, 2015) Extri nsic motivation occurs when an external reward on a specific task is required to get a certain grade such as homework. Active participation requires students to be in charge of creation of knowledge through active participation and to feel more confident a nd engaged with their learning (Abeysekera & Dawson, 2015) To motivate students, a curriculum will need to be develo ped with the technology that captures and motivates the student internally and externally through multimedia learning tools. Development of Multimedia Tools for Flipping The flipped classroom relies heavily on visualization with video and lecture presenta tion which creates conditions for creativity (Martin & Schwartz, 2014). The development of multimedia learning tools and applications to engage the learner are rapidly being developed with programs such as Khan Academy, Ted Ed, and other multi media ap plic ation tools for students. I n guidelines for designers of multimedia learning, Clark & Mayer (2016) online version of a video, lecture, reading, or graphic s that enhan ce s the content of the lecture. the same so does the learning, no matter how the delivery method of the content is Media that delivers flipping instruction must be eng aging but well designed using both words and graphics to show relationships and to achieve specific learning
43 gains and maintain student engagement. Butcher (2014) concluded that his results supports the multimedia principle that effective learning materials should combine visual and verbal materials to target learned concepts. These media tools can be helpful to maintain motivation, but the goal is to increase engagement of the learner to connect with the subject matter. To increase motivation and engagement in students, new technological tools including interactive video e learning systems are currently be ing develo ped for educational platforms. Zhang (2006), developed an interactive video e learning system using asynchronous learning networks and videotaped lectures to evaluate learning effectiveness. The system utilized a multimedia integrated e learning system that was self paced, and accessed online at any time. Student scores were significantly higher on learning performance and students had increased sa tisfaction with interactive video. The results indicate that the interactive group scores were significantly higher than the other three groups with respect to both learning outcomes and learner satisfaction with e learning. The non interactive group achie ved equivalent te st scores to those with video. Consequently, simply adding multi media video may not a lways result in learning gains. Rather, multi media content that is under individual control over random access to content can lead to higher satisfactio n and better learning outcomes. With more interactive and richer media available, the learner who prefers this learning style will be more flexible and at tentive to the media. Al Zahrani (2015) suggests that students must be provided with adequate e learni ng tools consisting of well designed video lecture to promote creative thinking in higher education students. It is the interactivity and control by the learner that can improve learning effectiveness (Zhang, 2006).
44 The National Research Council (2009) cha llenged instructors to meet the needs of students by moving from passive learning to active learning methods and incorporate technological advances. Agriculture education research studies that have used flipped classroom methods primarily focused on studen t perception and engagement. A review of these studies is important to direct the focus for new studies on the effectiveness of this active learning method. To research flipped classroom instruction and its effect on student perception and engagement, Con ner et al. (2014), examined the flipped classroom approach with undergraduate agriculture education majors used the flipped classroom approach in a teaching methods and principles course. The findings of the study were mixed. Some of the groups reported di ssatisfaction in the course consistent with Strayer (2007) observations. However, most focus groups reported overall satisfaction in pedagogical knowledge in the flipped teaching methods course McCubbins (2016) evaluated student perceptions and flipping u sing an active learning method of the flipped classro om called Team Based Learning. perceptions concerning an undergraduate capstone course. Overall, student perceptions were positive in relation to individual and te am learning and their ability to understand more difficult course concepts thus supporting the adoption of a student centered course design. McCubbins (2016) recommended that groups in agricultural education could design programs and trainings for faculty to meet specific competencies needed for effective flipped instructional design (McCubbins et al., 2016; Strayer, 2012). Therefore, there is a need for agriculture education teachers to be trained in flipped instructional method and specific training in ag riculture biotechnology
45 To continue the expansion of opportunities and possibilities through STEM, there exists a need for well prepared STEM workforce (National Academies of Sciences (2016). The challenge is to prepare citizens for this expanding workfor ce and use innovative tech nology through STEM education. Agriculture biotechnology education is an innovative way to increase science content literacy and motivate students to pursue careers in STEM fields through agriculture (Boone et al., 2006; Wilson et al., 2002; Zeller, 1994). Graham et al. (2013) explains that there needs to be a focused effort to motivate and retain students to study the STEM fields. Many talented students flee STEM majors because they find introductory courses uninspiring. This att rition can be corrected by incorporating classroom teaching practices such as active learning. He further explains that active learning improves understanding and retention of concepts and information which helps students identify as scientists. In educat ion, motivation is the driver of student engagement which is a powerful influence of confidence (self efficacy) and persistence. (Dwe ck, 1986; Graham et al., 2013). Faculty are reluctant to try active learning strategies such as flipping because they lack experience in applying these methods. It is essential to provide training in evidence based instruction. It is recommended that this should be accomplished through pre service and in service trainings, grants, workshops and other incentives to increase sci ence content literacy need in STEM courses. How does flipping relate to my proposed research? The teaching modules created will address the structure recommended by both Strayer (2012 ) and Zhang (2006). Step wise instructions in the form of video based co ntent lectures for flipping
46 content will be designed in a step by step method to include scaffolding of concepts of dividing plant tissue culture methods into meaningful segments to promote student understanding and independence. E mbedded assessments to me asure content knowledge learning gains important to examine the design of the lessons. Likewise, t he design of the lesson mo dules will be created to address motivational concerns by using active learning strategies and teaching methods focused on standard based instruction and assessment. These methods not only increase teacher content knowledge of plant tissue culture, but also motivate teachers to use the lessons with students. Thus, improving perceptions and attit udes about plant biotechnology. Developin g new innovative curriculum modules to teach biotechnology with active learning methods of flipping instruction can help educators integrate the knowledge they need to deliver the content to students and gain motivation and self efficacy. Th e flipped class room method ( Figure 1 1) can be incorporated as one of the innovative student centered methods to help deliver new content. Examining the flipped classroom approach in agriculture education professional development is a method to assess its efficacy and i ts effect on learning (Conner et al., 2014; Roach, 2014, McCubbins, 2016). Figure 1 1. Bloom s Taxonom y (revised). Reprinted with permission from Vanderbilt University Center for Teaching, https://cft.vanderbilt.edu/guides sub pages/blooms taxonomy/ (October 5, 2017).
47 Factors Affecting Implementation of Biotechnology Many high school teachers frequently must teach courses outside their major fields of st udy. Biotechnology is an area that is outside the scope and sequence of most current agricultural education courses offered at the university. Brown (1998) reported that 64 % of teachers surveyed in Kentucky thought that science teachers or a team of scien ce/agriculture/technology education teachers should teach biotechnology. The significant factors that will be investigat ed in my research study include lack of training, time, curriculum and equipment resources, and motivation to teach agriculture biotech nology at the high school level. Studies conducted by Borgerdin g, et al. (2012); Mowen et al. (2007 b ); Kwon and Change (2009); Bryce and Gray (2004) ; Reicks et al. (1996) recognized and address these concerns. Teachers are also concerned how agriculture biotechnology program s wi ll fit into current curricula. D evelopment of career academies leading high school reform were designed to engage students and better prepare them for the workforce by integrating academic and technical courses organized around career themes and work based learning opportunities (Estacion, et al. 2011) Agriculture Biotechn ology is a career themed academic endeavor consisting of courses in a nimal and plant biotechnology. These career pathways have created obstacles for current teachers that have only taught in other areas of ag riculture ing these new pathways include learning specific content, student understanding of the course content, motivation, time and budg et concerns. There is evidence (Kidman, 2009) that teacher attitudes may have an effect on science practice in general. These obstacles will need to be addressed for teachers to feel confident in delivering the content to their students.
48 Factors affecting the impl ementation of biotechnology include lack of teacher professional development and training in agriculture biotechnology. Traditional instructional methods used in teaching agriculture biotechnology are shifting in a new direction. The advances in technology are creating change in the physical classroom. These changes are transforming instructional methods to deliver science con tent infused with technology. The problem for agriscience teachers is to utilize the technology to help advance student content knowl edge of various biotechnology concepts in animal and plant based biotechnologies in agriculture. According to Thoron (2010) the National Academy of Sciences (1996) called for more student centered classroom and learning activities along with th e National Science Foundation (2014) Another problem with teaching biotechnology is the insufficient amount of instructional t ime allotted to teach biotechnology laboratory exercises. This time allotment is not sufficient to teach in a traditional 50 minute high school class period. Utilizing video lectures and virtual instruction at home or during warm up activities allowing mor e time for actual instruction to complete more rigorous biotechnology laboratories. T eacher views of biotechnology vary. Some teachers express positive views on teaching biotechnology (Kwon and Change 2009) while some express re servations (Michael, 1997) Other teacher related factors which may influence individual implementation of biotechnology. One of these factors include s teacher experience. Kwon and Change (2009) reported that teachers with more experience were less likely to teach biotechnology whi le Steele and Aubusson (2004) did not find a relationship between years of experience. Experienced teachers have the ability to
49 adapt curriculum more effectively if th eir content knowledge of the subject matter is sufficient. Bogerding et al. (2012) report ed that additional training and professional development positively impacted biotechnology teaching practices. This observation h ad previously been reported by both Steele and Aubusson ( 2004 ) and Zeller ( 1994). Other factors for further investigation are t he lack of teacher professional misconceptions of biotechnology, instructional methods, and lack of confidence. These are contributing factors as discussed by Wells & Kwon ( 2008) Other contributing factors include school setting, laboratory and equipment availability, cost and administration support as supplemental factors (Kwon, 2009) Zeller (1994) researched teacher attitudes towards biotechnology content areas and the required training in biotechnology. He determined ther e was a significant biotechnology content knowledge and i mplementation in the classroom. It was revealed that three major roadblocks prevent the implementation of biotec hnology into the curriculum. These include lack of money for equipment, lack of time to prepare and train, and problems with implementation of material into an already crowded curriculum. Current teacher views of the status of these previously recognized challenges w ill be investigated using the Teachers Belief About Biotechnology Survey administered to current agriculture education teachers in Florida (C ha pter 2) Increased teacher preparation and workshop trainings would directly improve the implementati on of biotechnology into the curriculum (Mowen et al., 2007 a ; Steele & Aubusson, 2004)
50 instruction will affect them personally with the amount of time needed to invest in changing the ir current practices as well as the extent to which biotechnology instruction may be personally interesting or (Borgerding et al., 2012 p.136 ) Purpose of the Study The purpose of this st udy is to develop plant biotechnology teaching modules for teachers to increase student and teacher content knowledge and change attitudes associated with teaching plant biotechnology concept s and techniques A goal of this research is to provide both inst ructional material s and professional development opportunities utilizing teacher plant biotechnology workshops. Secondly, the question whether teaching plant biotechnology laboratory modules can be improved by employing a blended instructional method calle d flipping the classroom to increase student content knowledge of the required for the plant shoot culture of Caladium bicolor will be examined. The basic question posed was: Is classroom flipping an instructional method that can help increase content know ledge for both agriculture education teachers and students? Teacher concerns about teaching biotechnology and the lack of equipment, time, and content knowledge will be addressed in this study by design ing more "teacher friendly" plant tissue culture lesso n modules. Finally, w ill addressing these concerns lead to more successful integration of plant biotechnology content in the agriculture education classroom? Statement of Objectives In this study, flipping methods used will include video lectures and asse ssments completed prior to classroom based plant biotechnology laborat ories. The research
51 method will address whether flipping can transform a biotechnology classroom from a teacher centered to a student center environment that results in biotechnology con tent knowledge gains The specific objectives are: 1. Determine the gender, grade level, degree level, subject area, biotechnology pre service preparation, current teacher beliefs on teaching plant biotechnology, and socio economic status of Florida high sch ool agriscience teachers utilizing a survey on attitudes and con cerns of teaching biotechnology (Chapter 2). 2. Develop comprehensive plant biotechnology teaching modules and lesson plans using blended learning methods for classroom use (Chapter 3) 3. Determ ine the content knowledge gain of teachers utilizing a flipped classroom lecture and lesson on plant shoot culture of Caladium bicolor during an in service training (Chapter 4) 4. Develop plant tissue culture lesson modules with blended learning laboratory activities for teachers to build content knowledge and efficacy in teaching plant biotechnology (Chapter 4) Statement of the Hypotheses For the purposes of statistical analysis, the research questions will be posed as null hypotheses. All null hypothese s were tested at the .05 level of significance. H O There is no significant difference in biotechnology content knowledge gain between agriculture biotechnology teachers who receive flipping instruction and those taught using a direct instruction (lecture ) method in a plant biotechnology module. H A Teachers who receive flipping instruction while completing a plant biotechnology module will achieve the same or higher content knowledge gain scores than teachers taught by a direct instruction (lecture) met hod. Definition of Terms The following terms are operationally defined for the purpose of this study: Agricultural Biotechnology: is a range of tools, including traditional breeding techniques, that alter living organisms, or parts of organisms, to m ake or modify
52 products; improve plants or animals; or develop microorganisms for specific agricultural uses (USDA, 2016). Content Knowledge: the subject matte r tested following the instruction that is measured by the level of correct responses from the p re and posttests. Content Knowledge Achievement: the level of correct responses following treatment from the content presented by the teacher. Direct Instruction (lecture) E xercise: the method of instruction that uses a traditional power point and lectu re before the laboratory exercise. Flipping Instruction E xercise: an instructional method that utilizes a video and audio with embedded questions. Pre Assessment: a multiple choice test that will used to measure responses before the instruction of the m odule. Pre Service Training: the education and training provided to student teachers before they have undertaken any teaching. Post Assessment: a multiple choice test that will be used to measure responses after the instruction of the module. In Servic e Professional Development: the workshop that will provide teachers training in instructional methods and laboratory exercises used in the study. Blended Learning: Blended learning environments combine face to face instruction with technology mediated i nstruction and will be used in the teacher workshop using the flipping instruction exercise. Potential Limitations of the Study The conclusions and implementation resulting from this study are subject to the following potential limitations: The results are limited to the extent that the instruction was restricted to a small group of teachers because of the size of the teaching laboratory facility. The data are limited to those obtained from purposively selected Florida agriculture teachers. Therefore, ge neralization of results of the study to other groups will be limited to the degree to which those groups match the population and sample in this study. Measured variables such as content knowledge are accurately identified.
53 The results are limited to the extent that they only apply to one module unit of instruction common to all participants in the study. Figure 1 2. Chronological o rder of r esearch t asks. Chronological Order of Research Tasks Teacher Survey Issued from November 2015 April 2016 Teacher demographics, attitudes and concerns about teaching biotechnology were collected from the survey. Develop Caladium Module Create Flipping Instructional Experiment 1 st Teacher Workshop November 2015 Three Teaching Modules Created and Tested Teacher Feedback on Challenges of Teaching Biotechnology 2 nd Teacher Workshop November 2016 Flipping Instructional E xperiment using caldium shoot culture teaching module Measured content knowledge gain
54 CHAPTER 2 TEACHER BELIEFS ABOUT TEACHING BI OTECHNOLOGY IN THE C LASSROOM Introduction The need to train a more diverse and well prepared STEM wor kforce has prompted develop ment of new innovative approaches in STEM education (National Science Foundation, 2017) T he STEM workforce is composed of many different sub groups that use STEM knowledge and skills directly or indirectly across diverse career fields. The acquisition of STEM skills and knowledge is vital for an i participate fully in the 21 st Century workforce (National Science Board, 2014) Agricultural, biological and enviro nmental life scientists, and Science and Engineering (S&E) related occupations including precollege teachers (e.g., science teachers, math teachers) are considered critical for preparing the STEM workforce to maintain innovation and competitiveness. Integration of biotechnology concepts and skills in the agriculture science education classroom can fulfill the needs of creating a STEM educated workforce ha ving the necessar y skills to succeed in these evolving career fields. Why then is biotechnology not widely taught in the secondary classroom? Brown, et al. ( 1998) reported that agriculture science teachers bel ieve there is a need for integrated biotec hnology curriculum which provides better integration of science concepts and applications spe cific to agriculture courses. Biotechnology instruction is interdisciplinary and requires close collaboration with teache rs from other disc iplines (Brown et al., 1998). There is agreement among educators that biotechnology education should be the vehicle to address the implementation of rigorous science content in the agriculture classroom and to prepare students for diverse career field in Science & Technology fields (Mowen et al., 2007 b ). A teacher
55 needs to believe that the curriculum will have a positive effect on the learning envir onment for students. Wilson et al ., perceived value of the benefits of an integrated biotechnology is a predictor for adoption of the curriculum because it will help students best prepare for their future and project a better image of the agriculture education p rogram. Considering the impact a teache r has on the curriculum and the learning environment, teacher attitudes and beliefs will have a strong influence on biotechnology teaching and learning in the classroom (Kidman, 2009) Statement of the Problem Barri ers exist in the implementation of biotechnology in the classroom T hese key factors include lack of teacher biotechnology content knowledge, availability and funding for equipment, and pre service and in service training as identified in studies by (Wilso n et al. 2002; Mowen et. al, 2007 b ). Pre service training is the education and training provided to student teachers before they teach. Wilson et al. (2002 ) reported that North Carolina teachers lacked the prerequisite knowledge needed to successfull y tea ch biotechnology skills. Aziz et al. (2009), found that the benefits of biotechnology teaching workshops for secondary school educators and subsequent integration into the school and attitudes about biotechnology. Kwon (2008) has pointed out that although the key factors affecting teachers integrating biotechnology have been id entified, large scale adoption and implementation of agriculture biotechnology curriculum has yet to be achieved. However, Mowen et al. (2007 b ) and Brown et al. (2006) identified the status of agriculture biotechnology instruction concluding that there was a need for further study. The identification and elimination of these perceived barriers and concerns in
56 Florida agriculture biotechnology will be important to help with broader implementation of plant biotechnology in the agriculture science secondary classroom. The theoretical framework for this study will be focus ed on the expectancy value motivational theory biotechnology (Wigfield & Eccles, 2000) Kwon (2008) and Wilson et al (2002) used a combinatio n of this theory and the motivational theory (Locke 1991) to study teacher motivati on as a major factor affecting perception of knowledge and decisions in the implementatio n of new educational initiatives such as biotechnology. According to the expectancy value theory, behavior is a function of expectations towards th e goal you are worki ng toward. Motivational theory is important in curriculum integration for teachers to have expectations as well as subjective task values the motivation to do an activity, and beliefs to integrate biotechnology with agriculture (Eccles & Wigfield, 2002) The re i s a need to determine the current beliefs, moti vations and concerns of Florida agriculture education teachers This will be accomplished in this study by using the Teachers Belief About Teaching Biotechnology (TBTTB) survey. Specifically, the survey, resul ts of which are described in this chapter are intended to highlight perceptions and attitudes of current Florida agriculture science teachers and their concerns for adoption and implementation of biotechnology in the agriculture education classroom. The i nformation generated will provide guidance in developing plant biotechnology teaching modules which will provide content knowledge and hands on technique experiences.
57 Purpose and Objectives The purpose of this survey is to identify Florida agriculture edu cation teachers perceived attitudes and concerns for teaching agricul ture biotechnology in Florida. To fulfill the purpose of this study, the followin g objectives will be addressed: 1. Determine the demographic characteristics of Florida agriculture science t eacher survey participants. 2. Determi ne agriculture science teacher attitudes and concerns regarding teaching biotechnology. 3. Determine if differences exist between teachers that currently teach biotechnology and those who do not teach biotechnology. 4. Determin e what types of pre service and in service biotechnology training Florida agriculture science teachers had completed. 5. Determine if differences existed between training and attitudes of agriculture science teachers currently teaching biotechnology. Method s General Procedures A survey questionnaire was developed to collect responses from agriculture education teachers in Florida. Specifically, t his research involved the creation and administration of the questionnaire, iotechnology (TBTTB), to collect data on demographic, attitudinal, and concerns of Florida agriculture science teachers. The results of the questionnaire were collected using Qualitrics software through the University of Florida after obtaining prior appr oval to conduct the study from the University of Florida Intern ational Review Board (IRB) ( Appendix A ). The survey res ponses were analyzed using the Statistical P ackage for the Social Sciences (SPSS) version 24.
58 Participant s The target population consisted of agriculture science teachers in Florida. The participants were derived from the UF Agriculture Education listserv consisting of all 429 agric ulture science teachers in the S tate of Florida (N= 429). Teachers were volunt arily aske d to participate in the survey. The survey was open from Novem ber 9, 2015 to April 22, 2016. Seventy eight teachers responded to the survey. They consisted of high school, middle school, agriculture science teachers and other categories including school administration R espondents were divided into two subgroups, teachers that currently teach agriculture biotechnology and those that were not currently teach ing agriculture biote chnology. Survey Instrumentation The IRB approved TBTTB survey instrum ent an d cover letter were transmitted by email via the University of Florida Agriculture Education listserv ( Appendix A) A link to the survey was provided for participants to anonymously participate in the survey. Follow up e mail reminders were sent out periodically during the study with the survey link, to remind participants to complete the survey The survey contained two multipart biotec hnology (five levels, twelve attitudes) and a section to collect demographic information. The survey was modified, with approval, from a version of the Beliefs Toward Biotechnology survey used by Kwon (2009) Written approval to revise the survey was granted by Dr. John G. Wells, Virginia Polytechnic Institute and State Universi ty Revisions were made to the demographic section, attitudinal questions were
59 developed, wording and question sequencing were revised, and open ended questions were eliminated. The first part of the TBTTB survey was designed to collect general demograph ic data pertaining to three categories : 1) general data: gender, years of experience, educational degree, teaching subject, teaching experience in agricultural science, and ex perience teaching biotechnology; 2) professional development; in service training and pre service courses ; and 3) school environment: school location, size and lab availability. A total of 15 demographic questions were developed within these categories. Part two of the survey consisted of eleven questions regarding teacher attitudes a nd concerns with t eaching biotechnology. Respondents used a 5 point Likert scale (1= Strongly agree, 2= Somewhat agree, 3= Neither Agree or Disagree, 4= Somewhat Disagree, 5= Strongly Disagree ) to record their agreement levels to each statement. The eleven questions were divided into two categories, attitude and concerns. (Table 2 2). Overall attitudes scores were calculated to determine if relationships existed between attitudes and selected demographics Data Collection This survey was open from November 9, 2015 April 22, 2016. Missing data in unanswered questions in the Likert responses, reduced the useable number of respondents to 62. Data were collected on line however; some participants did not fully complete the survey. Descriptive analyses of the attitude scores included measures of central tendency (mean, median or mode) and variability (frequ encies or standard deviation). Appropriate measures of association including a One Way Analysis of
60 Variance (ANOVA) were used to examine relationships betwee n selected teacher demographic variables and attitudes towards teaching biotechnology, along with Reliability, as a measure of internal consi stency for the attitudes summated mean score revealed an alpha coefficient of .66, indicating the summed scale was at the lower level of consistency. A significance level of p 0.05 was employed. Results Based on the results of the TBTTB survey, approximately 50% of the agriculture biotechnology science ed ucators agree d that th ey had concerns about the specific time required to teach biotechnology. Identifying these perceived needs and concerns is important to help with broader implementation of biotechnology in the agriculture science s econdary classroom i n Florida. Survey results specific to the four objectives are presented in Table 2 1 Demographics of the sample group Objective #1 : Determine the demographic characteristics of Florida agriculture science teacher participants Seventeen questions i n Part 1 of the survey were used to collect teacher demographic data. Twenty five respondents were male (40.3%), while thi rty seven (59.7%) were female. Of the teachers responding, twenty one (33.8%) were middle school teachers, thi rty two (51.6%) were high school, and nine (1 4.52%) were combined middle/ high school teachers. The highest degree earned included thirty two teachers es, and one (1.6%) with a PhD. The subject areas in which they taught consisted of fifty two
61 (83.9%) licensed in Agriculture Education, two (3.2%) in science, and twelve (12.9%) in other areas (i.e., Administration K 12, Biology, English, History, Gifted, and Special Education). Respon dents were asked to describe the type of c ommunity in which they taught. Twenty nine teachers (46.8%) taught in rural schools, ten (16.1%) were in urban schools, twenty three (37.1%) in suburban communities. Finally, teachers that taught biotechnology in their agriculture education classroom totaled 28 (45.2%), while the other thirty four (54.8%) did not currently t each biotechnology. The average years teaching agriculture biotechnology at the middle school level was 3.1 years. At the high school level tea chers taught agriculture biotechn ology an average of 5.4 years. Two teachers at combined middle/high schools did not provide information on the number of years teaching biotechnology. Attitudes and Concerns about Teaching Biotechnology Objective #2: Deter mine agric ulture science teacher attitudes and concerns toward teaching biotechnology ( Table 2 1 ) Agriculture science teachers were asked to express their attitudes and concerns toward ten biotechnology statements using a 5 poi nt Likert type scale (1=Strongly agree, 2=Somewhat agree, 3= Neither agree or disagree, 4 = Somewhat disagree, 5= Strongly disagree). Fifty four ( N=54) of the original 78 participants responded to the attitude questions. Descriptive data on the teacher res ponses to the statements are reported as frequencies and percentages ( Table 2.1 ). The responses were from the en tire group of teachers. There wa s moderate positive agreement to five of the questions (Statements 1, 3, 5, 7, and 9 ) Seventy eight percent (77.8%) of the fifty four
62 enhan High would like to know of several methods and (67.3%), I would like to know how the integration of biotechnology is better than the methods I presently use or plan to my employ when I do my job (66.7%). My present schedule is preventing me from learning too much about integrating biotechnolog y concepts (63.6%). There was moderate disagreement to statements (2, 6, and 10) with the concerns regarding job role change, methods and feedba ck from students about biotechnology. Objective #3 : Determine if differences existed between teachers that currently teach biotechnology and those who do not teach biotechnology Agriculture science teachers were divided into two groups, based on response s that required a yes or no response to Question #7 of the survey: Do you currently teach biotechnology? Based on the subgroups 47.4% of (N=54) teache r respondents currently teach biotechnology, versus 52.6% of (N = 54) teachers that did not currently teac h biotechnology. Creation of these two subgroups was necessary to compare differences in attitudes between the two groups. T teaching agriculture biotechnology are outlined in ( Table 2 2 ) There was stron g agreement in both groups on S tatement #10 regarding concerns on student atti tudes toward biotechnology Regarding S tatement #1: a concern about the amount of time to teach biotechnology 55% of the group that current ly teach biotechnolo gy and 59% in the group currently not te aching biotechnology regarding expressed concerns. There was stron g agreement between both groups; 74.4% and 57.1%, respectively. There was
63 some disagreement between groups regarding the current schedule in terfering with learning and the integr ation of biotechnology concepts (Statement #3). The group that currently is teaching biotechnology were in strong agreement (48%) versus the group that is not teaching biotechnology, which strongly dis agreed (56%) with the stat ement. Finally, regarding S tatement #8 teachers from the group that were not currently teaching biotechnology expressed agreement on the need for coordinating with fellow teachers to learn more about biotechnology. Bo th groups somewhat agreed with S tatem ent #5: I would like to know how the integration of biotechnology instruction is better than the methods I presentl y use or pl an to employ when I do my job. Agreement existed in both groups to the need for integration of biotechnology, (48.1 %) for teacher s currently teaching biotechnology versus (37.0%) that are not currently teaching biotechnology. Mean agreement levels for each of the statements are reported in Table 2.3 for both teacher groups. Agriculture science teachers t ( 2.11 0.97 ), and their students attitudes toward biotechnology ( 2.3 1.14 ). (1) how their job will change when they begin teaching biotechnology ( 1.96 1.29 ), and item (2) the time needed to teach biotechnology concepts ( 2.3 1.25 ). In addition, they wer e concerned with their student attitudes about biotechnology ( 2.44 1.31 ). A one way analysis of variance (ANOVA) was conducted using the attitude summated mean scores to determine if there was a difference between the two groups of teachers, those c urrently teaching biotechnology and those not currently teaching
64 biotechnology .701) and there were no outliers in the data, as assess ed by inspection of a boxplot. ANOVA results revealed no significant difference between the two groups F (1,53) = .41, p = .53) as reported in Table 2 4 Objective 4 : Determine what types of pre service and in service training Florida agriculture science teachers have completed in biotechnology and determine if differences existed between training and at titudes of agriculture science teachers teaching biotechnology. Regarding the types of pre service and in service training of Florida agriculture scie nce teachers in biotechnology, 77.4% of the teachers never participated in any formal pre service coursewo rk in biotechnology, while 16.13 % of teachers reported that they had only one college course and 6.5% of the teachers reported two or more courses in their pre service coursework in biotechnology. Likewise of the in service professional development traini ng experienced by these sixty t wo teachers, 50% of the teachers reported taking a professional development course in biote chnology while the other 50% had not. The 50% of teachers that did take professional development in biotechnology included 22 (62.9%) teachers that completed a one day workshop, 4 (11.4%) took a two day workshop, 2 (5.71%) took a 3, 4, or 5 day workshop, one (2.9%) a one week workshop, two (5.7%) a two week workshop while four teachers took other professio nal development opportunities. The lack of pre service courses in biotechnology were analyzed to determine if this wa s a factor limiting the implementation of biotechnology in the agriculture classroom and to determine if the
65 amount of background training in biotechnology influenced del ivering biotechnology scientific concepts to students ( Table 2 5 ). A one way ANOVA was conducted to determine if the mean attitude score towards teaching biotechnology was related to the level of teacher pa servic e and in service training. There was a statistically significant difference between groups in pre service training as determined by a one way ANOVA, F (22,32) =2.13, p = 0 .025 verifying that the lack of pre service training and coursework had a significan t ttitudes toward biotechnology. There was no statistically significant difference between teachers that had in service training versus those teac hers that did not have training ( p =0.386) and t he attitude mean score ( Table 2 6 ). Discussion The purpose of the survey was to identify and characterize current Florida agriculture teacher perceived attitudes and concerns for teaching agriculture biotechnology. In Florida, it is common for the Department of Educat ion to imple ment new educational programs. Agriculture biotechnology is one of these programs requiring the imposition of new standards and curricula i n agriculture science programs. It is often the case that teachers have to create or implement these new programs within their schools (Kidman, 2009) Yet, the TBTTB survey results indicate that it is critical to determine the attitudes and concerns of teachers that will be involved in teaching these new programs (i.e. agriculture biotechnology) and address the concerns about teaching biotechnology To understand teacher motivation, teacher behavior and attitudes toward the content areas in biotechnology were examined through the survey statements measuring the attitude s and concerns.
