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STUDIESONCUCUMBERGREENMOTTLEMOSAIC VIRUSINJAPAN* 1 NOUYE ,T.,1NOUYE,N.,ASATANI,M.andMITSUHATA,K. 1 . INTRODUCTION I nt h es p r i n go f1966 ,av i r u sd i s e a 田 o fcucumbernewt oJ a p a n田 cun 吋 w i d e l yi nv i n y l h o u 田 si nK i n k i ,Chugoku,ShikokuandKyushud i s t r i c t .S e r i o u s F1 Kurume ・ O c h i a iH 'cucumber damagewasdonet omosto ft h ev i n y l h o u s e so f‘ s u f f e r e d fromt h ed i s 回 s e .E s 戸c i a l l y ,i nTokushimaP r e f e c t u r e ,i twass a i dt h a t a b o u t60戸 r .c e n to fv i n y l h o u s e so fcucumber , a b o u t49hai n町 e a ,hadsuffered fromt h ed i 総a s e( 2 ) . Le a v e so fcucumberp l a n t si n f e c t e dw i t ht h ed i 田a s ee x h i b i t e dg r 民 nv e i n b a n d i n gandd i s t i n c td a r kg r e e nm o s a i csymptomswhichwere i cv i r u sandw a t e r m e l o nm凶 a i c d i f f e r e n tfromt h o s ei n d u c e dbycucumberm偲 a v i r u s . R e v e r s i b l ew i l t i n go fi n f ,配t e dp l a n t swaso b s e r v e dcommonlyu n d e rt h e v i n y l h o u 舘 c o n d i t i o n s . F r u i t swere田 v e r e l ym o t t l e dandd e f o r m e dbyc h l o r o t i c s p o t t i n gandr o u n d s h a 戸 ds w e l l i n gd a r kg r e e ni nc o l o r . Thea u t h o r smades t u d yont h ec a u 担 1 v i r u si s o l a t e dfromd i 蜘 s e dp l a n t岨 mp l e sc o l l 配 t e dfromv a r i o u sl o c a l i t i e s , andi d e n t i f i e dt h ev i r u sw i t hcucumb 町 g r e e n i cv i r u s(CG : M : M V )whichhadf i r s tb e e nd e s c r i b e dbyA i n s w o r t hi n m o t t l em佃 a 1935i nEngland( 1 ) . F u r t h e r ,t oa c h i e v et h ec o n t r o lm e a s u r e ,theauthors∞n d u c t e dsomeex 戸r i m e n t sont r a n s m i s s i o no fCG : M : M Vandoni n a c t i v a t i o no ft h e v i r u si ni n f e c t e dp l a n tt i s s u e sb u r i e di ns o i lbym e t h y lbromidef u m i g a t i o n . I I . IDENTIFICATIONOFTHEVIRUS 1 . HostRangea i 鈍 dS ymttoms ∞ulationt e s t swerec o n d u c t e di nt h eu s u a lmanneru s i n gcarborundum出 In ana b r a s i v e . Toc o n f i r mv i r u si n f e c t i o n ,backi n ∞ulationwasmadet ot h et 白t p l a n t ss u c ha scucumber,戸t u n i aandDalurastramonium. Fort h esamep u r p o s e, v i r u sp a r t i c l eo b s e r v a t i o nond i p p r e p a r a t i o n swasmadeu n d e rt h ee l e c t r o nm i c r o 舵o p e . S u r f a c eo fi n ∞ulatedleaveswass t e r i l i z e dbyNa a P O ,( 1 0-15m i n .i n3 %田lution)andwashedthoroughlywithdetergentandrunningwaterwhenback i n 田 u l a t i o nwasmadefromt h ei n o c u l a t e dl e a v e s . o s tr a n g eo fCGMMVandsymptomsi ns u 民e p t i b l ep l a n ts p e c i e s I nT a b l e1,h a r es u m m a r i z e d . Ass e e ni nt h et a b l e ,CGMMVi n f e c t ss y s t e m i c a l l ym佃 ys p e c i e sofω∞r b i tp l a n t s .I ti n f e c t sl o c a l l ytobaα0 , p e t u n i aandDalurastramonium. 0也e rp l a n ts p e c i e st e s t e dwerea l lf o u n dt ob en o ts u s c e p t i b l et oCGMMV. ・ τ o i sa r t i c l ei s∞m p i l e d ,bωedupont h er e s u l t si nt h ep a p e r sw r i t t e ni nJ a p a n e s ei n Nogaku ,51,( 4 )p u b l i s h e di n1967( 1 2 ,1 3 ,1 4 ) . Th i sworkwωsupportedbyt h eg r a n t i n a i d Kenkyu a t i v eR白 銅r c h,Sc i e n t j { i cR飴 e a r c h e so ft h eM i n i s t r yo fEd u c a t i o ni n1 9 6 5 . f o reo.唱戸r T .I n o u y e ,N. I n o u y e ,M. A鈍 ta凶 &K. Mit s u h a t a : 50 TABLE1 HostRangeo fCGMMV 悶 p t i b l ep l a n ts 陣 i 句 S u , , D i f f u s ec h l o r o t i cs p o 悦i n g ,m o t t 1 i ng L e a fy e 1 1 o w i n g ,n e c r o s i s ,l e a fd i s t o r t i o n ,mωaic,r e v e r s i b l e w i l t i n g Cucumismelo Y e l l o wd o t s C .m elov a r .conomon Ye 1 1 0 wd o t s C . melov a r .makuωa Y e l l o wd o t s C .s a t i v u s V e i n b a n d i n g ,s e v e r emo 悦l i n g ,r e v e r s i b l ew i l t i n g ,m o t t l e da n d d e f o r r n e df n i i t s C u c u r b i t amazima Ye J 1 0wmωaic ,s t u n t i n g C .m oschata F a i n tc h l o r o t i cb l o t c h C . teto Y e l l o wmωaic ,s t u n t i n g Lag蝿 a 1 ' i al e u c a f l t h av a r . G r e e nv e i n b a n d i n ginωmep l a n t s ,o t h e r 百 a r es y m p ωm1儲 c l a v a t a a r . gou1'da M L .l e u c a f l t h av o t t l i n g 1 ' i c a Luffac y l i f l d F a i n tm o t t l i n gi ns o m ep l a n t s ,o t h e r sa r es y m p t o m l e 鑓 1 ' d i c acha 1 ' a f l t i a Momo V e i n a lc l u o r o s i s ,m i l dm o t t l i n g ,s t u n 出g DatU1'as t1'amoflium L o c a lc h l o r o t i co rn 舵 r o t i cs 戸弘 n os y s t e m i ci n f e c t i o n a r . N i c o t i a n atabacumv L o c a ll a t e n ti n f e c t i o n Sa r n s u n N. tabacumv a r . Whit e L c 阻1l a t e n ti n f , 回i o n B u r l e y Pt u n i ah y b r i d a L o c a ll e s i o 凶 i nv a r i o ω f o r r n s 切 鉛m e t i r 悶 l 畑 1 l a t e n ti n f e c t i o n Bn i n c a s acr i f ' 1 ' a 1 ' u l l u sl Iu lga 1 ' i s C i t , , N o n s u 舵 e p t 1 " b l ep l a n ts 戸c i 白 Betav u J g a 1 ' i s,B r a s s i c a1'ata ,Callisteρhuschin'flsis,CatsIcumannuum, C h e f l o p o d i u mama 1 ' a n t i c o l o r ,Gomthrenaglobosa,Lactucasativa, LycoT' 1 ' s i C O f l s c u Z e n t u m ,Nicotiana glutinosa,N. 1'ustica,Phasolus 1 ' i s ,P.vulga1'l s,Rathan ss a t i v u s ,50 ,a ; maz,5 t i f l a c i ao l ' 1 ' aCla, angula T e t 1 ' agoniaeztansa,V i c i afaba,Vi帽 ca1'O S l l a e ,Ziflnia Z l Ig a f l s , “ , , 2 .P h y s i c a lP r o p e r t i e si nv i t r o P h y s i c a l pro 戸r t i e so f CGMMV i nv i t r o were examined ,using cucumber e d l i n g sa sthet 田 tp l a n t . CGMMVwasi n a c t i v a t e da ttemperatur ,白 o f80t o 900C f o r10mim 成 田 ex 伊s u r e . 