Organic Amendment to Reduce Nitrogen Loss in Sandy Soils

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Title:
Organic Amendment to Reduce Nitrogen Loss in Sandy Soils
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english
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Gu, Wen
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University of Florida
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Gainesville, Fla.
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Thesis/Dissertation Information

Degree:
Master's ( M.S.)
Degree Grantor:
University of Florida
Degree Disciplines:
Soil and Water Science
Committee Chair:
He, Zhenli
Committee Co-Chair:
Ma, Lena Q
Committee Members:
Silveira, Maria L
Wilson, Patrick C
Stoffella, Peter J

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Subjects / Keywords:
nitrogen
Soil and Water Science -- Dissertations, Academic -- UF
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Soil and Water Science thesis, M.S.
bibliography   ( marcgt )
theses   ( marcgt )
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Electronic Thesis or Dissertation

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Abstract:
Nitrogen (N) is a nutrient often associated with acceleratedeutrophication of surface water. The extremely low holding capacity fornutrients and moisture is the fundamental problem of sandy soil in Florida,which causes losses of nutrients into waterways. Soil organic amendment canimprove soil quality by increasing soil’s holding capacity for nutrients andmoisture and is, therefore, a promising approach to reduce loads of nutrientsand toxic metals in surface runoff water and/or leachate. Organic amendment canincrease or decrease mineral N in soil, depending on the C/N ratio of theorganic materials and N availability in the soil. Hydra-Hume is a commercial product, which shows the potential inenhancing soil N availability or reducing fertilizer N loss. In this study,incubation, column leaching, and laboratory analysis were conducted to examinethe effects of Hydra-Hume on transformation dynamics and leaching potential ofmineral N in two representative soils (Alfisol and Spodosol) under citrusproduction in south Florida. The obtained results indicated that Hydra-Humeamendment enhanced assimilationof mineral N, when it is applied to the soil, however,soil orginal C/N ratio also need to be considered. Hydra-Hume applicationreduced NO3-N leaching regardless of chemical fertilizer addition. Significantreduction in total NO3-N amount could be observed when Hydra-Humeapplied alone in both of the soils, but an application rate of 44.8 kg ha-1is required. If fertilizer was incorporated with the treatments, significantreduction of total NO3-N amounts and total mineral N amount could beobserved, but a minimal application rate of 44.8 kg ha-1 is requiredto accomplish significant results in Spodosol and 22.4 kg ha-1 inAlfisol.
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In the series University of Florida Digital Collections.
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Includes vita.
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Description based on online resource; title from PDF title page.
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This bibliographic record is available under the Creative Commons CC0 public domain dedication. The University of Florida Libraries, as creator of this bibliographic record, has waived all rights to it worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law.
Statement of Responsibility:
by Wen Gu.
Thesis:
Thesis (M.S.)--University of Florida, 2012.
Local:
Adviser: He, Zhenli.
Local:
Co-adviser: Ma, Lena Q.
Electronic Access:
RESTRICTED TO UF STUDENTS, STAFF, FACULTY, AND ON-CAMPUS USE UNTIL 2013-12-31

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lcc - LD1780 2012
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UFE0044801:00001


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1 ORGANIC AMENDMENT TO REDUCE NITROGEN LOSS IN SANDY SOILS By WEN GU A THESIS PRESENTED TO THE GRADUATE SCHOOL OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE UN IVERSITY OF FLORIDA 2012

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2 2012 Wen Gu

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3 To all who nurtured my intellectual curiosity, academic interests, and sense of scholarship throughout my lifetime, making this milestone possible

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4 ACKNOWLEDGEMENTS Fi rst of all, I would like to thank my esteemed advisor, Dr. Zhenli He, for his expert guidance and support. Not only was he readily available for me providing comprehensive instructions in the lab, but also he always read and responded to the drafts of my w ork in time and with patience. His comments are always perceptive and helpful, without which this dissertation would not have been finished. My thanks go out to my co advisor, Dr. Lena Q. Ma and colleagues from Biogeochemistry of Trace Metals group for the hospitality and support I had received during my first two pertinent suggestions for the research group as well as her class. I would also like to give my since re thanks to Drs. Peter J. Stoffella, Patrick C. Wilson and Maria L. Silveira for serving on my advisory committee. I am grateful for their advice, assistance I deeply express my appreciation to m y colleagues in Soil and Water Science Laboratory: Drs. Yuangen Yang, Shengke Tian, Zhigang Li, Suli Li, Ying Guo, Jinghua Fan, Ph.D. students Santanu Bakshi, Mr. Brian Cain, for providing useful academic and social assistance. We studied and relaxed toget her. Without their help and friendship, the successful completion of my Master learned, enjoyed and benefited from teamwork. I appreciate the people on the f aculty and staff at Soil and Water Science Department as well as Indian River Research Center of University of Florida for providing me wonderful environment for both studying and living during the two years. I am particularly thankful to Dr. Peter J. Stof fella, Ms. Laura McKeon, and Ms. Velma Spencer for their timely and efficient

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5 assistance and arrangement in the student dorm. I want to also thank Mr. Michael Sisk and Ms. Jackie White for all their helps with my course registration, defense arrangement, e tc Last but not least, I would like to thank my parents and all of my friends for their love and support. I will cherish all the happy moment and memory.

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6 TABLE OF CONTENTS page ACKNOWLEDGEMENTS ................................ ................................ ................................ ............. 4 LIST OF TABLES ................................ ................................ ................................ ........................... 8 LIST OF FIGURES ................................ ................................ ................................ ......................... 9 ABSTRACT ................................ ................................ ................................ ................................ ... 11 CHAPTE R 1 LITERATURE REVIEW ................................ ................................ ................................ ....... 13 Nitrogen and Plant Nutrition ................................ ................................ ................................ .. 13 The Role of Nitrogen in Sustainable Ag riculture ................................ ................................ ... 14 Soil Nitrogen ................................ ................................ ................................ ........................... 19 Nitrogen Transformation and Availability in Soil ................................ ................................ .. 21 Nitrogen Losses into the Environment ................................ ................................ ................... 22 Implication of C/N Ratio in Nitrogen Management ................................ ............................... 26 Conclusions and Per spectives ................................ ................................ ................................ 35 2 ORGANIC AMENDMENT EFFECT ON NITROGEN TRANSFORMATION IN SOILS ................................ ................................ ................................ ................................ ..... 37 Introduction ................................ ................................ ................................ ............................. 37 Materials and Methods ................................ ................................ ................................ ........... 39 Soil Collection and Characterization ................................ ................................ ............... 39 Incubation Study and Soil N Dyna mics Analysis ................................ ........................... 39 Data Analysis ................................ ................................ ................................ ................... 40 Results and Discussion ................................ ................................ ................................ ........... 40 Effe cts of Organic Amendment on Soil pH and EC ................................ ........................ 40 Effects on N Dynamics in Soils ................................ ................................ ....................... 41 Conclusion ................................ ................................ ................................ .............................. 43 3 ORGANIC AMENDMENT EFFECTS ON NITROGEN LEACHING ................................ 54 Introduction ................................ ................................ ................................ ............................. 54 Materials and Methods ................................ ................................ ................................ ........... 55 Results and Discussion ................................ ................................ ................................ ........... 57 Effects of Organic Amendment on Soil pH and EC ................................ ........................ 57 Effect on N Leaching and Soil N Pool ................................ ................................ ............ 58 Potential Effects on Heavy Metals ................................ ................................ .................. 59 Characterization of Leached Soil ................................ ................................ .................... 60 Conclusion ................................ ................................ ................................ .............................. 60

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7 4 SUMMARY, CONCLUSIONS AND PERSPECTIVE ................................ ......................... 73 Summary and Conc lusions ................................ ................................ ................................ ..... 73 Perspective ................................ ................................ ................................ .............................. 75 LIST OF REFERENCES ................................ ................................ ................................ ............... 76 BIOGRAPHICAL SKE TCH ................................ ................................ ................................ ......... 85

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8 LIST OF TABLES Table page 2 1 General properties of the two soils and Hydra Hume ................................ ....................... 51 2 2 General properties of the Hydra Hume ................................ ................................ .............. 51 2 3 Statistical analysis of KCl extractable NH 4 N in Alfisol ................................ ................... 52 2 4 Statisti cal analysis of KCl extractable NO 3 N in Alfisol ................................ ................... 52 2 5 Statistical analysis of KCl extractable NO 3 N in Spodosol ................................ ............... 53 3 1 Statist ical analysis of NO 3 N concentration and total amount of NO 3 N in leachate from Spodosol ................................ ................................ ................................ .................... 70 3 2 Statistical analysis of NH 4 N concentration and total mineral N in leachate from Spodosol ................................ ................................ ................................ ............................. 70 3 3 Statistical analysis of heavy metals in leachate from Spodosol ................................ ......... 71 3 4 Statistical analysis of NO 3 N and total amount of NO 3 N i n leachate from Alfisol .......... 71 3 5 Statistical analysis of NH 4 N concentration and total mineral N in leachate from Alfisol ................................ ................................ ................................ ................................ 72 3 6 S tatistical analyses of heavy metals in leachate from Alfisol ................................ ............ 72

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9 LIST OF FIGURES Figure page 2 1 Sampling site ................................ ................................ ................................ ...................... 44 2 2 Effect of organic amendment on Alfisol pH as a function of incubation time .................. 45 2 3 Effect of organic amendment on Spodosol pH as a function of incubation time .............. 45 2 4 Effect of organic amendment on Alfisol Electrical conductivity (EC) as a function of incubation time ................................ ................................ ................................ ................... 46 2 5 Effect of or ganic amendment on Spodosol electrical conductivity (EC) as a function of incubation time ................................ ................................ ................................ .............. 46 2 6 Effect of organic amendment on KCl extractable NH 4 N in Alfisol ................................ 47 2 7 Effect of organic amendment and fertilizer on KCl extractable NH 4 N in Spodosol ....... 47 2 8 Effect of organic amendment on KCl extractable NO 3 N in Alfisol ................................ 48 2 9 Effect of organic amendment and fertilizer on KCl extractable NO 3 N in Spodosol soil. ................................ ................................ ................................ ................................ ..... 48 2 10 Effect of organic amendment on KCl extractable NH 4 N in Spodosol ............................. 49 2 11 Effect of organic amendment and fertilizer on KCl extractable NH 4 N in Spodosol ....... 49 2 12 Effect of organic amendment on KCl extractable NO 3 N in Spodosol ............................. 50 2 13 Effect of organic amendment and fertilizer on KCl extractable NO 3 N in Spodosol ....... 50 3 1 Column leaching setup ................................ ................................ ................................ ....... 62 3 2 pH of leachate form Spodosol amended with fertilizer and different rates of Hydra Hume ................................ ................................ ................................ ................................ .. 63 3 3 pH of leachate form Alfisol amended with fertilizer and different rates of Hydra Hume ................................ ................................ ................................ ................................ .. 63 3 4 Leachate EC form Spodosol in relation to fertilizer and different r ates of Hydra Hume ................................ ................................ ................................ ................................ .. 64 3 5 Leachate EC form Alfisol in relation to fertilizer and different rates of Hydra Hume ..... 64

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10 3 6 Leachate N H 4 N concentration from Spodosol in relation to different rates of Hydra Hume alone ................................ ................................ ................................ ........................ 65 3 7 Leachate NH 4 N concentration from Spodosol in relation to fertilizer and different rates of Hydra Hum e ................................ ................................ ................................ .......... 65 3 8 Leachate NO 3 N concentration from Spodosol in relation to different rates of Hydra Hume alone ................................ ................................ ................................ ........................ 66 3 9 Leachate NO 3 N con centration from Spodosol in relation to fertilizer and different rates of Hydra Hume ................................ ................................ ................................ .......... 66 3 10 Leachate NH 4 N concentration from Alfisol in relation to different rates of Hydra Hume alone ................................ ................................ ................................ ........................ 67 3 11 Leachate NH 4 N concentration from Alfisol in relation to fertilizer and different rates of Hydar Hume ................................ ................................ ................................ .................. 67 3 12 Leachate NO 3 N c oncentration from Alfisol in relation to different rates of Hydra Hume alone ................................ ................................ ................................ ........................ 68 3 13 Leachate NO 3 N concentration from Alfisol in relation to fertilizer and different rates of Hydra Hume ................................ ................................ ................................ .................. 68 3 14 Soil pH after leaching in relation to fertilizer and different rates of Hydra Hume ............ 69 3 15 Soil EC after leaching in relation to fertilizer and different rates of Hydra Hume ........... 69