66 Comparing the TBTTB survey results to that of previous surveys, reveal s that there has not been much progress in changing the attitudes of teachers towards teaching agriculture biotechnology (Garrett, 2009; Kidman, 2 009; Kwon & Change, 2009; Mowen et al., 2007 a ; Mowen et al. 2007 b ; Wilson, et al. 2002) Mowen et al. (2007 a ) reported that a positive relationship existed between the knowledge of the teachers and attitudes toward biotechnology. Even though teachers expressed favorable attitudes toward biotechnology t he data were insufficient to fully explore the relationship between attitudes and kno wledge considering the lack of a true random sample at the state, regional and national level (Mowen et al 2007 a ). In the TBTTB survey, we followed the recommendation of Mowen et al. (2007 a ) to complete a truly random sampling at the state level with Florida agriculture science teachers and measured attitudes and concerns of teach ing agriculture biotechnology. The TBTTB survey results also indicate d that there were three major issues and concerns that teachers expressed about teaching biotechnology. These include d : 1) concerns about the time needed to teach biotechnology concepts; 2) teacher concerns about student attitudes toward biotechnology; and 3) concerns that deman ding teacher schedule s were preventing them from integrating biotechnology concepts into the classroom. To further examine these concerns demographics of current Florida agriculture education teachers were co llected. Over half of the teachers were fema le and taught high school. In contrast to the Mowen et al. (200 7 a ) study of Texas educators, 78 % were male. Less than half of the teachers responding (45.2%) were currently teaching biotechnology, the other teachers were not cur rently teaching biotechnolog y. T he
67 average years of teaching biotechnology in high school was only 5.1 years, and 3.1 years at the middle school. Currently in Florida, the agriculture biotechnology program is ap proximately 5 8 years old. P rogram age should be taken into consideration as it could be a factor in fluencing teacher attitudes and concerns about the implementation of biotechnology. The purpose of O bjective # 2 was to determine Florida agr iculture biotechnology teacher attitudes and perceived concerns (barriers) to integrati ng agriculture biotechnology in the classroom. In North Carolina, Kentucky, Tennessee ,Texas, and West Virginia the implementation of biotechnology has been documented (Aziz et al., 2009; Kwon, 2009; Mowen et al., 2007 b ; Boone et al ., 2006; Wilson et al., 2002). These studies were important to identify possible barriers that may be currently affecting the implementation of biotechnology in Florida classrooms. Over two thirds of the teachers responding to the survey in Florida expressed the need for enhanced instru ction, supplemental materials, instructional methods and specific laboratory acti vities to teach biotechnology. Sixty seven (67%) of teachers reported that they had major concerns for implementing biotechnology and the time requir ed to teach biotech nology. To determine if there were differences in attitude scores between teachers that currently teach biotechnology and those who do not teach bi otechnology, teachers were divided into two subgroups. Less than half (45%) of the teachers surveyed currentl y taught biotechnology. There was no significant difference between attitudes of teachers that currently teach biotechnology and those that do not. This is an important point as even those w ho are teaching biotechnology did not beli eve that they were suffi ciently prepared to teach biotechnology in their classroom. S imilar teacher perceptions were
68 reported by Wilson et al. (2002) and Boone (2006) While these studies were conducted in states other than Florida, our current results indicate that teacher perce ptions have not significantly improved. Additionally, agriculture education teachers, regardless of subgroup, indicated strong agreement on barriers of time constraints, methods of instruction and stude nt attitudes on biotechnology. The major difference b etween the group th at taught biotechnology and those that did not teach biotechnology was re garding instructional methods. Teachers that are currently teaching biotechnology have adapted instructional methods; project based learning, direct instruction, co operative learning, and hands on learning and laboratory exercises to the available biotechnology curriculum that is currently available in the marketplace. Therefore, those teachers with more experience in teaching biotechnology, may seek out additional t raining to refine their skills and methods as recommended by Kwon and Chang, (2009). This training will motivate teachers to develop and improve their competency level in biotechnology. To further investigate teacher attitudes on biotechnology content kno wledge in Objective # 4 it was determined that there were differences in the number of courses taken in pre service training and the number and length of in service training workshops. There was a significant di fference between groups, (p = 0 .025) in the a ttitudes of teachers that had pre service courses and those that did not. Only 16% of the teachers completing the survey had one course or more in their pre service training pertaining to biotechnology. There wa s a perceived need to have pre service course s in agriculture biotechnology for undergraduate students that were preparing to be agricul ture education teachers. This pre service training is necessary to fully prepare
69 them for implementing biotechnology as future educators as suggested by Scott et al. 2006 and Brown et al. ( 1998) Kwon and Chang (2009) reported that teach ers expressed a motivation to integrate current biotechnology curriculum through Career and Technical education classes ( e.g. Agriculture Education, Technical Education in Florida). Expectancy value theory explains that e fficacy expectations encompass the question whether or not one can effectively perform the behaviors necessary to produce the outcome (Eccles & Wigfield, 2002) It was suggested that teacher professional development programs emphasizing biotechnology content should include hands on activities implemented during professional development workshops at the dis trict, and state levels. The majority of Florida agriculture educa tion teachers indicated that they did not complete any pre service courses in biotechnology and less than half of the teachers had not participa ted in any in service training. Wilson et al. (2002) similarly reported that half of the teachers surveyed in No rth Carolina have never attended a training in biotechnology with less than half of teachers receiving training specific to agriculture biotechnology. It is important to investigate if training can increase content knowledge and skill and increase teacher motivation to integrate biotechnology. Aziz et al. (2009) noted an increase in teacher confidence relat ing to biotechnology after attending workshops with hands o n biotechnology train ing. Offering biotechnology in service workshops in Florida could have a similar positive effect on teachers finding biotechnology useful and build teacher efficacy However, Mowen et al. ( 2007 a ) reported th at only 36% of the experienced teachers surveyed completed a biotechnology workshop. This emphasizes the need to motivate teacher s to participate
70 in in service training by providing funding for travel and registration expenses as well as compensation for time. Increasing both pre service and in service training will facilitate the wide sp read adoption of biotechnology. The number of Florida teachers reporting receiving in service tr aining in biotechnology was 50% with this training mostly consisting of only one day workshop s Given the broad multi disciplined knowledge base n ecessary to teach biotechnology there is a need to develop and present multi day workshops focusing on specific content areas and skills. The training needs of Florida agriculture bio technology teachers parallel those of other agriculture educators around the country. Teachers need to attend training and complete more than one workshop to fully comprehend and teach b iotechnology Therefore, teachers should be receiving additional in service training to increase their confidence level and motivation to integrate biotechnology in to the classroom. The result s of the survey reveal that barriers continue to exist in teaching and integrating biotechnology in the agriculture science classroom. Development of comprehensive and detailed p lant biotechnology teaching modules consisting of a lecture overview, lesson p lan, a student hands on laboratory exercise, lab report guidelines and rubrics need to be developed to alleviate some of the concerns of time necessary to teach the concepts New instructional methods and change s student attitudes toward biotechnology. are likewise required. These modules must be designed for use by students and administered easily by teachers. Consequently, the availability of these teaching modules could remove some of the perceived barriers to teaching plant biotechnology in Florida.
71 Co nclusions Understanding of the relationships between teacher attitudes, knowledge, and confidence in developing strategies is important in motivating teachers to implement a biotechnology curricul um. Results of the TBTTB survey indicate that Florida agricu lture science teachers have concerns regarding the excessive amount of time needed to prepare for and teach plant biotechnology. They also have concerns about the methods needed to teach and implement biotechnology concepts and skills. The results of this survey indicate the need to address the concerns of time, equipment availability and content knowledge in biotechnology These concerns can be addressed by develop ed teaching modules which : 1) have a hands on component; 2) requir e minimum equipment ; 3) lim ited preparation time; 4) hav e a high probability of success; 5) can be taught in one or a se ries of 50 minute class periods; and 6) include all background content materials. Finally, to address training gaps and increase teacher efficacy and motivation to teach biotechnology, workshops specific to plant biotechnology are needed in which teachers are shown how to integrate these teaching modules into their curriculum These trainings should occur during pre service training and in service professional devel opment opportunities for teachers in agriculture education. Creating these opportunities will overcome teacher attitudes and concerns about teaching biotechnology and provide an opportunity to increase biotechnology concepts, content knowledge and skill A ddressing the gaps in teacher training will help build teacher content knowledge not acquired in pre service college courses or other in service biotechnology trainings.
72 Recommendations generated from this study include the following : Supervisors, teach ers, school district administration Florida Department of Education and UF/FAMU seek funding in the Florida legislative budget, and through federal educational grants for curriculum resources equipment and in service training that make teaching content a nd laboratory skills affordable for all teachers in agriculture biotechnology. Florida university and extension faculty, and biotechnology industry members, agribusiness trade organizations, can be advocates for direct funding Additionally, grants from t he National Science Found ation, can help provide in service training for all agriscience teachers on biotechnology concepts and curricula to help teachers integrate biotechnology into their classrooms. In addition, pre service education in biotechnology ne eds to be required for new agriculture teachers entering the field. Curriculum development should include standards based instruction specific to biotechnology infused with blended instruction methods to help increase content knowledge for both teacher and student. In addition, the materials need to be affordable and easily adapted to classrooms that lack traditional laboratories and are not fully equipped or funded. A study should be completed on the state, and national level on the mo st cost effective methods to train agriculture biotechnology teachers and the topics that need to be taught specific to plant and animal biotechnology. This could be accomplished by completing replicated studies of bio technology training workshops.
73 Table 2 1. Survey re sults of teacher beliefs about teaching biotechnology Statement SA% A% N% D% SD 1) I am concerned about the time needed to teach biotechnology concepts. 30.9 67.3 36.4 25.5 1.8 5.5 2) I would like to know how my job will change when I begin teaching biotechnology. 27.3 47.3 20.0 30.9 10.9 10.9 3) My present schedule is preventing me from learning too much about integrating biotechnology concepts. 29.1 63.6 34 .5 18.2 10.9 7.3 4) I would like more information on the time required to learn biotechnology 21.8 56.4 34.5 29.1 7.3 7.3 5) I would like to know how the integration of biotechnology instruction is better than the methods I presently use or plan to employ when I do my job. 24.1 66.7 42.6 20.4 7.4 5.6 6) I know of several methods and approaches of how to teach biotechnology. 9.3 50.0 40.7 12.8 12.8 9.0 7) I would like to know what biotechnology will require of me in the immediate future. 31.5 64.8 33.3 25.9 7.4 1.9 8) I would like to coordinate my efforts in learning about biotechnology with my fellow teachers. 25.9 63.0 37.0 25.9 7.4 3.7 9) I would like to know how to supplement and enhance biotechnology instruction. 42.6 77.8 35.2 16.7 3.7 1.9 10) I am concerned about my students' attitudes toward biotechnology. 29.6 57.4 27.8 22.0 14.8 5.6 omewhat agree combined. Note Likert Scale: SA= Strongly agree, A = Somewhat agree, N= Neither agree or disagree, D = Somewhat disagree, SD = Strongly disagree
74 Table 2 2. Teachers attitudes and concerns about teaching biotechnology subgroup per centages. Question #7: Do you teach biotechnology in your Agriculture Education classroom? Yes n=27(0) No n=28(3) Attitude Statement SA% A% N% D% SD% 1 I am concerned about the time needed to teach biotechnology concepts. Yes 25.9 48.5 18.5 3.7 3 .7 No 32.1 25.0 32.1 0.0 10.7 2 I would like to know how my job will change when I begin teaching biotechnology. Yes 3.7 18.5 48.1 14.8 14.8 No 50.0 21.4 14.3 7.1 7.1 3 My present schedule is preventing me from learning too much about integratin g biotechnology concepts. Yes 0.0 48.1 22.2 11.1 18.5 No 0.0 21.4 14.3 10.7 53.6 4 I would like more information on the time needed to teach biotechnology Yes 0.0 37.0 37.0 7.4 18.5 No 0.0 32.1 21.4 7.1 39.3 5 I would like to know how the i ntegration of biotechnology instruction is better than the methods I presently use or plan to employ when I do my job. Yes 0.0 48.1 22.2 7.4 22.2 No 0.0 37.0 18.5 7.4 37.0 6 I know of several methods and approaches to teach biotechnology. Yes 0.0 55. 6 14.9 11.1 18.5 No 0.0 25.9 22.2 25.9 25.9 Note : Likert Scale: SA= Strongly agree, A = Somewhat agree, N= Neither agree or disagree, D = Somewhat disagree, SD = Strongly disagree Yes = I currently teach biotechnology, no = I do not currently teach b iotechnology
75 Table 2 2. Continued Attitude Statement SA% A% N% SD% D% 7 I would like to know what biotechnology will require of me in the immediate future. Yes 0.0 40.7 22.2 7.4 29.6 No 0.0 25.9 29.6 7.4 37.0 8 I would like to coordinate my efforts in learning biotechnology with my fellow teachers. Yes 0.0 29.6 29.6 7.4 33.3 No 0.0 44.4 22.2 7.4 25.9 9 I would like to know how to supplement and enhance biotechnology instruction. Yes 0.0 37.0 11.1 0.00 51.9 No 0.0 33.3 22.2 7.4 37. 0 10 I am concerned about my toward biotechnology. Yes 29.6 25.9 29.6 11.1 3.7 No 29.6 29.6 14.8 18.5 7.4 Note : Likert Scale: SA= Strongly agree, A = Somewhat agree, N= Neither a gree or disagree, D = Somewhat disagree, SD = Strong ly d isagree Y es = I currently teach biotechnology, n o = I do not currently teach biotechnology
76 Table 2 3. Mean response of agriculture attitudes of teachers teaching biotechnology ot teaching biotechnology. Question #7: Do you teach biotechnology in your agriculture education classroom? Item Statement Teaching Biotechnology Not Teaching Biotechnology 1 I am concerned about the time needed to teach biotechnology concepts 2.11 0.97 2.37 1.25 2 I would like to know how my job will change when I begin teaching biotechnology. 3.19 1.04 1.96 1.29 3 My present schedule is preventing me from learning too much about integrating biotechnology concepts. 3.00 1.18 4.00 1.27 4 I would like more information on the time required to learn biotechnology 3.07 1.11 3.59 1.31 5 I would like to know how the integration of biotechnology instruction is better than the methods I presently use or plan to employ when I do my job. 3 .04 1.22 3.44 1.34 6 I know of several methods and approaches of how to teach biotechnology. 2.93 1.21 3.52 1.16 7 I would like to know what biotechnology will require of me in the immediate future. 3.26 1.29 3.56 1.25 8 I would like to co ordinate my efforts in learning about biotechnology with my fellow teachers. 3.44 1.25 3.15 1.26 9 I would like to know how to supplement and enhance biotechnology instruction. 3.67 1.44 3.48 1.31 10 I am concerned about my students' attitudes t oward biotechnology. 2.33 1.14 2.44 1.31 Note : Likert Scale: 1= strongly agree, 2, somewhat agree, 3 = neither agree or disagree, 4= somewhat disagree, 5 = strongly disagree
77 Table 2 4. Difference in attitude mean scores of agricult ure scie nce teachers that teach biotechnology and teachers that do not teach biotechnology. Statement Sum of Squares df Mean Square F Sig. Attitudes toward b iotechnology between g roups 0 .116 1 0 .116 0 .410 0 .525 Attitudes toward b iotechnology within g roups 15 .043 53 0 .284 Total 15.159 54 Table 2 5. Participant pre service and in service training experiences in teaching biotechnology. Category Trainings/Courses Total # of Teachers Pre Service Courses None 1 Course 2 C ourses 3 Courses 4 Courses 5 or more 48(77.4%) 10(16.1%) 1(1.6%) 0 1(1.6%) 2(3.2) % In Service Training/Course Length or Time in Training No Yes 1 Day Workshop 2 Day Workshop 3, to 5 Day Workshop 1 Week Workshop 2 Week Workshop Course: College/Univ ersity Other 31(50%) 31(50%) 22(62.9%) 4(11.4%) 2(5.7%) 1(2.9%) 2(5.7%) 0(0.0%) 4(11.5%) College Coursework specific to biotechnology No Yes 56(90.3%) 6(9.7%) Teaching Biotechnology No Yes 34(54.8%) 28(45.2%)
78 Table 2 6. Differences in attitude s of agriculture science teachers on pre service and in service training preparation Sum of Squares df Mean Square F Sig. In your pre service teacher preparation program, how many courses did you take to prepare for teaching biote chnology. Between Groups 32.82 22 1.49 2. 133 .025 Have you participated in any in service professional development for teaching biotechnology? Between Groups 5.950 22 .270 1.110 .386
79 CHAPTER 3 ASSESSMENT OF PLANT BIOTECHNOLOGY TEACHING M ODULES Introduc tion Biotechnology continues to move from the laboratory and i nto both the public sector and classroom. Biotechnology is technology based on biology and harnesses cellular and biomolecular processes to develop technologies and products to improve our live s and the planet (Biotechnology Innovation Organization, 2017) Agricul ture is being radically transformed by this technology through development of a vast array of products to increase production increase disease resistance and i ncrease shelf life in produce. However, there is a lack of understanding about biotechnology and biotechnological concepts in students teachers, and the general public (Aziz, et al., 2009) There is much fear and concern that comes from not knowing the science that underpins virtually all agriculture biotechnology. To prepare students for the biotechnology workforce and identify the critical content knowledge and skills in biotechnology, development of plant biotechnology educational materials and curriculum training is needed. Society is in desperate need to know how t hings work so they can critically evaluate the risks and benefits of any biotechnological application. Teachers that are presented with new curricula or content are faced with learning and delivering this content in the most effective manner to students (Dunham et al, 2002) This can be a daunting task to a teacher who is not fully prepared to teach a rigorous scient ific and technical discipline. There are several barriers in teaching biotechnology in the high school curriculum. These involve the lack of; 1) funding for equipment and laboratory materials, 2) time to complete the laboratory activities in a standard class period 3) develop and assess teacher content knowledge
80 in biotechnology and 4) in service tr aining specific to agricultural biotechn ology applications. In this chapter, these limitations are partially addressed by developing novel teaching modules that help to alleviate some of these barriers Teachers simply do not have sufficient time to const ruct, reflect and develop content and instructional methods for delivery. They rely on curriculum resources and training to integrate their knowledge and deliver relevant instr uction to their students. C urriculum and training for plant biotechnology is gre atly needed for agriculture teachers to integrate the curriculum in the high school setting and overcome these barriers. The Teachers Belief About Teaching Biotechnology (TBBTB) survey in Chapter 2 was used to investigate the availability of professional development. It was determined that 90% of teachers did not take any college courses or specific training in biotechnology in their pre service preparation, while 50% of the teachers did not attend any type of biotechnology in service training. Research st udies by Mueller et al. ( 2015) ; Brown, et al. (2014) ; Kwon (2009); Mowen et al. (2007 a ); Scott et al. (2006) all identified the need fo r professional development and training in biotechnology. Additionally, Kwon (2009) concluded that the lack of understanding of biotechnology concepts resulted from insufficient professional development (e.g. workshops). Wilson et al. (2002) reported that teachers that have high self perceived knowledge about biotechnology were more willing to teach biotechnology. Therefore, te achers must have high efficacy which is defined as the confidence in their capability to bring about student engagement and learning (Tschannen Moran and Hoy, 2001)
81 To address the barriers of limited time, equipment and funding as well as content knowledge training in biotechnology, novel t eaching modules need to be developed to provide hands on training using pl ant tissue culture techniques. In Florida, any new teaching module designed must adhere to Florida Common Core Standards and Next Generation Sunshine State Standards (Florida Departm ent of Education, 2017). More specifically, lesson plans, assessments, and laboratory exercises must be designed to meet these standards for use with students in the classroom. Purpose and Objectives The purpose of this study was to explore agr iculture bio technology teacher knowledge levels and attitudes toward plant biotechnology by participating in a plant tissue culture training workshop. The objectives were to: 1. Develop three plant tissue culture modules designed for use in a high school agriculture cla ssroom 2. Investigate changes in teacher content knowledge in plant biotechnology following completion of three plant tissue culture modules in a professional development workshop setting. 3. e ffectiveness of the teaching modules used in the in service workshop. 4. agriculture biotechnology in the classroom.
82 Materials and Methods Participants The participants in this in s ervice workshop were agricultural science teachers that taught biotechnology in secondary school s in Florida (N= 14 ). The sampling was derived from the University of Florida Agriculture Listserv and focused on teachers that have taught agricultural biotech nology and had some desire to improve their plant biotechnology knowledge to en hance their classroom teaching. An announcement was delivered through the agriculture education listserv at the University of Florida soliciting current agriculture biotechnolog y teachers who were asked to voluntarily participate in a pilot workshop using newly designed plant tissue culture tea ching modules. This sample population was not randomized, so caution should be used in generalizing the results beyond this population. Wo rkshop Date and Location The professional development workshop was held on November 13 th 2015 at the Plant Micropropagation Teaching Laboratory at the University of Florida, Gainesville, Florida. Workshop Design Three novel hands on plant tissue culture teaching modules consisting of lesson plans and laboratory exercises were presented in an 8 hour training and professional development opportunity for teachers at the University of Florida micropropagation laboratory. Teacher partici pants were given an age nda ( Appendix C ) and three teaching lesson modules ; Micropropagation of Carnation of Shoot Culture, Micr opropagation of Parrot Feather through Shoot Organo genesis, and Plant Biotechnology of Synthetic Seed Each teaching modu le included comprehensive lesson plans with PowerPoint
83 overviews of the concepts of the three laboratory exercises (Appendix I) laboratory activities, student labo ratory reports, rubrics and pretests and posttests in tissue c ulture methods. Workshop sessions consisted of lecture overviews, laboratory equipment and sterile technique, media preparation, and laboratory exercises. At the beginning of the workshop, 14 teacher participants completed a pre test consisting of 9 multip le choice que stions on tissue culture concepts and techniques In addition, before completing the modules a focus group discussion on perceived teacher barriers to teaching agricul ture biotechnology was conducted. The teacher comments were recorded on a w hiteboard by the researcher. A copy of the discussion responses and a list of perceived barriers can be found in ( Table 3 1 ) in the discu ssion section of this chapter. These comments were not statistically analyzed, rather they provided infor mation to help the researcher design the teaching module described in Chapter 4. Next, a PowerPoint lecture overview on plant tissue culture methods and techniques was presented to the teachers and a copy of the PowerPoint was provided to the teachers in their lesson m anual (Appendix I) This manual was intended for teachers to take back to their classroom for use with their students and as a resource for teaching these lessons to their students. The content in the presentations was signifi cantly revised from laboratory exercises and overviews designed by Kane et al. (1994) while other modules were newly created The individual laboratory exercises served as templates to create associated module s Teachers were taught the methods of aseptic technique, media preparation and performed three laboratory procedures during the workshop trai ning. After completion of the three laboratory exercises
84 teachers completed the posttest containing the same multiple choice questions that were on the pretest. The data on the pretest and posttest were analyzed using IBM SPSS Statistical Software, Version 24. Development and Design of Teaching Lesson Modules The plant tissue culture teaching modules were developed using the Understanding by Design (UbD) framewo rk (McTighe & Wiggins, 2012) This framework focuses on writing a curriculum by working backward at the end of a unit to focus on the desired results to achieve the goals or standa rds. McTighe & Wiggins (2012) explained that when designing curricula, the learning performance tasks, resources and instructional methods need to be strategically placed in the un it to meet the goals or standards and enhance learning. They also explain the UbD focuses on helping students come to an understanding of important ideas and then be able to transfer that learning to other situations. The UbD approach consists of three ge neral stages. The first stage is to identify the desired results what should students know, understand and do! This include s identify the transfer goals. Teaching for transfer is defined as giving learners opportunities to apply learning in new situation s with timed feedback on their performance, meaning making, and acquisition of skills (Wiggins & McTighe, 2005 ) These standards and essential questions help learners deepen their understanding of the content. The goals and standards for the plant tissue culture modules were derived from the Florida Next Generation State Stan dards in plant biotechnology. The second stage of UbD is to determine acceptable evidence. How will we know if students have achieved the desired results and concepts? The assessment evidence needs to document and validate that learning has occurred in Stage 1. The pretest and posttest
85 were the f ormative assessment in this research study, the laboratory reports and laboratory rubrics provide the performance task assessments. The third stage in t he UbD process is to plan learning experience s and instruction by transfer. The lesson plan, overviews a nd laboratory activities in the module design incorporated the three interrelated learning goals for UbD; acquire, make meaning and transfer. This was accomplished by the hands on laboratory exercises. Teachers were guided through each tissue culture metho d and had the opportunity to ask questions and assess their performance. The design of the teaching modules was very deliberate in focusing on standard driven lesson plans that focus on teaching, skill performance, assessing for understanding, and transfe r of learning, as outlined by Understanding by Design Model McTighe & Wiggins, (2012) Most importantly the design of the teaching modules was focused on the major barriers of lack of time, equipment, and content knowledge that teachers had expressed in the survey discussed in Chapter 2. A standard lesson plan format that is commonly used by high school teachers was used to deliver the lesson to the teachers participating in the wor kshop. The lesson plan included standards, objectives, essential questions, cross curricular integration, laboratory activity, summarizing strategies, and summative and formative assessments. I designed the lesson template to follow how I would normally te ach these concepts in my own classroom and the format that is required in Florida school districts. Each lesson consisted of Florida Sunshine State Standards in Plant Biotechnology, reading and writing strategies, activation strategies, lesson instruction and a hands on laboratory activity consisting of one of the three methods of tissue
86 culture production. A well designed PowerPoint lecture overview was used to build content knowledge of each tissue culture me thod presented in the workshop. Reading, writi ng or math exercises were included in each lesson that focused on specific standards and course objectives. For example, math calculations were included in the synthetic seed lesson to calculate percent and molar solutions of the sodium alginate needed to cr eate the artificial seed coat. High resolutions photos were created for the laboratory exercises to demonstrate step by step procedures of each tissue culture method. Most importantly a major focus was to develop accurate timelines and stopping points fo r teachers in the lesson, so they could continue the following day and still have successful results. To help minimize teacher preparation my goal was to ensure that a single lesson or concept cou ld fit into a traditional 50 55 minute class period. Most sc ience tools and equipment used in the laboratory exercises could be borrowed from the traditional science department or could be purchased at a low cost to fit within limited school budgets. Teacher instruction pages and directions for media preparation, m aterial lists and suppliers for chemicals and plant material were provided for ease in ordering correct materials needed for the laboratory activities Research Design A pre test and post test experimental design was used in this study The hypothesized rela tionship studied was to compare teacher posttest scores on the same subjects before and after completing the plant tissue culture teaching modules. The study was designed to investigate the following research question: 1. Was there an increase in agriculture teacher content knowledge based on pre test /posttest performance after completing the plant tissue culture training modules ?