1noneexperiment ,i twass t i l li n f e c t i v ea t850C. o 1 tw i t h s t o o dd i l u t i o nt o10-8 andagingf o r8monthsa t20C. 田 3 .T r a n s m i s s i o n CGMMVi se a s i l yt r a n s m i t t e dbyp l a n tj u i c e ,intβr-plant∞ntactandmanual h a n d l i n g s . I ti st r a n s m i t t e dthroughi n f e c t e d田 i landprobablythrough記 eds , butnotbyAρ h i sg o s y ρi i( ? )andcucurbitl e a fb e e t l e . R e s u l t so fas e r i e so fe x perimentsont r a n s m i s s i o no fCGMMVared白 c r i b e di nt h enextc h a p t e r . 4 .C r o s sP r o t e c t i o nT e s tw i t hCGMMVandTMV Mottledl e a v e so ff i v eSamsunt o b a c c 田 i n f e c t e dwithTMVwerec h a l l e n g e i n o c u l a t e dwithCGMMV. About20daysa f t e rt h ec h a l l e n g e i n 田 u l a t i o n ,back 10 ∞ulationsweremadetocucumberseedlingsfromchallenge-in∞ulatedleavesof 5 1 C u c u m b e rG r e e nM o t t l eM ω a i cV i r u s whichs u r f a c ew田 s t e r i l i z e dby3% Naa P < λ回 lutionandwashedthoroughlywith d e t e r g e n tandr u n n i n gw a t e r . A l lt h ecucumber配 e d l i n g sbecamei n f e c t e dw i t h ,thoughtheincubation戸 riodwasprolongedcomparedwiththe配吋CGMMV 白 l i n g si n o c u l a t e dw i t hCGMMVa l o n e . I na n o t h e rt e s t ,leav o ff i v eSa msun t o b a 田 op l a n t swereh e a v i l yi n o c u l a t e dwithCGMMVont h r e es u c c e s s i v ed a y s . Tend a y sa f t e rt h ef i n a li n o c u l a t i o n,i n ∞ulatedleav田 werechallenge i n o c u l a t e d 民 op l a n t sbe 回 mei n f e c t e dw i t h w i t hTMV. A l lt h ec h a l l e n g ei n o c u l a t e dt o b a ,althoughtheap戸 aranceofsymptomswasdelayed2-5dayscompared TMV ∞ulatedwithTMValone. witht h et o b a c c op l a n t si n 1 5 .E l e c t r o nM i c r o s c o ρy NumerousTMV-liker o d s hapedp a r t i c l 白 a r eo b s e r v e di nd i p p r e p a r a t i o n so f i n f e c t e dp l a n t s . Modall e n g t ho fCG お仏1Vp a r t i c l 回 i nd i p p r e p a r a t i o nmountedi n 侶 p ho 同n 肝s t a t eap 戸a r e dt obe3 mμinl e n g t hand18mμ ,i nd i a m e t e r . pb ∞ 6 .P u r i f i c a t i o n CGMMVi se a s i l yp u r i f i e dbyt h ef o l l o w i n gm e t h o d . FrozencucumberI 伺 V白 wered i s r u p t e di nab l e n d e ra d d i n gM/10ph 冊 p h a t eb u f f e r and1 : 1m i x t u r e o fc h l o r o f o r mandb u t a n o l . Themixturew錨 s q u e e z e dwithtwol a y e r so fg a u z e and偲 n t r i f u g e da t1 , 000gf o r10m i n u t e s . Aqueousl a y e rwasf u r t h e rc l a r i f i e d , 000g .A f t e rtwoo rt h r e ec y c l e so fh i g h byac e n t r i f u g a t i o nf o r10m i n u t e sa t10 andlow 司 問dc e n t r i f u g a t i o n s( 6 0m i n .a t70 , 000g and10mi n .a t1, 000g), p u r i f i e dv i r u ss u s p e n s i o nwaso b t a i n e d . 7 .S e r o l o g y S国 An t i 田r at oCG l¥仏1vwerep r e p a r e di nr a b b i t sbyi n j e c t i n gp u r i f i e dv i r u s p e n s i o ni n t r a v e n o u s l yandi n t r a m u s c u l a r l yw i t ht h eu田 o fF r e u n d ' sa d j u v a n t . TABLE2 C r o s sr e a c t i o n samongCGMMV ,TMVandORSV i nm i c r o a g g l u t i n a t i o nt e s t A n t i s e r u m A n t i g e n 256 H 一サ一側一 情一一士一冊 州 一一州一 市 H • •• 1 2 8 一 + ORSV 64 川 m 一一冊土州一骨一一州 H ORSV CGMMV TMV ORSV 32 H 側一川柑-一⋮州 州一一州 廿 τMV TMV ORSV CGMMV TMV 16 州一一州掛川m m-+州 川 一 H -⋮附-州川m 州一一州川附川m CGMMV CGMMV 健 r u md i l u t e d(1: A n t i 8 t I t 512 1 0 2 4 + 十 + + + ト + 朴 十 + 初4 8 So u r c eo ft h ea n t i g e n :CGMMV( c u c u m b e r ) ,TMV( t o b a ∞0),ORSV( c a ω e y a ) . H: Ex 位祖C旬 。f h e a l t h ycuωmb 凪 t o b a ∞0,a n dc a 凶. e y a ,f r o mt h et o p . " ' De u b U u lr 回 c t i ∞5 w e r eo b s e r v e d S a l i n e 52 T .I n o u y e ,N .I n o u y e ,M .A銅 tani & K. Mi匂叶l l l t a : I nm i c r o a g g l u t i n a t i o nt , 田t s , t h es p e c i f i ct i t e r so ft h ea n t i 民 r awere1 :1 , 024佃 d1 : 2, 0: 4 8 . Thea n t i s e r ad i dn o tr e a c tw i t ht h ej u i c eo fh e a l t h yc u c u m b e r . Cro 鎚 r 伺 c - ,TMVandORSV t i o n si nm i c r o p r e c i p i t i nt e s twerec o n d u c t e d .u s i n gCGMMV (印o n t o g l 舗 umr i n g s 伊 tv i r u s )andt h er e s 戸: c t i v ea n t 民r a . Table2showst h e 民 団l t s . An tiζGMMV民 間m r e a c t e dwi 出 ORSV ,butnotwithT恥1V. o ntheother hand ,CGMMVreactedwith釦 必TMV田 rum. ThereactionsbetweenCGMMV anda n t i G RSV記 r u na r es t i l li n ∞nclusive. Fur出. e rex 戸r i m e n t sont h e田 r o l o g i c a l r e l a t i o n s h i pamongt h ev i r u 錨 i nt h i sv i r u sgroupa r eb e i n gc o n t i n u e dby出e a u t h o r 百. 1 1 1 . SOMEEXPERIMENTSONVIRUSTRANSMISSION 1 . Ex ρ ' e r i m e n t so nSeedT r a n s m i s s i o nofCGMMV ( 1 )R e c o v e r yofCGMMVfrom‘ F1 Kurume-OchiaiH'S e e d sC o m m e r c i a / / y Producedi n1965 S i n c et h e∞c u r r e n c eo fCGMMVi ncucumberi nv i n y l h o u 錨 i nt h es p r i n go f 1966w田 a l m o s te x c l u s i v e l yl i m i t e dt oonev ,訂i e t y ,F1 Kurume-仇 hiaiH, 出e ,配t e ds e e d si nt h i sv訂 i e t yseemedt obew i t h i nt h eboundo fp c 鴎 i b i l i t y部 v i r u s i n f 出ef i r s t田 町c eo ft h ee p i d e m i c . Thes e e d so ft h ev a r i e t yc o m m e r c i a l l yp r o d u c e d ,therefore,weretestedf o rt h ep o s s i b i l i t yo f舘 edt r a n s m i s s i o no ft h ev i r u s . i n1965 swered i v i d e di n29g r o u p s ,eachcontaining10民 e d s . S田 dc 伺 t sand S田 d embry 団 o fe a c h田 edgroupwereg r o u n di n2and4mlo fw a t e r ,J)田P配 tively, andt h e錨 . pe x t r a c t swereu s e df o re l e c t r o nm i c r ,佃c opyandi n o c u l a t i o non白e ∞tyledonsofcucumberseedlings. In 回 meexperimentstheembryoswere田 wn i ns t e r i l i z e d釦 i lf o rt 田 t i n gt h es 民 dt r a n s m i s s i o no ft h ev i r u s ,resulting,however, i nnoi n f 民 t i o n s . I nt h e回 pe x t r a c to ft h