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11 Abstract of Thesis Presented to the Graduate School of the University of Florida in Partial Fulfillment of the Requirements for the Degree of Master of Sc ience ORGANIC AMENDMENT TO REDUCE NITROGEN LOSS IN SANDY SOILS By Wen Gu December 2012 Chair: Zhenli He Cochair: Lena Q. Ma Major: Soil and Water Science Nitrogen (N) is a nutrient often associated with accelerated eutrophication of surface water. The ex tremely low holding capacity for nutrients and moisture is the fundamental problem of sandy soil in Florida, which causes losses of nutrients into waterways. Soil organic amendment s and moisture and is, therefore, a promising approach to reduce loads of nutrients and toxic metals in surface runoff water and/or leachate. Organic amendment can increase or decrease mineral N in soil, depending on the C/N ratio of the organic materials and N availability in the soil. Hydra Hume is a commercial product, which shows the potential in enhancing soil N availability or reducing fertilizer N loss. In this study, incubation, column leaching, and laboratory analysis were conducted to examine the effects of Hydra Hume on transformation dynamics and leaching potential of mineral N in two representative soils (Alfisol and Spodosol) under citrus production in south Florida. The obtained results indicated that Hydra Hume amendment enhanced assimilation of mineral N, when it is applied to the soil, however, soil orginal C/N ratio also need to be considered. Hydra Hume application reduced NO 3 N leaching regardless of chemical fertilizer addition. Significant reduction in total NO 3 N amount could be observe d when Hydra Hume applied alone in both of the soils, but an application rate of 44.8 kg ha 1 is required. If

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12 fertilizer was incorporated with the treatments, significant reduction of total NO 3 N amounts and total mineral N amount could be observed, but a minimal application rate of 44.8 kg ha 1 is required to accomplish significant results in Spodosol and 22.4 kg ha 1 in Alfisol.

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13 CHAPTER 1 LITERATURE REVIEW Nitrogen and Plant Nutrition Numerous mineral elements are needed for plant normal growth and adequa te food and fiber production. Among these elements, Nitrogen (N) is an essential element for plant growth and a comparative larger amount is required by most agricultural crops than the other soil borne elements (Stevenson, 1982). Nitrogen is a major part of all amino acid and the component of nucleic acid and chlorophyll. Also, It is essential for carbohydrate use within plants. Therefore, almost all the vital biological processes are related to N, which is a basic constituent of the functional plasma (pr otein, nucleic acids). A good supply of N is necessary for normal plant growth, but also uptake of other nutrients (Brady, 2008). Many legumes and some other plant species can obtain N directly from the atmosphere, but in agriculture system, most N is obt ained from the soil and external sources, such as fertilizers, or through mineralization process. An optimal supply of N will bring optimal crop yields. Therefore, maintaining a balance of N availability according to the needs of the plant is a hot point f or sustainable agriculture. Plants respond quickly when supply of N is adequate in the soil. Their leaves turn dark green and the productivity can be dramatically stimulated. A deficiency of N in plants limits the synthesis of proteins and chlorophyll. It inhibits plants to assimilate CO 2 and synthesize carbohydrates, and consequently exhibits chlorosis (yellowish or pale green leaf colors) and stunted growth (Zhao et al., 2005). Timing of N application is also important. Late application of N may cause ear ly stage lack of N, which may have adverse effects on annual crop yield (Jansson et al., 1982). It is recognized that some crops are well adapted to unfavorable conditions, growing improver when soil has high nutrient levels. Excessive N promotes developm ent of aerial organs

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14 with relatively poor root growth. The presence of N in excess promotes development of the aerial organs with relatively poor root growth. This increase may cause excessive vegetable growth and the risk of lodging. It may also reduce th foliar diseases (FAO, 1984). To meet the food demands of a growing world population, global use of N fertilizers in intensive agricultural systems has increased dramatically in the last decades, espe cially for greenhouse vegetable production systems (Ju et al., 2005). Due to the relatively high economic value of the higher yields and difficulties in the precise management of nutrient supply, growers tend to use extra large amounts of fertilizers in or der to obtain maximum yield. However, the efficiency of fertilizer N is limited, with often less than 50% of the applied N taken up by the crop (Raun et al., 1999). Ju et al. (2007) mentioned, only in one of the vegetable production areas in China, fertili zer N is often applied at rates > 1200 kg N ha 1 per crop, and two or three crops are grown each year. The huge fertilizer load and extremely low crop recoveries of fertilizer nutrients may increase the risk of environmental pollution of both air and water and marked deterioration in soil (Wiesler et al., 2001) To improve N efficiency and achieve the optimal crop yield in agriculture, integrated N management strategies, which contain improved fertilizer, soil, and crop management practices, must be taken i nto consideration. The Role of Nitrogen in Sustainable Agriculture The global human population, currently 7.5 billion, is projected to reach 10.5 billion by 2050. The vast majority increase will occur in the developing countries of Asia and Africa, accordi ng to United Nations estimates. Increased urbanization and industrialization bring rapidly growing population, which result in greater food demands. Meeting the nutrition requirement and food demand of the growing population has become a big challenge due to decreased irrigational water supplies and other environmental concerns (Singh et al., 2011). Moreover, population

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15 growth and expanding demand for agricultural products constantly increase the pressure on land resources. The pressure on land may cause la nd degradation and intensification of the cultivation of arable lands, and also expansion of cultivation into marginal areas. Mismanagement of arable areas by farmers and grazing areas by livestock owners is one of the major causes of soil degradation (FAO 2000). Various sources suggest that 5 to 10 million hectares are being lost annually to severe degradation. If this trend continues, 1.4 to 2.8 percent of total agricultural, pasture, and forestland will have been lost by 2020 (Scherr and Yadav, 1996). L osses of N by land degradation are estimated to range from 1 to 100 kg ha 1 per year (White, 1986; Rose and Dalal, 1988), which makes land degradation a major cause of long term decline in the fertility of agricultural soils. Land degradation may also have negative effects on the farm, such as deposition of eroded soil in streams or behind dams, contamination of drinking water by agricultural chemicals, diversion of water sources from other users by irrigation, health problems caused by wind eroded soil or loss of habitat, which will affect the livelihood and economic wellbeing and nutritional status of over a billion people (Scherr and Yadav, 1996). Therefore, it seems sensible to consider alternative approaches, like sustainable agriculture. Continuous c ultivation with inappropriate farming practices may result in severe depletion of nutrients and soil organic matter. Henao and Baanante (2001) reported that soils of most countries in North Africa are being depleted of nutrients at rates ranging from 20 to 50 kg NPK/ha/year. Lal (2001) reported that the depletion of soil organic matter in tropical regions could be as high as 70% as a result of cultivation for 10 years. Sustainable agriculture is vital in agricultural needs. A switch from replacing chemical fertilizers with green manure. It attempts to require more information about

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16 environmental characteristics and the environmental impacts of agricultural practices in order to make sure the environmental resources can be fully utilized and no harm was done to it. The techniques need to be environmental friendlyto ensure safe and healthy agricultural products (S ingh et al., 2010; Hue and Silva, 2000). Erenstein et al. (2012) reported that sustainable agricultural practice, such as zero tillage (ZT) practice, could drastically reduce tillage intensity and loss of nutrients for the wheat crop from eight to a single tractor, save 15 16% on operational costs and boost 4% yield increase over average farmer reported conventional yields of 4.2 tons ha 1 in the rice wheat systems in South Asia. scale u se of chemical external inputs, such as fertilizer N, led to movements that searched for alternatives to sustainable agricultural practices. However, the use of N is mainly determined by economic incentives such as profitability and subsidies, and less by environmental costs (Spiertz, 2009). To manage nutrients and productivity of agro ecosystems, Drinkingwater and Snapp (2007) proposed an ecosystem based approach, which minimizes N loss and maintain soil reserves with nutrient inputs equal to harvested exp orts. To maintain long term fertility in sustainable agroecosystem, soil organic matter (SOM) is a key factor, which has great influences on many readily measurable soil functions or processes (Yilmaz, 2011). It drives biological processes involved in nutr ient cycling as a reservoir of nutrients and energy and also has a profound influence on soil basic properties, such as cation exchange capacity, chelation of metals and stabilization of soil structure. Agricultural production cannot be sustained if nutri ents removed during cropping are not replenished or if appropriate agricultural practices are not implemented to maintain or increase soil organic matter. Therefore there is a surge of research on organic farming in recent years and it is seen as

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17 a sustain able alternative to conventional intensive agricultural systems (Stockdale et al., 2001) Nutrient management in organic systems is based on fertility buildin combined with recycling of nutrients by organic materials, such as farmyard manure and crop residues, with only limited inputs of permitted fertilizers (Loes and Ogaard, 2001). Nitrogen also plays a very important role in deter mining crop yield. Rasmussen et al. (1998) reported that in the long term experiments in UK, wheat yields with fertilizers exceed those without external N input by a factor of 2 3. Dobermann and Cassman (2002) reported that grain yields of maize increased from 3 to 14 t ha 1 with an increase in plant N accumulation from 50 to 300 kg N ha 1 and rice yields increased from 2 to 8 t ha 1 with an increase in plant N accumulation from 25 to 200 kg N ha 1 It means with an increased amount of fertilizers, especial ly N, crop yield will be greatly enhanced. However, more external N inputs may bring more environmental impacts. The N inputs are among the first constituents to be removed by soil erosion because they are concentrated in the surface soil and are less den se than other soil constituents (DeBano and Conrad, 1976). Before 1960s, N fertilizer was used at a relatively low rate and crop N uptake was mainly dependent on manure applications, biological N fixation and indigenous N supply through mineralization of s oil organic matter. Since the early 1960s, the use of N fertilizers has dramatically increased and nowadays 30% 80% of N applied to farmland is lost to surface and groundwater, and to the atmosphere (Goulding et al., 2008). Goudriaan et al. (2001) reported that fertilizer N accounted for 60% of the net primary production in 1990, thus resulting in over application of fertilizer N from the economic response. Since 1980, the amount of fertilizer N has exceeded the amount taken up by crops globally. It means t hat only yield improvement was considered and not environmental sustainability.