87 H O There was not a significant difference in mean posttest scores after completing the plant tissue culture training modules. H A There was a significant difference in mean posttest scores after completing the plant tissue culture training modules. Dependent variable: Test scores (measured out of 9 points). Independent variable: Time, which has two levels: Time point #1: Immedia tely before the start of the workshop training modules. Time point #2: After the completion of the workshop training modules. Statistical Analysis Descriptive statistics using a paired t test was used to determine significant paired observation s in content knowledge scores before and after comple ting the three novel hands on plant tissue culture teaching modules. These descriptive methods were chosen as they best suited the targeted population, Florida agricultural biotechnolo gy teachers. Paper based instruments were used to collect data on the pretest and posttest scores during the training workshop. Results To answer the research que stion: Was there an increase in agriculture teachers content knowledge based on a pre/posttest performance after completing the plant tissue culture training modules? A paired samples t test was used to determine whether there was a statistically significant mean change in scores between the pretest and posttest when participants participated in a plant tissue culture training workshop. Two outliers were detected tha t were more than 1.5 quartiles from the edge of the box plot as seen in the histogram ( Figure 3 1). Inspection of the values did not reveal them to be extreme and they were kept in the a nalysis. The assumption of normality was not
88 violated, as assessed by Shapiro p=.827 ). Teacher participants scored higher on the posttest ( M =6.538, SD = 4.08) as opposed to the pretest ( M= 4.076, SD = 1.85), a statistically significant mean in crease of 2.146 points, 95% CI [1.620, 3.302] t (13) = 6.379 p <.001, d = 1.76 (Figure 3 1). The mean difference was significantly different from zero. Teachers that attended the plant biotechnology workshop received a significant increase in their biotechnology knowledge after completing the workshop training. Figure 3 1 Mean change in test scores on teacher content knowledge following completion of overviews and three tissue culture modules Discussion To prepare students for careers in agricu ltural biotechnology it is critical that current biotechnology concepts and techniques be taught in the agriculture science classroom. Aziz et. al. (2009) emphasized that current teacher technical knowledge was critical in making their pedagogical decision s, so students develop informed views about biotechnology. It also has direct social implications as teachers play a vital role in disseminating relevant information directly to their students and indirectly to society
89 ( Schibeci ) (2003). Aziz et. al (2009 ) also contended that teachers obtaining hands on biotechnology workshop experiences not only obtain enhan ced teacher knowledge content but also benefit by developing more positive attitudes toward the usefulness of biotechnology. Given this, te acher pre paration and continual training updates are also critical to the effective implementation of biotechnology in the classroom (Zeller, 1994). A s recommended by other researchers (Aziz et al., 2009; Borgerding et al. 2013; Mowen et al. 2007 a ; and Zeller, 1994) the need for teacher professional development training in agriculture biotechnology was addressed by inviting the teachers to participate in an in service workshop taught by trained experts in the field of pl ant biotechnology at the University of Florida. These results could be useful in providing insights regarding future development of teacher biotechnology training workshops and associated instructional materials A primary ob jective of this study was to f irst develop and the n assess the effectiveness of three plant tissue culture modules plant biotechnology classroom Teachers completing the three tissue culture modules: Micropr opagation of Carnation using Shoot Cult ure, Mic ropropagation of Parrot Feather using Shoot Organog enesis, and Plant Biotechnology of Synthetic Seed demonstrated a significant knowledge gain. The change in the pretest and posttest w as significa nt with a 2.2 point increase (23.8%) in mean score These results indicate that the workshop training was effective in increasing content knowledge of teachers completing the overview and plant tissue culture modules. Furtherm ore, results confirmed that an eight hour training could improve teacher content k nowledge in biotechnology. These results are supported by Bigler & Hanegan ( 2011) ; Wilson et al., 2002) ; and Aziz et al. ( 2009)
90 who predicted that teacher content knowledge gains would occur after hands on professional devel opment training. Teachers completing the three teaching modules during the workshop at the University of Florida, expressed more confidence in teaching biotechnology In addition, teachers commented on the training required to be successfu l l y teach plant t issue culture in the classroom Teachers stated that they were usually unsuccessful when they attempted similar technique s with their students as the cultures typically become contam inated with fungi or bacteria The teachers commented that learning how to properly clean and culture the explants using proper aseptic techniques was beneficial. The teachers agreed that the effectiveness of using hands on laboratory techniques demonstrated in the workshop would be helpful when repeating these laboratory exer cises with their students. Although a formal state wide teacher perception survey was ongoing (November 2015 to April 2016; ( Chapter 2) at the time of the workshop, to immediately determine current barriers that Florida agric ultural biotechnology teachers believe d prevented biotechnology from being integrated into the classroom a round table discussion was completed with the workshop participants prior to their completing the plant tissue culture teaching modules. The intention was to use the perceived ba rriers generated during this round table discussion to further refine the design of the current and future teac hing modules to increase their effectiveness and acceptability. During the round table discussion, teacher s shared numerous barriers that impede d teaching plant bio technology in Florida (Table 3 1 ) One of the major obstacles was the difficulty of obtaining startup funds to establish a new biotechnology teaching
91 program. The lack of funding from the school district, the cost of the biotechnology e quipment, the design and set up of the laboratory, and finding biotechnology curricula, including hands on teaching modules that appropriately address the Next Generation Florida State Standards Even though biotechnology curricula are available, teachers still find it necessary to combine several instructional resource s to meet the state standards. Teachers simply do not have the time to create new and/or adapt existing curricula Another important point mentioned during the group discussion was the lack o f teacher content knowledge and the concepts and skills needed to teach biotechnology ef fectively. This was later supported in the results of the state wide agriculture b iotechnology survey (Chapter 2) completed in April 2016 in which only 50% of the teach ers surveyed had any training in biotechnology. Teachers are searching for free workshops an d training and opportunities through the university extension programs, agriculture education or supporting industries Unfortunately school distric ts are not able to fund teacher biotechnology training Teachers emphasized th at these confounding issues made it difficult to be mot iv ated to teach this discipline. Clearly, a major challenge is the limited professional development workshop opportunities for teachers to gain hands on training in biotechnology techniques that is directed towards teaching agricu ltural biotechnology. Other issues reported included the use and cost of commercially available biotechnology k its C omments from the teachers indicated that kits such as those sold by Bio Rad, are sometimes unreliable in generating useful results making t hem difficult to use and leave students unmotivated. Teachers expressed concern with the additional
92 time needed to set up a standard bio technology laboratory exer cise. F or example biotechnology kits demonstrating PCR (polymerase chain reaction), are difficult to use for teac hing in a high school classroom as up to three hours are required to set up the components of the laboratory exercise. In addition, the follow up exercise cannot be completed by the students in a 50 minute class period. The resulting failure therefore leads to a lack of teacher and student frustration learning and interest in biotechnology Table 3 1 Teacher perceived barriers in teaching bio technology in the classroom expressed by the workshop participants in November 2015 Lack of materials, and proper laboratory space provided by school district to teach Agriculture Biotechnology. Difficult to start up a biotechnology program. There is a lack of communication and agreement to which materials, equipment and curriculum resources that need to be provided to manage a biotechnology laboratory. A lack of funding at the district level to set up a biotechnology teaching laboratory. Major time co nstraints to teach extensive laboratory exercises required for biotechnology. Many teachers are required teach other agriculture or science courses in addition to biotechnology. A standard 50 minute class period is not sufficient to complete an extensive biotechnology laboratory exercise. Lack of teacher content knowledge needed to effectively teach biotechnology. Laboratories take longer than expected and require multiple hours of preparation. Reliability issues of current biotechnology laboratory kit s. Student ability levels are mixed in the classroom, many times different levels of biotechnology are taught in the same class period (example: Agriculture Biotechnology level 2 and 3) requiring different standards to be addressed at each course level. The mixed ability level of students makes it difficult to teach the same laboratory exercises effectively. Administrative support at the district level does not offer teacher training in biotechnology and it is left to the individual teacher to find other sources of training needed to teach biotechnology. The teaching modules designed for this study addressed four of the top barriers reported by teachers ten years ago (Mowen et al. 2007 b ). As mentioned earlier, this included lack of: 1) equipment; 2) cl assroom/lab space; 3) time; and 4) instructional materials, and 5) teacher knowledge. Based on the 2016 Florida state wide survey
93 (Chapter 2), responses by Florida agriculture teachers clearly indicate these barriers still exist today. These barriers were specifically addressed in the teaching modules by providing background knowledge in the form of assigned readings, overview lectures, and basic hands on plant tissue culture laboratory exercises that can be completed in a few class periods. To address the issue of cost for equipment and tools, laboratory exercises were designed to be completed using inexpensive materials. These laboratory exercises offer the possibility of being modified to adjust for available equipment and budget constraints. However mi nimal items required would include beakers, scalpels, forceps, hot plates, tissue culture vessels and access to a well maintained transfer hood. These basic laboratory tools can be used to complete the steps of aseptic technique, media preparation and inoc ulation of the plants. The space requ ired of plant tissue culture could be integrated into a standard classroom with access to sinks and a n easily assembled plant growth bench. The time constraint barrier was addressed by subdividing each of the t issue cu lture modules into time defined exercises some of which were extende d over multiple class periods. The possibility exists that the overview lecture portion could be flipped as homework resulting in more face time with the instructor, thus saving more time for laboratory activities and skill building. The effectiveness of integrating flipping into an interactive lecture of a Caladium plant tissue culture teaching module was examined in Chapter 4. Limitations in the availability of informational teaching reso urces were also addressed by providing teaching strategies in the modules including readings, media preparation recipes, equipment and material supply lists, and the sources for
94 purchasing materials were provided in the teaching lesson module notebook. Oth er resources provided to workshop participants include d perspective. These were important to provide additional resources for teachers prepare and manage their laboratories and maintain sterile technique necessary for successful plant tissue culture. Given the lack of available plant biotechnology instructional materials as expressed by teachers, teaching lesson modules were designed for both teache rs to prepare the laboratories and provide student laboratory instruction. Further, t hese teaching modules address ed the S tate of Florida standards for plant biotechnology and were purposefully designed to be standard driven and provide opportunities for l earning transfer by providing teachers and students the opportunity to apply the knowledge and skills needed for job performance. Cross curricular activities were used in the lesson planning to incorporate, reading, technical writing, math and engi neering principles incorporated in STEM education. The learning transfer was created by using teaching strategies that addressed the transfer learning suggested by McTighe and Wiggins (2012). To address content knowledge barriers each module incorporated both for mative assessment by using laborat ory rubrics and summative assessments using pretest and posttests used to measure knowledge gains in the classroom. The fourth objective of this study was to informally determine whether the overview lectures and the spec ific plant biotechnology teaching modules, completed by the teachers, alleviated some of the preconceived barriers including time constraints, content knowledge, and equipment availability to teach plant tissue culture in the
95 classroom. T hese were addresse d by creating concise overview lectures and detailed hands on laboratories that were designed to minimize classroom time constraints by providing clear stopping points and minimal preparation time. Teachers expressed gratitude and excitement after learning about the th ree methods of tissue culture. They discussed how the hands on skills and techniques were helpful in performing the laboratories in the workshop and how they would be more willing to try tissue culture with their own students. The teaching mod ules reinforced the concepts learned in the training T eachers expressed that they were grateful to have not only the plant biotechnology teaching modules but also the resources offered by the university Following the workshop two teachers contacted the instructors to obtain materials and additional information, so they could implement one of the tissue culture modules in their classroom. in retrospect, having the participants complete th ree teaching modules during one workshop proved challenging for the teacher. It became apparent to the workshop instructor that decreasing the number of modules from three to two w ould a llow more time for participant reflectio n after completing each module Allowing more time for the teachers to complete fewer modules could also further increase gain s in teacher confidence to teach the modules in their agriculture biotechnology classroom s. Conclusion and Recommendations I ncreas e in teacher content knowledg e and confidence toward biotechnology are important for adoption of biotechnology curriculum. Workshops and training s that allow teachers to practice the basic hands on skills in biotechnology are the most effective use of training time. In addition, train ing workshops will have a positive effect on
96 motivating teachers to incorporate biotechnology in to the ir classroom s To overcome barriers including lack of content knowledge, time constraints, and other barriers faced by agricultur e teachers, as noted in Table 3 1 will require a curriculum that addresses these barriers. The three plant biotechnology teaching modules developed in this study address these issues and could serve as a model for development of additional biotechnolog y teaching modules. It is recommended that similar teacher professional development workshops are needed including hands on biotechnology in service trainings. A combined effort betwee n the school districts and land grant universities to provide training i n agriculture biotechnology is desperately needed. A combination of pre service courses in biotechnology and molecular biology, and in service training will be required to build s trong agricultural biotechnology programs in the Florida's secondary classroo m s and empower students to embrace these career fields. Providing adequate funding to develop and offer this training is the challenge. Teaching Lesson Module #1 Plant Biotechnology Module #1 Micropropagation of Carnation: Shoot Culture Wendy Vidor Univer sity of Florida Unit: Plant Tissue Culture Techniques Lesson Essential Question(s): 1. What is micropropagation? 2. What is s hoot tip culture and why is it the preferred method for micropropagation? 3. Why i s sterile technique critical for micropropagation? Les son Duration (dates): Two class periods (45 50 minutes) Introduction (30 minutes), Student Procedure (30 minutes for sterilization), Explant Transfer (15 20 minutes Lab Report (15 20 minutes) Lesson Goals(Objectives): The student will be able to . 1. D emonstrate the use of aseptic technique in the laboratory. 2. Explain the function of cytokinins in plants. 3. Identify the shoot meristem tissue of carnation.
97 4. Perform shoot culture on potted carnation plants. 5. Use surface sterilization procedures to establish no dal explants. 6. Review concepts related to establishing shoot culture s of carnation 7. Identify containments in tissue culture media. KNOW (Vocabulary, big idea, facts) UNDERSTAND (Important concepts, principles, ideas) DO (How will this be accomplished)? Big Idea: To know the difference in origin of shoots produced in plant tissue culture. Vocabulary: Apical dominance Apical meristem Aseptic technique Axillary shoots Cytokinin Culture medium Meristematic tissue Carnations are one of the most economica lly important global cut flower crops. There is high heterozygosity in carnation requiring that it be vegetative ly propagated by cuttings. Carnation production quality and yield is negatively impacted by various diseases. Meristem culture can be used to eradicate disease in carnations. Shoots are clonally produced via shoot culture by production of axillary shoots. Demonstrate the use of aseptic technique using a checklist. Prepare and mix media stock solutions for micropropagation. Prepare Carnat ion nodal explants for sterilization. Manipulate the growth of plants using the plant growth regulator (NAA) Naphthalene acetic acid. Measure adventitious shoot production developing from lateral buds and growth and then graph results. Activating Stra tegy: students and activate their prior knowledge? To help students activate prior knowledge for this lesson have students complete the Flipping Exercise #1 before class or as a warm up activity. They will need to answer the three questions on the last slide as pre lab questions. Day 1 : Flipping Exercise #1 : (15 20) minutes. PowerPoint overview with pre lab questions used as a flipping exercise. What Background Knowledge is Needed? Background Knowledge: Students will need to read the Introduction paragraph provided in the student lab sheet provided in the lesson.
98 Background Information: Tissue culture in the classroom can be successful if aseptic technique and the procedures are performed correctly. In this exercise, you will be performing shoot culture or shoot culture consisting of apical meristem tissue, leaf primordia and developing leaves from Carnation (Dianthus caryophyllus). This procedure is used for clonal propagation to enhance formation of axillary shoots. This procedure works by stimulating shoot growth from lateral buds. This process is performed by disrupting the apical dominance in shoots cultured on media supplemented with cytokinin. Cytokinin is a plant growth regulator that stimulates cell division i n cell roots and shoots. The addition of this growth regulator to the medium to stimulate rapid development of shoots. The advantages of shoot tip culture include the rapid production of pathogen eradicated plants. The purpose of this laboratory exercis e is to demonstrate procedures required for in vitro establishment of carnation shoot tips. Vocabulary: Meristematic tissue: cells that can divide to form new cells which make up the plant body. In vitro propagation: Propagation of plants in culture ve ssels under controlled conditions of temperature and light on a defined culture medium. Cytokinin : A class of plant growth regulators that promote cell division and shoot development. Culture medium : A medium containing water, mineral salts, micro and ma cronutrients, organic components, vitamins, sucrose, plant growth regulators and a gelling agent used for supporting growth of plant cells, tissues or plants.
99 Figure 3 2. The shoot tip is comprised of the apical meristem and subtending leaf primordi a and developing leaves Photo courtesy of author. Materials/Handouts: Class set of the student lab handout (attached) 2 pots of Dianthus caryophyllus (carnation) Laminar flow hoods or premade tissue culture boxes Laboratory supplies (provided i n lab handou t) Handout on Stages of Micropropagation Handout on preparing carnation multiplication media PowerPoint on Micropropagation Methods (Appendix I) Rubric on Aseptic Technique Pretest and Postt est Advance Preparation: 1. Prepare the carnation multiplication me dium per the directions in the handout. (You may have advanced students prepare the medium as an additional lesson). (This should be done after school. It will need to be sterilized in the autoclave or pressure cooker) 2. Copy the Teacher Handout : Tissue Cult ure Media for Carnation. (See attachments) 3. Copy the student laboratory handout, one for each student. (See attachments) 4. Prepare the lab equipment for sterilization of the cuttings. 5. Copy the pre and post test for the unit. (See attachments) 6. Copy the lab rub rics. One on sterile technique, the second on lab report. (See attachments) Procedure and Discussion Questions with Time Estimates: Day 1: Aseptic Technique in Tissue Culture (15 20 minutes) Flipping Exercise : Power Point Presentation : Micropropagatio n Methods in Appendix I. Students can view the PowerPoint for homework or teacher can present in class. Students can take notes using a lab notebook. (10 minu tes) a short pre test should be administered to the group and teacher should collect after the Ase ptic Technique PowerPoint.
100 (10 15 minutes) Review with students Stages of Micropropagation (See image located on last page of student laboratory sheet) Day 2: Washing the Shoot Explants Teacher Preparation (20 minutes): Teacher should prepare these lab materials for washing the nodal shoot explants. Plant Material: Dianthus caryophyllus (2 3 pots from local garden center) Lab Materials : (Prepare glassware for students, one beaker for each solution and one set of everything else). Note: Have students po ur and measure each solution and gather materials from a central location that you have prepared ahead of time! 1) 95% alcohol solutions 2) 1.2% sodium hypochlorite solution containing 2 drops Tween 20 per 100 ml 3) Sterile distilled deionized water 4) Standard and large size forceps, spatula and scal pel with number 11 blade 5) Sterile petri dishes 6) 10 culture tubes containing sterile agar solidified Carnation Shoot Multiplication Medium 7) Sterile "Disinfecting Bottle" 8) Sterile graduated cylinder 9) Cheesecloth square s and rubber bands for jars. Student Activity: Homework (The night before lab activity) Flipping Exercise : Carnation Shoot Culture PowerPoint Presentation. Students will answer Pre Lab Questions on Student Lab Handout. (5 minutes) Have students answer the essential question for the day. EQ : What is the function of the cytokinin in the medium and why do w e need to use this for shoot culture? The function of the cytokinin is required in the medium for cell division and induces growth of axillary shoots of carnation. Break students into lab groups and have them get the sterilized explants from the previous day. (5 minutes) Students should assemble in groups of 2. They will need to obtain the laboratory materials listed on the lab sheet and return to the ir lab tables. (25 30 minutes) Students will measure and pour solutions and label glassware. Students will have ten culture tubes containing medium. They will sterilize ten of the nodal explants of the Dianthus caryophyllus and sterilize according to t he directions in the student lab sheet. (This should be done in the laminar flow
101 hood or clean box). Teacher should observe and grade students using rubric on the proper use of aseptic technique. (See attachments) (5 min) Students will need to soak the ex plants in sterile water with plant preservative medium (PPM) at 1ml/L of distilled water and place lids securely on tissue culture cup covering the explants. (Students can then leave them in the hoods to soak overnight) Day 3: (5 minutes ): Have students answer the essential question. EQ: Why is it important not to touch the scalpel or forceps on any surface in the laminar flow hood? (5 minutes) : Have students assemble in groups of 2 and go to the laminar flow hoods or pre assembled clean box. Have them take their cleaned explants into the hood and place the tools in the glass bead sterilizer set at 260 C. Please remind them to take them out after a two minute and place them inverted in the rubber stoppers until they are cool. (20 30 minutes ): Student s should take ten culture tubes containing 12 ml medium to the transfer hood (Make sure they into the medium). Have students take one nodal explant from the jar and pick up by the stem se ction with sterilized forceps and scalpel cut off any bleached tissue at cut ends of explants. Gently inoculate the bottom of the explant slightly into the medium. Cap the culture tube immediately after inoculation and place the tools back into the glass b ead sterilizer. (5 minutes ): Clean work surface with water moistened towel then spray down hoods with 95% ethanol. Turn off glass bead sterilizers, remove scalpel blades and dispose of in container. Clean and sterilize all tools and glassw are. Students s hould take post test. Assessment Suggestions: Pretest and Postt est Flipping Exercise: Grade Questions Rubrics: Sterile Technique, Lab Report 1. What is the purpose of adding cytokinin to the medium? 2. Why do we need to include meristematic tissue on the nod al explants? Exit Ticket: Ticket out the Door 1. How can we use micropropagation to limit diseases in carnation? 2. Why would this be important in the commercial
102 production of carnations? Lab Report: Questions for Discussion at the end of the lab report. Students can continue the lab by transferring the shoots that are produced after 3 weeks into the greenhouse (Stage 4). They may also write up and test a hypothesis for an agriscience research project or proficiency. Students may also prepare the carna tion medium and experiment with different types of cytokinin light levels, etc. for an SAE or proficiency. Resources and References: 1. Ali, A., H. Afrasiab, S. Naz, M. Rauf and J. Iqbal. 2008. An efficient protocol for in vitro propagation of carnatio n ( Dianthus caryophyllus ). Pak. J. Bot. 40: 111 121. 2. Ahmed, A.A., E.A.H. Khatab, R.A. Dawood and A.M. Ismeil. 2012. Evaluation of tip culture and thermotherapy for elimination of carnation latent virus and carnation vein mottle viru s from carnation plants Interna t. J. Virology 8: 234 239. 3. Cacas, J.L. E. Olmos and A. Piqueras. 2010. In vitro propagation of carnation ( Dianthus caryophyllus L.). Methods Mol. Biol. 589:109 116. 4. Danial, G.H., A.N. Yousif, M.S. Omar. 2009. Clonal propagation of Dianthus caryo phyllus L. through tissue culture. 2 nd Kudistan Confer. Biological Science (special issue) J. Duhok Univ. 12: 91 95. 5. George, E.F. 1993. Plant Propagation by Tissue Culture. Part 1. The Technology. Exegetics, Ltd., Edington. 6. Jones, J.B. 1986. Determining markets and market potential of horticultural crops. In : Tissue Culture as a Production System for Horticultural Crops. pp. 175 182. R.H. Zimmerman, R.J. Griesbach, F.A. Hammerschlag, and R.H. Lawson (eds.). Martinus Nijhoff Publishers, Boston. 7. Mangal, M., S.V. Bhardwaj and A. Handa. 2004. Production of virus free carnation pla nts through heat therapy. Def. Sci. J. 54: 53 56. 8. Miller, L.R. and T. Murashige. 1976. Tissue culture propagation of tropical foliage plants. In Vitro 12:797 813. 9. Morel, G. and C Martin. 1952. Guerison de dahlias atteints
103 d'une maladie a virus. C.R. Acad. Sci., Paris 235:1324 1325. 10. Morel, G. 1964. Tissue culture: A new means of clonal propagation of orchids. Amer. Orch. Soc. Bull. 33:473 478. 11. Pierik, R.L.M. 1987. In Vitro Cultu re of Higher Plants. Martin Nijhoff Publishers, Boston. pp. 172 181. 12. Roest, S. and G.S. Bokelmann. 1981. Vegetative propagation of carnation in vitro through mult iple shoot development. Scient ific Hort. 14: 357 366. 13. Saher, S. A, Piqueras, El. Hellin, an d E. Olmos. 2005. Prevention of hyperhydricity in micropropagated carnation shoots by bottom cooling: implication of oxi dative stress. Plant Cell Tiss ue Org. Cult. 81: 149 158. 14. Styer, D.J. and C.K. Chin. 1984. Meristem and shoot tip culture for propagati on, pathogen elimination, and germplasm preservation. Hort. Rev. 5:221 277. 15. Yadav, M.K., A.K. Gaur and G.K Garg. 2003. Development of suitable protocol to overcome hyperhydricity in carnation during micropropagation. Plant Cell Tiss ue Org. Cult 72: 153 156. 16. Zhang, J., X. Wu, Y. Bi, Y. Wu, G. Lin, Y. He and Z. Mao. 2013. First report of Fusarium proliferatum infecting carnation ( Dianthus caryophyllus L.) in China. J. Phytopathology 161: 850 854. 17. Ziv, M., G. Meir and A.H. Halevy. 1983. Factors influenci ng the production of hardened glaucous carnation plantlets i n vitro, Plant Cell Tiss ue Org. Cult. 2: 55 65 ESE Student Accommodations/ Modifications for Lesson: What accommodations/ modifications/ differentiation did you use? Accommodation Type: Who/When Used or Offered: Think Pair Share Cooperative Groups Class Discussion Collaborative Lab Groups During Activation Strategy During Lab and Activation Lesson Reflection: Note changes to be made, reflections for future use of lesson, and other notes here. Florida 2016 17 CTE 15.0 Conduct scientific investigation and apply
104 Curriculum Frameworks and Standards Florida Standards: Reading and Writing Florida Next Generation Math and Science Standards results. 16.0 App ly genetic principles to agricultural production. 18.0 Demonstrate laboratory skills as applied to biotechnology 18.01 Maintain and interpret biotechnology laboratory and production records. 18.02 Operate laboratory equipment and measurement devices. 18.03 Demonstrate aseptic techniques in the biotechnology laboratory. 18.05 Prepare buffers, reagents, solutions and media. 25.0 Demonstrate proper tissue/cell techniques. 25.01 Prepare a lab using aseptic techniques for a tissue culture facility. 25.02 Descri be the effects of growth hormones on tissue/cell culture. 25.03 Demonstrate the use of sterile instruments and materials. 25.04 Produce plants using tissue culture methods and prepare a written report of data and results. Demonstrate methods of micropropag ation of plants. Demonstrate methods of plant production. 35.03 Prepare and mix stock solutions of media for micro propagation. 01.02 Determine the central ideas or conclusions of complex process, phen omenon, or concept; provide an accurate summary of the text. LAFS.910.RST.1.2 01.03 Follow precisely a complex multistep procedure when carrying out experiments, taking measurements, or performing technical tasks, attending to special cases or exceptions d efined in the text. LAFS.910.RST.1.3 01.02.01 Determine the meaning of symbols, key terms, and other domain specific words and phrases as they are used in a specific scientific or technical context relevant to grades 9 10 texts and topics. LAFS.910.RST.2.4 SC.912. L.14.10: Discuss the relationship between evolution of land plants and their anatomy. SC.912. N.1.7: Recognize the role of creativity in constructing scientific questions, methods and explanations.
