A spot disease of cauliflower


Material Information

A spot disease of cauliflower
Physical Description:
McCulloch, Lucia
United States -- Bureau of Plant Industry
G.P.O. ( Washington )
Publication Date:

Record Information

Rights Management:
All applicable rights reserved by the source institution and holding location.
Resource Identifier:
aleph - 29605509
oclc - 37307712
System ID:

Table of Contents
    Front Cover
        Page 1
        Page 2
    Letter of transmittal
        Page 3
        Page 4
    Table of Contents
        Page 5
    List of Illustrations
        Page 6
    Introduction and inoculations
        Page 7
        Page 8
    Description of the cauliflower leaf-spot organism
        Page 9
        Page 10
        Page 11
        Page 12
        Page 13
    Name of organism and Latin diagnosis
        Page 14
        Page 15
        Page 16
        Page 17
    Back Cover
        Page 18
Full Text
~A S'PO'll I)II~S1 1'C IP ( L 1 1 (111
ii29 1911 .


Ass'islao Chii)'ofPnrcuz, W\ILLIA A.1 TAY XLOR.
Chief ('1 r, J.%.I 1 jo" '
E vin F. -mnjth 1'i C11harge.
RI. E. BI. McI1ouney S cee-h JAj i!
Nellie A. Br uwn, Lucia ii uluha I Mary xKatlierinie B-N an, scicntijf A .,siants.

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11L 1. t" T' R AT'I I () S.
PL ATE I. Upper s-urfaces of aili flower leve,,, shoxinug nal uml infect in Nvi! hloe
B3acteriumu mouic~Iol uin .........................16......
11, IUndeq sud"f:q of ca"Hlowm-x e, showXing 1nidril), polleI hy jiallnda
iife(ii 11 vii h Bami Mn mn nmrcoel um ....................... 1
IBact ,iimn macueicolum u~~i............................................. 16

Di. 1'. 1 -

[III \' tlilt andx ai n t'tI-ill l it' PN Iit d iva l- c lt' v lto i t N vent io
mmlliltox the fWwwwi w oi f tiWit PaIhg oa. nt5140 11
lw w b ,p rd~ rysu s I tiim r in :H:nvtr (1 )
Thw wem ;Lst large ohe -~ area (in tob the o l'i ,o po