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18 Under the current global background, sustainable food security draws lots of attraction. Sustainable agriculture combines economic profitability, environmental health and ethic al soundness as three main objects. It focuses on meeting productivity, efficiency and efficacy aims. Eickhout et al. (2006) explored the role of N in future world food production and environmental sustainability, and concluded that despite improvements in N use efficiency of food production systems in developed countries, total reactive N loss would grow strongly to 2030 because of the intensification of crop production systems in developing countries. Therefore, there is a need in balancing the use of fer tilizer N and the key challenges are to develop global capacity to produce adequate food in a sustainable manner, and establish communication about the risks of N excess in agroecosystems and implement legislation (Spiertz, 2009). Current knowledge for ach ieving sustainable crop intensification through measuring both optimal use of external inputs and make conservations for natural resources within an agroecosystem is inadequate. In order to increase intensification and diversification of agricultural produ ction systems, innovative sustainable technologies, appropriate policies, and economic incentives will have to be developed (Keerthisinghe et al., 2003). In the case of N, there is a need to gain refined information on N cycling processes and to assess the value of the crop, soil or fertilizer management practices designed to improve overall N use efficiency of the agroecosystem, with the ultimate goal of enhancing sustainable intensification of agricultural production while conserving the natural resource (Chalk et al., 2002). Spiertz (2009) also put forward two strategies to meet sustainability goals in food production, with a safe and profitable use of N: One is developing low input, high diversity agricultural systems. Agricultural system diversity, such as crop choice and crop rotation, may minimize the risks of yield reduction by abiotic and biotic stress. The other one is developing

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19 high input, low diversity agricultural systems. Some crops with high yield and high N response are chosen to achieve a ma ximum productivity per unit of land. An optimized N management during the whole crop cycle may control N losses. Soil Nitrogen Most of the N in terrestrial systems is found in the soil. Normally, A horizons have about 0.02 to 0.5% N and a value of about 0 .15% N in cultivated soils (Brady, 2008). Nitrogen in soil is largely bound to organic matter and mineral materials that protect it from loss but make it unavailable to the plants. Generally, only a small amount of N exists in available mineral forms, such as NO 3 and exchangeable NH 4 + Therefore, N cycling in soil is closely related to organic matter turnover. Microorganisms are responsible for soil N transformations, which play a vital role in determining the availability of N for plant growth and crop p roduction (Stevenson, 1982; Powlson, 1998). Some studies have shown approximately 1.5 to 3.5% of the organic N mineralized in soils annually (Hue and Silva, 2000; Wilson and Jefferies, 1996). After the land is first placed under cultivation, the amount of N decline, and N removed by harvested crops must be compensated with an equivalent amount of fixed N into organic matter to form equilibrium with the characteristics of local weather, tillage and cultivated practice, and soil types (Stevenson, 1982). Since mineral N is highly mobile and limited in the soil, if it is cannot be taken up by plant roots and microorganisms, it may be lost through gaseous emission, mainly by denitrification and volatilization, or leaching to the groundwater, both of which may cre ate environmental hazards (Hauck, 1990; Jenkinson, 1990). The main method scientists applied to maintain or restore soil nutrients and increase crop yields is the application of N fertilizers (Hirel, 2011). Fertilizer N can greatly improve crop quality by enhancing the protein content of the grain and forage crops. Also, some researches showed the role of applied fertilizer N in facilitating crop

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20 uptake of other essential nutrients (Whitehead et al., 1986; Pederson et al., 2002; Fageria et al., 2005). For e xample, the physical association of NH 4 + with P can increase the uptake of fertilizer P. Murphy (1978) reported that the placement of anhydrous NH 3 with an ammonium phosphate solution is soil may significantly enhance uptake of the fertilizer P. Moreover, in calcareous soils, fertilizer may band with Zn and thus improve Zn uptake by plants. Tilman (2002) showed that with the amount of mineral N fertilizers applied in soils increased by 7.4 fold, the overall yield increased only by 2.4 fold. It means that the fertilizer N use efficiency has declined sharply. It is true that there is a genetic variability for both N absorption efficiency and for N utilization efficiency in most of the crops (Hirel et al., 2007). However, the main reason is probably the sever ely disturbed N balance due to the intensification of agricultural activities and the application of chemical fertilizers. Over 50% and up to 75% of the N applied to the field is lost by leaching into the soil (Hodge et al., 2000; Asghari et al., 2011). Wi th yearly applications of N fertilizer, excessive amounts of N are brought into the environment and have a number of undesirable impacts on water, terrestrial, and atmospheric resources (Ribaudo et al., 2011). Also, changes in the N cycle associated with e xcessive soil N loading may bring ecological impacts, such as changes in the structure of ecosystems and biodiversity, and economic impacts (Matson, 1998; UNEP, 2004). Similarly, the use of N fertilizer may also affect human health both in positive and in negative ways, depending on the rates of N fertilizer used in the ecosystem. Crop yield may be greatly increased by applying high rates of N fertilizer, while negative health effects may occur in direct (pollution of air and water) and indirect (ecological feedback to disease) ways (INI, 2004).

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21 Nitrogen Transformation and Availability in Soil Nitrogen is the element that controls most of the biological activity in soil. The movement of N between the land, water, and atmosphere defines the cycling of N on a global scale. Soil also has an internal N cycle where N is converted from one form to another (Rosewell, 1976). Mineral soils in the temperate regions of the world contain between 0.06 to 0.3% N and approximately 90% of the N is in organic forms (Matthews 1992). Inorganic forms exist primarily as ammonium (NH 4 + ) and nitrate (NO 3 ). Once in the mineral form, N is easily transported between air, water and soils. The internal N cycle in soils is constantly changing due to biological, chemical and physical pr ocesses. The rate at which N becomes available is determined by the complexity and stability of organic matter and by microbial activity. It may occur in days or, if the N is in a very stable form, it may take years. Microorganism cannot metabolize elemen tal N (N 2 ), which means N becomes biologically active when it is fixed or bound, such as incorporating into ammonium (NH 4 + ) and nitrate (NO 3 ). Fixed N flows through the food web (plant animal humans/predators). This fixation process distinguishes the N cy cle from the carbon (C) cycle. From a global environmental point of view, there is a serious concern about the excessive growing fixed N. Agriculture accounts the largest fraction (some 86%) of the total human released fixed N (Jordan and Weller, 1996). Ni trogen fixed by human activity now exceeds the amount by all terrestrial natural processes combined. Balance studies have reported that the ways of N loss from soil due to uptake, leaching, erosion, and denitrification are replenished to varying extents by biological N 2 fixation processes. When agriculture is introduced into these systems, this nutrient cycle is broken and the normal supply of nutrients for any agricultural activity comes mainly from the soil. Additional nutrients must be supplied as extern al inputs to maintain crop growth and yield. Organic wastes, such as human, animal, and crop wastes, are used an sources of nutrients added to the soil. There are many

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22 examples of the disappearance of whole civilizations due to diminished soil fertility an d productivity. Rivas et al. (2012) studied soil N dynamics after a severe forest fire and reported that the inorganic N concentrations were typically greater or equal to those of net mineral N in forest soil after fires can be attributed to higher soil pH, N release from dead roots and previously inaccessibl e N forms, increased anion exchange capacity and decreased N decrease in net N mineralization would be the result of increased C/N ratio of the remaining soil organ ic matter (SOM), a decrease in SOM quantity and quality occurred as a result of pyrolysis, and recalcitrant char production or an increase in N immobilization due to enhanced microbial activity and/or increases in organic C and phosphorus (P) concentration Therefore, changes in the composition or abundance of microbial and plant communities are cru internal N cycling in soil. Nitrogen Losses into the Environment In modern society, when the natural organic waste additions cannot meet the need of crop production, fertilizers and several other materials have been used as additional so urces of crop N requirements has been evaluated by growing a test crop under unfertilized conditions (Glendining et al., 1996; Bhogal et al., 2000; Sieling et al., 2006). The excessive N loads may cause an increase of N loss from the soils though denitrification (nitrous oxide (N 2 O) emission), volatilization, and leaching, etc It is estimated that the anthropogenic N 2 O emissions to the atmosphere are about 3 8 Tg N annually and agricultural systems impart a large portion of anthropogenic emissions (Mosier and

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23 Kroeze, 1998). Bouwman (1996) reported a simple linear relationship to relate the total annual direct N 2 O emission (E) from fertilized fields to the N fertilizer applied (F): E=1+1.25F. The emission factor, 1.25 1% of N fertilizer applied, given by Bouwman, has been adopted by the emission of N 2 O affected by N fertilizer applied. This emission factor makes the methodology for calculating emissions simple and transparent. However, there are still disadvantages on the large uncertainties and impossibility to distinguish various regions, crop type and weather pa tterns (Ruser et al., 2001; Kuikman et al., 2006). Smith (1998) reported that in Scotland the average emission factor was less than the default value of 1.25% of the mineral N applied, and suggested a lower temperature may account for this difference. Dob bie (1999) reported that the emission factor differed greatly among grassland, potato, broccoli, and grain cereal crops. Therefore, IPCC in 2001 suggests that the direct N 2 O emission from soil may require a better calculation based on different regions. Or ganic amendment, as a practical manner, has been widely used for improving soil fertility. The type of amendment is a vital factor affecting nitrous oxide emission (Shelp et al., 2003). Aulakh (1991) reported that the C/N ratio of the organic amendments g reatly affect the amount of N 2 O emission, by affecting mineralization and immobilization rate of N in soil. Higher C/N ratios (>30) may lead to immobilization of mineral N and microorganisms decompose organic residue and incorporate mineral N into their bi omass. A relative low C/N ratio (<20) in plant residue shows a high mineralization rate, which NO3 can be taken up by the plants. C/N ratios could be a good predictor for N 2 O emission. In the early stage, Bremner and Blackmer (1981) reported a negative rel ationship between these two factors. Huang et al. (2004) reported that the relationship between N 2 O emission and C/N ratio is linearly negative, with

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24 correlation coefficient (R 2 ) ranging from 0.783 to 0.986. Amendment of organic residues may significantly increase the rate of soil respiration and develop anaerobic environment more rapidly, which will trigger more N 2 O emission (Ding et al., 2006). Huang et al. (2004) and McKenney et al. (1993) reported that the emission rate of N 2 O decreased significantly b y adding wheat straw and sugarcane stalk, as compared with other materials in clay soils. Also, in sandy clay loams, animal manure and slurry generated more N 2 O emission than mineral N fertilizer (Laughlin, 2001; Khalil et al., 2002). By integrating N imm obilization data, Vigil and Kissel (1991) reported the break even point between net N immobilization and mineralization of organic residues was at a C/N ratio of 41. Therefore, some organic amendments, such as sugarcane stalk and wheat straw, which have C/ N ratio greater than 41, may stimulate NH 4 + immobilization and N 2 O consumption, and hence reduce N 2 O emission. Senbayram et al. (2012) and Jahangir et al. (2011) studied the relationships between N and C sources in reducing N 2 O emissions by pot incubations in the lab. A microscale for exploring processes that occur in deep soil horizons was used in both of their studies. Additions of different C sources to the soil via fertilization or crop residue incorporation may increase denitrification. Senbayram et al (2009) of organic matter with high labile C content to fertilized agricultural soils may lead to derived N 2 O emissions. Organic amendments used as alternatives to mineral N fe rtilizers may have contrasting effects on greenhouse gas emissions, depending on soil C/N ratio. Leaching and erosion are another major way for N loss in terrestrial systems, which are estimated to range from 1 to 100 kg ha 1 year 1 (Teixeira et al., 2004) Microorganisms play an important role in improving soil fertility by metabolizing N, which cannot be uptaken by plants

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25 directly. Once organic N is mineralized and converted into NH 4 + and subsequently into NO 3 it can be lost via leaching. The mineral f orms of N are very soluble and can be easily leached into surface run off and groundwater, especially nitrate (NO 3 ) and urea (CO(NH 2 ) 2 ) ( Redhaiman, 2000; Umar and Iqbal, 2007) ). However, the process of biological conversion is time consuming and the move ment of NH 4 + in the soil is limited by binding to the cation exchange complex (CEC) of clay particles and soil organic matter. Inappropriate N fertilizer application and management, which exceed the need of crop production, may trigger large N leaching los ses. During leaching, the losses of mineral N are often accentuated by an abundance of precipitation combined with low evapotranspiration losses, which enhance water saturation, leading to subsurface or surface water flow, especially in fallow period (Legg 1982). Leaching loss of N from organic material depends mainly on two factors. The first is the net amount of organic N that is mineralized and converted into NO 3 Secondly, once the organic N is nitrified to NO 3 sufficient water has to be present to allow downward movement (Legg, 1982). However, if there is too much water, the soil becomes waterlogged and NO 3 will be lost by denitrification. Soil texture is an important property that determines how much NO 3 is lost by leaching or denitrification. A clay soil with a heavy texture has a lower hydraulic conductivity than a lighter, sandy soil. Therefore, in a heavy textured soil, NO 3 is more susceptible to denitrification, whereas in a sandy soil, leaching is more likely to be the main mechanism of NO 3 loss. The leached nitrate from agricultural lands is considerable and contaminates groundwater, rendering it unfit for human consumption. The increase of N in surface water will have increased biological productivity and brings about eutrophication, whi ch generate undesirable changes

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26 including proliferation of algae and aquatic macrophytes, dissolved oxygen depletion, and a decrease in water clarity (Conley et al., 2009). While several real and potential health and environmental impacts of N exist, agric ulture and environmental practices have major effects only on those impacts which involve excessive nitrate in drinking water, eutrophication, and perhaps on O 3 depletion (Mahvi et al., 2005; Ju et al., 2007). Few studies did intelligent risk assessment a nalyses of these impacts, especially when human health as well as social goals and values are involved. The growing worldwide demands for food will involve a continued increase in the use of fixed N in agriculture, most of which must come from fertilizers. However, the great willing of making a better world with environmental safety tell us the importance in managing N both in agriculture and environmental quality (James et al., 2011; Stevenson, 1982). Implication of C/N Ratio in Nitrogen Management The ke y problem in managing N for agriculture is to achieve the balance between N supply and crop demand without excess or deficiency ( Cassman et al., 2002). A definition of nutrient RCS) practice standards: managing the amount, source, placement, form, and timing of the application of plant nutrients to the soil (USDA, NRCS, 2006). Several scientists studied N use efficiency (NUE, always assessed by C/N ratio) as an indicator to check the management of N in agriculture. They define NUE of a cropping system as the proportion of all N inputs that are removed in harvested crop biomass, contained in recycled crop residues, and incorporated into soil organic matter and inorganic N pools. Th e excessive N is lost from the cropping system and thus contributes to the reactive N load to the environment. (Hussain et al., 1996; Timsina et al., 2001; Zheng et al., 2007; Gaju et al., 2011).