eembryosb o t hv i r u sp a r t i c l e sandi n f 配・ t i v i t yc o u l dn o tbed e m o n s t r a t e dbye l 配 t r o nmic I<濁c opyandb i o a 闘 y . I nt h e回 p e x t r a c tp r e p a r a t i o n sfrom5g r o u 戸 o f記 edc o a t smanyr o d s hapedp e . r t i c l 田 we 問 d e t 配 t e dbye l 配 t r o nm i c r < 濁c opyandr e c o v e r yo fi n f e c t i v i t yw部 mades u ∞essfully. M倒 o ft h er o d s h a I 凶 p a r t i c l 回 w eres h o r tp a r t i c 1 e s , ando n l y8.3p e r白 n t .o ft h e u r e dp a r t i c l 白 h adt h el e n g t ho f300: t25mμ. m回 s 倍 dg r o u p sc o n t a i n i n gt h er o d s h a p e dp a r t i c l e s∞i n c i d e dw i t ht h es e e d Ast h es grou 戸 f romwhichi n f e c t i v i t ywasr e c o v e r dbyi n o c u l a t i o n ,i tc a nbec o n c l u d e d t h a ta tl e a s toneo u to ft e ns e e d si nt h es 伺 dg rouphadbeenc o n t a m i n a t e dw i t h e e dc o a t . Th 田, i tc a nbei n f e r r e d出a ta b o u t1.7 i n f e c t i o u sv i r u sp a r t i c l e si nt h es p e rc e n t .o ft h ecucumbers e e d su s e di nt h et 白 t s伺 nhavec a r r i e dCG~品1V in t h e i r記 e dc o a t s . However ,nos田 dlingsproducedsymptomswhen404蹴 dlings 缶 d回 m p l e swereg r o w n . o u to ft h esameb a t c ho ft h es ( 2 )l n f e c t i o n sCauseddyA r t i f i c i a / / yC o n t a m i n a t e dS e e d s Cucumbers 田 d sc o n t a m i n a t e da r t i f i c i a l l yw i t hCGMMVon出e i rs u r f a ぼ S were田 wnt ot 田 tw hethero rn o tt h es e e d l i n g sgrownfromthemwouldr e v e a l C u c u m h e rG r e e nM o t t 1 eM明 i cV i r u 9 53 anysymptomo fi n f e c t i o n .H e a l t h ys e e d swereimmersedi nt h es a pe x t r a c to ft h e i n f 配 t e dl e a v r i e donap i 配 eo ff i l t e rp a p e r ,andsowni nt h es t e r i l i z e d田 l i l , 回 ,d らー6記 e d sp e r15cmp o t . H a l fo ft h es e e d swerei n j u r e donap a r to ft h es e e d C倒 t b e f o r eimmersingi nt h e回 pe x t r a c t . Att h es t a g ewhent h ec o t y l e d o n sf u l l y 偲 p anded ,halfoftheseedlingsof伺 chplotwereremovedfromthepotsandre p l a n t e di nt h e臼 mep o t s . I n f e c t i o no fcucumberp l a n t swasd e t e r m i n e df i n a l 1 yon t h eb a s i so ft h ee l 配 t r o nm i c r 儲 ∞pyo fd i p . p r e p a r a t i o n sfromt h eyoung 白 tl e a v 白 from30t o40daysa f t e r回 w i n g . 1 TABLE3 Seed l i n gi n f e c t i o nt h r o u g hs e e d so fw h i c hs u r f a c ewasc o n t a m i n a t e d w i t ht h e組 pe x t r a c to fCGMMV ・i n f e c t e dcucumberl e a v e s E x p e r i m e n t I I I ・ 油 I n j u r e de e U瓜n j u r e d飽 眠i s A" ' * 。・ B A B 。 1 / β / 日 田 2 7 8 ν o 0 f / 弱 5 田 I s 7 6 4 2 1 0 / 5 5 5 5 l ν 0 4 / 5 田 3 S 7 0 I V V 舵 d sw e r ei n j u r e db yc u t t i n gas m a l lpo此i o no f概 d∞a t . 紳Seed l i n g swe r en o t回 n s . .s p l a n t e d( A )a n dt r a n s p l a n t e d( B ) . 糾噂 N u m e r a t o ri st h en u m b e ro fc u c u m b e rp ) a n 包 i n f 制吋; d e n o m i n a t o ri st h en u m b e ro fp l a n t sr a i s e df r o mt h et r e a t 国 民e d s .I n f e c t i o n a∞c u 口組 a s ω r l y a sa t也e1 s to r2 n dl e a f5旬 g e . τ h ed a t e so f鈎 叩ng,t r a n s p l a n t i n g ,a n df i n a le x a m i n a t i o n a r 官邸 f o l l o w s:E x p .1 ;1 .J 1 . ,ー ー ー , 6 . Ag . : Exp .1 I;2 5 .J l . ,1 .Ag . ,6 .A g .:Ex p .I I I; 3 .A g . ,9 .Ag . ,3 0 .A g . :E x p .I V ;1 . Sep . ,7 .S e p . ,3 . Oct.: E x p .V ;2 0 . Sep . ,2 9 . 缶p . ,3 0 . Oct. I I I R白 u l t sa r e時 eni nTable3 . Threeo u to f464s e e d l i n g sgrownfromt h eu n i n j u r e ds e e d swerei n f e c t e dwitht h ev i r u sa s回 r l ya st h es e c o n dl e a fs t a g e .S o me ,配t e ds 田 d l i n g sgrownfromt h ei n j u r e ds 田d sseemedt ohaveb 民 nc a u s e d o ft h ei n f byt h ei n f ,配t i o no fembryoduet ot h edeepi n j u r yg e t t i n gt ot h eembryo. A f t e r t h e民 cu ロe n c eo ft h e s es e e d l i n gi n f ,配t i o n ,new∞currenceofinfectionofcucum. o to b s e r v e du n t i lt h e6-7l e a fs 匂g e .l twasi n c o n c l u s i v ewhether b e rp l a n t sw部 n o rn o tt r a n s p l a n t i n ga tt h ec o t y l e d o n a r ys t a g ei n c r e a 記 dt h ef r e q u e n c yo fi n f e c t i o n . S e e d so b t a i n e dfroma r t i f i c i a l l yi n f e c t e dp l a n t swere回 wna f t e rt h es t o r a g eo f a b o u tonemonth. Ea chs e e d l i n gw出 t r a n s p l a n t e da tt h e∞t y l e d o n a r ys t a g ei n 出es t e r i l i z e d回 i li na15cmp o t . O b s e r v a t i o n swere∞n t i n u e du n t i lt h e6-8l e a f eo u to f102p l a n t sw出 i n f e c t e dont h e35thdaya f t e rs o w i n g . s t a g e . On 2 . Eゆ e r i m e n t so nt h eI n 戸c t i o nw i t hCGMMVt h r o u g hSoi / ( 1 )I n l e c t i v i t y0 1CGMMVA s s o c i a t e dw i t hR o o tD e b r i si nS o i l I n f e c t e dcucumber蹴~lings werer a i s e di n15cm戸 t s ,7seedlings戸 rp o t . Af t e rsymp 句 msa p p e a r e dont h r e eo rf o u rupperl e a v 白, a e r i a lp a r t so ft h es e e d l i n g swerec u to f f , andt h e nt h ep o t sc o n t a i n i n gt h eundergroundp a r t swereb u r i e d a tt h ed e p t ho f0cm( g r o u n dl e v e , ) l 25cm,and50cmi ns o y b e a nf i e l d( o nt h e 25tho fJ u l y ) . The回 i li nsomeo t h e rp o t swast u r n e do v e randr e p a c k e di nt h e T .I n o u y , e N .I n o u y e . M. A s a t a n l & K. M i t s 由a t a : 54 samep o t s ,whichwereωburiedthatthe回 i ls u r f a c eo ft h ep o t swasa tt h esame l e v e la st h a to ft h ef i e l d . l nanotherp l o t出. ep o t sweresubmergedi np l 酪 t i c∞ n . t e ra b o u t30 ,60,and70daysthefragmentsofcucumberr ∞t swith t a i n e r s . Af a d h e r i n g田 i lp a r t i c l e swerec o l l e c t e d ,crushedinmortarsinM/100ph伺 phate b u f f e ra tpH7 .O . Thesape x t r a c t swererubbedont h eo o t y l e d o n so fcucumber 配e d l i n g sandt 白 t e da l s oe l 配 t r o nmicro 駒 p i c a l l y . TABLE4 Agingo fCGMMVi ni n f e c t e dcucumberr ∞ ,t 5b u r i e di n5 0 i li n田 ybeanf i e l d * Dur圃也m ofbmying B u r i e da tt h ed e p t ho f 1mon 也 B柿 * ネ A判 * 2m o n t h s A B -JUE 生江一 A B 1佃 1判 + + 0 + 2 -p 5 4 S / / / 5 伊" " 十 0 1 / / / 3 25αn 0 1 / / / 4 3 + 50αn + + + Tumedo v e r ω i l ' + + 5 5 / / 5 3 / / 3 S u b m e r g e d鉛 i l " + 0 + • T h ea e r i a lp むt so ft h ei n f e c t e dc u c u m b e rp l a n t sr a i s e di n1 5cmp o t sw e r ec u to f fa n dt h 鎚e p o t sw e r eb u r i e di n飽 c hp l o to nt h e2 5出 o fJ u l y . 