sp)ots (b) not bec'4~mie nioreP linnl 2 to :1 miilliietel's ill I -est; diameter,
tlt 14 lug whrt cn11'i('NIN W ded t I 1' s)ts u1silial (.)lsce, fliii ieased
11"Ll Of1 (,( 41'ider'a lil '-ize.
ANliell wIilY the uppler' ()i'a' leave Was~ 'ubj eced ~ to~ t le iliocu-
lat ill." spraYc verY fe~w intfect i lis ro,"ulted While illu ithat ions oil tile
S4 S1111'ae --a\ -o miieisr jifet i4aiS (PII. 111). So far as (d)erved
ill ,.dect 1i 4ls tlit(, ttkc-,' p4411 ~s lace wlvl t1 r )iwhl stom11at a, and
llw4 51t v fr()i 4 the I d su rfac. Tile I111er aflid ile yervy x-()Ill leaves
"Wiear 0o b)e1 Ipalrialy l ()I.een (c41)IpleteIY imniitle, while thIos~e 4f
nt eriled iat e age ((m iites zime p~lanit) may' be serio usly affected.
The diseased leavc es 1 eclmi vue llow ale d fall off iin from three to five
wVee! ,. Lme~ vm~e ll ie Lavo alld 1ie (1iW )tl 4 i are healthy. II lider
ouer rallther dIry 11thmN(e cmid4lt~i1mN 1i1 114) case was there e-videnice of
IinfecG1441 mt1 tit.\ but1 t11 Moc41ulzited leaves.
From)14 s1)wts (dcvcl~ 11" 4411 Iezteinw 1.44ulatedl wXithi t Ii original
cuiltuire thle ()rga:li ,j!1 WilaS 11'" eate liv isolated I) y 111clls of petri-
dishi pmilvdh plat es, a i d si ucult irs frm C111 Q4 doiiis we-re '1g0ain used
b~riloctlai(11, awnvs ili.the prm'liuctl )f1144 the chlaract erist ic
infect 14011.
The Idclwc 4 ill ill c(,( rehniiiiied I'licatIy ; also4 numeri(iouis other
cauilillowv ] ) illill iIle salmte ".reeiillUse.
Cabbages" 1114 owtlat eII wXith tihis ()r-aillisill becllie inI feced ('il h le
S,11111( fl111fier'Is tie( (cailll b\X e with wl e ( \c(Tl ~j41, viz,, thle spots
were dwi rker. Fro sp( NI4s app.14 eaing 411 thle inlit ('at ed cabbages tile
on-amisin wi ns i,,( datc ('4 ad t ('sled (Ml Cauli1flower, with Ith rlesl'ult of
tlite produclitiml ()f' thle disQ("S
lDuriln. May '111d1 .Jiul 1 ()f 1! oUJ all ii 14o(culat im is 1'estilte( ill successfully.
lilld tqnci4al i11fecIt 011S.
()it .11ly 10, 19119, Seve'n pihlts m-c(re( inlocullated l ie usual mnanner'
zild, co4111intry~~ t44 explct at 14411, iw4 inifect im 1s resulted'I. The' ('(411it 1011,
were t114 5s11114 as Iil previl 4 iN 1110 cillat ins, c'XC4'l)t that tilie t empera-
ttire was hioigher at t his t imen, 26' to 34' C. (78' to) 9V8 F.). Sub)5e-
qun exlperlhulelits si f W t at tilie bacterium ca1llsim' the disease
refuses t4 ow ]i art ificial hia in t 290 C. (S~4' F.) or above. It is
Ip1n dbble, thleref ore, inht iai this case tie( high t empieratutre p~revenltedl
After J ill y no)i 1 1,t liuts wxer availalIe for inoculait ionl mt 11 1Jan11-
n r v 2 4, 19 10, NN i i i 10(1 lant it ; w c e I i ( i la ed. No) inifectiowis n4suilted.
0ii Februiar .v 2, 19 10, Mi mor,1e phlints Nvr ( Ifll~willatd and14 no illfec-
4,'l n-is I'4Nll ed.1 .All (- lmit iis se4'1i1ned fa \,,wablle, aidlos o05f \-irilenlce
ill tite cl tire '( s 1"leldi'E ans tit(e caulse' 4f failure to infect. mei
cuilture is Itldl I'm Ii' iil 'tat lol were d escenidedth Irouighi numerous
lausfels made tlw -wtl1 g114 V inter'. aui r-stm- Ik c'ult ure wh~i had1(
a gl 'a ii(lil -crs Weld miadeh fr'i' the1 li' ;eptcjniber stock and used
, 25

it I I I1 2 It' )li I I t I1. F l it I 't I I 2, Pitt I i o 1h w)L 1 i~'at1111 ta 1, ~'
Iwt vvvcr i zind t h i x e juii t I t Im w l it c,~ ti 11\ v- i t 1, co Iu it(,
I tj I to 'la I x it I t (' a ctck c f I lI Ix IIt x si 1) I'
Il il a. I t i I I t F I I I I i I, c \ I lIt It I I 1 I I I I )(411 1 1
I I .1 i l tz' s n I- If t 11t I t la t11 f I v ii I nx l Ii I u I F x u- 1, t'l
()it DecF I aaI I r I i l 1t' 11d t Ii lt a III' r it- I I I I 1iti 1F 11' 1i ; t Iz xxt -
I I 1 14 4 111, 1 it I c I II tI I t I t I t t' l A I* Ii
08Y I I. I II I I~uf- suIt ovil c d tit' :11c rtui~t t- [Ic CI ifit~-
rc I II

I)V 4 CIt I I "I'l I IN 4 W TI IF 1, VA )I' I t I,( I k %'I sNI.