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27 Since Calvert (2004) presented the idea of proportionality b etween C and N, C/N ratios have been considered an important soil characteristic. The C/N ratio of the organic material added to the soil influences the rate of decomposition of organic matter and this results in the mineralization or immobilization of soi l N. If the added organic material contains more N in proportion to C, then N is released into the soil from the decomposing organic material. On the other hand, if the organic material has a less amount of N in relation to the C, then the microorganisms w ill utilize the soil N for further decomposition and the soil N will be immobilized and become unavailable to plant. The soil C/N ratio is an important soil fertility indicator due to the close relationships between soil organic C and total N. Generally, t he C/N ratio of the surface soil falls within narrow ranges, usually about 10 to 12 for cultivated, agriculturally soils of temperate regions. However, there are many factors which affect soil C/N ratio, such as climate (Miller et al., 2004), soil conditio ns (Diekow et al., 2005; Galantini et al., 2004; Oudraogo et al., 2006; Yamashita et al., 2006), vegetation types (Franzluebbers et al., 2000; Puget and Lal, 2005), and agricultural managements (Raun et al., 1998; Dalal et al., 2011; Liang et al., 2011). Boberg (2009) stated that the C/N ratio of an organism in soil reflects how much C, which microorganisms requires for biomass production, in relation to N. For plants, C can be used as energy source and thereby lost as carbon dioxide. To produce biomass a nd balance the need, the C/N ratio of the substrate must be higher than that of the microorganism and the C/N ratio of the new organism will be lower than the substrate. Moreover, N is essential to produce the protein rich microbial cells. Inorganic N is i mmobilized into bacterial cell when the metabolized organic substrate has a high C/N ratio (Azim et al., 2008; Kirkby et al., 2011). The C/N ratios of microbial organisms are relative constant and the range of fungi and bacteria is in the range 8 25

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28 and 5 18, respectively (Chapin et al., 2002; Rousk, 2009). For a general soil, microbial biomass may vary with the fungal/bacterial ratio, while a range of 10:1 to 12:1 is recommended (Griffin, 1972). Therefore, when applying N fertilizers or organic amendments, such ratio range can provide a reliable basis with which to determine the extent of available C and the stabilizing nutrients and to consider implications for management strategies to optimize retention of stable available N in soils. Nitrogen mineralizat ion depends not only N concentration of the substrate, but also the decomposability (Janssen, 1996). Although plants can only take up the inorganic forms (NH 4 and NO 3 ), it may also return organic N compounds to the soil in litter fall. Vegetation therefore influences N mineralization and nitrification through competition with microbes for nutrients and through litter quality and quantity (Vitousek et al., 1982). The break down of organic residues by microbes is dependent upon the C/N ratio. Hoorman and Isla m (2010) conducted a study on comparing two separate feed sources, a young tender alfalfa plant and oat or wheat straw. Young alfalfa plant has more crude protein, amino acids, and a lower C/N ratio, which make it much easier for microbe to digest. However oat and wheat straw has more lignin, less crude protein, and less sugars in the stalk and a high C/N ratio. It needs more time and N to break down the high carbon source and be decomposed by microbes. Therefore, a relative low N content or a high C/N rat io is associated with slow soil organic matter decay and generally, immature or young plants have a higher N content, lower C/N ratios, and faster soil organic matter decay (Lewis and Papavizas, 1974). As well for composting, the initial C/N ratio is one of the most important factors (Michel et al., 1996). A C/N ratio less than 20 allows the organic materials to decompose quickly while a C/N ratio greater than 20 requires additional N and slows down decomposition. If a considerable

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29 amount of C is in the fo rm of lignin or other resistant materials, the actual C/N ratio could be larger than 20. Generally, initial C/N ratios of 25 30 are considered ideal for composting (Kumar et al., 2010). If a high C based material with low N content is applied to the soil, the microbes will tie up soil N, though it will be eventually released. Recently some researchers have successfully conducted composting at lower initial C/N ratios from 15 20 ( Huang et al., 2004; Zhu, 2007; Ogunwande et al., 2008; Kumar et al., 2010). Co mposting at lower initial C/N ratios can increase the amount of manure treated, but can also increase the loss of N as ammonia gas. As the soil organic matter decomposes, the C/N ratio of most plant residues tends to decrease due to the gaseous loss of CO 2 Therefore, the percentage of N in the residual SOM rises as decomposition progresses. The C/N ratio influences the two main decomposing groups, bacteria and fungi, in soils. Bacteria are the first microbes to digest newly input organic plant and animal residues in the soil. Bacteria have a high N content in their cells and are generally less efficient at converting organic C to new cells. The relative importance of decomposition by fungi is that it can help decrease the application of N fertilizer (van G roenigen et al., 2007), while organic matter with a high C/N ratio is believed to stimulate the fungal contribution to decomposition (Henriksen and Breland, 1999; Thiet et al., 2006). For most of the soil, the typical C/N ratio is around 10, which means th e N is available to the plants. The 10:1 C/N ratio of most soils reflects an equilibrium value associated with most soil microbes (Bacteria 3:1 to 10:1, Fungi 10:1 C/N ratio). The C/N ratio theory also applied in many agricultural practices in N management such as cover crops, crop rotation, organic amendments, etc Many crop production practices have the potential to create adverse environmental impacts with respect to N. Due to the intensified crop management, such as greater inputs of N fertilizers (Til man et al., 2002; Erisman et al., 2008),

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30 the global cereal production doubled in the past 40 years. However, not all of the applied N can be taken up by plants. When the natural capacity of a system to cycle N is exceeded in a given locality, the excessive N may be accumulated in the ground or surface waters and large amounts of NH 3 or N 2 O may be lost to the atmosphere. The sources of N are diverse. Usually, exogenous N is from the residues of plant, animal origin and fertilizers. Techniques are established for minimizing N losses and maximizing N efficiency, including efficient agricultural management, innovative applications of agricultural technology, regulations limiting fertilizer applications, and possible even changes in fundamental patterns of land u se and crop production (Stevenson, 1982) In general, practices which decrease soil erosion and surface runoff will also reduce the amount of N lost from croplands by these routes. As a nutrient management tool, the use of cover crops in cropping systems i s an important strategy. Cover crops, also called green manure, can be defined as close growing crops that provide soil cover, and soil improvement between periods of normal crop production (SSSA, 1997). They are not grown for market purpose and are well s uited for conservation agriculture. Growing cover crops in a rotation can reduce erosion through greater levels of soil cover as well as improve soil fertility and suppress weeds (Flower, 2011). Cover crops can be leguminous or nonleguminous. Legume cover crops fix atmospheric N into a form plants and microorganisms can use and thus they are used as a source of N (Smith et al., 1987). Non legume species recycle existing soil N and can reduce the risk of excess N leaching into groundwater (Meisinger, 1991). There is interest in growing leguminous cover crops, such as red clover (Trifolium pratense L.) and hairy vetch (Vicia villosa L.), particularly on erodible soils (Goss et al., 1995). These leguminous cover crops apply one more potential on N fixation thro ugh biological pathways, which can benefit in reducing the need of N fertilizer for

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31 succeeding crops (Singh et al., 2004). Nonleguminous cover crop can accumulate inorganic soil N and hold it as an organic form, which can reduce the mineral N leached from the soil and leave the soil bare. Cover crops can reduce the potential for percolation by extracting water from the soil, reducing the potential for percolation and consequent leaching (Chapot et al., 1990). The holding N will be released after decompositi on of the cover crops. Black oat ( avena strigosa Schreb.) is widely used as a cereal cover crop due to its rapid growth, high biomass production in southern Brazil and some area in USA. As a cereal, it has a high C/N ratio, which leads to slower decomposi tion rate and hence possible immobilization of soil N following some cereal cover crops (Flower, 2011). Rye ( S. cereale L.), a cereal, had great potential on accumulating the residual inorganic N after corn harvest (McCracken et al., 1994). In addition to fixing N, several studies reported that cover crop could reduce the potential for nitrate leaching from the farm fields (Owens, 1990; Brandi Dohrn et al., 1997; Staver and Brinsfield, 1998). Meisinger et al. (1991) conducted a study on the impacts of cove r crop on nitrate leaching and showed that cover crops reduced both the mass of N leached and NO 3 concentration of leachate by 20 to 80% are compared with no cover crop control. Francis et al. (1998), Shepherd (1999), and Rasse et al. (2000) reported that incorporating a nonleguminous cover crop in a cropping system has reduced nitrate leaching since the cover crop can reduce water percolation and also effectively use of nitrate. Cover crops are sometimes planted after the principal crop has been removed t o control erosion. Reduction in soil erosion by cover crops is associated with increasing in soil organic matter content, which improve soil water infiltration and holding capacity. However, there are still some possible disadvantages of cover crop. If cov er crop has a very high C/N ratio, it may create N deficiency for the succeeding crop if excessive N is immobilized and not released in a

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32 timely manner (Vyn et al., 1999). Moreover, adverse weather often prevents planting or growth of the cover crop. Crop rotation is a practice, which can reduce the requirement of N fertilizer. Crop rotation is to plant different crops on the same field in successive years and usually, the cropping sequences change with specialized farming. For example, corn soybean is a c ommon rotation group. Soybean can scavenge residual fertilizer N from the corn crop. John et al. (1975) estimate that about 40% residual N is removed by soybean. A recent study by Hons (2005) reported that the 60% increase in lint yield of unfertilized cot ton under rotation was attributed to increased soil organic matter (SOM), total N, residual nitrate N, and water content. Reddy et al.. (2006) also reported that the improved cotton growth in rotation with corn was attributed to increase SOM. The sequence of crop rotation is very crucial. Root exudates from plants cause change in microbial community composition and biomass and may depend on plant species specific differences in the quality of resources (e.g. carbon) input (Ladygina and Hedlund, 2010). Plant s that have a high C/N ratio may increase N immobilization and use up the residual N, which may decrease the yield of next crop. Organic amendments, including farmyard manures, slurries, sewage sludge, green wastes and composts, can increase soil fertility and quality to improve crop productivity and yields (Lupwayi et al., 2005; Fageria, 2007). Organic amendments can improve the physical properties of the soil, such as water retention capacity, bulk density and soil structure. Moreover, the application of organic amendments increases soil nutrients, which can reduce the need for inorganic fertilizers (Bellamy et al., 1995; Burgos et al., 1996; Barker, 1997). Several types of organic amendments have been used for improving crop production. Steffen et al. (19 95) discussed comparative tomato yields of conventional agriculture system and