105 SHOOT CULTURE OF CARNATION STUDENT LABORATORY SHEET Purpose: The primary purpose of this laboratory exercise is to demonstrate procedures required for the successful isol ation and in vitro establishment by using nodal explants. MATERIALS AND PROCEDURES Plant Material : Carnation ( Dianthus caryophyllus ) potted plants. Materials : 1) 95% alcohol solutions 2) 1.2% sodium hypochlorite solution containing 2 drops Tween 20 per 100 ml 3) Sterile distilled deionized water 4) Standard and large size forceps, spatula and scalpel with No. 11 blade 5) Sterile petri dishes 6) 10 culture tubes containing sterile agar solidified Carnation Shoot Multiplication Medium 7) Sterile "Disinfecting Bottle" 8) Sterile graduated cylinder SC.912. L.16.17: Compare mitosis and meiosis. S C.912. L.14.1: Describe the scientific theory of cells SC.912.14.7: Relate the structure of each of the major plant organs and tissues to physiological processes. MAFS. 912.N Q.1.2: Define appropriate quantities for descriptive modeling. MAFS. 912.A CED.1. 3 Represent constraints by equations or inequalities, and by systems of equations.
106 Figure 3 3 Materials used to complete exercise; (A) Sterile wash bottle;(B) Disinfecting bottle; (C) Tween 20 (D) Graduated cylinder with dilute bleach November 13, 2015. Photo courtesy of author. Figure 3 4 Tissue culture tools; (A) No.11 blade scalpel, forceps; (B) rubber stopper to hold scalpel and forceps, and glass bead sterilizer November 13, 2015. Photo s courtesy of author. Procedures : 1) Obtain several carnation shoots from the potted plants. On a cutting board use a scalpel to cut the shoots into nodal explants making sure to leave stem tissue above and below the nodes. A B
107 Figure 3 5 Carnation shoot node explants with leaves removed (far right) November 13, 2015. Photo cou rtesy of author. 2) Trim the leaf blades at each node leaving about 10 mm of the blade attached to the stems. Be sure to use forceps to handle the shoots. 3) Place carnation nodal segments into a plastic cup, cover with cheesecloth and fasten with a rubber ban d. 4) Remove surface debris on nodal segments by rinsing in flowing tap water for 10 minutes. ( PLEASE BE SURE TO USE COLD WATER ). Figure 3 6 Carnation shoot explant rinsing procedure November 13, 2015. Photo s courtesy of author. 5 ) Wipe down trans fer hood work space and side walls with 95% alcohol. 6) Make sure that the glass bead sterilizer is turned on. The core temperature of the glass bead steril izer should be at least 250 C. Place shoot explants into TC cup with water Cover with che ese cloth and secure with rubber band Rinse under flowing tap water
108 7) Sterilize scalpel and forceps in the glass bead sterilizer fo r two minutes. Insert instrument handle bases in the holed rubber stopper. Allow to cool. 8) Prepare the "Disinfecting Solution" by adding 85 ml sterile water and then 15 ml Clorox concentrated bleach (containing 8.25% sodium hypochlorite) into a gradua ted cylinder. Add two drops of Tween 20. This solution will contain ~1.2% sodium hypochlorite. 10) After the 10 minute rinsing period, remove cheesecloth and gently pour off water and bring the tissue culture cup into the transfer hood. Transfer each nodal s egment from the cup using the sterile forceps and place into a sterile "Disinfecting Bottle" and add enough "Disinfecting Solution" to completely cover the nodal explants. Recap and cover cap with the foil and then place bottle on the gyratory shaker for 8 12 minutes at 80 100 rpm. Check for bleaching of the cut ends of the nodal explants. Figure 3 7 Sequential steps to surface sterilize the Carnation shoot explants November 2013. Photo s courtesy of author. Transferring rinsed shoot tip explants into disinfection bottle Add Tw een 20 to diluted bleach Add diluted bleach to shoot explants Add 70% ethanol to sterilization bottle for 1 minute Drain off alcohol solution Rinse tissue with sterile water Shoot explants shaken in bleach solution for 8 minutes Shoot explants in dilute bleach
109 10) IN THE TR ANSFER HOOD decant the disinfecting solution into the cup and immediately add sterile water and soak with frequent gentle agitation for 1 minute and repeat twice. *You can stop at this point if there are not enough class times to inoculate the explants Make sure to add a drop or two of plant preservative medium (ppm) at 1ml/L (2 drops) to the sterile water and recap the jar with the explants. This can be held at this step until the following class period. 11) Take the culture tube with prepa red medium to the hood and disinfect the outside of the culture tube with alcohol before placing in the laminar flow hood. 12) Remove the sterile explants from the disinfecting bottle and place on a sterile petri dish half. Trim off bleached ends of the nodal explants. Take one explant with the long forceps and pick up by the leaf bases. Place the explant into the media so that the end of nodal explants is submerged into the medium. The cut off bases should be above the medium. 13) Seal culture tubes with one layer of sealing film, label, date and then place in racks on the culture bench. 14) Turn off glass bead sterilizer and wipe off hood work surface with a paper towel moistened with water then spray work surface and sidewalls with 95% eth anol. Turn hood light off. Drain off bleach solution Rinse bleach tissue with sterile Surface sterilized shoot ready water 3x ready for final trimming Figure 3 8 Rinsing procedure for Carnation shoot node explants November 13, 2015. Photo s courtesy of author.
110 RESULTS Record weekly observations on your cultures for 6 weeks. Be sure to compare differences in the g rowth responses between nodal explants from the potted plants. At the number of shoots. Note the presence of abnormal growth. Construct a well designed table or graph o f your data. QUESTIONS FOR DISCUSSION What sources of error occurred in this experiment? What other plants can be produced using shoot culture? What are some of the advantages of producing plants using micropropagation? Scientific Method Laboratory R eport NAME: _________________________________ DATE: _______________ Purpose or Problem What is your experiment about or what question are you trying to answer? ______________________________________________________________________ __________________ ____________________________________________________ ______________________________________________________________________ __________________ Hypothesis Materials: (Include all materials needed and the quantity). 1. 2. 3. 4. 5. 6. 7. 8. 9. 10.
111 Procedur es (Write out simple step by step procedures of the experiment). 1. 2. 3. 4. 5. Results and Data Collection Record weekly observations on your cultures for 6 weeks. Be sure to measure the number of shoots produced from lateral buds. Depending on explan t response percentage of surviving explants; 2) the mean number of shoots produced per nodal explants in each culture tube. Note the presence of abnormal growth. Construct a well designed table or graph of your data. DATA TABLE Date Measured: Culture Tube # Total # of axillary shoots per nodal explant Average # of total axillary shoots per explant. (Divide total in 1 st column by 10) Observations: Abnormal Growth, contamination, etc. Week #1: (Dates) 1 2 3 4 5 6 7 8 9 10 Week #2: 1 2 3 4 5 6 7 8 9 10
112 Week #3: 1 2 3 4 5 6 7 8 9 10 Week #4 1 2 3 4 5 6 7 8 9 10 Week #5 1 2 3 4 5 6 7 8 9 10 Week #6 1 2 3 4 5 6 7 8 9 10
113 Conclusion: What happened during the experiment? Did y ou prove or disprove the hypothesis? What sources of error did you find and why do you think the error occurred? According to the data, describe how the control group and variables responded. Student Name: ________________________ Class: ____ ___________________ Aseptic Technique Rubric: Not Acceptable 1pt Needs Improvement 2 pts Acceptable 3 pts Clean/Sterilize Hood Does not clean hood in a manner that is acceptable. Seems lost or confused. Cleans hood to acceptable degree. Hood is not co mpletely sterilized based on criteria described in lecture/reading. Cleans hood exactly as learned. Hood is sterilized and ready to use. Gathers Equipment Does not gather needed equipment. Needs reminding of materials needed for skill, seems lost. Gathers most materials. Identifies items are needed or excess materials gathered. Gathers all necessary equipment to correctly perform skill. Place items in flow hood correctly Improper placement of items in flow hood. Student asks questions regarding the proper placement of items in the flow hood. Recognizes improper placement of items in flow hood. Identifies problem, corrects problem or explains proper procedure. Places all items in the flow hood correctly for use. Disinfect tools Places tools on surface of hood and not in the holder or sterilizer. Places tools safely in holder sterilizer between use. Places tools safely in holder and sterilizes equipment as instructed. Clean Hood after use Hood is left unclean after use. Hood is partially cleaned but instructions completely. Hood is cleaned completely according to instructions. Gowns/Attire (P.P.E.) Improper preparation, attire, or hygienic standards. Preparation, attire, hygienic standards noted to instruc tor is corrected or excused Appropriate preparation, attire, and meets hygienic standards Comments:
114 Carnation Shoot tip Initiation and Multiplication Medium Simplest Method for Medium Preparation by Instructor Note: Order media components from Phytotechnology Laboratories (product numbers included). (L467) Linsmaier & Skoog Basal Medium W/Sucrose Agar. Add 41.43 g per liter water. (K483) Kinetin liquid (1mg/ml) Add 2 ml to 1.0 liter of prepared medium. Napthalenaeacetic Acid (NAA; 1mg/ ml). Add 0.1 ml to 1.0 L of prepared medium. Adjust pH to 5.7. Heat to completely melt agar & dispense 12 ml per 150 X 25 mm glass culture tube. Sterilize by autoclaving. Cool tubes on 45 slant (for culture initiation). Note: If you leave the nodal e xplants sit overnight in the disinfection bottle you will need to order Plant Preservative Mixture from Plant Cell Technologies. http://www.plantcelltechnology.com/pl ant preservative mixture ppm 100 ml/ Order: 100ml bottle and use 1ml/L or approximate 2 drops to the disinfectant bottle with the nodal explants. (This is not a component of the media preparation)
115 PowerPoint Overview of Carnation Module
119 Teaching Lesson Module #2 Student Laboratory Sheet MICROPROPAGATION OF PARROT FEATHER ( MYRIOPHYLLUM AQUATICUM ) BY SHOOT ORGANOGENESIS FROM STEM INTERNODES INTRODUCTION The most commonly used and reliable micropropagation method for the in v itro propagation of most horticultural crops is shoot culture (enhanced axillary branching). The primary feature of this pathway is that shoots are produced from explants such as shoot tips, or nodes, which possess pre existing, shoot meristems Plants are generated following subsequent multiplication via enhanced axillary branching and rooting of these shoots (5). Shoot regeneration can also occur adventitiously from tissues with no pre existing meristems. This process is called shoot organogenesis There are two types of shoot organogenesis: indirect and direct Indirect shoot organogenesis involves the development of shoots indirectly from an intervening callus derived from the explant. Indirect shoot organogenesis may be less desirable for commer cial clonal propagation because the plants produced by this method may exhibit greater genetic variation (1,2,4). During direct shoot organogenesis, shoot meristems develop directly from the explant. Adventitious meristems arise from the epidermis or sub jacent layer and usually are of single cell origin (5). Direct shoot organogenesis is more desirable because the probability of genetic variation among the plants produced is lower. However, one limitation to this method is that chimeras cannot be propag ated true to type through production of adventitious shoots (5,8). Many ornamental genera including Saintpaulia (African violet), Gloxinia Episcia (Carpet plant) and Begonia can be readily propagated in vitro by shoot organogenesis from pieces of interno de, leaf blade or petiole explants (3,9). In these genera, shoots can arise directly from the epidermis, but indirect shoot organogenesis may occur simultaneously in the same culture if callus is promoted. This problem usually occurs following inappropri ate selection of the type or relative level of cytokinin or auxin added to the medium. In this laboratory exercise, shoot organogenesis will be demonstrated using stem internodes of Parrot feather, ( Myriophyllum aquaticum ), a popular ornamental aquatic pl ant used in aquaria and water gardens (6,7). The influence of the cytokinin 2 isopentenyladenine (2 iP) on shoot organogenesis will be examined.
120 MATERIALS AND PROCEDURES Plant Material : Sterile shoot cultures of Myriophyllum aquaticum Materials : 1) 5 shoot cultures of Myriophyllum aquaticum 2) 95% alcohol 3) Forceps and surgeon's scalpel with No. 10 blade 4) Sterile petri dishes 5) 5 vials of the control medium and +2 iP medium ( green color medium ). 6) Sterile distilled deionized water Procedures : 1) Wipe down transfer hood workspace and sidewalls with 95% alcohol. 2) Turn on glass bead sterilizer and allow to heat to 250 C. 3) Obtain holed black rubber stopper; place in hood and spray with 95% alcohol. Allow alcohol to evapora te. 4) Sterilize scalpel and forceps in the glass bead sterilizer. Insert base of tool into the holed rubber stopper or place on a piece of clean foil in the laminar flow hood. 5) Obtain 5 shoot cultures of Myriophyllum aquaticum 6) Spray each tube with 95% alcohol and wipe dry and in the transfer hood remove the closure from one of the tubes. 7) Using sterile technique remove the shoot from the tube by gently grasping it about halfway down the stem with sterile forceps and then pulling it out o f the tube. 8) Place the shoot in a sterile petri dish. Do not be concerned if the ends of the shoot extend over the edge of the dish. We will not be using these areas of the shoot. 9) Add enough sterile water to the dish to partially cover the stem. 10) With the sterile scalpel, cut above a nd below each node ( Figure 3 13 ). This should yield 5 to 6 internode explants, each about 1 cm in length. 11) Transfer the internodal segments into a petri dish containing a small volume of sterile water. 12 ) Repeat Step 6 through 11 using with the remaining shoot cultures. Use a new petri dish half as a cutting surface to divide each shoot culture.
121 13) Starting with the control medium, transfer one of the internodal explants into each of vials. Be sure t o place the explants horizontally in firm contact with the medium. 14) Place the cap on each vial but keep it slight loose. Seal with one layer of para film. Figure 3 9 Cuts required to obtain stem internode explants November 13, 2015. Drawing cour tesy of author. Figure 3 10 Formation of shoots from internode explants in the presence of cytokinin 2iP. Shoots originate from the epidermal layer on the internode explants November 13, 2015. Photo s courtesy of author.
122 RESULTS Score the total n umber and length of adventitious shoots produced on each stem response for shoot number and length on each medium. QUESTIONS FOR DISCUSSION 1. Based on your results, what role might play in the control of direct shoot organogenesis? 2. Provide an explanation why adventitious shoots also formed on the internode explants cultured on the medium not containing cytokinin?
123 Scientific Method Laboratory Report NAME: _________________________________ DATE: ___________________________ Purpose or Problem What is your experiment about or what question are you trying to answer? _________________________________________________________________ _____ ______________________________________________________________________ ______________________________________________________________________ _________________ Hypothesis Materials: (Include all materials needed and the quantity). 1. 2. 3. 4. 5. 6. 7 8. 9. 10. Procedures (Write out simple step by step procedures of the experiment). 1. 2. 3. 4. 5. Results and Data Collection (In the form of a table, include drawings or photos). Score the total number and length of adventitious shoots produced on each stem inte rnode explant after three weeks culture on each medium. Calculate the mean response for shoot number and length on each medium.
124 Data Table: Number and length of adventitious shoots of parrot feather. Date Measured: A dventitious shoo ts per explant Adventitious shoot length per explant (mm) Control + 2iP Control +2iP Week #1: (Dates) Week #2: Week #3: Total : Mean : Calculate the mean shoot number and length on each medium. (T he mean is the average) Conclu sion: What happened? Did you prove or disprove the hypothesis? What sources of error did you find and why do you think the error occurred? Please describe how the control group and variables responded according to the data.
125 Student Vocabul ary Page Student Name ________________________ Title__________________________________________________________________ HOW WELL DO I KNOW THESE WORDS? Directions: As you read this selection on shoot organogenesis, place bold faced or highlighted word s in the appropriate column. You should then write a brief definition or a synonym for that word next to the word you have written in each of the columns. Words I still NEED HELP in Understanding I THINK I understand this word I KNOW a meaning for t his word
126 Shoot Organogenesis Pretest and Posttest True/False Indicate whether the sentence or statement is true or false. ____ 1. The scientific name for parrot feather is Myriophyllum aquaticum. ____ 2. P arrot feather is considered a terrestrial plant. ____ 3. An internode is located between a set of nodes on a stem. ____ 4. Shoot organogenesis uses explants taken from Stage II multiplication cultures. Multiple Choice Identify the letter of the choice t hat best completes the statement or answers the question. ____ 5. The method of micropropagation that uses the production of shoots directly from explants is called ______________? a. aseptic technique c. direct organogenesis b. Indirect shoot organogene sis d. non zygotic embryogenesis ____ 6. The production of adventitious shoots followed by rooting of individual shoots is called ________________? a. shoot culture c. tissue culture b. non zygotic embryogenesis d. shoot organogenesis ____ 7. What type is of plant growth regulator is 2iP? a. auxin c. gibberellin b. cytokinin d. abscisic acid ____ 8. What type of explant is needed for direct shoot organogenesis? a. callus c. stem internode b. meristem tip d. none of the above ___ 9. What is NOT a method of micropropagation ? a. Shoot culture c. non zygotic embryogenesis b. Shoot organogenesis d. tip cuttings Short Answer 10. What role might cytokinin s play in the control of direct shoot organogenesis?
127 Key for Pretest and Postt est: Shoot Organogenesis True and False 1. True 2. False (Aquatic Plant) 3. True 4. True Multiple Choice 5. B: Indirect Shoot Organogenesis 6. D: Shoot organogenesis 7. B: cytokinin 8. C: stem internodes 9. D: tip cuttings 10. The cytokinin 2iP promotes adventitious shoots regeneratio n within 7 days and at a faster rate with greater shoot production than internode explants cultured on medium without 2iP. Cytokinins promote cell division and meristem formation in plant root and shoot tissues
128 Teaching Modu le Lesson #3 Plant Biotechnology: Synthetic Seed #3 Wendy Vidor Matanzas High School Plant Biotechnology STEM Academy Unit: Plant Propagation Types Lesson Essential Question: What is synthetic seed? Lesson Duration (dates): Two: 90 minute blocks (4 d ays) Lesson Goals By the end of this lesson, this student will: KNOW (Vocabulary, facts) UNDERSTAND (Important concepts, principles, ideas) DO (How wi ll this be accomplished ?) Embryo, zygote, synthetic seed, somatic, clone, in vitro, cryogenics Under stand the difference bet ween natural seed formation and synthetic seed s formed from somatic embryos What are some of the potential uses of synthetic seed in agriculture and horticulture? Informative Reading: Text Coding Math: Percent and Molar Solu tions Laboratory Exercise: Synthetic Seed Encapsulation (Includes Math Strategy for Percent Solutions.) Writing: Inquiry Lab Report Activating Strategy: students and activate their prior knowledge? Use your learning managemen t system, online discussion tool or similar software for group discussion. (Assign groups before discussion and have recorder post answers after a 5 minute discussion) Use On line timer. Post this Question: What is a zygote? (Discuss for 2 minutes and group post) Post next question: What is the function of the seed coat? (Give 2 more minutes and have group recorder post response) Teacher will explain and can begin discussion if needed. What Background Knowledge is Needed? List any pieces of bac kground knowledge students will need to learn during the activating strategy to be successful Students will need to have background knowledge of: How is a zygote formed? Background on seed structure (embryo, endosperm) Mitosis vs. Meiosis (somatic cells vs. sexual reproductive cells)
129 in completing this lesson. Clarifications of Misconceptions/ Generalizations: Seeds can only form because of sexual reproduction. Vocabulary: What vocabulary will students learn during this lesson? How will they lea rn and apply these words/phrases/concepts? Zygote a fertilized egg cell that resulting from the union of a female gamete (egg) with a male gamete. Embryo a bipolar structure that develops in the seed after fertilization. Somatic embryos an adventitio us embryo derived from vegetative cells consisting of shoot and root meristem. Synthetic seed somatic embryos processed in some way to be handled like seed. Students can use a Frayer Diagram to complete vocabulary. Instructional Strategies: How will m aterial in this lesson be presented? Include research based strategies. Day 1: Text Coding : Present the article and have students use specified text coding to incorporate technical text. Incorporate vocabulary strategies using the following graphic o rganizers. Using the CIS Lesson Sheet students will read the text and use the coding system of N= New Information, I =I know this. (Reading Strategy) Day 2: PPT: Display and review key concepts in the PPT presentation on what is synthetic seed. Also a basic demonstration on how to perform the lab activity. Math Review : Using an overhead projector, review with students on how to perform percent solutions for alginic acid and calcium chloride solutions. (In the classroom, the students, will use la boratory notebooks to perform calculations.) Day 3: Lab Procedures: Review personal protection equipment (P PE ): Gloves, goggles and a prons. Follow lab procedures as specified on lab sheet.
130 Students will only mix calcium chloride solutions as specifie d by teacher. Teacher will need to premix the sodium alginate solution overnight on a magnetic stir plate. (Have each student group make up calcium chloride solutions and pipette into glass or plastic vials.) Seedlings must be started ahead of time an d be about 4 days old. Day 4 : Prepare seeds for planting: Create an Inquiry Based Lab on grow out of synthetic seed. Write Formal Lab Report in Lab Notebook. Day 5 : Assessment: Laboratory Report Discussion Questions and Math Calculation Worksheet. Assessment Prompt: How will you assess student understanding? What is the purpose of the alginic acid in the lab procedure? What types of seed would this be useful for in horticulture? Summarizing Strategy: How will students summarize what they have learned? Ticket out the Door: (Assessment prompt) What are the uses of synthetic seed in agriculture? Extended Thinking Strategy: How will students extend their knowledge beyond this lesson? Students will design an Inquiry Based Lab to grow out the pla nts they encapsulated in the lab. They will need to set up a formal lab inves tigation and report on how the synthetic seed vs. traditional seed perform. Resources: What resources (textbooks, materials, audio/visual aids, technology, etc.) were used in this lesson? Laboratory Manual Copy of Agriquest article on Synthetic Seed http://www.agriquest.info/synthetic_seeds.php Highlighters or pens for text coding Copies of Ticket Out the Door Student Laboratory Notebooks to record lab Copy of Math Worksheet for Solutions Computer access for TodaysMeet.com Response PowerPoint Presentation on Micropropagation Methods (Appendix I) References 1. Ammirato, P.V. 1983. Embryogenesis. In : D.A. Evans, W.R. Sharp, P.V. Ammirato and Y. Yamada (eds.), Handbook of Plant Cell Culture, Vol 1. Techniques for Propagation and Breeding, pp. 82
131 123. Macmillan, New York. 2. Brown, D.C.W, K. Finstad and E.M. Watson. 1995. Somatic embryogensis in herbaceous dicots. In : T. Thorpe (ed.) In Vitro Embryogenesis in Plants, pp. 345 415. Kluwer Academic Publishers, Dordrecht. 3. Dunstan, D., T.E. Tautorus and T. Thorpe. 1995. Somatic embryogensis in woody plants. In : T. Thorpe (ed.) In Vitro Embryogenesis in Plants, pp. 471 538. Kluwer Academic Publishers, Dordrecht. 4. Hacci us, B. 1978. Question of unicellular origin of non zygotic embryos in callus cultures. Phytomorphology 28: 78 81. 5. Halperin, W. 1995. In vitro embryogenesis: some historical issues and unresolved problems. In: T. Thorpe (ed.) In Vitro Embryogenesis in Plants, pp. 1 16. Kluwer Academic Publishers, Dordrecht. 6. Krishnaraj, S. and I. Vasil. 1995. Somatic embryogensis in herbaceous monocots. In : T. Thorpe (ed.) In Vitro Embryogenesis in Plants, pp. 345 415. Kluwer Academic Publishers, Dordrecht. 7. N ishimura, S. T. Terahima, K. Higashi and H. Kamada. 1993. pp. 175 181. Bioreactor culture of somatic embryos for mass propagation of plants. In : K. Redenbaugh (ed.), Synseeds: Applications of Synthetic Seeds to Crop Improvement. CRC Press, Boca Raton. 481 pp. 8. Redenbaugh, K., J.A. A Fujii and D. Slade. 1993. pp. 35 46. Hydrated coatings for synthetic seeds. In : K. Redenbaugh (ed.), Synseeds: Applications of Synthetic Seeds to Crop Improvement. CRC Press, Boca Raton. 481 pp. 9. Steward, F.C., M .O. Mapes and K. Mears. 1958. Growth and organized development of cultured cells. II. Organization in cultures grown from freely suspended cells. Amer. J. Bot. 45: 705 708. 10. Steward, F.C., L.M. Blakely, A.E. Kent, M.O. Mapes. 1963. Growth and organ ization in free cell cultures.
132 In : Brookhaven Symposium No. 16, Meristems and Differentiation, p. 73. ESE Student Accommodations/ Modifications for Lesson: Accommodation Type: Who/When Used or Offered: Think Pair Share Cooperative Groups During act ivation strategy During lab Florida 2016 17 CTE Curriculum Frameworks and Standards for Plant Biotechnology Science Standards: Grades 11 & 12 (Florida Next Generation Sunshine State Standards) Mathematics Standards: Grades 11 & 12 (Florida N ext Generation Sunshine State Standards) 18.02 Operate lab equipment and measurement devices. 18.05 Prepare buffers, reagents, solutions and media. 20.03 Synthesize information from a range of sources (e.g. texts, experiments, simulations) into a cohere nt understanding of process, phenomenon, or concept, resolving conflicting information when possible. LAFS 1112.RST 3.9 20.02. Determine the meaning of symbols, key terms, and other domain specific words and phrases as they are used in a specific scientifi c or technical context relevant to grades 11 12 texts and topics. LAFS.1112RST.2.5 34.05 Propagate plants using tissue culture techniques for producing synthetic seed culture. SC.912. L.14.10 : Discuss the relationship between the evolution of land plant s and their anatomy. SC.912. L.16.17 : Compare mitosis and meiosis and relate to the processes of sexual and asexual reproduction and their consequences for genetic variation. SC.912. N.1.7 : Recognize the role of creativity in constructing scientific q uestions, methods and explanations. 03.01 Make sense of problems and persevere in solving them. 22.04 Model with mathematics. MAFS.K12.MP.3.1 22.06 Attend to precision. MAFSK12.MP.6.1 Lesson Reflection : Note changes to be made, write reflections for f uture use of lesson and adjustments to plan. Teacher will note any changes or adjustments needed after the lesson.