means of o-ne to five polar flagella. which are two to three times the
length of the rod. (Stained by Van Ermengenm's method; also by
lfugh Williams's method.) Motility occturs in most artificial media.
Ti l eef-bouillon cultures grown and kept at 0.50 to 1.5' C. for four
months the oroauansm is still motile. Involution forms were found in
alkalnue beef bouillon (- 17 on Fuller's scale). Petuidozoogloex
occur ill Uschlinskv's solutitoni and in acid beef bouillon.
The organism dloes nut stain bv Gram. Modified Gram, using
amvl alcohol, gives a deep blue stain. It stains readily and strongly
wNith carlbol fuielsin, with an alcoholic solution of gentian violet, and
with a, stain obtained from )r. Kinvoun which contains inithylene
lblue silver nitrate, azure I, and azure I. It is not acid fast.
Agar plates (+ 1.5 pPl(-pon *Z d bcef bouillon qith I pcr cent agar).-
The colonies are visible on the second day as tilly white specks
temperaturee 23' (.).
TI tllree to four (I \-s the coloniies are 1 to 3 millimeters in (ialneter,
white (op:alescent in transillitted light), rointl, smooth, flat, shining,
and translucent, with edges entire. Structure, uner han lens,
coarsely granular with internal reticulations. Buried colonies small,
lens shaped. With age the colonies become dull to dirty white,
slighifiv iireglar in shape, the edges undulate, slightlI crinkled, and
with itidistitict radiat -ilg marina lines. The internal reticulationis
(lisap)wear aIl(] the loarsel Y gratuitlar al)earance chuaniges to finely
grat1nullar. InI thinlY sown )plates 7-day --old colones are 6 to S milli-
licter's ill lialneter; I5-day-ld colonies are 12 to 15 millimeters in
Aaj(Ir stabs.-Tlhe surface of the agar is covered in two days (220 to
24' C.) by a thin white gro\-th. or several dayx s the stab shows a
moderate growth iin the upper S to 10 mI llimeter;s, but this does not
con tiinue. Finally, te stab is alunostift ot quite, invisjible. Crystals
appllear in the stab and on the surface.
ArpirP slants.-Tn smear ctultulrs the sir-face is covered in two (lays
temperaturer, 190 to 210 C.) with a thini white growth, glistening,
coarselY anw irregularlY pitted. Wile sediment in the V.
Tu streak: cultures ii two lays temperaturee 190 to 21' C.) the
Streak is 3 to 5 millimeters wide, wiit, mrgins slightit undulate.
Tlie internal reticulations seen in colonies oil plates are present in.
Hie st break cultures. At right anwgles to tte streak ar firine lines
cxt e ling, from center to mu arhn.
kfrllr cli t ures become slig-htlv greenlish.