柿Dep t ha t也eωpo ft h ep o tb u r i 叫 meas 町 e df r o mt h e鈎 l is u r f a c eo ft h ef i e l d . 本市喰 E I 伐 釘o nm i c r 凶∞p yo ft h ee x t m c tf r o mt h e r o o tf m g m e n 包 w i t ha d l 町 i n g鉛 i l p a 此i c l 四. 材 料 B i o a 蜘 yonωαlIIl~町民edlings w i 出 t h e e x t r a c tu s e df o re l e c t r o nm i c r 閣∞p y . 紳柿*N u m e r a t o ri st h en u m b e ro fc u c u m b e rp l a n 旬 i n f e c 総 da n dd e n o m i n a t o ri st h en u m b e ro fp l a n t si n o c u l a t e d . 'Thei n f e c t e d鈎 l iw a sl o c 田 n e d o tf i l l e dw i t ht 匝 i n f e c t e ds o i lw a si m r n e r s e di nw a t e ri np l a s t i c∞n t a i n e r . a n dr e f i l l e d . P 11 Asshowni nTable4 ,thedecompositionoftheremainingr ∞ ,tfragmentshad n o ts oe x t r e m e l yp r α: e ededa f t e ronemonthb u r y i n g ,andthevirusparticlesand , i n f e c t i v i t yc o u l dbee a s i l yd e m o n s t r a t e d . Buryingf o rtwomonthso r70days however ,r田 ultedinalmostthoroughdecompositionoftheroots,andi tw錨 haro t op i c kupt h e記 : g m e n t so fr , ∞t sfromt h es o i li nt h ep o t s出a thadbeent u r n e d t h eo t h e rhand ,inthesubmerged o v e ro rb u r i e da tt h es u r f a c eo ft h ef i e l d . m p o s i t i o no ft h er ∞ ,t swaso b s e r v e d ,andalargenumberof p l o tnoa p p a r e n td民 o v i r u sp a r t i d 白 a nds t r o n gi n f 民 t i v i t ywered e m o n s t r a t e d . Althought h edemonli nt h ep o t st h a thadbeentumedo v e r s t r a t i o no fafewv i r u sp a r t i c l e sfromt h e田 i o rb u r i e da tt h es o i ls u r f a c eo ft h ef i e l dwasmades u c c e s s f u l l y ,c o n s i s t e n ti n f e c t i v i t yc o u l dn o talwaysbe r 配o v e r e d , 伊s s i b l yb e c a u s et h enumbero fcucumber s e e d l i n g su s e dwasi n s u f f i c i e n t . o n ( 2 )I n f e c t i o nofCucumberR o o t sw i t hCGMMV I n o c u l a t i o n01CGMMVt oCucumberR o o t s Cucumbers e e d l i n g sr a i s e d ,t h e i rr o o t swerewashedi nt a p p i n gwater ,and i ns t e r i l i z e ds o i lwereremoved 戸 df o roneminutei nt h e回 pe x t r a c to ft h ei n f e c t e dl e a v ' 田・ Then , 回c ho f d i p r a n s p l a n t e di na10cmp o tf i 1 l edwiths t e r i l i z e d鉛,i l . Asseen t h es e e d l i n g sw部 t h esymptomsbegant oapp 伺 ra se a r l ya s6daysa f t e ri n ∞ulationin i nTable5,t 回 mep l a n t s ,whilei tt ∞kaboutonemonthfor田 meotherplantstodevelopthe symptoms. Whent h ee x p e r i m e n t swerec l o 関 d ,30-35perぼ n t .o ft h ep l a n t s C u c u m b e rG r e e nMot t 1 eM制 i cV i r u s 55 TABLE5 I n f e c t i o no fcucumberp l a n t sbya r t i f i c i a li n ∞ulationtor ∞t s witht h e回 pe x t r a c to fCGMMV-infectedcucumberl e a v e s , d G o r f 泊 o s ∞ w 田 t 叫 h d a 1 1 F 凶 M U F B N o.of No.of I n 四 b a t i o n Emrimnt i n : 2 2 1 e d 訪 問 吋 I I l 1 s tl e a f 2 n dl e a f 1 9 4 5 1 4 6-18d . 3 5 1 1-3 0d . ds m y m i α E 司 J∞ 司D 自 V s阻p l d a n b 旬M ・ 副y N o .o f R+・・ R+ R p l a n t s L+ L- Le x a r n i n e d 日 艇 位E 叩 5 1 0 。 。5 。5 5 ホ・事 *s 戸c i m e r 凶 o fe l , ぽt r o nm i c r o s ∞pyw e r ep r e p a r e df r o mt h ey o u n g 田 t l e a v e sb yd i pm e t h 吋 a n d f r o mt h ee x t r a c to fr ∞t s . F i n a l l ye x a m i n e d3 0d a y sa f t e ri n ∞u l a t i o ni nExp .1 ,a n d3 2d a 1 8 i nE x p .I l . * *R+;r ∞ 包a r ej n f , 制ed:R-;r ∞t sa r eu n i n f e c t e d : L+;l e a v e sa r ei n f ぽt 吋: L-;l e a v 飽 a r eu n i n f e c 旬d . ホ ホ 岬 A lt h o u g h3o u to ft h e5p l a n 包 w e r e(R+.Lー}ぉ ar e s u l t o fe l ぽt r o nm i c I'C剛泊'P y ,a l lt h ep l a n 包 w e r es h o w nt ob e(R+.L+)b yi n ∞u l a t i o nt e s t . remainedh e a l t h yi nap 戸a r a n c e .o u to fwhich5 p l a n t si nex 戸r imentI Iwere shownbye l e c t r o nmicr ω ,∞ pyandb i o a s s a yt obev i r u s i n f e c t e dbothi nt h eyoung白 t l e a v e sandr ' ∞t s ,but5plantsinex 戸r iment1釦 d5 p l a n t si nex 戸r imentI I proved旬 bev i r u s f r e e . l n f e c t i o nt h r o u g hCucumberR o o t si nt h eS o i lPermeatedw i t ht h eSゆ E x t r a c to fCGMMV ・ l n f e c t e dCucumberL e a v e s Cucumber田 e d l i n g swere r a i s e di ns t e r i l i z e ds o i li n10cmpo 凪 3揖 e d l i n g s戸 rpot ,andatthr偲 different s 匂g 田 o fgrowth ,10mlof1/10dilutionofi n f 配 t e dplantj u i c ewasd r i p 戸 di n t o 伺,c hp o tbyas y r i n g es oa s旬 a v o i dt h ec o n t a c twitht h ea e r i a lp a r to ft h e配 e d l i n g s . Halfo ft h ep l a n t si neachp l o twerei n j u r e da tt h e i rr ' ∞t sbyp r i c k i n gt h e田 i lwith ap i n c e t t eb e f o r ed r i p p i n gt h ei n f e c t e dj 凶伺. G r o w t hs 包g eo fs e e d l i n g sa t位e a t m e n t * Co t y l e d o n 1 s tl e a f 2 n d1 伺 f R∞t sw e r e I n j u r e d 0 / 3 0 帥 0 / 3 0 ⋮ 一 語 TABLE6 I n f e c t i o no fcucumbers e e d l i n g 百 w ithCGMMVt h r o u g hr ∞t si nt h e田 i l permeatedw i t bt h e鵠 pe x t r a c to fi n f e c t e dcucumber1 伺 v e s 1 / 1 8 " 'T h e鈎 l io f伺 c hp l o tw a s戸 r m e a 句dw i t ht h e1 / 1 0d i l u t i o no ft h ee x t r a c to fi n f e c t e dc u c u m b e rl e a v e so nt h e4 t ho fOc t o b e r ,a n di n j u r i e sw e r eg i v e nt or ∞t sw i t hp i n c e t t e∞ t h ep r e v i o u s d a y .