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33 an ecologically oriented system, which emphasized the building up of organic amendments. The latter system hada greater soil water holding capacity and sharply reduced irrigatio n requirements. Moreover, the yield of NO.1 fruit was 55% 60% greater than the other one. Webb and Biggs (1988) examined the effects of adding humate as organic amendment on the growth of citrus. Trees treated with humate exhibited higher water uptake and produced more vigorous growth flushes. Foley et al. (2002) studied the paper mill residue applied as an organic amendment to increase vegetation production. Paper mills waste are the residues combining the primary residues from paper making process with th e secondary wastewater treatment sludge and have a C/N ratio less than 20:1. It was applied in the Central Sands region of Wisconsin increased the water holding capacity and plant available water by 33% 80%. The utilization of organic wastes as amendments had an efficient and low cost effective method of disposal these products. Sharifi et al. (2008) compared the field with or without a history of organic amendment (Solid beef (Bos taurus) manure) in potato (Solanum tuberosum L.) production. The results sho wed that historically amended soil had 35% higher values of potentially mineral N and 8% higher proportion of mineral N partitioned to the stable mineral N pool. The long term addition of amendments results in important changes in active and stable soil or ganic C and N fractions that can influence soil N dynamics. Single applications of organic amendments contribute a small amount to mineralized N in the subsequent year, the combined contributions of organic N from repeated applications can lead to a subst antial increase in soil N mineralization potential (Eghball et al.., 2004; Flavel and Murphy, 2006; Mallory and Griffin, 2007). The increased soil N mineralization potential can lead to higher soil N supply and subsequently reduce the need for fertilizer N

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34 Organic amendments may also affect crop yield through disease suppression. In some cases, organic amendment may release toxic compounds to directly inhibit the pathogens, such as isothiocyanates released from degradation of brassicaceous material (Lazze ri and Manici, 2001), or increase microbial biomass and activity to suppress disease (Bailey and Lazarovits, 2003). Ayongwa et al. (2011) studied the organic amendment impact on host parasite dynamics of sorghum bicolor and striga hermonthica and high qual ity amendment will have a negative impact on S.hermonthica severity and incidence. C/N ratio describes the organic matter quality; where in high quality organic matter has a low C/N ratio and low quality organic matter a high C/N ratio. Some organic wast es that are rich in N can release large amounts of mineral N through mineralization and improve fertilizer efficiency. When mineral N exceeds the amount needed to fill the gap between crop uptake needs and the supply from theses sources, Excessive N may be lost by leaching and denitrification to the environment. Therefore, the change of N dynamics after applying organic wastes need to be recorded to access effective use of the material, minimizing the losses of nitrate in leachates and avoiding negative env ironmental effects that it may cause in groundwater. Burgos et al. (2006) compared 3 organic materials, municipal solid waste compost, non composted paper mill sludge, and agroforestry compost on the N dynamic of a sandy soil. All of these three amendments had different immobilization N periods followed by positive mineralization. Due to the different C/N ratios, the first amendment had a higher nitrate leaching rate. Nevertheless, the nitrate losses represented a low amount compared with the total N added to soil. Radersma and Smit (2010) evaluated the effect of paper pulp as organic amendments on the N loss from fodder radish and found that adding paper pulp lead to a strong decrease in denitrification and N leaching.

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35 Conclusions and Perspectives Nitrogen is an essential macronutrient for plant growth. A deficiency of N may cause symptoms in plant leaves and stunt plant growth, while an adequate supply of N in the soil greatly promotes rapid plant growth and crop yield. Nitrogen undergoes a wide variety of transformations in soil, most of which involve the organic fractions. An internal N cycle exists in soil apart from the overall cycle of N in nature. Even if N gains and losses are equal, the N cycle is not static. Continuous turnover of N occurs through m ineralization and immobilization. Soils vary greatly in their N contents. Nitrogen content of the soil decline greatly during continued cultivation. Human activities, particularly fertilizer N use, may enhance biological N fixation and increase mass flow o f fixed N. Excessive accumulations of N may bring adverse effects from the global environmental point of view. Therefore, agricultural practices in the future will focus more on conserving energy and minimizing adverse environmental effects arising from th e use of N fertilizers. Appropriate N management and fertilizer application should be concerned so as to protect N from leaching and denitrification. Conventional N fertilizer will continue to be used but in a efficient method, and new fertilizers will be developed to achieve a specific release rate with active uptake by the plants. Nitrogen in organic materials is gradually mineralized after applied to the soil, and part of that plant available N is taken up by crops, immobilized in the SOM pool, or lost i n gaseous form or through leaching. Release of a substantial amount of N from medium to low quality organic resources often continues beyond a growth season, potentially resulting in significant residual effects. Many techniques, such as erosion control, improved irrigation, increased use of legume crops, cover crops, and organic amendments, can be applied to improve the efficiency of N use.

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36 Their effective utilization involves application of systems analysis approaches to farming, and also alters biologi cal activity by changing C/N ratio in the soils. More studies are needed to identify effective amendment to soils in order to reduce N loss, and other techniques will be developed as the social and economic pressure for efficient N management increases. Wh ile several real and potential health and environmental impacts of N exist, agriculture and environmental practices have major effects only on those impacts which involve excess nitrate in drinking water, eutrophication, and perhaps on O 3 depletion (Mahvi et al., 2005; Ju et al., 2007). Seldom researches indicated intelligent risk assessment analyses of these impacts, especially when human health and social goals and values are involved. However, the growing worldwide demands for food will involve a contin ued increase in the use of fixed N in agriculture. Most of this increase must come from fertilizer. Moreover, the great willing of making a better world with environmental safety tell us the importance in managing N both in agriculture and environmental q uality (James et al., 2011, Stevenson, 1982).

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37 CHAPTER 2 ORGANIC AMENDMENT EF FECT ON NITROGEN TRA NSFORMATION IN SOILS Introduction Nitrogen ( N) is a major nutrient element for citrus production. In United States, about 75% of the citrus production comes f rom Florida and about 28% of Florida citrus acreage is on sandy Entisols along the central Florida ridge (Mattos et al., 2003). In sandy soils, application of N in excess of tree requirements, inadequate placement and timing, or irrigation scheduling may r esult in leaching of nitrate (NO 3 ) below the root zone (Alva and Tucker, 1999). For citrus growth, the mineralization of N from soil organic matter, crop residues, N reserve. The application of organic amendments increases soil nutrients, thus reducing the need for inorganic fertilizer s (Bellamy et al.. 1995; Baker, 1997 ). Moreover, the use of organic amendment is a common practice to improve soil conditions and physical properties, such as water retention capacity, bulk density and soil structure (Entry et al., 1997). For environmental management of N from organic amendment, factors and processes that affect the extent and rate of conversion of organic N to plant available N or loss to the environment should be accounted for (He et al., 2000). Plant available N is defined as the sum of the mineral N, mainly as initial nitrate nitrogen (NO 3 N) and ammonium nitrogen (NH 4 N) content in soil, plus the organic N mineralized (Gilmour and Skinner, 1999). During the mineralization process, soil microorganisms transform organic N to inorganic for ms (NO 3 N and NH 4 N). At the same time, a process of immobilization of inorganic N takes place with the synthesis of proteins by microorganisms (Brady, 1990). This N dynamic system has been described as a mineralization immobilization turnover (Jansson and Persson, 1982). This turnover can be used to estimate a net mineralization rate or a net immobilization rate under

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38 different soil conditions or types. Better description and knowledge on N dynamics can provide a more efficient use of organic and inorganic fertilizers and minimize the N losses. Different characteristics of the organic materials, soil texture and properties, environmental factors, can influence the rate of N mineralization in soils (Sims, 1995). The C/N ratio of the organic matter is invers ely proportional to the net N mineralization (Appel and Mengel, 1990). The optimum C/N ratio has been reported to be between 15 and 40 (Cabrera et al., 2005). For the influence of soil texture, Scott et al. (1996) suggested that small pores protect organic matter from microbial attack whereas large pores facilitate N mineralization. The soil microbial biomass is another important component of the soil organic matter that regulates the transformation and storage of nutrients (Martens, 1995). K 2 SO 4 extractabl e organic C increased after soil fumigation with chloroform (CHCl 3 ) suggested that a procedure developed for evaluation of biomass C could be used to measure the dynamics of N in soils (Sotta et al., 2007). The organic amendment used in this study is Hydra Hume, a commercial product from Helena chemical company. Webb and Biggs (1988) studied the effects of humate amended soils on the growth of citrus. Humate, a humic acid, implies these substances are acidic, while they actually are colloidal and behave lik e clays in soil. Humate can be a source of inorganic nutrients that are held by exchange matrices and has a fundamental effect on water holding capacity. However, the effect of this new commercial product to the N dynamics in soil is still unknown. The ma in aim of this study is to investigate how Hydra Hume affects N dynamics and determine the decomposition and nutrient release kinetics after the organic amendment is applied to different soils. An incubation experiment was conducted to estimate the N miner alization.

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39 Materials and Methods Soil Collection and Characterization Two soils (one Alfisol and one Spodosol) were collected from representative commercial citrus groves in the Indian River area, south Florida. They were selected based on soil properties. For each location triplicate samples were randomly collected at 0 15 cm depth, with 3 soil cores being combined to make a composite sample. After removing plant material and stones, the soil was dried and then sieved (<2 mm). General characteristics of th e study sites are presented in Table 2 1 and Figure 2 1. The soil samples were air dried, ground, and passed through a 2 mm sieve prior to physical and chemical analyses. Soil pH (1:1 soil: water ratio) and electrical conductivity (1:2 soil: water ratio) w ere measured in deionized water using a pH/ion/conductivity meter (Denver Instrument, CO). Total soil carbon (C) was determined by combustion using a C/N analyzer (Vario MAX CN Macro Elemental Analyzer; Elemental Analysensystem GmbH, Hanau, Germany). Incub ation Study and Soil N Dynamics Analysis Portions of soil sample (each 500g oven dry basis) were weighed into Ziplock plastic bags (28 26.7 cm, 3.74 L capacity). The pellet organic amendment was ground to pass a 1 mm sieve before it was mixed into the soi l. The application rates of organic amendment were: 0, 112, 224, 448, 560 kg ha 1 soil, respectively. The mixtures were incubated at 25 C in after its moisture was adjusted to 70% of its field water capacity. Water losses were compensated by the addition of distilled water during the experiment. At the intervals of 0, 7, 14, 21 and 28 d, subsamples were taken from the incubation bags and analyzed for pH, electrical conductivity (EC), soil available N and microbial biomass. For each treatment, there were th ree replications and the experiment was followed completely randomized design, in order to reduce the other influences.