Wf ho"ilUn. Ap"HAWffi1f I"A IM"A IffIif 10 hfill ft 21 to 25 Q,
if inoculiated froln~ V igorous~' bouiIlon cjlrc- clo' fudsII thbink
it) 6S h~ours' :i is JntOlk, wt hv~1 faXl AIM infflf 1f2 1 hoursf''. rw f
gn'fi1i is' Imi~ at the suni-facv, mX wre a Ivfle iaw vr is fwn"Aftl 17is. is-
Ifot af true IpP(HIN, as Ai dli,-inlv f;rc wh Iell On cuhrc ar1fle e hanfdled"
No zoo)gdovea, are' 1wefe- There is I'- ff inf1 II f iX \ o la d It f \ ; th r ;I-
latfe. After several Xi"A th'~le prMf'fI~lalifff A XLlt' an shlin I,, 0
(list e i (tifaflit X, Unl wXit If ill:11 I r ,l \'sl if) it, '111 oi i n'll
iM41ICS S iiQ Ig M4*eliAi AMtf seeralfZf Ifflofi hs* 1 JIfh peIpitle 'i"
Ivrou-tifh~ell ffwti 1 until wi achit v l of K! 1 IN~f oxafic' nidl afffI + f 36 W.
10 N' d ~e In1f IPIff' IV If, Sof~ SI11 asl' If o 1ff sl'~JIvridd. ift I hs
XXvi no'eft mlf h hfIf int ions Uns provjf' ed hyf phltingf outf andH by test on'~Vf
nowl v' I in af toxi hnfv ihty Imffffilhuf ( \ ()I I us'ed I I h14
or ( fafi fff :rc\X Htl in 7 19, 22. hss- in 21, andfl wff A~ Af if)
- 25 -'i 26,'f~f nnd 21 "I 's If I M.~f X i wofI X, Xu Ition fwilfadi ian il
IBoo~i/bwf Whd/ x(AI/Im chlophl.-A bf bouilon I dusiIfffi~l' 2 Iwr ('('nt NXf I
then -rowthX Iis as goo n, I in J4ai 101' NOIfuL Wf.\\lihI 1110 MUS [lt fl Of
Moret' Nn~ I(he gifii' 'XI gfiffdindX lewsenfs until it is *scarM vI lnoticlef1I1(
in1 I, A [wr' ('nit sidic" M1 gIr 'lef n in a I [Iw'r cenit s'(luft IhI, I hff
(ffmv f.ivi i', ff111 ffn l. fn af 2 fIw' (wff sohobiif 1f the ("jf nkniff'f 6;
fifnt"l Im f! t"s' M, thanff in If('f iMM~fhji W W X t I(I >' a(T'
W i' f bW on oty' r' c/w vo -f/ r the 11 f irst 2 1 luirs 1ff" hie g"f~foX- t i-I
stffff( hat r etard1 ed BA-11 endf ( df1If1 I' is 1ffr no' ffH1qn (imb befe Iel f l
c.v.for l(inwr I iiIfn fwl sh lf; i)f I ndfl those~( inf jlinf wor( Ihouff if.
.After' three' Iffoht ile WKSIII f1 luediim waf' shlit lfy)X lkr'MI(.
pe f'le or~~lfI iff) 1ll fOWN1141f IV' Afivr mIffh'nel 1AIS kihm I
zolto I Aridanlift, XNil' flocculenf'ft. 1WfiliceIf %%bile. tender'f' OAK--if
inI "'rin andf~' wf w w f qIf"4 I''i flIv thef' f f di' i i s c ~ ff~hn andIffI I IIfl I ff



pale 2rci n-luorecten t, (betwe Nvcci ater green anld greenish glaucous.,
Ree 'rti ei de C'ouleurs, P~aris, 1905) ; more precipitate than ill beef
X01-1' 4 hlitoyatn ( + 10 onl Fuller's scale). -Ple stabl cultures li(Ile-
lied inl S to 10 daYs (temperature 17' to IS' C.). Girowti from mtir-
(Tel cilorin. Slight, whilte, granular ptrecipitat e. Slight green
1i114wescen ce.
The plate04 cultuiires sho iwed no11 signts Of (rrowtlh inl 24 hours at 17' t~o
I s C. Ill three d nys -,vell-i ;olatvd colonies va ry from mnere points to
1,01ii1(l, grow tlt- 2 iiill iji0ta ""'Sil diameter. The gelatin is liquefiedl inl
cuik o hollow'. Aarm) "I nashu 1r colollic, vlntire, of larger colonies
fiinrwitte. IThickl sow pla\ i htecs enltirelY liqlt&iod ill I w(x days at 15,
I C) C.
L d~u as i 1.-lThe mod ium ib1ecomnes dark lue at ti te surface inl
1 2 to 24 h ours>. TIw a 1a rken in pr tcceds1~(1 dox inard ill definite layers
it IN tl 1n i (o ) d I a te w ,-%]i o Ie i idi u i i iIs la rk WA u c wx t li a sliA it, whlite
pre tIpita ate. I utr i ix 11n milt'~ ob,,ervx'ot tti te iedium remtained
(1,11k blhue (reflerf ed liaJlt ) t md liq uid. liialY I) evzixporation tile
Incrit'l ltil becomIies l thieed, but,1 there ;-. at it() t 10 MV ti YSep~arationl
A fewv culture" ,]1owecd a trace of reduction of litmul at t le bottom.
Jilk. As ill tile lit mus-iltilk elultures, growthi andt color-chiange inl
1he milk beginu ati the su4rface, proceeding dIownIward iii (lelitnite( layers.
al 1 5 to 20. days t le wleI( tube (10 c. c. Of milk) is yellow (necar
1)idgnvay's N;iple, x eihtW: bitt somewhat, duller. and Wvitlt a grevni,.i
t ingre) and tnrinsluccnt. 'No sepitrat ion into curdi and whey. Fat
lot, clam)ged. Ill fou)tr months tlte mtedium is quiite dark (reddish-
roiid -m(1siliewliat, thick (ev aporated to about 5 c. c.). Small
t ,vro- 11 cryst'll' are forlnedl. These tre dist inct ly visile only With
a lens>.
[ ~C~ti 14 f~ ~OUI )1. (i~ ~v hiiiodlrat e to cop ious pellicle -whiteC,
tender, breankinfr anid siiikiin easly. IPseud~ozowgloea, are p resent.
Th ere is a g reenishli lttorescnice. The Old cu1 ltuores are much like
th osv inl Feriis soluit in.
LI 111 ttitH,, it1 li ogi tis 5aerobi( andi (oesniot fortn~ras.
It wns t c,,-;te ill fertiei t at ioul ltubeS ill file I)rTeec Of dIC-IIrose,
snalrt ,v, lact se, imt ose, glvccrflin, and 1mann11it l each of tese
c(otImnl cmloiltltids Iwiilg adtdedl to at bx".11 solution co liistinzg of I
per ce tt Or ittv's peptmit d vsed inl x aler. It did( ntot grow ini
tlie en'~d 1( of tilie ferilleil at loll tubiles ill filie pre'Cl'ie of aniy of
Amm, ol, f; /~lU~Hi prduho" er1tc.