133 CIS: Synthetic Seed (Reading/Writing) Goals: What do you want students to learn (content) from this article? What do you want student s to be able to do? Title of Text or Article: Agriquest: (Synthetic Seeds): http://www.agriquest.info/synthetic_seeds.php Step 1 Tasks: Teacher asks hook question, reads aloud to students while students mark text, they read the text and participate in directed note taking. Purpose: To bring world relevance to text reading, establish a purpose for reading, model fluent reading, provide opportunities for students to become interactive with the text, and think critically about information in the text. Hook Question: What are synthetic seeds and why are they important to agriculture? Predictive Written Response to Essential Question Paragraph # Specific Vocabula ry Word Part of Context 1 Synthetic Seed 1 Somatic embryogenesis 2 Zygotic embryos Vocabulary Instruction Review necessary vocabulary words and then direct students to words introduced in the text by paragraph number. Teacher may include effe ctive vocabulary strategies at this point. Teachers add a brief definition on chart paper. Variations for Vocabulary Instruction: Word Study Guide, Frayer Model, graphic organizers, word wall interaction.
134 Reading #1 Text Marking N = New Information I = I know this Model for students by reading the text aloud and coding a portion of the text. Students follow along and mark their copy. Students proceed to code the rest of the text independently. Students share text markings with table group or partn er. Reading #2 Directed Note Taking Record notes containing the most important information relevant to the guiding question. Present the guiding question to direct students thinking while taking notes. Teacher models note taking using an example state ment from the text, then selecting the category or categories that support the statement. Students complete note taking collaboratively or independently. Conduct small and whole group discussions on the reading notes. Ask groups to come to consensus o n which category is the most impactful according to the support from the text. Name: _____________________________ Class Period: _________________ Directed Note Taking: Guiding Question : What are some characteristics of synthetic seeds? Paragraph # NOTES Check relevant categories below 1 2
135 Synthetic Seed Lab Math Calculations Worksheet Directions: Calculate the percent and molar solutions below needed for the Syn thetic Seed Lab. (Please show your work below). Sodium Alginate is mixed at 10g/L to generate a 1% solution. 1) Calculate the amount of alginic acid powder needed to make 1L of 2% alginic acid solution. 2) What if I want to make only 250ml of a 2% soluti on? Molar Solutions : 3) What is the formula weight of CaCl 2 ? 4. How many grams of CaCl 2. are needed to make 1L? 5. What if I wanted to make only 500ml of a 100mM of CaCl 2 ? M ath Worksheet: Answer Key Florida Mathematics Standards: MACC.912.S IC.2: Un derstand and evaluate random processes underlying statistical experiments; MACC.912.N Q.1.3: Choose a level of accuracy appropriate to limitations on measurement when reporting quantities MACC.912.N Q.1.1: Use units to understand problems and to guide the solution of multi step problems; choose and interpret units consistently. 1. 10g x2 = 20g/L 1L 2 20g = x___ 1000ml 250ml X = 20g x 250ml 1000ml X = 5 g of Sodium Alginate 2. Atomic Mass of CaCl 2 Ca = 40.02 Cl = 35.45 x 2
136 3. Calculate the formula weight by adding the Atomic mass of each atom. FW = 40.02+ 35.45 + 35.45 = 110.98g/ Mol 4. = 0 .1M CaCl 2 ( aqueous ) =11.098g CaCl 2 5. = 5.549 g CaCl 2 Ticket Out the Door: Why would synthetic seed be useful in agriculture? Why or why not?
137 STUDENT LABORATORY SHEET: SYNTHETIC SEED P urpose : The primary purpose of this laboratory exercise is to simulate procedures required for encapsulating non zygotic embryos to produce synthetic seed. INTRODUCTION: In higher plants, sexual reproduction involves fusion of gametes to form a single cel l zygote which develops into a complete plant through the process of zygotic embryogenesis Thus, the plant is a new individual prod used from a single cell and is also genetically distinct. Since the mid 1950s (9,10) it has been demonstrated that cultur ed vegetative (somatic) plant cells have the potential to act like zygotes and develop into what are termed non zygotic (NZ) embryos which possess both shoot and root meristems (Figure 1). These bipolar non zygotic embryos can either develop directly on cu ltured tissue ( direct non zygotic embryogenesis ) or directly on an intermediary callus stage ( indirect non zygotic embryogenesis ). Non zygotic embryogenesis has been observed in vitro in many angiosperm and gymnosperm species. (1,2,3,5,6). Non zygotic em bryos are structurally like zygotic embryos found in seeds and possess many of their practical features, including the capacity to develop into complete plants without the requirement for separate shoot development and rooting phases (9). Haccius (4) has defined a plant embryo (zygotic or somatic) as a new individual arising from a single cell and having no vascular connection with the maternal or explant tissue. Since NZ embryos develop from individual vegetative cells or groups of cells, the potential f or very efficient clonal propagation exists. This efficiency has been exploited through development of liquid culture NZ embryogenetic production systems using bioreactors (7). However, one major obstacle is that the NZ embryos do not have a protective se ed coat as zygotic seed and therefore are very susceptible to damage during handling and planting. Consequently, research has been conducted to develop procedures to process NZ embryos as synthetic (artificial) seed for commercial clonal plant production. This application is often referred to as s ynthetic seed technology and encompasses processing NZ embryos as synthetic seed and the subsequent development of a delivery system for mechanical planting (8). Numerous synthetic seed delivery systems have be en examined using NZ embryos. These include: 1) uncoated hydrated; 2) uncoated desiccated; 3) encapsulated desiccated; 4) encapsulated hydrated; and 5) hydrated in fluid drilling gel NZ embryos. One of the more frequently used delivery systems involves e ncapsulation of hydrated NZ embryos in alginate gel. This gel coating serves as a protective covering and may be multi layered to enclose a synthetic endosperm core containing growth regulator, carbohydrates and/or antibiotics (Figure 3 15 ). Encapsulatio n requires embedding the NZ embryos in a drop of viscous alginate and allowing the outer surface of the alginate to gel following submersion in a calci um containing solution (Figure 3 16 ). The encapsulated NZ embryos are then rinsed in water and handled l ike seed. In this exercise the procedures used for production of synthetic seed though encap sulation will be demonstrated.
138 To eliminate the need to use NZ embryos generated from in vitro culture, Arabidopsis seedlings will be used to demonstrate the conc epts. MATERIALS AND PROCEDURES Plant Material : Figure 3 11 Four day old Arabidopsis seedlings germinated on filter paper November 13, 2015. Photo courtesy of author. Materials : Small tapered Spatula and/or inoculating loop Nalgene Squ ibb 250 mL polypropylene funnel (Catalog # 4300 0250) containing 2% sodium alginate (Alginic acid; Sigma Chemical Product# A 0682) Support stand (Fisher Catalog# 14 (Fisher Catalog# 14 052B) 100 mM aqueous calciu m chloride (CaCl2) solution 20 mL glass scintillation vial with screw cap
139 Figure 3 12 Polypropylene funnel with alginic acid on ring stand. November 13, 2015. Photo courtesy of author. Figure 3 13 (A) Sprouted seedling placed into alginate b ead: (B) encapsulation of Arabidopsis seedling in alginate solution November 13, 2015. Photos courtesy of author. Procedures : 1) Pour 100 mM CaCl 2 solution into a scintillation vial until about full. 2) Adjust the stopcock on the alginate containing funnel so th at flow rate allows for a large alginate bead to slowl y form before it drops (Figure 3 16 ). A B
140 3) Place a seedling on the tip of the tapered spatu la or inoculating loop (Figure 3 17 ). 4) Embed the seedling in the developing alginate drop. 5) Allow the embedd ed seedling to drop into the scintillation vial containing the aqueous solution of 100 mM CaCl 2 (Fig. 3 17 ) The outer al ginate layer should immediately gel. 6) Repeat step 2 with using additional seedlings. 7) After 5 minutes, rinse the encapsulated seedlings in tap water and store in water in scintillation vial containing water. QUESTIONS FOR DISCUSSION 1) What are the advantages of producing synthetic seed by encapsulating N Z embryos? 2) What would the benefits be of producing encapsulated syntheti c seed with concentric gel layers containing various energy reserves such as sugars and plant hormones?
141 Teacher Plant Tissue Culture Workshop Pretest/Posttest 1. In vitro culture of plant cells and tissue is called? A. propagation B. cross pollination C. plant tissue culture D. micropropagation 2. What bacterium is commonly used in plant genetic transformation? A. Agrobacterium B. Bacillus thuringiensis C. Escherichia coli D. None of the above 3. The production of axillary shoots followe d by rooting of individual shoots is called? A. shoot culture B. non zygotic embryogenesis C. shoot organogenesis D. meristem tip culture 4. How does the plant growth regulator class called cytokinin s function in a plant? A. S timulate s cell division an d shoot formation B. S uppress the growth of side buds and stimulate root development. C. S timulate shoot elongation. 5. What piece of equipment in a tissue culture lab is used to sterilize media, glassware and tools? A. a utoclave B. m icrowave C. b lea ch D. h ot water 6. What plant growth regulator class does 2iP belong to? A. auxin B. cytokinin C. gibberellin D. abscisic acid 7. What type of explant is needed for direct shoot organogenesis? A. callus B. meristem tip C. stem internode D. none of the a bove
142 8 What component is NOT found in tissue culture media? A. water B. gelling agent C. mineral salts D. enzymes 9 What is the pH range recommended for tissue culture medium? A. 4.5 6.0 B. 5.0 6.0 C. 6.0 7.0 D. 7.0 8.0
143 CHAPTER 4 EFFECTS OF FLIPPING ON INCREASE IN TEACHER CONTENT K NOWLEDGE Introduction Biotechnology is having major impacts on many sectors of society including the g eneration of jobs and careers. There is a need to train and educat e students and tea chers in biotechnology careers (Borgerding et. al., 2012) Because of this need there have been an increase in the number of biotechnology curriculum programs in high schools. In Florida, there are several challenges in teaching biotechnology in the high school classroom that were revealed in the Teachers Belief About Teaching Biotechnology survey that was administered to agriculture biotechnology teachers and conducted during this research (Chapter 2). The objective of this survey was to evaluate teachers attitudes and beliefs about the major issues that teachers face in teaching the biotechnology curriculum and standards (Florida Department of Education, 2017) The teachers, and administrators in agriculture education reported four major concerns that contributed to the difficulty in teaching biotechnology including the lack of; 1) pre service college course work in biotechnology (85.3%), 2) pre service training of biotechnology course content (77. 5%), and 3) equipment and laboratory facilities to teach biotechnology (58.3%). 4) the need to supplement and enhance biotechnology instruction ( 77.8%) Mowen et al. (2007 a ) used the survey instrument Agriculture Teachers Attitudes and Implementation of Bio technology to determine perceived barriers and attitudes to tea ching biotechnology and teachers' beliefs about their roles in teaching biotechnolog y. The self perceived knowledge levels of teachers about specific biotechnology topics were found to have a s ignificant relationship between their knowledge of the topics and
144 teach ing the topic in the classroom. Regarding years of teaching experience, agriculture biotec hnology teachers that were well prepared in their pre service programs in traditional agricultu re courses and had knowledge of emerging technologies, such as biotechnology, had developed and maintained new knowledge. It was reported that only 36% of this group had ever attended a biotechnology workshop (Mowe n et al. 2007 a ) Based on survey results, a griculture education teachers lack background knowledge and un derstanding of biotechnology concepts to properly integrate the concepts into th e curriculum. Traditional instructional metho ds used in teaching biotechnology are shifting in a new direction and these changes are transforming instructional methods to deliv er science content infused with technology. Agriscience teachers need help to effectively utilize technology in the classroom to help advance student content knowledge of various biotechnology concepts in animal and plant based biotechnologies in agricultu re Another problem encountered in teaching biotechnology is the insufficient amount of instructi onal time allotted to prepare and teach individual biotechno logy laboratory exercises. Often the required time allotted is not sufficient to teach in a traditi onal 50 minute high school class period. Utilizing video lectures and virtual instruction at home or during warm up activities may provide more time for actual lab instruction to complete more rigo rous biotechnology laboratory experiences. According to (Thoron, 2010) the National Academ y of Sciences has recommended more stu dent centered classr oom and learning activities to address this issue. Development of plant biotechnology laboratory exercises which can be completed in a class period are needed.
145 There is a need to utilize more effective teaching methods (Abeysekera & Dawson, 2015; Bergmann & Sams, 2012; Strayer, 2012) In the l ast five years there has been a paradigm shift using a new pedagogical model known as the flipped classroom in which the typical lecture and homework are reversed by delivering instructional content usually online, outside the classroom (Ove rmyer. 2014). I n a flipped classroom, traditional content delivery methods, such as lectures, pres entations, videos, and readings are completed as homework and the practice problems normally completed at home are solve d in the face to face classroom (Mason, Shuman, & Cook, 2013; McCubbins, 2016; Tucker, 2012) The Flipped Learning Network definition of flipped learning (Flipped Learning Network, 2014) states that it is the face to face component of stud where the educator guides students to apply concepts and engage creatively in the (McCubbins, 2016) The most meaningful learning outcome in a flipped classroom is the additional class time that inverting the classroom provides for practice of more complex problems. (Szpara gowski, 2014; Tucker, 2012) There has been a call for quantitative research on the effects of the flipped model on student achievement. Stevenson & Harris, ( 2014) states that even with the rising popularity of the flipped classroom, traditional lecture is still the no r m in higher education teaching. Advancements in instructional technology incorporating learning management systems provides instructors the opportunity to improve and restructure course delivery methods in higher education (Con ner & Stripling, 2014 ) C ourses designed with embedded technology including video, assessm ent, and interactive
146 content have a greater probability of being viewed positively by students and administrators if they are desig ned for students to be active learners. A successful course design will enhance the use of flipping with blended learning techniques using a mix of technology and face to face instruction. Gardner (2012), inverted a n agriculture economics classroom specifi cally using the flipping met hod by incorporating recorded lectures including online audio and video files for students to complete before class Students viewed online videos that were assessed through homework, quizzes, tests and assignments using a learn ing management system. He measured student grades and time spent on the online content items and discovered a correlation betw een effort and time spent on the topics compared to overall performance resulting in a positive impact on student grades. He found that low performing students simply did not take the actions needed to be high performing students, possibly due to time constraints. that st udents found satisfaction in this flipped lectur e method and that it helped students learn course concepts. The effects on learning outcomes were unclear due to lack of student effort (motivation). Gardner emphasized that teachers should use enforcement mechanisms within the online video segments to ensure student engagement in completing the videos and class activities. Conner et al. (2014), like Gardner (2012), studied student perceptions by flipping an agricultural education teaching methods course Students reported mixed perceptions that the online video modules were ineffective and d id not impact learning. Also, students noted that improvements and quizzes need ed to be more challeng ing and that in class time more focus ed on specific content in the module. The
147 recommendations generated from this study suggest that instructors of simila r courses attempt using the flipped classroom approach to test the efficacy of the model in other content areas but focus on measuring content knowledge and skill acquisition and not just student perception s McCubbins (2016), b ased on the recommendations of Conner et al. (2014), observed student perceptions in an undergraduate capstone inverted course. Overall, the student perceptions were positive and supported the adoption of a s tudent centered course design. Con ner et al. (2014), recognized the need to provide workshops for faculty members within the College of Agriculture that specifically embrace student centered instruction and specific competencies needed for effective flipped instructional design. In general, it has been demonstrated that a flipped classroom approach does result in increased student engagement, perception and learning gains (Conner et.al., 2014; Deslauriers et al., 2012; McCallum et al. 2015; McCubbins et al. 2015). The recommendations generated from these previous studies encour age additional research to determi ne the efficacy of flipped classroom instruction by measuring content knowledge gain and skill Examining the flipped classroom method in agriculture biotechnology education training is ne cessary to assess its efficacy Con ner et al. ( 2004) and its effect on learning challenging scientific concepts (McCubbins 2016). to apply these concepts and new technologies utilized in STEM careers is cr itical for survival in the workpla ce. To fill the need for technology infused learning environments in high schools training must be provided to enable educators to effectively apply existing and emerging technologies to improve learning experiences and ca pture the attention of students f or
148 more positive learning outcomes, and depth of knowledge retention. The flipped classroom strategy provides an opportunity to ad dress integration of emerging technologies used in biotechnology with teacher training Training instructors how to use the em bedded technology within a flipped classroom is vital for co ntent areas like biotechnology. These recommendations guide my research study Purpose and Objectives The purpose of this chapter is to further investigate the influence of the flipped instructio nal method on content knowledge gain of Florida agriculture biotechnology teachers completing a laboratory module during a plant biotechnology teacher training workshop. The objectives were to : 1) D etermine if flipping a narrated lecture overview of shoot cul ture methods of Caladium bicolor will result in content knowledge gains in agriculture biotechnology teachers 2) E xamine the use and effectiveness of a detailed hands on biotechnology laboratory module during a teacher in service workshop. Methods The indep endent variable in this study was the teaching method used in the lecture overviews during the beginning of the workshop training. The two types of teaching methods consisted of a flipped (traditional overview lecture) ( Appendi x J ) versus a flipped narrated video lecture of shoot tip culture of Caladium bicolor (Object 4 1) The dependent variable in this study w as content knowledge posttest achievement scores. Covariates were used to adjust grou p means to reduce bias and compe nsate for previous k nowledge of the subject matter. The covariate was the pretest for the unit
149 of instructions. The researcher of the course administered the Pretest and Posttest as ined by the researcher and the maximum possible score on the instrument was 100. Workshop Setting and Participant Selection The study was conducted as a one day teacher professional development biotechnology laboratory workshop. The focus was on shoot tip culture methods for the micropropagation of the landscape plant Caladium bicolor The one day workshop was held on November 18, 2016 in the Micropropagation Teaching Laboratory in the Environmental Horticulture Department at the University of Florida, Gain esville, Florida (Appendix G) Florida agriculture e ducation teachers who teach high school biotechnology were invited to participate in the professional deve lopment workshop. The workshop consisted of ten teacher participants completing a novel hands on teaching module developed specifically for teachers that teach plant biot echnology concepts to students. All ten teacher participants taught biote chnology. Research Integrity and Compliance Research including the proposal for the pre/posttest assessment was approved by the Institution Review Board (IRB ) at the University of Florida ( Ap pendix D ). Teachers invited to participate in the workshop signed an informe d consent protocol ( Appendix E ). In the informed consent, the objectives and experimental procedures were clearly outlined The protocol for the pre/posttest assessment was IRB approved. This study fell under the category of exempt. Teacher participants did not know which group or method of instruction they would receive when they registered for the workshop
150 Experimental Procedures A novel module lesson plan was developed using the understanding by design framework by (Wiggins & McTighe, 2005) The laboratory module lesson plan, Caladium Shoot Culture, was formulated using the Next Ge neration Science Stand ards Integrated Instructional Sequence BSCS 5E Model (NSTA, 2016) Teachers were randomly assigned to one of two groups by placing the names in an Microsoft Excel spreadsheet and dividing the group in half after randomization. The con trol group (non flipped) group was given the traditional overview lecture while the experimental group (flipped) received the flipped interactive instructional video. Teachers assigned to either group were administered the same written pre test which consis ted of 10 true false a nd 15 multiple choice questions before the lecture was delivered The pre test question topics consisted of basic concepts of aseptic technique, tissue culture methods an d basic laboratory procedures for cleaning explants of Caladium b icolor. The two groups sep arated, and the pretest was administered in separate classrooms. To test the efficacy of constructivist (active learning) versus traditional lecture the control group receive d the traditional lecture overview. The control group received a traditional 45 minute lecture overview of the Caladium bicolor module and micropropagation techniques presented by the instructor using Microsoft PowerPoint slides. The lecture overv iew was delivered without any embedded media or questions, as it would be delivered in the traditional teacher centered classroom The flipped group receiv ed the flipped lecture overview. The overview was created and presented using Microsoft Power Point Mix 2007, and consisted of the concepts related to using me ristems shoot culture for disease eradication, surface
151 sterilization techniques to establish caladium shoot explants in culture, and tips in isolating caladium shoot explants. The online lecture overview was accessed on a computer as an online learning exp erience. The experimental group received about 5 minutes training on how to navigate through the flipped instructional video. The video was accessed by a link to the Microsoft Power Point Mix web site galley and was created specifically for this research study Object 4 1. Interactive lecture overview ( https:// http://ufdc.ufl.edu//IR00010128/00001 ) The interactive lessons offer a blended learning experience utilizing video clips and assessment. The in tegration of the quiz questions and the video clip were important in the design of the flipped lecture to engage and motivate the learner to connect with the lesson lear ning objectives of the lesson. This software application was chosen to deliver the flip ped lecture overview because of the ease to create a high value interactive lesson that can be easily integrated and published on many learning platforms. The participants in the flipped group were given 45 minutes to view the online interactive video lec ture and complete the embedded assessment questions intended to engage the learner (Zhang et al., 2006) The video was developed based on the concepts of the Cognitive Theory of Multimedia Learning (Clark & Mayer, 2016) T he flip ped lecture video consisted of two parts, an interactive lecture of PowerPoint slides, embedded video and assessment questions. Part one of the video, consisted of an overview of caladiums a nd micropropagation techniques. Additional media components added included a video clip on aseptic technique and three multiple choice q ues tions on the content presented. Part two of the video lecture consisted of an overview of
152 caladium production and specific instructions regarding the sterilization and preparation of the plant material for the laboratory exercises. During the lecture overview participants could take notes, but they were not allowed to discuss the content of the notes with each other This helped eliminate any influence that peer interac tion might have on individual posttest performance on the exam. There was one multiple choice question embedded in this section regarding surface sterilization to reinforce prior knowledge and for learners to engage with the content Pre test Posttest Instrument The pret e st and posttest instrument were designed to assess content knowle dge gain. The data from the pre test and posttest were collected during the Shoot Culture of Caladium Teaching Workshop held in November of 2016. The pretest and posttest assessment were creat ed to measure the skills and competencies of shoot culture through the Caladium bicolor laboratory experience using a randomized control group pre test posttest design. Instrument reliability was measured including normality by Shapiro Wilks test (p > 05 ), h omoscedasticity, variances, and testing for ou tliers (Kutner, et al., 2004). All data w ere collected on a paper based form. The researcher administered both t he pre test and posttest and graded the exams. The pretest and post test consisted of 10 true/false questions, and 15 multiple choice questi ons The first data collection point wa s the administration of the pre test and was completed before the intervention. The posttest was issued at the end of the 7 hour workshop. The pr etest and posttest were scored downloaded and imported into IBM SPSS Statistics, Version 24. At the end of both sessions, participants were issued the written posttest e xam The questions on the pretest and postte st were the same
153 After administering the lecture overviews both groups returned to the laboratory to be gin the shoot culture exercise. The learning outcome s were measured by the estimated between group means on the pre and post test scores using an analysis of covariance or ANCOVA using I BM SPSS Statistics, Version 24. A workshop questionnaire was used at the end of the intervention to prov ide open comments on the effectiveness of the traditional lecture overview and the flipped lecture overview. Workshop Questionnaire After completing the labor atory exercise each participant filled out a workshop questionnaire ( Appendix F ). T eachers were then asked to comment on the specific instructional treatment method (flipped vs. non flipped) they participated during in the lec ture overview. Caladium Laboratory Module In addition to the assessment, the instructional plans were created for the Caladium Shoot Culture Module The l esson plan and laboratory exercise were developed as the product to achieve the content standards and objectives for the workshop and to be used in the classroom as a complete module lesson plan. This lesson module was developed incorporating backward design as described in the Understanding by Design Framework by Wiggins et al. (2005). The goal of back ward design is to plan the learning exercises with the summative assessment in mind. The assessment portion is planned in the beginning rather than at the end of the module. The purposeful layout of the lesson module is designed around the assessment of th e pretest and posttest and measured for content gain knowledge. The laboratory activities on shoot culture production follow the assessment reinforcing the learning
154 objectives while building content knowledge and skills needed to perform the teaching labor atory. Research Design As described above, a r andomized control group pre test posttest desig n (Fisher, 1960) was used in thi s study This design was chosen because of the randomization of individ uals to control confounding variables that may influence the study (Campbell & Stanley, 1963) The confounding variables that were controlled include eliminating bias with random samples and counterbalancing by testing under two condit ions, treatment versus control. The hypothesis tested in this study was that the intervention (flipping) would increase content knowledge gain. The treatment in this experimental design was presented using an interactive flipped classroom lecture as the learning activity in an online learning environment. The dependent variable that was evaluated in this study was achievement of content knowledge gain. This was determined by achievement on the posttest following the completion of the lecture overview ( face to face overview lecture v ersus interactive video). A pre test was used to statistically control for teacher prior knowledge on the achievement test. The study was designed to investigate the following research question. 1. Is there a statistical diffe r ence between mean group scores in content knowledge of biotechnology teachers who received the traditional lecture method compared to teachers taught using the flipped classroom method? The resea rch question was posed as a null hypothesi s The null hypot hesis was tested at the p 05 level of significance.