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40 Soil pH was measured in slurry with deionized water and 1 M KCl solution at a solid: solution ratio of 1:1 using a pH/ion/conductivity m eter (pH/ Conductivity Meter, Model 220, Denver Instrument, Denver, Co). Electrical conductivity was measured in slurry with deionized water at a solid: water ratio of 1:2 using the same pH/EC meter. Extractable NH 4 N and NO 3 N was determined by shaking a 5 g air dried sample in 25 ml 2 M KCl for 1 h. Concentrations of NH 4 N and NO 3 N in the filtrate were analyzed with a N/P Discrete Auto analyzer (EasyChem, Systea Scientific LLC, Oak Brook, IL). The net N mineralized due to the applied organic amendment wa s calculated by the difference between inorganic N in each treatment and the control soil. Data Analysis All data were analyzed using SAS program (version 8.2, SAS Institute, 2004). Differences in the soils with different organic amendment applications wer e tested by means of a one way analysis of variance (ANOVA). Significant statistical differences of the mean values of the P <0.05). Results and Discussion Effects of Organic Amendment on Soil pH and EC The Alfisol is calcareous and contained slightly less available N than the Spodosol (Table 2 1). The Spodosol is acidic in nature, but the pH was raised by liming to close being neutral. Both soils are very sandy, with more than 90% sand, and had low organic matter (<1% organic C). Application of organic amendment slightly increased soil pH initially due to the release of alkaline substances from decomposition of the product (Butterly et al., 2010; Xu et al., 2006). The soil pH d ecreased with the incubation time, probably due to release of organic acids with further decomposition of the organic matter (Fig. 2 2, Fig. 2 3). In addition, Ca and Mg from the organic amendment can replace hydrogen ion (H + ) on the soil exchange complex, thus

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41 decreasing soil solution pH. However, even at the highest application, soil pH was about 7.3 to 7.5, which is suitable for most crops. Fertilizers containing NH 4 + can induce soil acidification due to nitrification of NH 4 + to NO 3 which produces H + ( Chien et al., 2010). Therefore, for the treatments with chemical fertilizer, pH decreased more than those receiving Hydra Hume alone. The pH changes in Spodosol had similar trends. ity. Organic 4 and 2 5). The soil EC values greatly increased with addition of fertilizer, because of the increased amounts of mineral ions. Even at the highest application ra te, the EC values of the amended soil were still Hydra Hume at the rates <560 kg ha 1 should not have any significant negative impact on crop growth based onsoil pH and EC. The EC changes in Spodosol had similar trends. Effects on N Dynamics in Soils In the first two weeks, application of organic amendment slightly increased KCl extractable NH 4 N in the Alfisol (Fig. 6). Compared with other organic materials, Hydra Hu me had a relative small N content (1.31%), thus, no significant difference was observed among the different treatments. The increase in NH 4 N is probably attributed to the mineralization of organic amendment and release of NH 4 N from decomposition of organ ic matter. The concentration of NH 4 N in the third week decreased markedly initially and continued to decrease with time to almost zero for all the treatments. This behavior has been reported in aerobic incubation experiments (Sanchez et al., 1997; Madrid et al., 2001; Burgos et al., 2005). The decrease in NH 4 N was generally accompanied by a corresponding increase in NO 3 N, indicating that the NH 4 N released from the organic amendment mineralization was nitrified into the NO 3

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42 N (He et al., 2000), as was sh own in Figures 2 6 and 2 7. Nitrate contents in the treatments tended to increase until the end of incubation period. The difference in original C/N ratio between the two soils affected the mineralization rates of organic amendment in the soils. The C/N r atio of Spodosol is 13.58, which is below the critical level for net mineralization process, while the C/N ratio for Hydra Hume is 42.13 (greater than 30), which may induce net immobilization of mineral N in soil. Therefore, in the first two weeks, applica tion of organic amendment in Spodosol slightly decreased KCl extractable NH 4 N (Fig. 7). With the incubation time, application of organic amendment slightly increased NH 4 N content, due to the mineralization of organic material and release of NH 4 N from de composition of organic matter. The general trend of NH 4 N change in Spodosol is similar to that in Alfisol. The organic amendment appeared to have a minimal effect on NO 3 N, as there was no difference in NO 3 N concentration between the amended soil and th e control (without amendment) when it was measured on the first day (measured immediately after being mixed). For Alfisol, treatments with only Hydra Hume tended to decrease NO 3 N content in the first two weeks at an application rate of 224 kg ha 1 and abo ve, because of net immobilization induced by addition of Hydra Hume (Fig. 8). However, when the soil was amended with fertilizer and Hydra Hume, the additional mineral N from fertilizer may promote mineralization, thus offsetting the effects of organic ame ndment, while the control still had the highest NO 3 N contents, indicating a net immobilization during the incubation period (Fig. 9). The difference of soil C/N ratio itself may have impacts on N dynamics in soils. For Spodosol, application of Hydra Hume only markedly decrease NO 3 N contents, while when amended with Hydra Hume

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43 together with chemical fertilizer, a net mineralization process occurred. Statistical analyses are listed in Tables 2 2 and 2 3. Conclusion Application of Hydra Hume, an organic amen dment with a C/N ratio around 42.13, induced immobilization when applied alone in both of the test soils. For Alfisol, the immobilization process only occurred in the first two weeks with the application rates of Hydra Hume of 224 kg ha 1 or above. Soil or iginal properties have an effect on N dynamics. The difference of C/N ratios in the two soils determined the different N dynamic trends when fertilizer and organic amendment applied together. The N dynamics in Alfisol, which had the C/N ratio around 22, sh owed a net immobilization, while in Spodosol, there was a net mineralization curve instead.

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44 Figure 2 1 Sampling site

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45 Figure 2 2 Effect of organic amendment on Alfisol pH as a function of incubation time Figure 2 3 Effect of organic amendment on Spodosol pH as a function of incubation time With fertilizer Without fertilizer HH rate (kg ha 1 ): With fertilizer Without fertilizer HH rate (kg ha 1 ):

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46 Fi g ure 2 4 Effect of organic amendment on Alfisol Electrical conductivity (EC) as a function of incubation time Figure 2 5 Effect of organic amendment on Spodosol electrical con ductivity (EC) as a function of incubation time With fertilizer Without fertilizer HH rate (kg ha 1 ): With fertilizer Without fertilizer HH rate (kg ha 1 ):

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47 Figure 2 6. Effect of organic amendment on KCl extractable NH 4 N in Alfisol Figure 2 7. Effect of organic amendment and fertilizer on KCl extractable NH 4 N in Spodosol HH rate (kg ha 1 ): HH rate (kg ha 1 ):

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48 Figure 2 8. Effect of organi c amendment on KCl extractable NO 3 N in Alfisol Figure 2 9. Effect of organic amendment and fertilizer on KCl extractable NO 3 N in Spodosol soil HH rate (kg ha 1 ): HH rate (kg ha 1 ):

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49 Figure 2 10 Effect of organic amendment on KCl extractable NH 4 N in Spodosol Figure 2 11 Effe ct of organic amendment and fertilizer on KCl extractable NH 4 N in Spodosol HH rate (kg ha 1 ): HH rate (kg ha 1 ):

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50 Figure 2 12 Effect of organic amendment on KCl extractable NO 3 N in Spodosol Figure 2 13 Effect of organic amendment and fertilizer on KCl extractable NO 3 N in Spodosol HH rate (kg ha 1 ):

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51 Table 2 1 General properties of the two soils and Hydra Hume Properties/ Soil type Alfisol (Sandy,siliceous,hyperthermic,Arenic Glossaqualf) Spodosol (Sandy,siliceous, Hyperthermic, Alfic Alaquods) Cropping history Citrus Citrus Soil texture (g/kg) San d 908, Silt 42.8, clay 9.6 Sand 902,silt 53,clay 45 pH (H 2 0) 7.8 6.8 Electrical conductivity 150.9 61.59 Organic C (g/kg) 1.03 2.68 Total N (g/kg) 0.05 0.2 C/N ratio 22.05 13.58 Table 2 2. General properties of the Hydra Hume Properties pH EC N% C% C/N ratio Hydra Hume 2.83 2977 1.31 54.98 42.13

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52 Table 2 3 Statistical analysis of KCl extractable NH 4 N in Alfisol KCl extractable NH 4 N (mg kg 1 ) in Alfisol Without fertilizer With fertilizer HH rate (kg ha 1 ) 0 112 224 448 560 0 112 22 4 448 560 Sampling time (Day) 0 2.43 2.41 2.44 2.3 2.43 7.41 7.09 6.77 7.3 7.95 7 0.06 b 0.08b 0.10a b 0.11a b 0.19 a 0.91 b 1.03a b 1.42 a 1.62 a 1.51 a 14 0.1 0.1 0.06 0.1 0.09 0.07 0.08 0.08 0.09 0.1 21 0.05 b 0.11a b 0.13a 0.14a 0.14 a 0.13 0.05 0.04 0. 07 0.06 28 0.53 a 0..42 b 0.30c 0.30c 0.29 c 0.36 a 0.38a 0.14 b 0.16 b 0.14 b *Means followed by different letters within the same row indicate significance level at P<0.05 by the Duncan multi range test. Table 2 4 Statistical analysis of KCl extractable NO 3 N in Alfisol KCl extractable NO 3 N in Alfisol Without fertilizer With fertilizer HH rate (kg ha 1 ) 0 112 224 448 560 0 112 224 448 560 Sampling time (Day) 0 3.47 3.18 2.76 2.61 2.58 8.07 ab 8.35a 7.72 ab 7.72 ab 6.70 b 7 5.58 a 5.63 a 4.73 b 4.73 b 4.73 b 13.66 a 14.00 a 12.09 b 13.47 a 14.21 a 14 5.49 5.44 5.19 5.18 5.06 16.18 18.75 19.36 13.18 16.69 21 7.61 7.57 7.47 7.73 7.07 18.75 a 16.89 ab 14.77 b 16.63 ab 17.69 ab 28 7.21 7.29 6.53 7.44 7.34 16.90 15.74 15.17 17.01 17.51 *Means follow ed by different letters within the same row indicate significance level at P<0.05 by the Duncan multi range test.

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53 Table 2 5 Statistical analysis of KCl extractable NO 3 N in Spodosol KCl extractable NO 3 N in Spodosol Without fertilizer With fertilizer HH rate (kg ha 1 ) 0 112 224 448 560 0 112 224 448 560 Samplin g time (Day) 0 1.08 1.09 1.11 1.23 1.13 4.92 ab 4.68 b 5.20 ab 5.92a 5.85a 7 2.00 ab 2.05 a 1.93 ab 1.75 ab 1.29 b 8.44b 8.38b 9.59a 10.11 a 10.29 a 14 2.56 ab 2.68 a 2.43 ab 2.31 ab 2.00 b 9.48b 9 .75b 13.98 a 15.51 a 13.63 a 21 2.75 a 2.72 a 2.59 a 2.43 ab 1.93 b 12.76 ab 9.47b 13.43 ab 14.96 a 14.92 a 28 2.81 a 2.85 a 2.37 b 2.35 b 2.25 c 9.8b 10.11 b 14.09 a 15.54 a 15.65 a *Means followed by different letters within the same row indicate significance level at P <0.05 by the Duncan multi range test.

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54 CHAPTER 3 ORGANIC AMENDMENT EFFECTS ON NITROGEN LEACHING Introduction Nitrogen (N) is a nutrient often associated with accelerated eutrophication of surface waters. It is estimated that each year, more than 1,000 ,000 kg N is transported to the St. Lucie Estuary (SLE) (St. Lucie River Issue Team Interim Report, 1998) and the resultant increased nutrient loads have been suspected to cause declining water quality and decreased grass coverage of the SLE and Indian Riv er Lagoon (IRL). In South Florida, high temperature and the sandy nature of soils provide favorable conditions for rapid mineralization of organic materials. As soils in this area have a coarse texture (sand content often >90%) and a relative low nutrient retention capacity, citrus trees cultivated on these soils receive high rates of fertilizer application, which could increase the potential risk of nitrate pollution to groundwater (He et al., 1999). Martin et al. (1994) reported that light textured soils and intensive production of crops under irrigated conditions could lead to considerable nitrate losses by leaching. holding capacity for nutrients and moisture and is, therefore, a very promising approach to reduce loads of nutrients and metals in surface runoff. Environmental management of N from organic amendment requires an understanding of the factors and processes that influence the extent and rate of conversion of organic N to forms plant available N or loss to the environment (He et al., 2000). In Chapter 2, it mentioned that the influence of soil texture on N mineralization is related to clay content (Breland and Hansen, 1996). Small pore spaces protect organi c matter from microbial attack whereas large pores facilitate N mineralization. Moreover, other soil