* *N u m e r a t o ri st h en u m b e ro fc u c u m b e rp l 阻旬 i n f e c t e da n dd e n o m i n a t o ri st h en u m b e r o fp l a n 包 t 児 a 飽d . Asshowni nTable6 ,asmallpercen 匂g eo fi n f e c t i o nc o u l dbeo b t a i n e d . AI s o , inanother関 tofexperiment,2outof30plantswerei n f ,民健dwhent h ei n . f 配 t e dj u i c ewasd r i p 戸 di n t ot h es o i lb e f o r et h ep r i c k i n g .1 twasnotc l e a rwhether orn o tt h er a t i oo fi n f e c t i o nc o u l dbei n c r e a s e dwhent h er ∞tswereinjured,becau 田 t h epe 詑 e ntageo fi n f ' 田t i o nwasn o thighenought od i s c 凶 S . 56 T .I n o u y e ,N .I n o u y e ,M . Asa t a n i&K .M i t s u h a t a : I n l e c t i o n0 1Cucumberi nt h eS o i lC o n t a m i n a t e dw i t hCGMMV-Inlected C u c u m b e rR o o tF r a g m e n t s . Thea e r i a lp a r t so ft h ei n f 配 t e dcucumberp l a n t s ,andt h e釦 i lc o n t a i n i n gt h el i v er o o t swasp i l e dona r a i s e di np o t swerec u to f f 油 田to fs l a t eandt u r n e do v e rt h o r o u g h l y,t h e nr e p a c k e di np o t s ,whichwasused a st h e‘ i n f e c t e d田 i l '. I noneexperiment,g e r m i n a t i n gcucumbers e e d sweresownont h ed a ywhen i n f e c t e ds o i l 'wasp r e p 訂 e d . Threeo u to f126田 e d l i n g sgrownwerei n f e c t e d t h e‘ ,butthea d d i t i o n a lnewi n f e c t i o n s a se a r l ya sa tt h e i rp r i m a r yl e a fs t a g eo fgrowth ru n t i lt h e5 t ho r6 t hl e a fs t a g eo fgrowthwhent h ee x p e r i m e n twas d i dn o t0∞u c l o s e d . I na n o t h e re x p e r i m e n t ,t h e‘ i n f e c t e ds o i l 'wasc o v e r e dw i t ha b o u t1cm l a y e ro fs t e r i l i z e d回 i lt oa v o i dt h ed i r e c tc o n t a c tbetween‘ i n f e c t e ds o i l 'ands e e d s 回 w n. J u s ta f t e rt h ep r e p a r a t i o no ft h e s ep o t sc o n t a i n i n gt h e‘ i n f e c t e d田 i l 'c o a t e d l ,228cucumbers e e d sweresownont h el a y e ro fs t e r i l i z e d釦 i l , w i t hs t e r i l i z e d釦 i and1 3 0g e r m i n a t i n gs 田 d sweresownont h edayn e x t . Therewas ,however, noe a r l yi n f e c t i o ni nb o t ho ft h ep l o t s . Al 田, 8 6s e e d l i n g sweret r a n s p l a n t e di n t ot h esamei n f e c t e ds o i l8d a y sa f t e r u tnoi n f e c t i o n swereo b s e r v e di nt h ee a r l ys t a g e so fg r o w t h . t h ep r e p a r a t i o n,b 3 . Ex ρerimentsont h eT r a n s m i s s i o n0 1CGMMVbyP l a n t t o ・ ρl a n tC o n t a c t andManualHandlings ( 1 )I n l e c t i o nC a u s e db yP l a n t t o ・ ρl a n tC o n t a c t F i v ecucumbers e e d l i n g swerer a i s e di na15cmp o t ,4healthys e e d l i n gwere h el a t t e ro fwhichwasi n o c u l a t e dby a tt h er i mando n ed i s e a s e da tt h ec e n t e r,t o挺 ew h e t h e ro rn o t r u b b i n gw i t hani n f e c t e dl e a fa tt h ep r i m a r yl e a fs t a g e,t i n f e c t i o n sc o u l db ec a u s e dbyn a t u r a lc o n t a c te i t h e rbetw 配 nl e a v e so rr o o t s . To ∞t s, i n f e c t e dp l a n twasi s o l a t e dfrom a v o i dc o n t a c te i t h e rbetweena e r i a lp a r t so rr t h eo t h e rh e a l t h yp l a n t s,u s i n gav i n y lt u b e . Ex 戸r i m e n t swerec a r r i e do u tu n d e r t h eg l a s s h o u s ec o n d i t i o n s . 配 nt h ea e r i a lp a r t so f S e v e no u to f20p l a n t swerei n f e c t e dbyc o n t a c tbetw ,whilenoinfectionwasobtainedbycontactbetweenther ∞t s . Ont h e t h ep l a n t s n f e c t i o n st ∞kplacebycontactbetweenr∞t swhencucumberp l a n t s o t h e rhand,i 田 di nh y d r o p o n i 田 i np l a s t i cc o n t a i n e r s . However ,i ti sp佃 s i b l et h a t田 me werer a l 町 i e smayhaveb e e ng i v e nt ot h er ∞ ,t sa tt h er e n e w a l so ft h en u t r i e n t回 l u t i o n , i n j whichmaya f f e c tt h er e s u l t . ( 2 )A r t i f i c i a lI n o c u l a t i o nb yP l a n tC o n t a c tandManualHandlings S o mes o r t so fa r t i f i c i a li n o c u l a t i o nbyc o n t a c tandmanualh a n d l i n g sa sshown i nT a b l e7weremadeoncucumbers e e d l i n g s . Thev i r u swase a s i l yt r a n s m i t t e d u g g e s t i n gh i g hf r e q u e n c yo ft r a n s m i s s i o no ft h ev i r u s bya l lt h emethodsl i s t e d,s bymanualh a n d l i n g sd u r i n gc r o p p i n g . ( 3 )D i s i n l e c t i n gE . 工f e c t s0 1SomeChemicalsagainstCGMMV I n f e c t e dl e a v e swerer u b b e dw i t hf i n g e r s ,andt h ef i n g e r swered i p 戸 df o r5 5 7 C u c u m b e rG r e e nM o t t l eM o s a i c. V i r 色 ‘ TABLE7 T r a n s m i s s i o no fCGMMVbyv a r i o u swayso fc o n t a c tandh a n d l i n g Wayso f∞n t a c ta n dh a n d l i n g I n f e c t i o n 1 . Cotyledo~s o fh e a l t h ys e e d l i n g sw e r er u b b e dw i t ht h ec u te n do fi n f e c t e dl e a f 2 . Co t y l e d o n so fh e a l t h ys 偲 d l i n g sw e r er u b b e dg e n t l yw i t hi n f e c t e dl e a f 3 .I n f e c t e dl e a v e sw e r er u b b c : 姐 w i t hf i n g e r s,a n dt h e nc o t y l e d o n so fh e a l t h ys e e d - l i n g sw e r er u b b e dg e n t l yw i t ht h e m I n f e c t e dl e a v e sw e r ec u tw i t hs c i s s o r s ,a n dt h e n, ∞t y l e d o n so fh e a l t h ys e e d l i n g s w e r ec u tw i t ht h e m 5 .S a pe x t r a c to fi n f e c t e dl e a v e sw a sd r o p p e do nt h es u r f a c e so fc o t y l e d o n so f 吋l i n g s h e a l t h ys e 4 . 1 0 / 1 0事 1 0 / 1 0 1 2 / 1 2 3 / 2 0 1 / 1 2 *N u m e r a t o ri st h en u m b e ro fc u c u m b e rp l a n t si n f e c t e da n dd e n o m i n a t o ri st h en u m b e ro f p l a n t si n o c u l a t e d . s e c o n d si nwater,196LiponF( a na n i o n i cd e t e r g e n t ),1096Teepol,o r396t r i s o o ra moment i nt a p p i n g water ,and then dium o r t h o p h o s h a t e( t s o p ),washed f rubbedt h eprimaryl e a v e so fcucumbers e e d l i n g s . 