155 H O There was no significant diff erence in mean pretest and post test scores of the expe rimental group versus the contro l group. H A There was a significant diff erence in mean pretest and post test scores of the control versu s intervention (flipped) group. 1. Control gr oup (Non Flipped): Teachers receiving the traditional in class lecture overview of Caladium bicolor shoot culture Experimental group (Flipped Group): Teachers receiving the flipped classroom interactive video lec ture of Caladium bicolor shoot culture. Rationale for Research Design and Statistical Analysis In r esearch Question # 1 it was knowledge gain ed between biotechnology teachers who received the traditional le cture method compared to teachers taught using the flipped classroom method ? To answer the posed research question, researchers suggests the use of analysis of covariance (Field, 2005, Ary et al. 2002). Quantitative data were described using descriptive statistics and analyzed using IBM SPSS Statistics 24 a software common for use in stati stical ana lysis in the social sciences (Field, 2013; Dimitrov et al. 2003). The hypothesis was tested using an ANCOVA with pretest scores as the covariate. This an alysis was chosen to reduce the error variance because of the random assignment of subjects to groups while guarding against systematic bias (Dimitrov & Rumrill, 2003) ANCOVA was executed to determine if there were any significant differences in the adjusted means between the two groups (flipped classroom video lecture) versus non flipped or traditional lecture on the t eache r content knowledge
156 (Ary, et al 2002). SPSS. Treatment levels consisted of the two caladium module lab overview lecture types randomly assigned to the groups of teachers attending the plant biotechno logy tissue culture workshop. Due to laboratory equipment limitations, only a relatively small number of participants (16 maximum) could be accommodated. Also, t he use of the pretest scores as opposed to just comparing posttest scores between gro ups help to reduce error variance and the power of the probability of detecting differences between the groups being compared (Campbell and Stanley, 1963) The randomized control group pretest and posttest experimental design can be depicted as: Table 4 1 Rando mized subjects, pretest posttest control gr oup d esign Group Pretest Independent Variable Posttest (R) E Y 1 X Y 2 (R) C : Y 2 __ Y 2 The first observation (Y 1 ) consisted of a pretest given to each participant in the control group and experim ental group prior to the treatment. The second observati on (Y 2 ) consisted of a posttest that occurred following the treatment and was given to both the control and experimental group immediately after the lectures were completed. The flipped interactiv e video overview lecture (X) represented the treatment. Res ults In this study the influence of teaching method on teacher content knowledge gains w as examined using a flipped video embedded online lecture overview before completing a plant biotechnology teacher training module. Due to limited space in the
157 teach ing laboratory facility, t he population in this study consisted of 10 teachers participating in a teacher plant biotechnology workshop held in November 2016. The first data collection consi sted of the Caladium Module Pre tes t with data collected from all te n participa nts with a 100% response rate. The second data collection consisted of the Caladium Module posttest (n =10) also with a 100% response rate ( Table 4.2 ). Table 4 2 Pre test and posttest response rate t otals Treatment Group # of Participants % No n Flipped (Control) 5 100 Flipped Group 5 100 The assumptions were tested separately and included normality, homoscedasticity, homogeneity of variances and testing for outliers (Kutner, et.al, 2004). There was a linear relationship between pre test and p osttest for each intervention type, as assessed by the visual inspection of the scatterplot that was not statistically significant ( Figure 4 1 ). There was homogeneity of regression slopes as the interaction term was not statis tically significant, F (1,6) =.001 p = .972 Standardized residuals for the interventions and for the overall model were normally distributed, as assessed by Shapiro p >.05) There was homoscedasticity and homogeneity of variances, as assessed est of homogeneity of variance ( p =.429) (Table 4 3 ) There were no outliers in the data, and there were no values with standardized residuals greater than 3 standard deviations. Tabl e 4 3 est of equality of error v ariances for posttest. F dF1 dF2 Sig. (p value) .694 1 8 .429
158 Teachers that taught using the flipped classroom video lecture method achieved posttest scores that were statisticall y different (83.74 3.6) compared to the contr ol group receiving the traditional lecture (80.26 3.6) ( Table 4.4 ). Table 4 4 Adjusted and unadjusted intervention mean scores and variability for post intervention of Caladium module e xam Unadjusted Adjusted N M SD M S E Control Non Flipped Group Posttest 5 80.8 9.5 80.3 3.6 Intervention Flipped Group Posttest 5 83.2 9.9 83.7 3.6 Note : N = number of participants, M = Mean, SD = Standard Deviation, SE = Standard Error. After adjustment for pre test scores, there wa s not a statistically significa nt difference (p=0.514) in post test mean scores between the teachers taught b y the two teaching methods and th ere was no significant difference observed from the flipped classroom interactive lecture on content knowledge gain score at the p<.05 therefore, the null hypothesis was accepted ( Table 4 5 ) The prete st and pos tt est doc ument are provided in Appendix H Table 4 5 ANCOVA test of Caladium module e xam Source Sum of Squares d f Mean Square F Sig. Group 29.941 1 29.941 .473 .514 Error 442.996 7 63.285
159 Figure 4 1. Scatterplot comparison of teacher pretest and posttest performance in a non flipped (control) and flipped plant biotechnology classroom Discussion and Summary In this study the effect of content knowledge achievement using a flipped method of instruction incorporating an online interactive video lecture overview was examined Results indicated that there was no significant diffe rence in pre test and posttest scores between the f lipped and non flipped teacher groups T h e accessible population consisted of high school teachers that have taught biotechnology in agriculture ed ucation, or biology in Florida. Research Question # 1; Is th ere an overall difference in content knowledge gain between biotechnology teachers who received the traditional lecture method (control group) compared to teachers taught using the flipped classroom method (experimental group) ? Analysis showed that the tr aditional group mean scores were slightly lower than the mean of the flipped group. However, this difference was no t stat istically significant different ( p = 0.514) between groups. No significant differences in content knowledge gains were observed from th e flipped classroom interactive lecture on content knowledge gain score Due to the small sample size in the workshop, the
160 effect size could not be determined to reach statistical separation. There still could have been a small positive and meaningful tr eatment effect that could not be accessed with such a small population. The difference observed can be attributed to two factors the years of teaching experience and completion of pre service and in serv ice training. Some teachers had been teaching biotech nology in their classro om for five years or could have affected the gain score between pretest and posttest. Another factor influencing gain scores in the pre/posttest scores could have been the differences in training level of the biotechnology teachers Teachers that had attended in service workshops and training may have achieved increased gain scores between pretest and posttest due to the effect of previous training This could reflect an increase in content knowledge and skills achieved in these previ ous trainings. Also, the number of pre service courses taken by teachers in molecular biology and chemistry could be a contributing factor in increasing understanding of biotechnology concepts. The results of this study support the null hypothesis. H O There was no significant diff erence in mean pretest and post test scores of the expe rimental group versus the contro l group. Threats to internal validity were addressed in the design of th e study as defined by Campbell and Stanley (1963). There are eight threats to internal validity; history, maturation, testing, instrumentation, regression, selection, experiment mortality, and an interaction of threats (Campbell and Stanley, 1963) The ba sic threats to internal validity of the randomized control group Pretest / Posttest design include history, maturation, selection and maturation.
161 1. History is the events that occurred between the first and second measurement of a study (Campbell & Stanley, 1963) History, in this study was controlled by minimizing the time between the two measurements. The testing was immediately given before and after flipping and non flip ping. 2. Testing was the second internal threat if the effect of the pre test affected the scores on the posttest (Campbell & Stanley, 1963) This threat was addressed by using identical questions on the pre test and posttest. 3. Selection is a threat to internal validity if there is bias during selection of pa rticipants (Campbell & Stanley, 1963) S election a nd testing can be minimized by making the groups comparable by randomizing the groups before treatment. 4. Maturation threat can occur due to biological or psychological can effect differences between pre and posttest (Cam pbell and Stanley, 1963) This threat was addressed External threats to this design i nclude population validity, how representative was the sample population and the reactive effects of experimental arrangements, the effect that is due because participants know they are experiencing the novelty, also known as the Hawthorne effect. To minim ize these internal and external threats, it was important that teachers were chosen to participate on ly if they were high teachers who taught biotechnology, specifically a griculture biotechnology. The selection of these teachers helped to make groups com pa rable, minimizing the threat validity of the study. Teachers were specifically chosen for their conte nt knowledge in biotechnology.
162 Research Q uestion # 1, the effects of teachers flipping method were addressed in results of the ANCOVA test analysis which d id not support our alternative hypothesis that content learning gain would occur in teacher participants in the flipped lecture compared to the tradit ional lecture control group. The findings do support an increase in individual test scores due to the work shop training as observe d in the posttest score gains in pre/posttest within groups outcome Inclusion of an overview lecture, regardless of teaching method results in significant knowledge gains within groups. Previous studies have shown that the flipped model improved student performance and perceptions (Andrews et. al, 2011; Conner et. al, 2014; Davies et. al, 2013; Deslauriers et. al, 2011; McCubbins et. al. 2016; Strayer, 2012). A common theme in these studies is that the most important aspect of the flipped model is not the videos, but the increase in face to face class time resulting in increased student achievement. This study suggests that instructors using flipped instructional methods may observe significant gains in content knowledge, but studen t motivation and engagement influenced individual pre and posttest score gains. To further investigate that the interactive video may improve higher learning satisfaction a workshop questionnaire was completed by teache r participants ( Appendix F ) Teacher participants in both groups experienced content lea r ning gains as reported in the data of estimated means on the posttest. Teachers in both groups experienced learning gains in the overall content of the workshop, but incorpo rating instructiona l video may not be sufficient to improvi ng e learning effectiveness. These results confirm those of Zhang et al. ( 2006) who concluded that use of e lear ning video in education may not always be sufficient to improve learning outcomes unless the individual can
163 control the pace of learning. It is the use of the interactive video that provides individual control by the learner that may lead to better learning outcomes of the objec tives of the lesson and lead to higher learner satisfaction and improved e learning environments. Students who could control movement in the form of reviewing and moving backward and forward demonstrated better achievement of learning outcomes and satisfac tion (Brame, 2016). Most participants in the experimental (flipped) group reported on the workshop questionnaires that the interactive video lecture (flipped) provided them the opportunity to go back and review content they may have previously missed and w as beneficial to have that option. The interactive video provided an opportunity to review content that was unfamiliar due to the interactive features in the video format. This should have resulted in a content knowledge gain in the flipped group, but tha t was not apparent in the results even though tests for reliability were assumed to be correct. A possible reason for the lack of significance in the posttest score between groups could be shortcomings in test questions generated using the classical test t heory (Lord & Novick, 1968). In the classical test theory, the standard error of measurement is assumed the same for all examinees. Hambleton et al. (1991), explains the scores on any test are unequal precise measurements for examinees of different abiliti es and the assumption of equal errors of measurement is not plausible. Another shortcoming of classical test theory is that it cannot assist in making predictions of how well an individual or a group of examinees might do on an individual test item. Fin ally, the length of the flipped video instruction could have a negative effect on learner motivation and attention span. Brame ( 2016) suggested from studies in biology courses, that for video to improve student l earning and enhance engagement they must
164 contain three key components. These components include reducing cognitive load by keeping videos brief and targeted on learning goals and that audio and visual elements should be used to explain appropriate concepts Finally, the use of enthusiastic conversation in the video will enhance engagement, and using embedded videos for active learning in the form of guiding questions along with interactive elements as well as associated homework. In fact, many of the teache rs in the flipped group suggested that the video lecture was too long and should be divided into two sections to increase motivation and retention which may have resulted in decreased content learning gains. This research shows that the flipped classroom m affect learners, but may have potential benefits when combined with interactive video lectures and laboratory experiences as seen in the Shoot Culture of Caladium bicolor teaching module. The implications of this resear ch based study may contribute to a positive change in education regarding the use of technology and media applications for conte nt achievement in the classroom for students and teachers of the future, both of which will know how to better learn in an onlin e environment. Limitations and Implications The most significant limitation of the study was that, due to space limitations, only a small number of participants could be accommodated in the micropropagation teaching laboratory This created a small sample p opulation to conduct the study. Another limitation was that the indivi know ledge varied. In the Teachers Belief About Teaching Biotechnology Survey (Chapter 2), 77.4 % of teachers reported they did not take any pre service courses to prepare for teaching biotechnology. When asked if these teachers participated in any in service professional development, 50% of the teachers reported they had participated in
165 some type of training in biotechnology. Teachers who pos sess extensive k nowledge in biotechnology score better overall on both the pre and posttests when comparing in dividual scores within groups. The teachers that use flipped learning in their classrooms may have been better suited to e learning than other tea chers. Because of the randomization of the groups in this study to provide homogeneity this limitation was not investigated. A third limitation of the study consisted of having only a single worksho p session and not a longitudinal study in which data were gathered from the same subjects, repeatedly over time such as that described by McLaughlin et al. (2013) Results of this stu dy demonstrated that the flipped classroom had no significant improvement in examination performance, but the flexibility of the model demonstrated enhanced learning experien ces for teachers in engagement. Further studies would need to be conducted in futu re teacher workshops to examine if the identified effects of interactive flipped video instruction would result in cont ent knowledge gains with a larger sample size. Results could then be generalized across a larger population of agriculture biotechnology teachers. The findings of this study indicated tha t using flipping methods do not always result in content learning gains. However, using blended learning techniques offer s the learner some flexibility in how they can individualize their learning to imp rove engagement and satisfaction in the learning process In previous studies of teaching methodologies, use of flipped video lect ures resulted in mixed results. Conner et al (2014) ; Fre eman et al. (2014); and Zhang et al. (2006) discuss the opportuni ties available to train teachers using n ew instructional methods while enhancing mot ivation
166 for learning and improving perception. Under some circumstances, interactive e l earning can have more positive res ults than traditional methods. The most important aspect of teac hing in a flipped model is not to just present video lectures but to use the face to face classroom time for dynamic and active learning opportunities (Overmyer, 2014). This is not an easy task. T eacher s must be trained in the flipped classroom method. Teachers need to be expert s in the subject matter, student facilitation and laboratory management to successful ly flip the classroom Conclusion Researchers have reported mixed results with using flipped multi media systems. My study demonstrated that using flipped interactive inst ructional videos may not alw ays be sufficient for improving learning gains utilizing summative assessment The interactive video can provide better individual control and result in some increa sed satisfaction by the learner but there may better methods to assess content learning gains in a flipped classroom. Further studies will be needed with a select group of teachers to investigate the flipped model of instruction for content learning gains and assessment development in agriculture biotechnology. Questi on selection and assessment types on formal assessments could have a significant impact on content learning. The question selection on the pre posttests could have a significant impact on the content knowledge gains. The benefits of using a variety of asse ssment methods in evaluating flipped classroom gains including formative, summative and self assessment in a flipped lesson module can promote deeper learning as reported by McLean (2016) by utilizing a variety of assessment formats to capture student learning in a flipped environment. Using different forms of formative, summative, and self asse ssment styles in the flipped
167 classroom can support better communication and collaborative problem solving and constructive feedback. Trainings and additional workshops will be needed to investigate the overall effectiveness in teaching biotechnology cours e content in the e learning environment. Most importantly, the type of assessments needed to effectively measure content learning gains in the flipped classroom must be developed. Teaching Lesson Module #4 Shoot Culture of Caladium bicolor Wendy Vidor Un iversity of Florida Unit: Micropropagation: Shoot Culture Lesson Essential Question(s): What is shoot culture and why is it a preferred method for in vitro propagation? Why is sterile technique important in micropropagation? How does the shoot cultur e of caladium differ from other methods of production? Lesson Duration (time estimate): 4 5 class periods (45 50 minutes) Lesson Goals(Objectives): 1. Demonstrate the use of aseptic technique in the lab. 2. Explain the function of cytokinins in plants. 3. Identify the shoot meristematic tissue in caladium. 4. Perform shoot tip culture on potted caladium plants. 5. Sterilize the explants using aseptic technique. 6. Explain the process of shoot culture for micropropagatio n. Standards: These frameworks support the following Next Generation Sunshine State Standards: SC.912. L.16.10; SC.912. N.1.3, 4, 6, 7; SC.912. N.2.1, 2., LAC.9.1. W.1.2, 2.6 Florida CTE Curriculum Frameworks: Plant Biotechnology 4 25.03: Describe the effects of growth hormones i n tissue/cell culture. 32.01: Evaluate the advantages and disadvantages of using the micropropagation techniques. 32.02: Demonstrate aseptic/sterile technique. 32.03: Prepare and mix stock solutions of media for micropropagation 32.04: Produce a crop using clonal tissue culture methods and prepare a written report of results.
168 KNOW (Big Ideas) UNDERSTAND (Important concepts, principles, ideas) DO (How wi ll this be accomplished ?) Big Idea : Shoots of all angiosperms and gymn osperms grow from their apical meristems The apical meristematic dome plus one or two accompanying leaf primordia. comprise the meristem tip The shoot tip consists of the apical meristem plus numerous leaf primordia. Larger shoot tips or lateral bu d primary explants are used for clonal propagation when pathogen eradication is not a concern. This micropropagation procedures is called shoot culture. The apical meristematic regions of caladium are used because they remain uninfected by systemic pathog ens. (virus, fungi, bacteria). This method is called meristem culture Larger shoot tip or lateral bud primary explants are used for clonal propagation when pathogen eradication is not a concern. This procedure relies on stimulation of shoot buds by disr upting apical dominance in shoots cultured in media by using a cytokinin. Shoots first established in vitro by meristem tip culture for disease eradication are then used for clonal propagation of axillary shoots. This micropropagation technique is called shoot culture The purpose of this laboratory exercise is to demonstrate procedures required for in vitro establishment of Caladium meristem tips. Students will perform shoot culture on potted caladium plants after performing a u sing a narrated ppt with instructions on the demonstration of the in vitro aseptic procedures. St udents will take a pre and post test of the concepts before and after the flipping lesson. Students will record observations by comparing differences in gro wth response for 6 weeks and calculate the percentage of surviving explants, the mean number of new shoots produced per original shoot tip,
169 and graph the data. Students will write a conclusion based on the data they collect from the experimentation. Activating Strategy: students and activate their prior knowledge? Flipping Exercise: Narrated PowerPoint video. What Background Knowledge is Needed? Tissue culture in the classroom can be successful if aseptic technique and proc edures are performed correctly. In this exercise, you will be performing shoot culture consisting of apical meristem tissue, leaf primordia and developing leaves from Caladium. This procedure is used for clonal propagation to enhance formation of axillar y shoots. This procedure works by stimulating shoot growth from lateral buds. This process is performed by disrupting the apical dominance in shoots cultured on media supplemented with cytokinin. Cytokinin is a class of plant growth hormone that stimul ates cell division in cell roots and shoots. The addition of this hormone to the media stimulates rapid development of shoots. The advantages of shoot tip culture help in the rapid production of pathogen eradicated plants that are first produced by using meristem tip culture. Clarifications of Misconceptions/ Generalizations: What misconceptions or generalizations might students come into this lesson with? One misconception that students may have about tissue culture techniques is that it is t he same pro cedure works for all plants. Vocabulary: What vocabulary will students learn during this lesson? How will they learn and apply these words/phrases/concepts? Meristematic tissue : cells that can divide to form new cells which differen tiate to form comp lex cells, tissues or intact shoots In vitro: in glass (Latin). Experimentation carried out on plants or tissues maintained in clear glass or plastic
170 vessels under artificial and usually sterile conditions. Cytokinin: a class of plant growth regulators t hat promote cell division and shoot development. Culture medium: medium used in tissue culture to promote growth of cells or plants containing water, mineral salts, micro and macronutrients, organic components, vitamins, sucrose, plant growth regulators an d a gelling agent used for support. Shoot culture: A micropropagation technique for culturing shoot tips and lateral buds of a plant stem to induce the formation of microshoots. Apical meristem: The dome like structure at the tip of a shoot or root consis ting of rapidly dividing cells. Instructional Strategies: How will material in this lesson be presented? Advance Preparation: 1. Prepare the caladium multiplication medium per the directions on the handout. (You may have advanced students prepare the me dium as an additional lesson). (This should be done after school. It will need to be sterilized in the autoclave or pressure cooker) Handout: Tissue Culture Media for Caladium. 2. Copy the student laboratory handouts one for each student. 3. Prepare the lab equipment for sterilization of the cuttings. 4. Prepare the lab equipment for inoculating explants including preparation of hoods and sterile equipment. 5. Copy the pre and post test for the unit. 6. Copy the lab rubrics. Day 1: (Pre Learning) 1. Ha ve the students take the pretest on the Caladium module. (10 minutes) 2. Individually assign the narrated PowerPoint to students as an activating strategy. (30 minutes) 3. The teacher should review the big idea concepts. a. What is an apical meristem? b. What is a meristem tip? c. What is a shoot tip? 4. The questions above may be assessed by using an Exit ticket. (see attachments at end of lesson. )
171 Day 2: Activity : Teacher Preparation (20 minutes): Teacher should prepare these lab materials for cleaning the e xplants. Plant Material: Caladium bicolor: Potted plants from greenhouse. Lab Materials: (Prepare glassware for students, one beaker for each solution and one set of everything else). Note: Have students pour and measure each solution and gather mate rials from a central location that you have prepared ahead of time! Cleaning shoot explants of potted Caladium bicolor plants. (The teacher should demonstrate how to cut the shoot tip.) See lab sheet. 1. Essential question : Why is using sterile technique important in micropropagation? (5 minutes) 2. Using the student laboratory sheet, instruct students on the proper technique to clean the caladiums in preparation for shoot culture. (20 30 minutes). 3. Follow the materials and procedures attached to the student l aboratory sheet. (See attached) 4. Students can complete the vocabulary graphic organizer as they are rinsing their stock plants. (10 minutes) Make sure that students transfer the clean explants into a sterile disinfecting bottle in the laminar flow hoods. 5. St op here if this is a 40 50 minute class. Students will need to place explants in sterile water with a drop of plant preservative if leaving overnight. Day 3: Transferring Explants to Media (5 minutes) : Have students assemble in groups of 2 and go to t he laminar flow hoods or pre assembled clean box. Have them take their cleaned explants into the hood and place the tools in the glass bead sterilizer set at 260 C. Please remind them to take them out after a few minutes with a hot glove and place them inverted in
172 the rubber stoppers until they are cool. Note: Students can complete purpose, hypothesis, procedures in the lab template. 1. (20 30 minutes ): Have students go to transfer hood and complete steps 13 17 on the Student Laboratory Sheet. 2. Have stud ents write up the laboratory report in their student notebook or on the lab template provided. 3. Students should take 10 culture tubes filled with media to the transfer hood (Make sure they inoculate the explan t into the media). Have students take one cutting from the jar and with sterilized forceps, gently inoculate the bottom of the cutting slightly into the medium. Cap the culture tube immediately after inoculation and place the tools back into the glass bea d sterilizer. 4. (5 minutes ): Clean and spray down hoods with alcohol. Turn off glass bead sterilizers, remove razor blades and dispose of in container, shut off fans in hoods. Clean all tools and glassware. Day 4: 1. Review of technique and concepts. Dis cuss errors in aseptic technique or procedures with students as needed. (10 minutes) 2. Postt est: Record results of pre and posttest. (20 minutes) 3. Review with students on how to take measurements and record data. ( 10 minutes) 4. Remind students they will need to measure explant growth weekly for 6 weeks. 5. Grade Lab Report with rubric at the end of the 6 week period. Assessment Prompt: How will you assess student understanding? Including informal (checks for understanding) and formal Wh at is micropropagation? What is shoot culture and why is it a preferred method for in vitro micropropagation? Why is sterile technique important in micropropagation?
173 (assignments/tests). See copies of Pre and Posttest. Rubric for Lab Report Summarizing Strategy: How will stu dents summarize what they have learned? Students will analyze the data and write a conclusion on their lab sheet. Extended Thinking Strategy: How will students extend their knowledge beyond this lesson? How will they apply what they have learned? Stude nts can set up their own variables with this experiment. They can add 2iP, change the amount of light, etc. This can be written up as an Agriscience Fair Research Project. Students can go to Stage 4: Acclimatization Phase Place microcuttings in container s in greenhouse and set up a growing experiment using different media, etc. This would be an appropriate Research Supervised Agriculture Experience. Resources: What resources (textbooks, materials, audio/visual aids, technology, etc.) were used in this l esson? 1. Aamir, A., A. Munawar, and S. Naz. 2007. An in vitro study on micropropagation of Caladium bicolor Intern. J. Agric. and Biol. 5:731 735. 2. Ahmed, A.A., E.A.H. Khatab, R.A. Dawood and A.M. Ismeil. 2012. Evaluation of tip culture and thermotherapy for elimination of carnation latent virus and carnation vein mottle virus from carnation plants. Internat. J. Virology 8: 234 239. 3. Ahmed, E.U., T. Hayashi, and S. Yazawa. 2004. Auxins increase the occurrence of leaf colour variants in Caladium regenerated fro m leaf explants. Scientia Hort, 100: 153 159. 4. George, E.F. 1993. Plant Propagation by Tissue Culture. Part 1. The Technology. Exegetics, Ltd., Edington. 5. Hartman, R.D. 1974. Dasheen mosaic virus and other phytopathogens eliminated from Caladium, Tara, and Cocoyam by culture of shoot tips. Phytopathology 64:237 240. 6. Jones, J.B. 1986. Determining markets and market potential of horticultural crops. In : Tissue Culture as a Production System for Horticultural Crops. pp. 175 182. R.H. Zimmerman, R.J. Griesbach, F.A. Hammerschlag, and R.H. Lawson (eds.). Martinus Nijhoff Publishers, Boston.
174 7. Miller, L.R. and T. Murashige. 1976. Tissue culture propagation of tropical foliage plants. In Vitro 12:797 813. 8. Morel, G. and C. Martin. 1952. Guerison de dahlias atteints d' une maladie a virus. C.R. Acad. Sci., Paris 235:1324 1325. 9. Morel, G. 1964. Tissue culture: A new means of clonal propagation of orchids. Amer. Orch. Soc. Bull. 33:473 478. 10. Pierik, R.L.M. 1987. In Vitro Culture of Higher Plants. Martin Nijhoff Publishers, B oston. pp. 172 181. 11. Styer, D.J. and C.K. Chin. 1984. Meristem and shoot tip culture for propagation, pathogen elimination, and germplasm preservation. Hort. Rev. 5:221 277. Shoot Culture of Caladium Overview ( Appendix J ) ES E Student Accommodations/ Modifications for Lesson: What accommodations/ modifications/ differentiation did you use to satisfy IEPs, 504 plans, individual student learning styles, and cultural differences? Accommodation Type: Who/When Used or Offered: Co operative Groups : Students will work in pairs during lab activities. Differentiated Instruction : 1. Complete an interactive assignment on line. ( Narrated PPT.) 2. Vocabulary Match All Students paired per ability level as necessary per individualized plan. Lesson Reflection: Note changes to be made, reflections for future use of lesson, and other notes here.
175 Student Laboratory Sheet SHOOT CULTURE OF CALADIUM Purpose: The primary purpose of this laboratory exercise is to demonstrate procedures re quired for performing aseptic technique using surface sterilization and to demonstrate the proper procedures for performing shoot tip explants of Caladium bicolor. INTRODUCTION Shoots of all angiosperms and gymnosperms grow from their apical meristems The apical meristem is the layered dome of actively dividing cells located at the extreme tip of a shoot and measures about 0.1 mm in diameter and 0.25 mm to 0.3 mm in length. Below the apical meristem dome there are ridges of progressively increasing si ze which represent newly formed leaf primordia ( 1). The apical meristematic dome plus one or two accompanying leaf primordia comprise the meristem tip (0.2 1.0 mm long). The shoot tip (1.0 20 mm long) consists of the apical meristem plus numerous le af primordia. When cultured in vitro excised apical meristems, meristem tips and shoot tips exhibit the capacity to regenerate plants that are genetically identical to the donor plants (4,5,7,8,11). Many horticultural crops become infected with pathog ens (virus, bacteria, fungi), which reduce vigor, yield and salability. In 1952, Morel and Martin (8) demonstrated that virus eradicated plants could be regenerated from cultured shoot tips of virus infected dahlia. The recovery of pathogen eradicated pl ants in vitro is because pathogen concentration often is not uniform throughout the infected plant. Frequently, the apical meristematic regions of rapidly elongating shoots remain uninfected by endogenous systemic pathogens. Morel (9) also demonstrated th e effectiveness of shoot tip culture as a rapid in in in vitro propagation procedure. His efforts to recover virus eradicated Cymbidium orchids from infected plants resulted in exclusion of virus as well as a clonal increase of plants at rates substantial ly higher than those achievable by traditional propagation methods. The isolation and culture of apical meristems for the specific purpose of pathogen eradication is called meristem culture Low survival rates of isolated apical meristems and the increase d chance of genetic variability following callus formation usually requires that disease eradication be accomplished using relatively larger meristem tip explants (2,11). This procedure is called meristem tip culture Meristem or meristem tip culture usu ally results in the production of a single unbranched shoot which is then indexed (screened) F igure 4 2. The shoot tip is comprised of the apical meristem and subtending leaf primordia and developing leaves. Photo courtesy of author.