1ll).C-1) I lld l-mlcil i1 f cler. lh Ii XiI~C1~ 1 i
IHydrmil s/ tlpi duuIly. tep 4ulpll1 i' not ,ii it culun ( olm
bf-px o ephi wr arC plitol \cvlllde- tllunip cIlde. orr Iutlin bec
ke ho ml ( It poil i le t The 'I l.1 CI) 81 11 'C (] 1ll plil ] -1W)U C Theuuii
temper)C:itut-e (2W1 to 2 1 C.) Vir-4, tempen)i'tI1lC ur1, fll-ml Ir .t1()-
to 50 C. wereC tied,1 l1ul, theC Ilel-111:1 (1(.:Il 1 ImIllt cc llgv In l10
about.1 lulIfXvay be \vveXXCCI, trinlls \v cre tou 11luide,~ of 45, 4W and 47'
C. Mmp I111 tha lf oft ile cultu res vxoc to 45' C n- 7 for I0 m i inn
c(midv)d in t 5 da ( vs. Of cuhmv WN'cxpo~cd to 46 C. I out1
ofi 12 cimildedI tiftcr II I dtiy The ol lier"s iieve c (LimidIe. ( )f 201 culi-
tur~es (\jp')w to) 17" C., 1lf"110 Cluded.
01h'11111~ (I/ ii u ( IjIllu 1 1 1 peral18 111v' fm.i grnm-11) is
2.1 t 25' C.
Jlxm al'/uUl tul aic t n .-Thie 11 I ili 11 ll l f'8II eh ~(w -noX iIs
v'er hm viz, 2WD" C.
bloxvI 0 C.
The) o1'allisin1 NvX vld (1 a11c1 'Xihlle hItm (hys nt :,)", 0);b1'
ill beef bImillmlt.
ANilell von11 ,,WI Nvel-clide bme(4 lbouil)ll (clltllv Nver dried (I oil
( (rt'&Il, Se, nild) kep~t in n ark place at t1jcmpera tmY-- (if 22" to 25'
C., 75) perI centI \ecC ki\lledl ]i 21 ho)Uu and1( 90( perP cent in 48 hiours.
All were dead inl five days~.
Fouri iniute's' exp)o-urIe to k.it g il led lC ill orgn isms, inl t hIiIv
pl~aic hinig cmX cred n, a check.
Free'(zill-~li Il) lwn lil of ldt :1md) Iund li fi~l~ hir I~v mid) f2(ive hours
b eforI e and1( after'iIC'i
'Beef bltilloll (imtC. ci i21lli lle n1ld) n1 h'ut)' frozen :11)( kept :0,l
tempe J)'Fat 1I'(es of --V to I'~ C. la1 Xcrage -L C.) I'iic daIC lV~s