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55 characteristics, such as heavy metal content and salinity, may also affect organic material decomposition and N mineralization. For soil quality, microbia l biomass is an important indicator (Elliott et al., 1996). Organic C from the organic material may enhance the growth of microorganism, thus increasing microbial biomass. In Florida, many soils are extremely sandy with low adsorption capacity for ions; th erefore, microbial biomass can be a sensitive indicator to the change of N dynamics in soils. The major objectives of this study were to 1) investigate the effect of organic amendment on N leaching in soils, 2) evaluate the effect of organic amendment on N availability in soil and 3) assess potential environmental risk of heavy metals from the soil amended with Hydra Hume. Materials and Methods The tested organic amendment is Hydra Hume, provided by Helena chemical Company, a commercial product showing pot ential for improving citrus growth in South Florida. However, its effectiveness for reducing fertilizer N loss is not well documented. The soils used in this leaching study were collected from a representative commercial citrus grove (citrus) and a vegetab le farm (vegetable). One is Alfisol and the other is Spodosol, representing two major soil types for citrus and vegetable production in the Indian River area. The relevant properties of the tested soils are listed in Table 2 1. The organic amendment was gr ound to pass a 1 mm sieve before it was mixed into the soil. Organic amendment was applied at the rates: 0, 22.4, 44.8, 89.6, 112 kg ha 1 soil, respectively. Fertilizer was applied at a rate of 168 N kg ha 1 (N P 2 O 5 K 2 O=12 4 15). The soil samples receiving no organic amendment or inorganic fertilizer were used as the control. Plexiglas leaching columns (30.5cm long, 6.6cm inner diameter), with several 5mm diameter holes at the bottom were used (Fig. 1). About 1.5 kg soil was packed into each column to the

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56 b ulk density of approximately 1.45 mg cm 3 with amendment mixed in the top of the soil column. There are three replications for each treatment. The soil columns were saturated with deionized water for three days to allow soil compact naturally and equilibri um of chemical and biological reactions prior to leaching. Leaching was conducted at 1, 3, 7, 14, 28, 42 day, 302 ml of rainfall in South Florida. The deionized water was applied using a peristaltic pump and the leachates were then collected from each leaching event and analyzed for pH, electrical conductivity (EC), NH 4 N, NO 3 N and metals. At the end of leaching, the soils were removed from the column and subsam ples were taken after through mixture, air dried, and measured for pH, EC, available N, Mehlich III extractable metals and microbial biomass. Leachate pH and EC were measured using a pH/ion/conductivity meter (pH. Conductivity Meter, Model 220, Denver Inst rument, Denver, Co). Available N (NH 4 N, NO 3 N) was analyzed using a N/P Discrete Autoanalyzer (EasyChem, Systea Scientific LLC, Oak Brook, IL). Available metals were determined using an inductively coupled plasma optical emission spectrometry (ICP OES, Ul tima, JY Horiba, Edison, NJ). Electrical conductivity and pH of soils after leaching were measured in slurry with deionized water at a soil: water ratio of 1:1 and 1:2, respectively, using a pH/ion/conductivity meter. Available N (NH 4 N, NO 3 N) was determi ned by shaking a 2.5 g air dried sample in 25 ml 2M KCl for 1h and the concentrations of NH 4 N and NO 3 N in the filtrate were analyzed with a N/P Discrete Autoanalyzer. Available metals were determined by extracting the samples with Mehlich III solution (M ehlich, 1984), and measuring the metal concentrations using the ICPAES, following EPA method 200.7 (EPA, 1998).

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57 Results and Discussion Effects of Organic Amendment on Soil pH and EC Incorporation with chemical fertilizer decreased leachate pH for both soil s, result from the nitrification of added NH 4 N (Fig. 3 2). Organic amendment generally increased leachate pH of the Spodosol for the first three leaching events, as compared with the control (Fig. 3 2), whereas the difference diminished and all leachate p H tended to approach 7.4 with increasing leaching events. The decrease in leachate pH by soil amendment with increasing leaching events was attributed to organic acids released from the decomposition of the organic matter during the leaching process (Fig. 3 2). In addition, Ca and Mg from the organic amendment can replace H + on the soil exchange complex, and thus decreasing soil leachate pH. However, relatively stable leachate pH with leaching times was observed in the Alfisol because of its calcareous natu re. With increasing organic amendment rates, pH of Spodosol increased, whereas that of Alfisol slightly decreased after six leaching events (Fig. 3 3), probably due to their different mechanisms affecting soil pH. om Spodosol were measured due to its low holding capacity and low salts (Fig. 3 4). The EC of leachates from Alfisol was higher as compared to the Spodosol, indicating higher salts contained in the Alfisol (Fig. 3 5). However, e between the control and treatments with organic amendment only. Application of chemical fertilizer increased leachate EC in both soils (Figs. 3 4 and 3 5). Obviously, fertilizer contributed to EC or salt concentration in the amended soils. In the first t wo leaching events, leachate from both soils amended with Hydra Hume and fertilizer had lower EC than the controls, implying that less mineral ions was leached from the soils amended with fertilizer and organic amendment. This behavior probably resulted fr om the retention of salts by

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58 the organic materials, which agreed with the results obtained in the incubation experiments in which processes of nitrate immobilization were observed. Effect on N L eaching and Soil N Pool The volume of leached water was very similar and independent of the treatments. However, the concentration of nitrate in the leachate varied depending on the treatments. Application of organic amendment tended to increase NH 4 N concentration in leachates, indicating that mineralization of org anic material in the amendment may have contributed to soil NH 4 N, which is subjected to leaching. Leachate NH 4 N concentrations decreased drastically in both soils in the first four leaching events and reached almost zero after the sixth leaching event (F igs. 3 6, 3 10). The decreased concentration of NH 4 N in leachates with time or leaching events may be related to depletion of NH 4 N because of nitrification to NO 3 N and microbial incorporation into organic fractions. Lower NH 4 N concentrations were obser ved in leachate from the Alfisol than the Spodosol, probably due to lower available N and organic matter in the former than in the latter (Table 2 1). Similarly, the total amount of NH 4 N recovered in leachates was much less for the Alfisol than the Spodos ol, while same trend and conclusion can be draw n from Alfisol. Nitrate leaching from soils amended with organic material depends on the nature of material, the applied dose, the time of application, the quantity of water applied as well as the type of soil (Burgos et al., 2006). Compared with the control, application of organic amendments drastically decreased NO 3 N concentration in leachates in the first leaching event (Fig. 3 8). The mechanisms of NO 3 N leaching reduction by organic amendment is likely re lated to the enhanced microbial transformation of NO 3 N into organic N such as microbial biomass N, with the increased input of organic C. Leachate NO 3 N concentrations decreased rapidly with increasing leaching events and were below 10 mg L 1 regardless o f treatments. Since most of

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59 NO 3 N in the soils was leached in the first two leaching events, application of organic amendment appeared to be effective in reducing NO 3 N in sandy soils (Fig. 3 8). A significant difference in the total amount of NO 3 N from t he six leaching events occurred between the control and the treatments amended with Hydra Hume, when the application rate of Hydra Hume reached 44.8 or above (Table 3 1). When fertilizer was incorporated, NO 3 N contents increased at the beginning, but decr eased in the subsequent leaching events, particularly for the soil amended with Hydra Hume, as compared with the control, indicating the potential of organic material in reducing NO 3 N leaching in soils (Fig. 3 9). An application rate of 44.8 kg ha 1 or ab ove is required for Hydra Hume to accomplish a significant reduction in NO 3 N leaching from the soils (Table 3 2). In addition, total mineral N (NO 3 N and NH 4 N) was calculated to understand the N pool in the amended soils. No significant difference in tot al amount of mineral N was observed between the control and treatments amended with Hydra Hume only, whereas a significant reduction of total mineral N in leachate was obtained for the fertilized soils when the application rate of Hydra Hume reached 44.8 k g ha 1 in the Spodosol and 22.4 kg ha 1 in the Alfisol. In general, organic amendment was effective in reducing N leaching in sandy soils when it was applied at 44.8 kg ha 1 in Spodosol and 22.4 kg ha 1 in Alfisol with fertilizer. Significant reduction in total NO 3 N amount was observed when organic amendment was applied only at the application rate of 44.8 kg ha 1 or above. Moreover, leachates from the Alfisol had a lower NO 3 N concentration than from the Spodosol (Tables 3 4 and 3 5). Potential Effects on Heavy Metals Trace metals, such as Cu, Cd, Co, Cr, Ni and Pb, are usually present at very low concentrations in soils, but are toxic to plants and soil microorganisms if at a high concentration. Controlling the input of toxicants is crucial for sustainabl e agriculture. Therefore, a critical

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60 evaluation for concentrations and potential toxicants in organic amendment was carried and statistical analyses were calculated between control and the treatments. The amounts of extractable heavy metals (Cd, Co, Cr, Ni Pb, Zn) were not affected by the amendment, mainly because of their extremely low concentrations (Tables 3 3, 3 6). The application of organic amendment does not increase bioavailability of heavy metals in soil or transport of these elements from soil to the environment. Characterization of Leached Soil With increasing organic amendment rates, pH of Spodosol amended with Hydra Hume alone decreased slightly, but increased slightly for the fertilized soil (Fig. 3 14). However, the changes were small (within 0.2 pH units), indicating that this organic material has a limited influence on soil pH. 1 ), although it had different trends with or without fertilizer (Fig. 3 15). The diffe rence between treatments narrowed gradually with increasing leaching events (Figs. 3 14 and 3 15). No significant difference in trace metals occurred between treatments, indicating that there was a minimal influence on the transport of heavy metals from th e organic material or fertilizer to the environment. Conclusion In this chapter column leaching was conducted to examine the effects of Hydra Hume on the transformation dynamics and leaching potential of mineral N in the Alfisol and Spodosol. The obtaine d results indicate that significant reduction in total NO 3 N amount can be obtained when Hydra Hume applied alone in both of the soils, but a minimal application rate of 44.8 kg ha 1 is required. If fertilizer is incorporated with organic amendment, signif icant reduction of total NO 3 N amount and total mineral N amount can be achieved at a minimal application rate of

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61 44.8 kg ha 1 for the Spodosol and 22.4 kg ha 1 for the Alfisol soil. The influence of organic amendment on the leaching of heavy metals from t he soils appears to be insignificant.

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62 Figure 3 1 Column leaching setup

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63 Figure 3 2 pH of leachate form Spodosol amended with fertilizer and different rates of Hydra Hume Figure 3 3 pH of leachate form Alfisol amended with fertilizer and differ ent rates of Hydra Hume HH rate (kg ha 1 ): HH rate (kg ha 1 ):

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64 Figure 3 4 Leachate EC form Spodosol in relation to fertilizer and different rates of Hydra Hume Figure 3 5 Leachate EC form Alfisol in relation to fertilizer and different rates of Hydra Hume HH rate (kg ha 1 ): HH rate (kg ha 1 ):

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6 5 Figure 3 6 Leachate NH 4 N concentration from Spodosol in relation to different rates of Hydra Hume alone Figure 3 7 Leachate NH 4 N concentration from Spodosol in relation to fertilizer and different rates of Hydra Hume HH rate (kg ha 1 ): HH rate (kg ha 1 ):

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66 Figure 3 8 Leachate NO 3 N concentration from Spodos ol in relation to different rates of Hydra Hume alone Figure 3 9 Leachate NO 3 N concentration from Spodosol in relation to fertilizer and different rates of Hydra Hume HH rate (kg ha 1 ): HH rate (kg ha 1 ):

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67 Figure 3 10 Leachate NH 4 N concentration from Alfisol in relation to differen t rates of Hydra Hume alone Figure 3 11 Leachate NH 4 N concentration from Alfisol in relation to fertilizer and different rates of Hydar Hume HH rate (kg ha 1 ): HH rate (kg ha 1 ):

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68 Figure 3 12 Leachate NO 3 N concentration from Alfisol in relation to different rates of Hydra Hume alo ne Figure 3 13 Leachate NO 3 N concentration from Alfisol in relation to fertilizer and different rates of Hydra Hume HH rate (kg ha 1 ): HH rate (kg ha 1 ):

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69 Fig ure 3 14 Soil pH after leaching in relation to fertilizer and different rates of Hydra Hume Figure 3 15 Soi l EC after leaching in relation to fertilizer and different rates of Hydra Hume 0 22.4 44.8 89.6 112 0+fer 22.4+fer 44.8+fer 89.6+fer 112+fer