1 n o c u l a t i o n byrubbing was a l s oa p p l i e dons e e d l i n g sbyt h ef i n g e r st h a twerecontaminatedwitht h ev i r u sb u t a t e r . 1 na n o t h e rexperimenteacho f396t 回 pa nd c l e a n e dby田 apandrunningw 1096Teepols o l u t i o n swasmixedi nv a r i o u sp r o p o r t i o n switht h esape x t r a c to ft h e i n f e c t e dl e a v e s ,andtheresultantsolutionswereusedforthebioassayoncucumb e rs e e d l i n g sande l e c t r o nm i c r o s c o p y . TABLE8 Dis i n f e c t ' o no fCGMMVc o n t a m ' n a t e dhandsand むf e c t i o u scucumbers a pbyc h e m i c a l s 田 E x p e r i m e n tU n t r e a t e d V i r u s < : o n t a m i n a t e dh a n dw a s i m m e r s e di n * w 五er1蒐--w完了秘 1 1 1 6 / 1 6 1 2 / 1 6 1 0 / 1 6 9/10 一 一 一 0 / 1 0 。 咽P 材 S O 1 6 0 / 1 6 / / 1 0 0 / 1 0 0 L i p o nFT e e p o lNS3PO~ 1 6 / 1 6 *紳 1 0 / 1 0 N i n ev o l u m e so fs a pe x t r a c t w e r em i x e dw i t ho n e刊 l u m eo f 10%1・館p o l 3%Na3PO~ J ./ 1 6 ' 0 / 1 0 ' 0 / 1 0 " 8 / 1 0 *F i n g e r st h a tr u b b e di n f e c t e dl e a v e sw e r ed i p p e df o r5 s e c o n d si ne a c ho ft h e 回,l u t i o n s, w a s h e df o ramomenti nt a p p i n gw a t e r ,a n dt h e nt h ef i r s tl e a v e so fh e a l t h yc u c u m b e rs 配 d l i n g s i n g e r sw e r es u f f i c i e n t l yc l e a n e dw i t h回 a pa n d w e r er u b b e db yt h e mo nt h e i rs u r f a c e s . 榊 F 旬p p i n gw a t e r . >1<事事 N u m e r a t o ri st h en u m b e ro fc u c u m b e rp l a n t si n f e c t e da n dt h ed e n o m i ∞u l a t e d. ' T r e a t e df o r5m i n u t e s . " T r e a t e df o r1h o u r . n a t o ri st h en u m b e ro fp l a n t si n . Waterand196L i ponFs o l u t i o nc o u l dn o t R e s u l t sa r ep r e s e n t e di nTable8 e l i m i n a t et h ev i r u sp a r t i c l e se f f e c t i v e l y fromt h ef i n g e r s . Thee f f e c to f1096 Teepol s o l u t i o nwasn o talwaysc o n s i s t e n t ,inonee玄 perimentgivingapositive r e s u l tbuti nanotherl i t t l e,i fany ,effect. Both396 tsopsolutionandwashing withsoapproved t obee f f e c t i v ef o rt h ed i s i n f 配 t i o no ft h ev i r u si nbothex 戸r i 回 p ouredi n t ot h e1/10 mentsr e p e a t e d . When 396 t s o pand 1096 Teepol we d i l u t i o no ft h ee x t r a c to fthei n f e c t e dl e a v 田 a ndr e a c t e df o r5minut 田, t h eformer 社t l ee f f e c t . gaveac o n s i d e r a b l ee f f e c tbutt h el a t t e r1 5 8 T .I n o u y , eN .I n o u y e ,M .Asa 旬 以 &K . Mit s u h a t a : As3 労t 回 p回,l u t i o nwasmixedi nv a r i o u sp r o p o r t i o n si n t ot h e1 / 2 0d i l u t i o n n i n u t e s ,aconsiderableinactivatinge日記t o ft h ei n f e c t e dl e a v e sandr e a c t e df o r5r W酪 o b s e r v e di nt h ep r e p a r a t i o nc o n t a i n i n g20v o l .p e rcen t .o f3%t s o ps o l u t i o n , butnon o t i 白 a b l ee f f e c ti nt h ep r e p a r a t i o n sc o n t a i n i n gl e s sthan20v o l .p e rc e n t . o ft h es o l u t i o n . However ,mixingof10vo. l percen t .o f3% t s o ps o l u t i o nand r e a c t i o nf o r30minutesbroughtaboutac o m p l e t ei n a c t i v a t i o no ft h ev i r u s . F i g .1 .Disi n t e g r a t i o no fCGMMV p a r t i c l e si nt h e回 pe x t r a c to fi n f e c t e dl e a v e sm i x e di n v a r i o u sp r o p o r t i o n sw i t ht r i & 泊i u m0此h o p h 田 p hatι3% s o l u t i o no f回 pw a sm i x e di n t o1 / 2 0 d i l u t i o no ft h es a pe x t r 百c to fi n f e c t e dc u c u m b e rl e a v 田 i nt h ep r o p o 此i o 凶 o f0( A ),2 . 5(8), 5 . 0(C),1 0 . 0(D),a n d2 0% (E). 1 0 %回l u t i o no fT e e p o lw a sm i x e di nt h ep r o p o r t i o no f 2 0 %f o rc o m p a r i ω n( F ) . Figure1showst h ee f f e c to f飴oponCG 恥品1Vp a r t i c l e s . Mosto ft h ev i r u s t a i n i n g5vo l . perc e n t .o f3労 t 回 p回 l u t i o nwere p a r t i c l e si nt h ep r e p a r a t i o n s∞n brokendown ,andonmixingmorethan10v o l .p e rc e n t .o ft h es o l u t i o nnov i r u s p a r t i c l e sc o u l db edemonstratedbyelectronmicr ,偲c o p y . WithN110NaOHi n C u c u m b e rG r e e nM o t t l eM ω a i cV i r u s 59 釦 nw i t ht 5O P,pHso ft h es a pe x t r a c t so ft h ei n f e c t e dl e a v 白 w eread j u s t e d c o m p a r i t o9.0 ,10.0,11 .0 ,and11.5,andv i r u sp a r t i c l 倍 i nt h es a pe x t r a c t sweree x a minede l e c t r o nm i c r o s c o p i c a l 1 y . AtpH11.0and11.5c o n s i d e r a b l eand∞m p l e t e c o l l a I 間 o ft h ev i r u sp a r t i d 田 ∞C町 民d ,r e s p e c t i v e l y . Fromt h er e s u l t s ,t h e r e f o r e , i tmayb ei n f e r r e dt h a tt h ea c t i v ep r i n c i p l eo f匂opa g a i n s tt h ev i r u s∞n s i s t si ni t s 白 o fpH. h i g hv a l u I V . INACTIVATIONOFCGMMVININFECTEDPLANTTISSUES BURIEDINSOILBYMETHYLBROMIDE S i n c e , 路 mentionedi nt h ep r e v i o u sc h a p t e r ,CGMMV戸 ぉi s t sf o ral o n g戸ト r i o do ft i m ei ni n f e c t e dcucumbert i s s u e sandi ti sim p c : 踊i b l e , i np r a c t i c , et or e movet h er e f u s e so fi n f ,民t e dp l a n t se n t i r e l yfromt h e回 i l ,theremaybether i s ko f c a u s i n gt h e配 conde p i d e m i ci nt h ef o l l o w i n g提 出onwhencucumberi sp l a n t e d , sv i n y l h o u 錨, o rt h ee q u i p m e n t sf o rg r a v e lc u l t u r et h a th a v e a g a i ni nt h ef i e l d b記 na t t a c k e dbyt h i sv i r u s . Ef f , 配t i v ec h e m i c a la g e n t sf o rt h ep r a c t i c a lp u r p 佃 e t oe r a d i c a t eCGMMVa r en o tknownh i t h e r t o . However ,afewchemicals,such a sm e t h y lb r o m i d e( 1 5 ,31)andethyleneoxide(25),have50farbeenreportedt o bee f f e c t i v ea g a i n s tTMV. Al 回, S a i t oe ta 1 .