showed no growth (luring, this p~eriod1, but cloudled moderately three
days after retimal to t einperatutre of IS' to 20' C., and TAates from
thlis tube gav-e pure cult ures of the cauiliflower organismn. Plates
poured before and after 10 days' freezing showed considerable reduc-
tlon in the nunibers of organisms, but the grrowth of the living, ones
w i ,- not retarded. Some tubes of beef bouillon, inoculated with a
3-itiiltimneter tool) irom a 48-hour bouillon culture andl kept frozen for
sev-ei days,, cloudeat within 48 hours after removal to a temperature
of IS' to 20' C. Another frozen for 22 days did not cloud after
reliloval to temnperatures of IS' to 20' C. The check clouded. The
or0*.alln gTrows readily at low temperatures, e. u., beef-bouillon
This organismn remains alive for six to eight, mionthis at temp~era-
ture,,. Avarl-il g from 18' to 24' C. on beef agar, Loetller's blood( sertum
an~d potato cylinders, andl in peptonizedl beef bouillon ( 15), beef
yelatil( 1) milk, Uschiinsky's, Fermi's, awl Colin's solutions.
EN-wpo)(ration was not prevented in these cultures and the media
I)ecd tille col'ic en t rat ed1, oft en dry, and vet tile organisin was frequently
ali\ve. Beef agar and b)eef-gelatin cultures tit temperatures of 12' to
15' C. and stdilject to Ic ,s evaporation (nrefrig-erator) were dead after
e-it ;n olts II ll edia 1(- favorable for t le growth of this organis,i
jw(4 b c ou' llo i liS sail, 1 a1kn;i i oraiI i teria, 1,vo but tw to)11
three 111oliths.
TInw group ii-iiw~r, accordhiw, to the dlescrip~tiv e chart of the Society
of American Bite ril )o(,]t,, is 211.3332023.
This, orga ilil a11pears to b e anl undlescribed form, andl lecaulse of
th e (chairact eri:, Ic, s.polt iiil- (of thli affected leaves thle name Bacteim
Mlhicu ihs i II~ osp ite b lvv iI us, cylii'(11 ri, a picibius rotutildatis, soli-
sa( ]w SIj) III., (III W" Iff-War (1luandue 10-30 bacutis in fianenta,
cmijiuiictis) ; bactili ; 1 .5- 1.0 !t x 0.S-0.9 If; 1-5 flagellis polaribus mno-
Iiii aeroIbiis, ashporis.
I I iilit at ill folii-, \-i s iBrassicae oleraceac in mnactis 1-3 mm. latis,
P I 811tTo -(?iTSV04 04 oF 0. (Coh liAe gelatillaum li(II-efacientes. Colo-
11MOe Ill Wgr ar rot lihalb nijteiites. i baili in mer-aiu
1',1e S011,111t,1, ill Spt iiiio die colonac 6- 111111. diani. sunt Baculi

S T'NI N1 R '

lI ) I t I~.it I K'-1 I) I ~t~iux I Il'tilt I It I I'tc i I~
a' jI i n z I IIII I :IcIIij t I l ~ h -Ii I Ix I I III I Ix I I-. tlm I ) t I II I III t II jp cI tI In
29' Cuh. ( e )i'ait ram -e V C. cait ur~ ar c it. St I'l t I I it(~'\ l e
sililIl l w iu c al-11. (lcc li)I illi 1- 1 lII r I n-cf '.lti I beent ai o itn i.
. >" l *iii. S 1cc141 Ic li ). ; exiti Ie lit' 111" ii c nl~ ( nr I I I I I1 al 11 11t
v agI v I \. Brl Jin l nt (1re .1 1 4 ) 1 i !il [,I liac Ia Ill (' li- ii
prodiciit uii im In t lml till m ii01m l tuit Ing~iltciittj
Te l nf- pt aiiiinhla caiul h 1am e otl Ing cr l ph In a the- preodil
the iictall spotand -, cut ed i l t()l hcIthli i v~ -8 cm il e pjallc 4 I t
ll Il liiltcri m ii cIIIC O.s l bem. of -

l11 26211111 08928II1II1II 111 32291111II I IIII lill 11
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