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70 Table 3 1 Statistical analysis of NO 3 N concentration and total amount of NO 3 N in leachate from Spodosol Without fertilizer With fertilizer HH rate (kg ha 1 ) 0 22.4 44.8 89.6 112 0 22.4 44.8 89.6 11 2 Leachin g time (day) 0 12.42 ab 14.8 4a 12.4 9ab 10.1 9b 11.2 6ab 36.0 0d 40.71 c 45.6 0b 41.8 4c 51.20 a 3 6.74 6.64 6.68 6.52 6.8 32.0 6a 30.63 a 19.9 7b 17.7 3b 13.13 c 7 2.46a 2.05 b 1.95 b 1.94 b 2.12 b 15.8 4 14.6 13.5 3 16.0 8 15.76 14 2.52 2.05 2.06 2.13 1.91 7.39a 2.81b 3.24 b 3.13 b 2.59b 28 1.45a 1.39 a 1.33 a 1.07 b 0.99 b 2.76a 2.52a 1.72 b 1.51 b 1.16b 42 1.52 1.88 2.56 2.23 2.57 1.70a 0.98b 1.06 b 1.01 b 1.01b Amount of NO 3 N (mg) 27.14 a 28.8 9a 27.1 ab 24.1 13b 25.6 83b 95.7 7a 92.03 a 85.15 b 81.3 2b 84.46 b *Means followed by different letters within the same row indicate significance level at P<0.05 by the Duncan multi range test. Table 3 2 Statistical analysis of NH 4 N concentration and total mineral N in leachate from Spodosol Without fertilizer With fertilizer HH rate (kg ha 1 ) 0 22.4 44.8 89.6 112 0 22.4 44.8 89.6 112 Leaching time (day) 0 3.32b 3.80 ab 4.00a b 3.96 ab 4.95 a 6.46 b 6.64 ab 7.29 ab 7.28 ab 7.78 a 3 1.39c 1.45 bc 1.74a bc 2.05 a 1.90 ab 7.75 b 8.21 ab 8.59 ab 9.32 a 8. 80 ab 7 1.32b 1.32 b 1.44b 1.66 ab 1.87 a 7.87 d 9.51 a 8.57 b 8.11 cd 8.44 bc 14 0.68c 0.79 bc 0.88b c 0.83 ab 1.05 a 3.32 b 3.83 a 3.87 ab 3.86 a 4.09 a 28 0.54b 0.69 ab 0.77a b 0.97 ab 1.18 a 1.18 c 1.13 bc 1.32 ab 2.11 a 2.22 a 42 0.15 0.12 0.21 0.16 0.31 0.52 0.37 b 0.37 b 0.55 a 0.57 a Total mineral N 34.56 a 37.0 9a 36.16 a 33.7 8a 36.9 7a 122. 8a 121. 7a 115. 2b 112. 5b 116. 7ab *Means followed by different letters within the same row indicate significance level at P<0.05 by the Duncan multi range test.

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71 Table 3 3 Statistical an alysis of heavy metals in leachate from Spodosol Without fertilizer With fertilizer HH rate (kg ha 1 ) 0 22.4 44.8 89.6 112 0 22.4 44.8 89.6 112 Heavy metal (mg k g 1 ) Cd 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Co 0.00 0.00 0.00 0.00 0.00 0 .00 0.00 0.00 0.00 0.00 Cr 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Ni 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Pb 0.10 0.09 0.10 0.13 0.13 0.15 0.15 0.13 0.13 0.14 *Means followed by different letters within the same row indicate significance level at P<0.05 by the Duncan multi range test. Table 3 4 Statistical analysis of NO 3 N and total amount of NO 3 N in leachate from Alfisol Without fertilizer With fertilizer HH rate (kg ha 1 ) 0 22.4 44.8 89.6 112 0 22.4 44.8 89.6 112 L eaching time (day) 0 12.16 13.6 14.1 14.3 11.79 19.59 19.2 8 21.48 29.71 24.85 3 8.77 8.34 8.58 6.30 5.96 62.17 46.0 9 50.96 42.93 46.56 7 5.36 5.06 5.03 4.09 3.77 23.41 29.2 6 21.52 30.64 20.75 1 4 3.11 2.92 2.26 2.43 2.86 16.00 13.1 6 10.08 13.62 16.68 2 8 3.13 3.02 2.19 2.93 2.83 3.46 3.05 3.06 3.77 3.77 4 2 1.32 1.30 1.26 0.88 0.69 1.05 1.15 1.15 1.15 1.05 Amount of NO 3 N (mg) 112.0 b 113.1 a 109.2 c 102.0 d 95.2 e 419.7 a 402. 9c 375.0 d 374.6 b 364.5 e *Means followed by different letters within the same row indicate significance level at P<0.05 by the Duncan multi range test.

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72 Table 3 5 Statistical analysis of NH 4 N concentration and total mineral N in leachate from Alfisol Without fertilizer With fertilizer HH rate (kg ha 1 ) 0 22.4 44.8 89.6 112 0 22.4 44.8 89.6 112 Leachin g time (day) 0 6.11 5.46 5.42 5.94 5.98 6.65b 7.10a b 7.44a 7.68a 7.89a 3 10.71 a 10.89 a 9.94b 9.74b 9.57b 15.07 a 15.75 a 14.19 b 14.19 b 13.63 b 7 8.38a 8.15a 7.18b 7.00b 6.55b 9.9 9.42 9.61 9.16 9.60 14 8.43a 8.55a 7.27b 6.95b 6.98b 12.52 11.11 9.11 9.26 9.10 28 5.58 5.17 5.00 5.02 5.06 7.41 6.36 6.58 5.77 5.77 42 3.12 1.98 2.24 2.40 2.35 4.28 4.12 3.48 3.88 3.58 Total mineral N 154.3 a 153.4 b 147.1 c 139.1 d 131.7 e 474.5 a 428.9 c 427.1 b 425.0 d 414.1 e *Means followed by diffe rent letters within the same row indicate significance level at P<0.05 by the Duncan multi range test. Table 3 6 Statistical analyses of heavy metals in leachate from Alfisol Without fertilizer With fertilizer HH rate (kg ha 1 ) 0 22.4 44.8 89.6 11 2 0 22.4 44.8 89.6 112 Heavy metal (mg k g 1 ) Cd 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Co 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 Cr 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Ni 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Pb 0.06 0.06 0.06 0.06 0.08 0.09 0.09 0.07 0.07 0.08

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73 CHAPTER 4 SUMMARY, CONCLUSIONS AND PERSPECTIVE Summary and Conclusions Nitrogen (N) is an essential nutrient for plant growth and crop production. N deficiency may hinder and stunt plan t growth, and therefore decrease crop yield. Large amounts of fertilizer N was applied into soils to obtain the maximum crop yield. If application of fertilizer exceeds the gap between crop uptake needs and the supply from the external sources, excessive N may be brought into the environment and contributed to environmental pollutions, such as water eutrophication, nitrogen deposition, etc Different from C cycle, elemental N (N 2 ) cannot be utilized by plants and microbial organisms directly, which means N becomes biologically active when it is fixed or bound, such as incorporating into ammonium (NH 4 + ) and nitrate (NO 3 ). However, m ost of the N in terrestrial systems is largely bound to organic matter and mineral materials that protect it from loss but make it unavailable to the plants. A mineralization process may be observed to convert N from organic forms into plant available forms, when fertilizer was added to soil. Therefore, to maintain a balance between N supply and crop demand without excess or defici ency is the main obligation of N management. For citrus yield and quality, N plays a very import role. However, the low use efficiency of fertilizer (usually from 30% 50%) with current practices not only represents a significant cost to the grower but als o contributes to environmental pollution, such as water eutrophication of St. Lucie River and ecological degradation of Everglades National Park in Florida. Soils in the major citrus and vegetable production area in Florida are composed largely of sand (sa nd content >90%) and have a low nutrient retention capacity. High summer temperatures and rainfall in Florida provide favorable conditions for rapid leaching loss of N from fertilizers, which may pose adverse impact to the environment. Agricultural practic es were applied to increase N use

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74 efficiency and reduce inorganic fertilizer N use, such as cover crops, crop rotation, organic amendment, etc Among these agricultural practices, soil amendment of humic substances may improve N use efficiency by crops by stimulating transformation of available N (NO 3 N and NH 4 N) into less mobile forms, such as microbial biomass and organic N pool. One of the main objectives therefore of this study was to quantify the effect of Hydra Hume, as an organic amendment, on N tra nsformation dynamics in both original and fertilized soil. Two representative soils (Alfisol and Spodosol) in South Florida were selected to test. The N dynamics in soil was affected by the C/N ratio of Hydra Hume as well as the original C/N ratio of the s oil itself. Hydra Hume, which had the C/N ratio around 42.13, induced a net immobilization process when applied alone in both soil types. For Alfisol soil, the immobilization process only occurred in the first two weeks with an application 224 kg ha 1 or h igher. However, fertilizer contains large amounts of mineral ion, which is an N reserve for microbial biomass. When fertilizer was incorporated with the treatments, different N dynamic trends were observed in the two soils, probably due to the difference o f C/N ratio in soils. The N dynamics in Alfisol soil, which had the C/N ratio around 22, induced net immobilization, while in Spodosol, a net mineralization curve occured. The other objective was to examine the effects of Hydra Hume on leaching potential of mineral N in Alfisol and Spodosol soils. Significant reduction in total NO 3 N amount could be observed when Hydra Hume applied alone in both of the soils, but at an application rate of 44.8 kg ha 1 If fertilizer was incorporated with the treatments, bo th of significant reduction of total NO 3 N and total mineral N amounts occurred, but a minimal application rate of 44.8 kg ha 1 is required to accomplish significant results in Spodosol soil and 22.4 kg ha 1 in Alfisol soil. In

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75 addition, no apparent influe nce on the leaching of heavy metals from the organic amended soil was measured. In general, for both soils, the applied rate of Hydra hume do not appear to pose an environmental risks with respect to the amounts of nitrate leaching and the concentration of heavy metals in leachate. Analysis of the leachate data confirmed that Hydra Hume had the potential in reducing mineral N leaching, but certain application rates need to be required if significant difference was needed. Perspective This study provided so me new information to soil and environmental science in the following aspects: (1) N transformation dynamics was affected by the properties of both organic material and the soil. (2) The application of Hydra Hume showed the potential in reducing mineral N leaching in both Alfisol and Spodosol. (3) No apparenet influence was observed on the leaching of heavy metals from organic material and fertilizer. However, future research is still needed to address concerns with attention to the following aspects. All t he experiments were conducted in the labtory; therefore, field trials are needed to confirm the effects of Hydra Hume on reducing NO 3 N leaching in the real circumstances. Moreover, potential contributions of irrigation to the N dynamics NO3 N leaching ne ed to be considered. The types of irrigation as well as the amounts of irrigation play an important role in affecting NO 3 N leaching. However, the effects of plants growth need to be taken into consideration. Ecosystem is an integral unit, which soil, wat er and plants interact with each other. Plants compete with microbes to take up mineral N, and during this process, plants take part in N transformation dynamics in soils. Therefore, the influence of plants is a key factor in N management.

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85 BIOGRAPHICAL SKETCH Wen Gu was born in Jinan, China. She received a B.S. in e nvironmental e ngineering from Ocean University of China, in 2010. She took an internship in Shandong Academy of Environmental Science and took part in the environmental impact assessment program for Chengbei wa stewater treatment plant in Shandong Province, China. She began her m aster study in the Soil and Water Science Department, University of Florida, in 2010 and received her degree in 2012. Since 2010, she has worked as research assistant at the University Soil and Water Science Laboratory (SWSL) at the Indian River Research and Education Center in Ft. Pierce. She conducted research on the n itrogen management in s andy soils, especially for reducing n itrogen loss by applying organic amendments. S he is a member of the American Society of Agronomy Crop Science Society of America Soil Science Society of America (ASA CSSA SSSA) and Gamma Sigma Delta.