( 2 2 )r e p o r t e dt h a tm e t h y lb r o m i d e wasf o u n dt obeane x c e l 1 e n tc h e m i c a la g a i n s t5Oi l b o m ewheatmωaicv i r U 8 . But , ont h eo t h e rhand ,Broadbente ta . 1( 9 )r e p o 抗e dt h a tm e t h y lb r o m i d ewasn o t e f f e c t i v ea g a i n s tTMVi nt o m a t ou n d e rt h eg l a s s h o u s ec o n d i t i o n si nw i n t e r . rn o tm e t h y lb r o Thea u t h o r smadeas e r i e so fe x p e r i m e n t st o民 ewhethero midec o u l di n a c t i v a t eCGMMVi np l a n tt i s s u 回 i n5Oi . 1 1 . M a t e r i a l sandMethods Methylb r o m i d e ,packedi na m p o u l e so rc a n s ,wass u p p l i e dfromKunoshima Chemic a 1l n d u s t r yC o . , L t d . E x p e r i m e n t swerec a r r i e do u td u r i n gfromJ u l yt o 仁)c t o b e ri n1 9 6 6 . Twos e r i e so fe x p e r i m e n 包 w ere made. 1 nt h ef i r s ts e r i 田, 2 01p l a s t i c b u c k e t swereu民 da sc o n t a i n e r sf o rt h ef u m i g a t i o no fm e t h y lb r o m i d e ,and,i nt h e 田 ∞n dsed 白, e x p e r i m e n t swered o n ew i t hs e m i f i e l d記 a l e . ,ora i r d r i e dl e a v e s ,stems,andr ∞t so fCGMMV-inf , 配 缶d Onegramo fg民 en cucumberwaswrap 戸 di nc o t t o ng a u z eandb u r i e di n卸 i li nb o t h配 r i e so fe x p e r i m e n t s . C l ,関e dp l a s t i cc o n t a i n e r swerep l a c e di nt h ea i r ys h a d eo u to fd ∞B M a t e r i a l sw e . 民 b u r i e da tt h ed e p t ho f20cmfromt h e5 0 i ls u r f a c ei nap l a s t i c c o n t a i n e randanampoulew i t hag i v e nd o s a g eo fm e t h y lbromidewasp l a c e don t h es u r f a 目. T hec o n t a i n e rw田 c o v e r e dw i t hav i n y ls h e e tands e a l e dt i g h t l yw i t h h e nb r o k e nw i t hp i n c ぽ s u n d e rt h ev i n y l油田t . F i v e v i n y lt a p e . Ampoulewぉ t f5 0 i lwerea p p l i e da sac o n t r o lc h e m i c a l, h u n d r e dgramso fc h l o r o p i c r i np e rm8o i n j e c t i n gi n t o釦 i lbyas y r i n g e . 1 no n eex 戸r i m e n t ,τ 'MV i n f e c t e dg r e e n,o rd r y t o b a c c ol e a v 白 w erea l s ob u r i e di nt h ep l a s t i cc o n t a i n e rt o g e t h e rw i t hCGMMVi n f 配 t e dcucumbert i s s u 同 a ndt r e a t e dw i t hm e t h y lb r o m i d e . ,v i r u sm a t e r i a l swereb u r i e da tt h r e ed i f f e r e n td e p t h s ,1,20, 1 nf i e l dt e s t s n o u y , e N .I n ∞y e,M. Asat a n i& K. Mit s 吐 1 8 . t a : T .I 60 and30cm ,in伺,chofthepJotstobetreated,andintheuntreatedcontrolplo臨 Ea cho ft h ep l o twas∞v e r e dwitha5ml v i n y l由民t ,andthecenterspaceunder 田 t wass e tbetw 田 n1 5and30cmi nh i g h t . Theedg 回 o ft h es h e e twere t h esh t i g h t l yc o v e r e dwith釦 i l . Ag i v e nd o s a g eo fmethylbromidewasi n t r o d u c e di n t o , ande v a p o r a t e di n ,thespaceunderthe吋nylsheetbyarubbertubeconnected witha回 nfromt h eo u t s i d eo ft h ep l o t . M a t e r i a l sweret r e a t e df o r24o r48hour 百i nt h et 田 t so fc o n t a i n e rs e r i 同 a nd 48hou ぉ i nt h ef i e l ds e r i e s . Thei n f e c t e dp l a n tt i s s u e st r 伺 t e dwithmethylbr o mideweregroundi namortari n10mlofM/100pH7.0p h o s p h a t eb u f f e r ,and 出 es a pe x t r a c t swereu s e df o ri n ∞ulationoncucumber,Daturastramonium, orpe 旬 mas 田d l i n g sf o rCGMMV , 組dN .g / u t i n o s af o rTMV. 2 .E / / e c t0 /Methy/Bromideont h eI n / e c t i v i t y0 /CGMMVi nI n / e c t e d i /i nT i g h t l yS e a / e dP / a s t i cC o n t a i n e r CucumberT i s s u e sB u r i e di nt h eSo 1 nTable9 t h er e s u l t so ft h et r e a t m e n twithmethylbromideo fCGMMVi n f e c t e dcucumbert i s s u 田 a sw e l la sTMV-inf 配 t e dt o b a c c ot i s s u 白 b u r i e di nt h e 回 i Ji nc l o s e dp l a s t i cc o n t a i n e r sa r eshown. TABLE9 l n a c t i v a t i n ge f f 民 to fm e t h y lb r o m i d ea g a i n s tCGMMVandT MV in i n f e c t e dcucumberandt o b a c c ot i s s u e sb u r i e d li ns 伺 l e dp l a s t i cc o n t a i n e r( 1 ) i n回 i 。 。 2 4 4 8 T i meo ft r e a t m e n t( h r . ) 3 1 伺 ) 1 伺 i c r i n Dωage( g / m 3 2 0 6 4 0 3 2 0 6ωCIU p CGMMVi nc u c u m b e r G r e e nl e a v e s 1 1 0 1 0 0 * 1 0 1 0 1 0 D L i r v y el s e も a e v m a , s 1 0 1 1 0 1 0 TMVi nt o b a α m 6 1 1 3 6 1 > G r e 回 l e a v 白〉> S ∞ 帥 947 9 〉 > 7 6 却 ∞ 〉S 8 0 ∞ 0 4 4 2 1 0 1 5 Dryl e a v e s >750 1 . ,N umbero fi n f e c t e dc u c u m b e rp 1 a n 包 o u to f1 0p 1 a n 包 i n o c 由凶.紳 T o 凶 n um l : 同 : ro fα . 1沼l 飴 o f N. g/lIt 印o s a .T e s t e do nt h e2 5 2 6 .J u l y . Maximumt e m p e r a t u 児; l e s i o n so n3l e a v 3 3 .7C: Minimumt e m p e r a t u r e ; 23.7C :A v e r a g et e m 戸r a t u r e s ;2 9 . 1a n d27.9C . 。。 。。 。 。 。。 。 0 0 。。 。 。。 。。。 。 。 。 。 旬 0 配 t i v eont h ei n a c t i v a t i o no fb o t hCGMMV Methylbromidewasfoundt obee f f andT島N,w h i l ec h l o r o p i c r i nc o u l dn o ti n a c t i v a t et h ev i r u 民s . B TMVwasc o m p l e t e l yi n a c t i v a t e dwhent r e a t e da tt h ed制 geo f640g/m ( c a . 8 41b/1 f t) o fmethylbromidef o r48h o u r s . Thisr e s u l ti si n∞n s i s t e n 白 w ith . 1( 1 5 ) . L o c a l l e s i o n sprodu 白 do nt h el e a v 田 o fN .g / u t i n o s a t h a to fJohnsone ta oni n ∞ulationwithTMVtreatedatdifferentd個 略 目 areshowninFig. 2 . CGMMV ,ontheotherhand,was∞mpletelyinactivatedbymethylbromide a tt h ed 明 g 白 o fmoret h a n160g/m'( 阻 . 11b/100げ)f o r24o r48h o u r si na 1 1 t h ep l a n tt i s s u e s ,exceptf o rt h ei n ∞mpJeteef f . 配ti nt h et i s s u e so fl i v es t e m st r 回 t s . edf o r24hour Toknowwhethero rn o tCGMMVc o u l dbei n a c t i v a t e dwiths m a l l e rd団 a g . 