176 for specific contaminants before being clonally multiplied by repeated proliferation of axillary shoots. When pathogen eradication is not a concern, relatively la rger (1.0 20 mm long) shoot tip or lateral bud primary explants, consisting of an apical meristem, leaf primordia and developing leaves are used for clonal propagation by repeated enhanced formation of axillary shoots. This micropropagation procedure is called shoot culture (7). This procedure relies on stimulation of shoot growth from lateral buds following disruption of apical dominance in shoots cultured on media typically supplemented with a cytokinin. Often shoots, first established in vitro by me ristem tip culture, serve as shoot tip or nodal explants for shoot culture. Similar to progress made in orchid production, the application of meristem or meristem tip culture followed by clonal production by shoot culture has had a positive impact on the rapid production of pathogen eradicated ornamental plants (4). One such example is with caladiums ( Caladium bicolor ). Caladium, with about 30 major varieties, is an economically important ornamental plant noted for its production of diversely colored vari egated leaves. Lake Placid, Florida with 50 to 70 million caladium tubers produced yearly is the "Caladium Capital of the World". Caladium varieties are vegetatively propagated by division of underground modified stems called tubers and are susceptible to pathogen infection resulting from unsanitary propagation practices. Consequently, caladium yield and quality are often severely impacted by various bacterial and fungal pathogens and viral pathogens such as Dasheen Mosaic Virus or Konjac Mosaic Virus. The application of meristem tip culture has been reported effective for caladium viral disease eradication (5). In vitro propagation procedures for disease eradicated caladium production had been described by Hartman (5). Tissue cultured caladium shoot cultu res are often impacted by a genetic variation resulting in mutations in plant size, leaf shape, coloration and variegation pattern (3). This variation results in both plant loss and reduced marketability. Purpose: The primary purpose of this laboratory exercise is to demonstrate procedures required for the successful isolation of caladium shoot buds and in vitro establishment of shoot cultures. The sequential micropropagation stages are outlined in Figure 4 9 Shoots produced can be successfully rooted under greenhouse conditions (Figure 4 8 ). MATERIALS AND PROCEDURES Plant Material : Caladium ( Caladium bicolor ) potted plants. Materials : 1) 70% and 95% alcohol solutions 2) 2.1% sodium hypochlorite solution containing 2 drops Tween 20 per 100 ml 3) Sterile distilled deionized water 4) Standard and large size forceps, spatula and scalpel with No. 11 blade 5) Sterile petri dishes 6) 10 culture tubes containing sterile agar solidified Caladium Shoot Establishment Medium
177 7) Stereomicroscope (15x mag nification) 8) Sterile "Disinfecting Bottle" 9) Sterile graduated cylinder Procedures : 1) Obtain a potted caladium plant. Cut off leaves leaving only the base of the petiole. Rinse off potting soil from the roo ts under running tap water (Figure 4 3 ). On a cutting board use a scalpel to cut the shoot tips off ( 4 3 ). Be sure to use forceps to handle the shoots. 2) Place caladium shoot tip into a plastic cup, cover with cheesecloth and fasten with a rubber band (4 3 ). 3 ) Remove surface debris on the shoots by rinsing in flowing tap water for 10 minutes ( PLEASE BE SURE TO USE COLD WATER ). Figure 4 3 Rinsing off potting media from roots, trimming off roots and leaves and cutting shoot explants Photo s courtesy of author. 4 ) Wipe down transfe r hood work space and side walls with 95% alcohol. 5 ) Thoroughly wipe off the dissecting microscope and stage with 70% alcohol. 6 ) Make sure that the glass bead sterilizer is turned on. The core temperature of the glass bead sterilizer should be at lea st 250 C. 7 ) Sterilize scalpel and forceps in glass bead sterilizer. Insert instrument handle bases in the holed rubbe r stopper. Allow to cool. (Figure 4 4 )
178 Figure 4 4 Tools used to complete Caladium bicolor shoot c ulture experi ment:(A) Tissue culture medium (B) Dissecting microscope; (C) Disinfecting bottle;(D) Sterile water; (E) Graduated cylinder with dilute bleach (F) Scalpel and forceps;(G) Sterile petri dishes; (H) Glass bead sterilizer November 18, 2016. Photo courtesy of author. 8 ) Prepare the "Disinfecting Solution" by adding 75 ml sterile water and then 25 ml Clorox Concentrated bleach (containing 8.25% sodium hypochlorite) into a sterile graduated cylinder. Add two drops of Tween 20. This diluted solution will contain ~2.1% sodium hypochlorite. 9 ) After the 10 minute rinsing period, remove cheesecloth and bring cup into the transfer hood. Transfer each shoot tip from the cup using the sterile forceps and place into a sterile "Disinfecting Bottle". 10 ) Cove r the shoot tips with 70% ethanol and soak for 1 minutes. 11 ) Decant off the 70% ethanol and immediately add enough "Disinfecting Solution" to completely cover the shoots. Recap and cover cap with the aluminum foil and then place bottle on the gyratory shaker for 12 minutes. Make a note of the starting time and remove from shaker exactly after 12 minutes. 12 ) IN THE TRANSFER HOOD decant the disinfecting solution into the cup and immediately add sterile water and soak with frequent gentle agitatio n f or 1 minute and repeat twice (Figure 4 5 ) A B C D E F G H
179 Figure 4 5 Step by step procedure for Caladium shoot explant sterilization November 18, 2016 Photo s courtesy of auth or. 13) Place a petri dish half onto the dissecting microscope stage. Using the scalpel with #11 blade, remove the overlying leaf base exposing the underlying lateral bud ( 2 1). Attempt to excise a 2.0 4.0 mm long shoot tip from each shoot ( 2 1) using the dem onstrated procedure (Figure 4 6 ). Figure 4 6 Use stereomicroscope (left) t o trim shoot explants by removing outer leaf base (right); comparison of trimmed and untrimmed shoot explant N ovember 18, 2016. Photo courtesy of author. 14 ) Using a sterile pair of long forceps, transfer a single surface sterilized shoot tip explant int o each of the ten (10) culture tubes containing Caladium Shoot Establishment Medium Be sure that the base of each shoot tip explant is in firm contact with the medium. Be sure to label your tubes Caladium and include the d ate and your initials (Figure 4 7 )
180 Figure 4 7 Inoculation of shoot explants (left) and inoculated culture (right) Photo s courtesy of author. 15 ) Seal each culture tube with one layer of sealing film then place in racks on the culture bench. 16 ) Turn off glass bead sterilize r and wipe off hood work surface with a paper towel moistened with water then spray work surface and sidewalls with 95% ethanol. Turn transfer hood light off. 17 ) After 6 weeks remove plants from culture tubes and collect the growth data as listed un 18 ) To acclimatize plants, divide shoot clusters into individual shoots (micro cuttings) and rinse any residual media off under flowing tap water. Plant micro cuttings in soilless potting mixture in plug tray sheets ( Figure 4 8 ) and cov er with clear vinyl propagation dome. Place in indirect sunlight for two weeks and then gradually remove cover to decrease humidity over a period of 1 week and then move the plants into higher light.
181 Figure 4 8 Acclimatization of rooted Caladium bic olor shoots Photo s courtesy of author. RESULTS Record weekly observations on your cultures for 6 weeks. Be sure to compare differences in the growth responses between sized shoot tip explants from the potted plants. Depending on explant response at the the percentage of surviving explants; 2) the mean number of new shoots produced per original shoot tip explant. Note the presence of abnormal growth. Construct a well designed table or graph of your data. QUESTIONS FOR DISCUSSION You isolated caladium shoot tips from large actively growing shoot tips. If your donor plants were virus infected, do you believe that you will recover any virus eradicated plants? Based on your own experience, is a scalpel equipped wit h a #11 blade the correct instrument to precisely excise caladium shoot tips?
182 Figure 4 9. Micropropagation stages.
183 CULTURE MEDIUM COMPONENTS COMPONENTS CAL A DIUM SHOOT ESTABLISHMENT MEDIUM MG/LITER NH 4 NO 3 1650 KNO 3 1900 CaCl 2 (anhydro us) 332.2 MgSO 4 (anhydrous) 180.7 KH 2 PO 4 (monobasic) 170 FeSO 4 7H 2 O 27.8 Na 2 EDTA 37.26 H 3 BO 3 6.2 MnSO 4 H 2 O 16.9 ZnSO 4 7H 2 O 8.6 KI 0.83 Na 2 MoO 4 2H 2 O 0.25 CuSO 4 5H 2 O 0.025 CoCl 2 H 2 O 0.025 Sucrose 30,000 Myo inositol 100 Thiamine HCL 0.4 2 iP 1 2.0 Agar 8.000 1 6 (gamma, gamma Dimethylallylamino) purine, a cytokinin
184 Teacher Preparation Sheet Caladium Shoot Culture Medium Simplest Method for Medium Preparation by Instructor Note: Order media components from PhytoTechnology Laboratories at: https://phytotechlab.com Product numbers are included in parentheses () below. 1. Add 41.43 g Linsmaier & Skoog Basal Medium W/Sucrose Agar (L467) per liter of distilled water. 2. Add 2 ml 2iP (D217, 6 Dimethylallylamino) purine 1 ) solution (1 mg/mL) per 1.0 liter of prepared medium. 3. Adjust pH to 5.7 with 0.1 KOH. 4. Heat to completely melt medium agar & dispense 12 ml per 150 X 25 mm glass culture tube. Cap each tube and place in a tube rack. Lab el the rack with the medium contained in the tubes. 5. Sterilize by autoclaving. Cool culture tubes on a 45 slant (for culture initiation). 1 2iP is a naturally occurring plant hormone that promotes shoot production. This solution should be stored in the d ark in a refrigera tor.
185 Student Laboratory Template: Scientific Method Laboratory Report NAME: __________________________ DATE: __________________________ CLASS: _________________________ Purpose or Problem What is your experiment about or what question are you trying to answer? ______________________________________________________________________ ______________________________________________________________________ _____________________________________________________ _____________ ____ Hypothesis: Materials: (Include all materials needed and the quantity) 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. Procedures (Write out simple step by step procedures of the experiment). 1. 2. 3. 4. 5. 6. 7. 8. 9. 10.
186 Results and Data Collection (I n the form of a table, include drawings or photos). Score the total number and length of adventitious shoots produced on each stem response for shoot number and length on ea ch medium. Date Measured: Tube Number Number of contaminated explants Shoot number per explant Abnormal growth Week #1 1 2 3 4 5 6 7 8 9 10 Week #2 1 2 3 4 5 6 7 8 9 10 Week #3 1 2 3 4 5 6 7 8 9 10 Week #4 1 2 3 4 5 6 7
187 8 9 10 Week #5 1 2 3 4 5 6 7 8 9 10 We ek #6 1 2 3 4 5 6 7 8 9 10 Total (1) Calculate the percentage of surviving explants. (2) The percentage of surviving explants (3) The mean number of shoots produced per original shoot tip explant. (4) Create a graph in excel of your data and calculations. Note: Students Should Insert Graph from Excel spreadsheet at the end of the laboratory report. Conclusion: (Write a detailed conclusion) What happened? Did you prove or disprove the hypothesis ? What sour ces of error did you find and why do you think the error occurred? Did any contamination occur? If so, was it fungal or bacterial?
1 88 QUESTION S FOR DISCUSSION You isolated caladium shoot tips from large actively growing shoot tips. If your donor plants we re virus infected, do you believe that you will recov er any virus eradicated plants? Based on your own experience, is a scalpel equipped with a #11 blade the correct instrument to precis ely e xcise the caladium shoot tips?
189 CHAPTER 5 CONCLUSIONS AND RECOMMENDATIONS T he purpose of this quantitative study was to : 1) identify Florida agriculture teachers perceived attitudes concerns and barriers for teaching agric ulture biotechnology in Florida; 2) create STEM based plan t biotechnology modules to address th e se concerns; and 3) assess the impact of incorporating flipping instruction on content learning gains in Florida agriculture teachers. In C hapter one the justification for identify ing barriers to implementing biotechn ology in the classroom were described based on published literature Previous survey results have identified that t eacher concerns about teaching biotechnology focused on the lack of equipment, time, and content knowledge This was addressed in this study dules. Secondly, the research question investigated in the study was whether teaching plant biotechnology laboratory modules can be improved by employing a blended instructional method call ed flipping to increase stude nt content knowledge. W ill addressing these concerns lead to more successful integration of plant biotech nology content in to the classroom? In Chapter 2 the need to train agriculture science teachers to effectively integrate ST EM biotechnology curricula in the classroom was examined A descriptive survey, was used to measure the demographics of current Florida agriculture teachers and their attitudes and concerns about biotechnology and determine if differences existed between training and the attitudes of Florida teachers. The survey results revealed that there are three major issues and concerns about teaching biotechnology. These included teacher concerns :
190 1) about th e time needed to teach biotechnology concepts; 2) about student attitudes toward teaching biotechnology; and 3) about demanding teacher schedules which prevent them from integrating biotechnology concepts into the clas sroom. These concerns were similar to the results of previous surveys on attitudes and concerns about teaching biotechnology (Garret, 2009; Kidman, 2009; Kwon & Change, 2009; Mowen et al. 2007a, Wilson et al., 2002). Additionally, it was found that there was a significant lack of pre service and in service training in biotechnology of the teachers surveyed. This training is critical for the development and implementation of the teaching module lessons for the workshops. In Chapter 3 the effectiveness of novel hands on, plant tissue culture mo dules developed by the researcher to address the concerns of time, equipment, and lack of content knowledge were assessed An in service professional development workshop was created to investigate the modules effectiveness in addressing the se concerns. A pretest/postte st experimental design measured content knowledge gain in teacher participants (N=14 ) in the workshop using a paired samples t test. Results indicated that there was a significant gain in posttest score (M = 6.54, SD = 4.08) as opposed to t he pretest, (M= 4.076, SD = 1.85) after completing the lecture overviews and tissue culture laboratory activities resulting in teacher content knowledge gain. In addition, a group dis cussion was conducted with the teacher s participating in the workshop Re sults of the round table discussion were recorded. Teachers reported concerns of time, equipment availability, funding, curric ulum resources and training as the barriers to integrating biotechnology in the ir individual classroom
191 Finally, in Chapter 4 th e following research question examined : Is there a statistical difference between mean group scores in content knowledge of biotechnology teachers who received the traditional lecture method compared to teachers taught using a flipped classroom method ? The research question was posed as a null hypothesis. H O There was no significant difference in mean pretest and posttest scores of the experimental group versus the control group. H A There was a significant difference in mean pretest and posttest sc ores of the contro l versus intervention (flipped) group. After adjustment for pretest scores, t he results of the ANCOVA indicated that there was no significant difference (p = 0.51) in the mean posttest gain scores between the two teaching methods and th e null hypothesis was accepted. The differences that could have contributed to the lack of content knowledge gain in the teachers included the number of years of teaching experience and the amount of biotechnology training acquired by the teache r particip ants. Also, the population sample size was small due to limitations in the teaching laboratory. Teachers in the flipped group, and non flipped group experienced overall learning gains of the content of the workshop, but the incorporation of instructional l ecture in an online format gave teachers the opportunity to individualize the learning process and increase motivation and engagement with the material. Recommendations In this dissertation the barriers in teaching biotechnology in the secondary classro om were addressed As reported almost 10 years ago, the same barriers were still preventing teachers from fully implementing the biotechnology curricula This study provides an effective design of curricula that address the barriers of time, equipment
192 and limited teacher content knowledge gain in plant biotechnology It also provide s a model to develop plant biotechnology curricula in high school based agris cience biotechnology education. Using standard based, blended learning instructional methods in cre ating professional development in service workshops for teachers can provide opportunities for teachers to integrate biotechnology into the classroom. Based on the findings of this study, t he following recommendations for teacher educators in curriculum and training of secondary school educators are presented below. Recommendations for Teacher Education and Curriculum I n service training for agriculture biotechnology teachers should be developed as they can result in content knowledge gains of teachers i n plant biotechnology. Using flipping methods do es not always result in content learning gains. However, teachers should employ blended learning techniques as they offer the student learner increased flexibility in how they can individualize their learnin g and improve engagement. Plant tissue culture modules designed specifically for high school teachers should be developed to provide a clear structure for teaching biotechnology content knowledge and the application of skills in the agriculture biotechnol ogy classroom B oth pre service courses and in service professional training should be develo p ed for Agriculture biotechnology teachers and to promote biotechnology curricula development and implementation Recommendations for Further Research The recomme ndations generated from this study for further research include: Replicated studies using a select group of teachers of a larger sample size to investigate the usefulness of flipped model of instruction for content learning gains and assessment developmen t in agriculture biotechnology. To overcome the barriers of time, equipment availability, and content knowledge, further studies need to be completed using similar in service workshop designs
193 Funding for p re service and in service training to prepare tea chers for biotechnology instruction will need to be increased through national education grants or funding in the Florida legislative budget to increase the integration of agriculture biotechnology in the secondary school classroom. In the present study tw o one day teacher in service workshops were completed a year apart Future studies should investigate the effectiveness of biotechnology workshop teaching methods covered over several days or weeks to fully investigate the effectiveness of the treatment s. This study could be replicated for training students utilizing the teaching lesson modules provided in this study.
194 APPENDIX A IRB APPROVAL FOR SURVEY
195 APPENDIX B INFORMED CONSENT SURVEY LETTER
197 APPE NDIX C TEACHER PLANT BIOTECHNOLGY WORKSHOP AGENDA 2015 Teacher Plant Biotechnology Workshop November 18, 2015 Wendy Vidor and Michael Kane Environmental Horticulture Department University of Florida, Gainesville, Florida
198 Teacher Plant Biotechnology Workshop Wendy Vidor and Michael Kane Environ mental Horticulture Department University of Florida Gainesville, FL WORKSHOP OVERVIEW The overall objective of the workshop is to provide instructional resources, conceptual background information and hands on laboratory experiences to facilitate the incorporation of plant tissue culture into plant biotechnology curricula in the most cost efficient man ner. INSTRUCTIONAL OBJECT IVES Upon completion participants will: Understand the principles and concepts of plant tissue culture, specifically micropropagation Be familiar with the laboratory procedures and equipment used to propagate plants using micro propagation Have instructional materials including PowerPoint lectures, laboratory exercises and other informational resources that can be used in the classroom Know where to obtain supplies to economically and successfully teach plant tissue culture in th e classroom
199 WORKSHOP AGENDA 2043 IFAS Research Drive, Rm. 111 November 13, 2015 Lecture Sessions 8:00 8:30 a.m. Sign in and obtain notebook 8:30 8:40a.m. Welcome & Introductions and Workshop Objectives (Pretest ) 8:40 9: 00 am. Teachers Per spective Ag/Plant Biotechnology. (Survey) 9:00 9:30a.m. Plant Tissue Culture: A Historical Perspective (PowerPoint) 9:30 10:20 a.m. Micropropagation Methods: Shoot Culture, Organogenesis and Non zygotic Embryogenesis (PowerPoi nt) 10:20 10:35 a.m. Break (refreshments provided) 10:35 10:50 p.m. Plant Tissue Culture: Instructional Resources 10:50 12:00 p.m. Laboratory Equipment and Sterile Technique 12:00 12:30 Lunch (Jimmy Johns) Laboratory Ses sions 12:30 1:45 p.m. Media Preparation Shoot Culture of Carnation / Flip 1:45 2:10 2:10 3:00 p.m. Shoot Organogenesis (Parrot feather) ) Already in culture 10 vials 3:00 3:15 Break 3:15 4:00 p.m. Synthetic S eeds 4:00 5:00 p.m. Questions & Discussion (Posttest ) HOTEL ACCOMMODATIONS Participants are responsible for their own lodging and meals. Numerous hotels a re available in the Gainesville. CONTACT S Wendy Vidor email@example.com Dr. Michael Kane Office: 352 273 4500 Cell: 352 359 3565 E mail: firstname.lastname@example.org
200 APPENDIX D IRB APPROVAL TEACHER PLANT BIOTECHNOLOGY WORKSHOP Behavioral/Nonmedical Institutional Review Board FWA00005790 PO Box 112250 Gai nesville FL 32611 2250 Telephone: (352) Facsimile: (352) Email: email@example.com DATE: 10/11/2016 TO: Wendy Vidor 164 Rolling Sands Drive Palm Coast, Florida 32164 FROM: Ira Fischler, Ph.D., Professor Emeritus Chair IRB 02 IRB#: IR B201601613 TITLE: The Effect of Flipping Instruction on Teacher Plant Biotechnology Content Knowledge Approved as Exempt You have received IRB approval to conduct the above listed research project. Approval of this project was granted on 10/11/2016 by IRB 02. This study is approved as exempt because it poses minimal risk and is approved under the following exempt category/categories: 1. This research will be conducted in established or commonly accepted educational settings, involving normal educationa l practices, such as research on regular and special education instructional strategies, or research on the effectiveness of or the comparison among instructional techniques, curricula, or classroom management methods. Principal Investigator Responsi bilities: The PI is responsible for the conduct of the study. Using currently approved consent form to enroll subjects (if applicable) Renewing your study before expiration Obtaining approval for revisions before implementation Reporting Adverse Events Re tention of Research Records Obtaining approval to conduct research at the VA Should the nature of the study change, or you need to revise the protocol in any manner please contact this office pri or to implementation.
201 Study Team : The Foundation for The Gator Nation An Equal Opportunity Institution Confidentiality Notice: This e mail message, including any attachments, is for the sole use of the intended recipients(s), and may contain lega lly privileged or confidential information. Any other distribution, copying, or disclosure is strictly prohibited. If you are not the intended recipient, please notify the sender and destroy this message immediately. Unauthorized access to confidential information is subject to federal and state laws and could result in personal liability, fines, and imprisonment. Thank you.
202 APPENDIX E INFORMED CONSENT LETTER FOR THE EFFECT OF FLIPPING INSTRUCTION ON TEACHER PLAN T BIOTECHNOLOGY CONTENT KNOWLEDGE Informed Consent Protocol Title The Effect of Flipping Instruction on Teacher Plant Biotechnology Content Knowledge Please read this consent document carefully before you decide to participate in this study. Purpose o f the research study The purpose of this research is to determine if Agriculture Biotechnology instruction can be improved by the learner centered flipping instructional method. What you will be asked to do in the study You will be asked to view a video presentation using a flipping instructional method on tissue culture of Caladium. You will be asked to answer embedded questions that will be on the video content of the Caladium shoot culture lesson. You will take a pretest and posttest after the modul e is completed. You will also be asked some open ended After the exercise you will return to the workshop and continue the lab exercises. Time Required 1 hour Risks and Benefits There will not be any discomfort or risks involved in the participation in this study. The possible benefits of this research study will allow participants the opportunity to use instructional materials to teach biotechnology concepts to your students. Compensation There will be no compensation for participating in this research. Confidentiality Your identity will be kept confidential to the extent provided by law. You will be randomly chosen and there will not be any coding associated with your name. The respons Your name will not be used in any report.
203 Voluntary participation Your participation in this study is completely voluntary. There is no penalty for not participating. Righ t to withdraw from the study You have the right to withdraw from the study at any time without consequence. Who to contact if you have questions about the study Wendy Vidor, Graduate Student, Department of Environmental Horticulture, P.O. Box 110675, Ph one 386 569 1055. Michael Kane, PhD, Department of Environmental Horticulture, P.O Box 110675, Phone: 352 273 4500. Who to contact about your rights as a research participant in the study IRB02 Office Box 112250 University of Florida Gainesville, FL 32 611 2250 Phone 352 392 0433 Agreement I have read the procedure described above. I voluntarily agree to participate in the procedure and I have received a copy of this description. Participant: _____________________________________ Date: ______________ Principal Investigator: ____________________________ Date: _______________
204 APPENDIX F THE EFFECT OF FLIPPING WORKSHOP EVALUATION QUESTIONNAIRE How would you rate the overall workshop? What instructional method did you receive? Flippin g or Traditional Lecture? Were the PowerPoint presentations of the lecture materials effective? If you were in the flipping instructional group, did you think this method increased your content knowledge of plant tissue culture? If so, how? If you wer e in the traditional lecture group, did you think this method increased your content knowledge? If so, how? Was the information provided in the lesson plan adequate? Were the hands on experiences using the lab equipment beneficial? How can we improve th is workshop to make it more applicable to your students and instructional needs? Do you think professional development workshops are necessary to teach biotechnology? Why or why not? PLEASE HAVE A SAFE TRIP HOME
205 APPENDIX G TEACHER PL ANT BIOTECHNOLOGY WORKSHOP AGENDA 2016 Teacher Plant Biotechnology Workshop November 18, 2016 Wendy Vidor and Michael Kane Environmental Horticulture Department University of Florida, Gainesville, Florida
206 Teacher Plant Biotechnology Workshop The Effect of Flipping Instruction on Plant Biotechnology Content Knowledge Wendy Vidor and Michael Kane Environ mental Horticulture Department University of Florida Gainesville, FL WORKSHOP OVERVIEW The overall objective of the workshop is to provide instructional resources, conceptual background information and a hands on laboratory experience to effectively facilitate the incorporation of plant tissue culture into plant biotechnology curricula in the most cost efficient manner. INSTRUCTIO NAL OBJECTIVES Upon completion participants, will: Understand the general principles and concepts of plant tissue culture for plant propagation specifically micropropagation Be familiar with the laboratory procedures and equipment used to propagate cal adium by shoot culture Have instructional materials including PowerPoint lectures, laboratory exercises and other informational resources that can be used in the classroom Determine if agriculture biotechnology instruction can be improved by using the lear ner centered flipping instructional method. Know where to obtain supplies to economically and successfully teach plant tissue culture in the classroom
207 WORKSHOP AGENDA 2043 IFAS Research Drive, Rm. 111 November 18, 2016 Lecture Sessions 8:30 9:00 a .m. Sign in and be randomly assigned to a teacher work group. 9:00 9:30 a.m. Welcome & Introductions and Workshop Objectives and Overview (Pre test) 9:30 10:15 am. Work Groups will rotate to complete instructional overview, (groups will be separat ed by instructional method) 10:15 10:30 a.m. Break and Refreshments ( provided) Laboratory Sessions 10:30 11: 00 a.m. Media Preparation for the Shoot Culture of Caladium 11:00 12:15 p.m. Cleaning and Disinfecting Caladium Shoot Exp lants 12:15 1:00 p.m. Lunch ) 1:00 2:00 p.m. Cutting and Transferring Caladium Shoot Explants 2:00 2:30 p.m. Post Test and Survey Questions 2:30 3:30 p m. Questions and Discussion Session 3:30 p.m. Adjournment HOTEL ACCOMMODATIONS (IF NEEDED) Participants are responsible for their own lodging and meals. Numerous hotels a re available in the Gainesville. CONTACT S Wendy Vidor firstname.lastname@example.org email@example.com Cell: 386 569 1055 Dr. Michael Kane, Office: 352 273 4500 Cell: 352 359 3565 E mail: firstname.lastname@example.org
208 UNIVERSITY OF FLORIDA AREA MAP Lecture and lab w ill be held in Rm 111, Building 68 located at 2043 IFAS Research Drive
209 APPENDIX H PRETEST AND POSTTEST SHOOT CULTURE OF CALADIUM Pretest and Posttest Shoot Culture of Caladium True/False Indicate whether the statement is true or false. ____ 1. A cytokinin is added to medium to promote shoot production from lateral shoots by disrupting apical dominance. un der sterile conditions. ____ 3. Tween 20 is a surfactant which promotes penetration of bleach into plant tissue during surface sterilization. ____ 4. Bacteria and virus usually infect the cells of shoot apical meristems of plants. ____ 5. Stage IV acclimatization is the process in which plants produced in the lab are adapted to survive and grow under greenhouse conditions. ____ 6. Microcuttings of all species can be readily rooted in a greenhouse. ____ 7. Shoot culture is the most widely used commercial micropropagation method. ____ 8. Chlorine bleach kills bacteria by changing the structure of bacterial proteins. ____ 9. The exp lant is the piece of tissue used to start a culture. ____ 10. Inoculation is the process of inserting the shoot tip into the medium. Multiple Choice Identify the choice that best completes the statement or answers the question. 11. Shoot Culture is a method for ___________? a. Rapid clonal plant propagation b. Disease eradication c. Low temperature plant storage
210 a. In a sterile environment b. On a defined culture medium c. Under contr olled conditions of light and temperature d. All the above 13. The three methods of micropropagation are _____, _____, _____. a. Shoot culture, root culture, and organogenesis b. Meristem tip culture, shoot organogenesis, and non zygotic embryogenesis c. shoot culture, shoot organogenesis, and non zygotic embryogenesis 14. The purpose of Stage 1 is to? a. Promote rooting b. isolate and sterilize donor tissues c. promote flowering 15. In which stage would the proliferation of axillary shoots be ob served? a. Stage IV b. Stage III c. Stage II 16. The primary purpose of Stage lll is? a. Shoot proliferation b. Microcutting rooting c. Surface sterilization d. Plant ex vitro acclimatization 17. A mother block is_______? a. A set of slowly growin g shoot cultures used as a reliable source of explants for Stage ll cultures. b. A set of donor plants maintained in a greenhouse c. Both A and B 18. What is the primary purpose of a transfer hood? a. It maintains a constant temperature b. Filters out dust particles creating a sterile work environment to decrease contamination. c. It provides adequate light to cut tissue d. All the above 19. A zone of rapidly dividing cells in which is the source of cells making up a plant? a. Flower bud b. Shoot apical meristem c. Root apical meristem d. Leaf tip
211 20. Process of transitioning in vitro produced plants to the ex vitro greenhouse environment. a. acclimatization b. microcutting rooting c. surface sterilization Caladium Procedure 21 a. Tampa, Florida b. Athens, Georgia c. Lake Placid/Sebring, Florida d. Apopka, Florida 22. Caladiums are commercially propagated by? a. Leaf cuttings b. Seed c. Production and division of tubers (underg round modified shoots) 23. One reason for rinsing and cut shoot tips in flowing tap water before surface Sterilization is? a. To prevent shoot explants for drying. b. To remove residual clinging of particulate matter and surface contaminants to increase effective surface sterilization. c. To decrease genetic variation 24. Surface sterilization of cut caladium shoot tips is achieved by? a. Soaking in 70% ethanol for 1 minute followed by soaking in dilute Clorox bleach for 12 minutes and rinsing three times in sterile water. b. rinsing shoot tips in tap water for 10 minutes followed by soaking in dilute Clorox bleach and rinsing three times in sterile water. c. Rinsing shoot tips in tap water for 10 minutes foll owed by soaking in 70% ethanol for 1 minute and rinsing three times in sterile water. 25. Acclimatization of the rooted cutting in Stage IV includes: a. Misting b. Heat c. Fertilization
212 Shoot Culture of Caladium Answer Section TRUE/FALS E 1. ANS: T Cytokinins are a class of growth hormones that promote shoot production in tissue culture 2. ANS: T 3. ANS: T 4. ANS: F 5. ANS: T 6. ANS: F 7. ANS: T 8. ANS: T 9. ANS: T 10. ANS: T MULTIPLE CHOICE 11. ANS: A 12. ANS: D 13. ANS: C 14. ANS: B 15. ANS: C 16. ANS: B 17. ANS: C 18. ANS: B 19. ANS: B 20. ANS: A 21. ANS: C 22. ANS: C 23. ANS: B 24. ANS: A 25. ANS: A
213 APPENDIX I MICROPROPAGATION METHODS OF SHOOT CULTURE, ORGANOGENENSIS AND NON ZYGOT IC EMBRYOGENENSIS POWERPOINT
235 APPENDIX J SHOOT CULTURE OF CALADIUM OVERVIEW POWERPOINT LECTURE
250 LIST OF REFER ENCES Abeysekera, L., & Dawson, P. (2015). Motivation and cognitive load in the flipped classroom: definition, rationale and a call for research. Higher Education Research & Development, 34(1), 1 14. Al Zahrani, A. M. (2015). From passive to active: The i mpact of the flipped classroom British Journal of Educational Technology, 46(6), 1133 1148. Andrews, T. M., Leonard, M. J., Colgrove, C. A., & Kalinowski, S. T. (2011). Acti ve learning not associated with student learning in a random sample of college biology courses. CBE Life Sciences Education, 10(4), 394 405. Ary, D., Jacobs, L.C., & Razavieh, A. (2002). Introduction to research in education (6th ed.). Belmont: Wadsworth/Thomson Learning. Association for Career and Technical Education. (2017). What is CTE? Retrieved July 21, 2017, from https://www.acteonline.org/cte/#.WcsTAciGNPY Aziz, A. N., Tegegne, F., & Wiemers, R. (2009). Benefits of hands on biote chnology training workshops for secondary school educators and college students. Journal of Biotech Research, 1, 72 79. Bandura, A. (1997). Self efficacy: The exercise of control. (C. Hastings, Ed.). New York: W.H. Freeman and Company. Barrows, H. S. (1986 ). A taxonomy of problem based learning methods. Medical Education, 20(6), 481 486. Bergmann, J., & Sams, A. (2012). Flip your classroom: reach every student in every class every day (1st ed.). Eugene: International Society for Technology in Education; ASC D. Bigler, A. M., & Hanegan, N. L. (2011). Student content knowledge increases after participation in a hands on biotechnology intervention. Journal of Science Education and Technology, 20(3), 246 257. Biotechnology Innovation Organization (Ed.). (2017). W hat is biotechnology? Retrieved from https://www.bio.o rg/what biotechnology Bishop, J. L., & Verleger, M. (2013). The flipped classroom: A survey of the research. 120th ASEE Annual Conference & Exposition. Atlanta: American Society of Engineering Educati on.
251 Blair, E., Maharaj, C., & Primus, S. (2016). Performance and perception in the flipped classroom. Education and Information Technologies, 21(6), 1465 1482. Boone, H., Gartin, S., Boone, D., & Hughes, J. (2006). Modernizing the agricultural education c knowledge, and understanding of biotechnology. Journal of Agricultural Education, 47(1), 78 89. Borgerding, L., Sadler, T. D., & Koroly, M. J. (2012). Teachers concerns about biotechnolo gy education. Journal of Science Education Technology, 22, 133 147. Brame, C. (2016). Effective educational videos: Principles and guidelines for maximizing student learning from video content. CBE Life Science Education, 15(4), 6. Brame, C. (2013). Flip ping the classroom. Vanderbilt University Center for Teaching. Retrieved (October 5, 2017) from https://cft.vanderbilt.edu/guides sub pages/flipping the classroom/. Brown, D. C., Kemp, M.., & Hall, J. (1998). On teaching biotechnology in Kentucky. Journal of Industrial Technology Education, 35(4), 44 60. Brown, J. C., Bokor, J. R., Crippen, K. J., & Koroly, M. J. (2014). Translating current science into materials for high school via a scientist teacher partnership. Journal of Science Teacher Education, 25(3 ), 239 262. Bruner, J. (1985). An historical and conceptual perspective. Culture, communication, and cognition: Vygotskian perspectives. New York, NY: Cambridge University Press. Bryce, T., & Gray, D. (2004). Tough acts to follow: The challenges to scien ce teachers presented by biotechnological progress. International Journal of Science Education, 26(6), 717 733. Bureau of Labor Statistics (2017). The employment situation. September 1,2017, U.S Department of Labor News Release USDL 17 1177, Washington, D .C. Butcher, K. R. (2014). The Multimedia Principle. In R. E. Mayer (Ed,),The Cambridge Handbook of Multimedia Learning (2nd ed.) 174 205. New York: Cambridge University Press. Campbell, D. T., & Stanley, J. C. (1963). Experimental and quasi experimental designs for research. In. Boston: Houghton Mifflin. Center for Precollegiate Education and Training. (2017). Retrieved August 13, 2017, from https://www.cpet.ufl.edu/ Clark, R. C., & Mayer, R. E. (2016). e Learning and the Science of Instruction (4th ed.). Hoboken, N.J.: John Wiley & Sons, Inc.
252 Conner, N. W., Rubenstein, E. D., DiBenedetto, C., Stripling, T., Roberts, G., & Stedman, N. L. P. (2014). Examining student perceptions of flipping an agricultural teaching methods course. Journal of Agriculture Edu cation, 55(5), 65 77. Cole, M., Feild, H., & Harris, S. (2004). Student learning motivation and psychological Academy of Management Learning and Education, 3(1), 64. Conner, N. W ., Rubenstein, E. D., DiBenedetto, C., Stripling, T., Roberts, G., & Stedman, N. L. P. (2014). Examining student perceptions of flipping an agricultural teaching methods course. Journal of Agriculture Education, 55(5), 65 77. Conner, N. W., & Stripling, C. T. (2014). Flipping an agricultural education teaching methods course. Journal of Agricultural Education, 55(2), 66 78. Cornell University, U.S. Agency for International Development, Agricultural Biotechnology Support Project II, & Program for Biosafety Systems. (2004). What is Agricultural Biotechnology. Cornell University. Curriculum for Agricultural Science Education. (2017). Retrieved from http://case4learning.org/index.php/curriculum Davies, R. S., Dean, D. L., & Ball, N. (2013). Flipping the classro om and technology integration in a college level information systems spreadsheet course. Educational Technology Research and Development, 61(4), 563 580. Davis, E. A., & Krajcik, J. S. (2005). Designing educative curriculum materials to promote teacher lea rning. Educational Researcher, 34(3), 3 14. Day, J. A., & Foley, J. D. (2006). Evaluating a web lecture intervention in a human computer interaction course. IEEE Transactions on Education, 49(4), 420 431. Deslauriers, L., Schelew, E., & Wieman, C. (2011). Improved learning in a large enrollment physics class. Science, 332(6031), 862 864. Despain, D., North, T., Warnick, B. K., & Baggaley, J. (2016). Biology in the Agriculture Classroom: A descriptive comparative study. Journal of Agricultural Education, 57( 1). Dimitrov, D. M., & Rumrill, P. D., Jr. (2003). Pretest posttest designs and measurement of change. Work, 20(2), 159 165.
253 Doolittle, P. E., & Camp, W. G. (1999). Constructivism: The career and technical education perspective. Journal of Vocational and Technical Education, 16(1), 1 21 Dunham, T., Wells, J., & White, K. (2002). Biotechnology Education: A multiple instructional strategies approach. Journal of Technology Education, 14(1). Dweck, C. S. (1986). Motivational processes affecting learning. Ameri can Psychologist, 41(10), 1040 1048. Eccles, J. S., & Wigfield, A. (2002). Motivational beliefs, values, and goals. Annual Review Psychology, 53, 109 132. Ejiwale, J. A. (2012). Facilitating teaching and learning across STEM fields. Journal of STEM Educat ion: Innovations and Research, 13(3), 87. Executive Office of the President National Science and Technology Council. (2013). Federal Science, Technology, Engineering, And Mathematics (STEM) Education Strategic Plan. Washington, D.C. S., & Bozick, R. (2011). Characteristics of career academies in 12 Florida school districts. Issues & answers. REL 2011 No. 106. Regional Educational Laboratory Southeast. Ewing, J., & Whittington, S. (2009). Describing the cognitive levels of professor d iscourse and student cognition in college of agriculture class sessions. Journal of Education. 50, 49. Fenstermacher, G. D. (1994). The knower and the known: The nature of knowledge in research on teaching. Review of Research in Education, 20, 3. Field, A ndy. (2013). Discovering Statistics Using IBM SPSS Statistics (4th ed.). London: Sage Publishing. Fisher, R. A. (1960). The Design of Experiments (7th ed.). London: Oliver and Boyd Flipped Learning Network. (2017). Flipped Learning Definition. Retrieved fr om http://flippedlearning.org/definition of flipped learning/ Florida Department of Education. (2016). Defining STEM. Tallahassee: Florida Department of Education. Retrieved from http://www.fldoe.org/academics/standards/subject areas/math science/stem/defi ning stem.stml Florida Department of Education. (2017). Agriculture Food and Natural Resources. Florida Department of Education. Retrieved from http://www.fldoe.org/academics/career adult edu/career tech edu/agriculture food natural resources.stml
254 Freeman, S., Eddy, S., McDonough, M., Smith, M. K., Okoroafor, N., Jordt, H., & Wenderoth, M. P. (2014). Active learning increases student perfo rmance in science, engineering and mathematics. JSTOR, 111(23), 8410 8415. Gardner. (2012). The inverted agricultural education classroom: A new way to teach? A new way to learn? Paper presented at the Agriculture and Applied Economics Associations Annual Meeting, Seattle, WA. Garrett, S. (2009). Professional development for the integration of biotechnology education. Queensland University of Technology, Australia. Gokhale, A. (1995). Collaborative learning enhances critical thinking. Journal of Technical Education, 7(1). Graham, C. (2004). Blended Learning Systems: Definition, Current Trends and Future Directions. San Francisco, CA: Pfeiffer Publishing. Graham, M., Frederick, J., Byars Winston, A., Hunter, A. B., & Handelsman, J. (2013). Increasing persis tence of college students in STEM. Science, 341(6153), 1455 1456. Hambleton, R. K., Swaminathan, H., & Rogers, H. J. (1991). Fundamentals of item response theory. London: Sage. Hanegan, N. L., & Bigler, A. (2009). Infusing authentic inquiry into biotechno logy. Journal of Science Education and Technology, 18(5), 393 401. Hord, S. M., Rutherford, W. L., Huling Austin, L., & Hall, G. E. (1987). Taking charge of change. Alexandria, VA: Association for Supervision and Curriculum Development. Jamaludin, R., O sman M.D. S. Z. (2014). The use of a flipped classroom to enhance engagement and promote active learning. 5, 124 131. Kane, M. E., Philman, N. L., & Jenks, M. A. (1994). A laboratory exercise to demonstrate direct and indirect shoot organogenesis using Internodes of Myriophyllum aquaticum. HortTechnology, 4(3), 317 320. Kidman, G. (2009). Attitudes and interests towards biotechnology: The mismatch between students and teachers. Eurasia Journal of Mathematics, 5(2), 135 143. Kuenzi, J. J. (2008). Science technology, engineering, and mathematics (STEM) education: Background, federal policy, and legislative action. Retrieved from http://digitalcommons.unl.edu/crsdocs/35/ Kutner, M. H., Nachtsheim, C. J., Neter, J., & Li, W. (2005). Applied linear statistic al models (5th ed.). New York, NY: McGraw Hill.
255 Kwon, H. (2009). Key factors affecting the implementation of biotechnology instruction in secondary school level technology education classrooms. Virginia Polytechnic Institute and State University, Blacksbur g, VA. its instruction in South Korea. Journal of Technology Studies, 35, 67 75. Locke, E. A. (1991). The motivation sequence, the motivation hub, and the motivation core. Organizational Behavior and Human Decision Processes, 50(2), 288 299. Lord, F. M., Wainer, H., & Messick, S. (1983). Principals of modern psychological measurement: A festschrift for Frederic M. Lord. Hillsdale, NJ: L. Erlbaum Associates. Martin, A.., & Schwartz, E. (2014). Making space for active learning: The art and practice of teaching. New York: Teachers College Press. Mason, G. S., Shuman, T. R., & Cook, K. E. (2013). Comparing the effectiveness of an inverted classroom to a traditional classroom in an upper division engineering course. IEEE Transaction on Education, 56, 430 435. McCallum, S. Schultz, J., Selke, K., & Spartz, J.(2015). An examination of the flipped classroom approach of college student academic involvement. International Journal of Teaching & Learning in Higher Education, 27(1), 42 45. McCubbins, O. P., Paulsen, T. H., & Anderson, R. G. (2016). Student perceptions concerning their experience in a flipped undergraduate capstone course. Journal of Agricultural Education, 57(3), 70. Mc Laughlin, J. E., Griffin, L. M., Esserman, D. A., Davidson, C. A., Glatt, D. M., Roth, M. T., Mumper, R. J. (2013). Pharmacy student engagement, performance, and perception in a flipped satellite classroom. American Journal of Pharmaceutical Education, 77( 9), 196. McLean, S. S. (2016). Flipped classrooms and student learning: Not just surface gains. Advances in physiology education, 40(1), 47 55. McLeod, S. (2015). Jean Piaget. Retrieved from https://www.simplypsychology.org/simplypsychology.org Jean Piaget.pdf McTighe, J., & Wiggins, G. (2012). Understanding by design framework. Alexandria, VA: Association for Supervision and Curriculum Development. Michael, J. (2006). Physiology Education, 30(4), 159 167.
256 Minhas, P. (2017). The effects of passive and active learning on student preference and performance in an undergraduate basic science course. Anatomic al Science Education, 5(4), 200 207. Moreland, J., Jones, A., & Cowie, B. (2006). Developing pedagogical content knowledge for the new sciences: The example of biotechnology. Teaching Education, 17(2), 143 155. Mowen, D., Roberts, G., & Wingenbach, H. J. ( 2007a). Biotechnology: An assessment Education, 48(1), 42 51. Mowen, D. L., Wingenbach, G. J., Roberts, T. G., & Harlin, J. F. (2007b). Agricultural rs, roles, and information source preferences for teaching biotechnology topics. Journal of Agricultural Education, 48(2), 103 113. Mueller, A. L., Knobloch, N. A., & Orvis, K. S. (2015). Exploring the effects of active biotechnology and genetics instruction. Journal of Agricultural Education, 56(2), 138 152. Myers, B. E., & Dyer, J. E. (2004). Agriculture teacher education programs: A synthesis of the literature. Journal of Agricultur al Education, 45(3), 44 52. National Academies Press. (2017). A Framework for K 12 Education: Practices, Cross cutting concepts and core ideas (p. 401). Washington D.C. National Council for Agricultural Education. (2015). Biotechnology systems career path way. Retrieved from https://www.ffa.org/SiteCollectionDocuments/council_biotechnology_systems_ca reer_pathway.pdf National Research Council. (2000). How people learn: Brain, mind, experience, and school: Expanded edition. Washington D.C: National Academies Press. National Research Council. (2009). Transforming agricultural education for a changing world. Washington, D.C.: National Academy of Sciences. National Research Council. (2012). A framework for K 12 science education; Practices, crosscutting concepts, and core ideas. Washington, D.C.: National Academies Press. National Science and Technology Council. (2013). Federal Science, Technology, Engineering, And Mathematics (STEM) Education Strategic Plan. Washington, D.C.: Executive Office of the President Nat ional Science and Technology Council. Retrieved from https://www.whitehouse.gov/sites/whitehouse.gov/files/ostp/Coordinating_Federa l_Science_Technology_Engineering_and_Mathematics.pdf
257 National Science Board. (2014). Revisiting the STEM workforce: A companion to science and engineering indicators. Na tional Science Foundation. Retrieved from https://www.nsf.gov/pubs/2015/nsb201510/nsb201510.pdf National Science Foundation. (2014). CTE pathways to STEM education. National Science Foundation. Retrieved from http://www.successfulstemeducation.org/sites/s uccessfulstemeducation.org/files/ CTE%20Pathways%20to%20STEM%20Occupations_0.pdf Overmyer, G. R. (2014). The flipped classroom model for college algebra: Effects on student achievement. Colorado State University, Fort Collins, CO. Pelech, J., & Pieper, G. W (2010). The comprehensive handbook of constructivist teaching: From theory to practice. Retrieved from http://onlinelibrary.wiley.com/doi/10.1111/teth.12016/abstract Piaget, J. (1952). The origins of intelligence in children. New York: International Univ ersities Press. Piaget, J. (1959). The language and thought of the child. Psychology Press: Chicago. Pollock, S. J., & Finkelstein, N. D. (2008). Sustaining educatio nal reforms in introductory physics. Physical Review Special Topics Physics Education Research, 4(1). Price, M. (2012). Pushing students towards STEM. Retrieved from http://www.sciencemag.org/careers/2012/07/pushing students toward stem Reicks, T., Hasse l, C., & Carr, T. (1996). A Case Approach to Food Biotech, 28(1), 33 38. Roach, T. (2014). Student perceptions toward flipped learning: New methods to increase interaction and active learning in economics. International Review of Economics Education, 17, 7 4 84. Roehl, A., Reddy, S. L., & Shannon, G. J. (2013). The flipped classroom: An opportunity to engage millennial students through active learning strategies. Journal of Family and Consumer Sciences, 105(2), 44 49. Rothwell, W. J. &, & Kazanaz, H. C. (19 92). Mastering the Instructional Design Process: A systematic Approach. San Francisco: Jossey Bass Publishers. Russell, J. (2003). The assessment of student learning is a pivotal component of effective teaching and learning. The Technology Teacher, 63(2), 28 32. of biotechnology. International Journal of Science Education, 30(2), 167 183.
258 Rich Lear ning, 70(6), 16 20. Sanders, M. (2001). New paradigm or old wine? The status of technology education practice in the United States. Journal of Technology Education, 12(2), 35 55. Sanford, J. C., Klein, T., Wolf, E., & Allen, N. (1987). Delivery of substanc es into cells and tissues using a particle bombardment process. Journal of Particle Science Technology, 5, 27 37. Savage, E., & Sterry, L. (1995). A conceptual framework for technology education. Technology Teacher, 50(2), 7 11. Savery, J. R., & Duffy, T. M. (2001). Problem Based Learning: An instructional model and its constructivist framework. Retrieved from http://citeseerx.ist.psu.edu/viewdoc/summary?doi=10.1.1.35.4403 Schunk, D. H. (2004). Learning Theories: An educational perspective. (4th ed.). Upper Saddle River, New Jersey: Pearson/Merrill/Prentice Hall. Scott, D., Washer, B., & Wright, M. (2006). A Delphi study to identify recommended biotechnology competencies for first year/initially certified technology education teachers. Journal of Technology Education, 17(2), 44 56. Shimamoto, D. (2012). Implementing a flipped classroom: An instructional module. PowerPoint presented at the Technology, Colleges, and Community Worldwide Online Conference, Honolulu, HI. Shulman, L. S. (1986). Those who understand : knowledge growth in teaching. Educational Researcher, 15(2), 4. Splan, R., Porr, S., & Broyles, T. (2011). Undergraduate research in agriculture: constructivism and the scholarship of discovery. Journal of Agricultural Education, 52(4), 56 64. Steele, F., & Aubusson, P. (2004). The challenge in teaching biotechnology. Research in Science Education, 34(4), 365 387. Stevenson, C. D., & Harris, G. K. (2014). Instruments for characterizing instructors' teaching practices. NACTA Journal, 58(2), 102 108. St rayer, J. F. (2012). How learning in an inverted classroom influences cooperation, innovation, and task orientation. Learning Environments Research, 15(2), 171 193. Stuart, L., & Dahm, E. (1999). 21st century skills for 21st century jobs. Federal Publicati ons, 151.
259 Stubbs, E. A., & Myers, B. E. (2016). Part of what we do: Teacher perceptions of STEM integration. Journal of Agricultural Education, 57(3). Szparagowski, R. (2014). Exploring the effectiveness of the flipped classroom. Retrieved from Bowling Gre en State University, Ohio: Thoron, A. C. (2010). Effects of inquiry based agriscience instruction on student argumentation skills, scientific reasoning, and student achievement. University of Florida. Retrieved from http://etd.fcla.edu/UF/UFE0041468/thoron_a.pdf Tschannen Moran, M., & Hoy, A. W. (10) Teacher efficacy: capturing an elusive construct. Teaching and Teacher Education, 17(7), 783 805. Tucker, B. (2012). The Flipped Classroom: Online instruction at home frees class time for learning. Education Next,12, 82 83. Turpen, C., & Finkelstein, N D. (2009). Not all interactive engagement is the same: Variations in physics professors and implementation of peer Instruction. Physics Education Research, 5(2). University of Florida. (2017). Center for Precollegiate Education and Training; University of Florida. University of Florida. Retrieved from https://www.cpet.ufl.edu/ University of Florida Center of Regenerative Health Biotechnology. (2017). Biotility. Retrieved from http://biotility.research.ufl.edu/academic programs in biotechnology.html U.S. Agency for International Development, Agricultural Biotechnology Support Project II, & Program for Biosafety Systems. (2004). What is Agricultural Biotechnology. Cornell University. Vasil, I. K. (2008). A history of plant biotechnology: from the Cell Theor y of Schleiden and Schwann to biotech crops. Plant Cell Reports, 27(9), 1423 1440. Vygotsky, L. (1978). Mind in Society: The development of higher psychological processes. Harvard University Press. ed.). New York: Longman. Wells. (1994). Establishment of a taxonomic structure for the study of biotechnology as a secondary school component of technology education. Jour nal of Technology Education, 6(1), 58 75. Wells, J. G., & Kwon, H. (2008). Inclusion of biotechnology in US standards for technological literacy: Influence on South Korean technology education Technology Conference.
260 Wilson, E., Kirby, B., & Flowers, J. (2002). Factors influencing the intent of North Carolina agricultural educators to adopt agricultural biotechnology curriculum. Journal of Agricultural Education, 43(1), 69 81. Wiggins, G., & McT ighe, J. (2005). Understanding by Design Framework (2nd ed.). Alexandria, VA: Association for Supervision and Curriculum Development (ASCD). Wigfield, A., & Eccles, J. S. (2000). Expectancy value theory of achievement motivation. Contemporary Educational Psychology, 25(1), 68 81. Zeller, M. (1994). Biotechnology in the high school biology curriculum: The future is here! The American Biology Teacher, 56(8), 460 464. Zhang, D., Zhou, L., Briggs, R., & J., N. J. (2006). Instructional video in e learning: As sessing the impact of interactive video on learning effectiveness. Science Direct, 43, 15 27.
261 BIOGRAPHICAL SKETCH Wendy Vidor was ra ised in Tinley Park, Illinois. After high school, Wendy attended Northern Illinois University in Dekalb, IL. Starting her career as a physical therapy major, Wendy switched her major to secondary education and earned a Bachelor of Science degree We ndy worked in the Horticulture i ndustry and studied h orticulture at Joliet Junior College, in Joliet, IL. After several years in the nursery and landscape industry, Wendy returned to teaching secondary education and taught physical e ducation and health education. Wendy and her family deci ded to move to Florida and she taught agriculture education in middle school. In 2008, Wendy de cided to pursue a Master of S cience degree in agricultural education at the University of Florida, Gainesville through the ir online program. During that time, she moved to the high school level and taught agriscience education, horticulture and agricul ture biotechnology. In 2013, after working on a grant project with her high school students and the Environmental Horticulture Micropropagati on Laboratory to grow sea o ats she became interested in learning more about tissue culture. Wendy decided to pursue he r doctoral degree in the Environmental Horticulture Department at the University of Florida while working as a full time teacher a nd graduated in December 2017. In her spare time Wendy enjoys spending time with her family, going to the beach and gardening.