白 8 o fn i e t h y lbromidethan160g/m,CGMMV-infectedgr 白 nl e a v 白 a ndr ∞t swere ∞ 6 1 C u c u m b e rG r e e nM . o t t l eM闘 i cV i r u s F i g .2 . I n a c t i v a t i o nb ym e t h y lb r o m i d eo fCGMMV( A )a n dTMV(8) i np l a n tt i s s u e sb u r i e d i ns o i l( c f .T a b l e9 ) . (A) L e f t :D. stramoniuml e a fi n o c n l a t e dw i t hCGMMVf u m i g a t e da tt h ed o s a g eo f55 g / m 3f o r48h 口u r s . R i g h t:D.stramonium1 白 , fi n ∞u l a t e dw i t hCGMMV f u m i g a t e da tt h ed ω a g eo f 3f 480g /m o r48h o u r s . R i g h th a l f l e a v e sw e r ei n o c u l a t e dw i t hu n t r e a t e dc o n t r o l . ( 8 )F romt h el e f tt ot h er i g h t ,N .g l u t i n o s al e a fi n o c u l a t e dw i t h TMV f u m i g a t e da tt h e B, d 関 a g eo f0,160,320,a n d640g/m ' r 白 戸c t i v e l y . t r e a t e da tf o u rd o s a g e s,55,110 ,160,and480g/mB,for24or48hours. Table 10 shows t h er e s u l t so b t a i n e d . A l lt h ed o s a g e swere s u f f i c i e n tt oi n a c t i v a t e CGMMVa f t e r48hoursa sb i o a s s a y e doncucumbers e e d l i n g s . 24hourst r e a t ,however,completeinactivation could notbe obtained at the minimum ment dosageo f55g/m3 . Althought h r e ed o u b t f u ll o c a ll e s i o n s wereproducedon3 h a l f l e a v e so fD aturastramoniumoninoculation with the 阻 p extract of the 3f i n f e c t e dgreenl e a v e st r e a t e da t160g/m o r48hours ,theymighthavel i t t l e ,i f o n T .I n o u y e ,: N .I n o t i y ' e ,' M .A 回 匂n i& K .M i t s u h a t a : 62 TABLE10 I n a c t L v a t i n ge f f e c to fmethylb r o m i d ea g a i n s tCGMMVi ni n f e c t e dcucumber t i s s u e sb u r i e di ns o i li ns e a l e dp l a s t i cc o n t a i n e r( 2 ) T i m eo ft r 伺 t m e n t( h r . ) 3 ) D o s a g e( g / m 2 4 55 1 1 0 1 回 B i 関 s s a y e do n 1 事 r C u c u m b e r G r e e nl e a v e s~1 D a t u r a O L i v eroo包 0 。 。 Ma t e r i a l !~u四mber 1 D a t u r a 0 一一面一 o 1>900糾 3 o ∞o 0 / : > 1 110 ~ 1 回 4 加 0 0 0 0 / : > 3 1 0 0 0 / : > 1 1 5 0 0 0 0 01> 900 01 >1200 01>32佃 0 1>41叩 O 1 0 1 0 *Numbero fi n f e c t e dc u c u m b e rp l a n t so u to f10p l a n 包 i n o c u l a t e d . 柿 N u m e r a t o ri st h esum o ft h en u m b e ro f1 ∞a ll e s i o n so n3 h a l f l e a v e so fD a t u r as t r a m o n i u mi n o c u l a t e dw i t ht h e f u m i g a t e dm a t e r i a l ;d e n o m i n a t o ri st h esumo ft h en u m b e ro f1 ∞a ll e s i o n so n 3h a l f l e a v e s i n ∞u l a t e dw i t hu n t r e a t e dm a t e r i a . 1 T e s t e do nt h e8-9. A u g u s t . Maximumt e m p e r a t u r e ; 34.5C : Minimumt e m p e r a t u r e ; 24.6C :A v e r a g et e m p e r a t u r e s ; 28.7a n d29.4C . 0 0 0 ,e f f e c t ont h er e s u l tb e c a u s eDaturas tramonium a st e s tp l a n t wasshown any t ob emored i v e r s eandloweri ns u s c e p t i b i l i t yt oCG l ¥ 品1Vthancucumber. I tmustben o t e dt h a ta 1 1t h er 白 u l t sd e s c r i b e dabovewereo b t a i n e dunderh i g h t e m p e r a t u r ec o n d i t i o n sduringsummer. Undert h ec ∞lercondjtionsinautumn, however ,theminimumd田 ageofmethylbromideforthecompleteinactivationo f 6 0g/m3 on 48hours treatment,which i sa b o u tt h r e e CGMMVi n c r e a s e dt o1 t i m e sa smuchdosagea so b t a i n e di nsummer( T a b l e1 1 ) . Also,TMVcouldnot 3 40g/m f o r4 8h o u r s . beanyl o n g e ri n a c t i v a t e da t6 TABLE11 I n a c t i v a t i n ge f f e c to fmethylb r o m i d ea g a i n s tCGMMVandTMVi n i n f e c t e dcucumberandt o b a c c ot i s s u e sb u r i e di n s o i li nseal~d p l a s t i cc o n t a i n e r( 3 ) om 川町一 m一 om 必一 m一 1m 。 T i m eo ft r e a t m e n t( h r . ) 3 ) D o s a g e( g / m 制0 0 1 5* CGMMVi nc u c u m b e rl e a v e s e a v e s 1 5 0 1 * * 4 1 TMVi nt o b a α:0 l Numbero fi n f e c t e dc u c u m b e rp l a n t so u to f15p l a n t si n ∞u l a t e d . 料 T o t a ln u m b e ro f1 ∞a l l e s i o n so n4l e a v e so fN.g l u t仰 o s a . T e s t e do nt h e18-19.Oct o b e r . Maximumt e m p e r a t u r e ; 21 .8C : Minimumt e m p e r a t u r e ; 8.7C: A v e r a g et e m p e r a t u r e s ; 14.1a n d16.0oC. 事 0 0 3 .E f f e c tofMethylBromideont h eI n f e c t i v i t y of CGMMVi nI n f e c t c d CucumberT i s s u e sBuriedi nS o i li naF i e l d A s , intheformerexperimentsmadebyusingtheclosedcontainers,methyl bromideprovedt oi n a c t i v a t eCGMMV ,twoex戸 rimentswerecarriedoutunder t h ef i e l dc o n d i t i o n si nsummer. I nt h ef i r s texperimenti n f 配 t e dcucumberl e a v e s wereb u r i e da tt h r e ed e p t h so f1 ,20,and 30cm from the ground level,and t r e a 制 民p a r a t e l ya tt h ed o s a g e so f1 8,3 6,and54g/m ぺfor48hours. CGMMV i na 1 1t h ei n f 配 t e dcucumberl e a v e sb u r i e di ns o i la tanyd e p t hwasc o m p l e t e l yi n a c t i v a t e da tt h eminimumdosageo fmethylbromide,1 8g/m2• 聞 1M o t t l eM ω a i cV i r u s C n c u m b e rG r 63 TABLE1 2 I n a c t i v a t i n ge f f e c to fm e t h y lb r o m i d ea g a i n s tCGMMVi ni n f e c t e d c u c u m b e rt i s s u e sb u r i e di ns o i li ns o y b e a nf i e l di nsummer 2 日 瑚g eg/m t i o no ft h et r e a t i n gm a t e r i a l L∞a 1 8 36 54 凸UAUAUnUAU AVAVAUAUAU 0 . 6m 0.2 m 0.3 m O.Olm 0.2 m 0.3 m 。 V0000 0.3 m B u r i e da tt h ed e p t ho f 一-mum D i s t a n 目 f r o mt h en o z z l e •N u m b e ro fi n f 出国 c u c u m b e rp l a n t so u to f1 0p l a n t si n o c u l a t e d . F u m i g a t e df o r48 h O U I S . .3C :M i n i m u mt e m p e r a t u r e ; T e s t e do nt h e1 8 1 9 .A u g u s t . Maximumt e m p e r a t u r e ; 31 :A v e r a g et e m p e r a t u r 四; 2 8 . 9a n d2 8 . 0oC . 2 4 . 9C 0 0 o n dex 戸r i m e n t ,t o examinet h ed i s t r i b u t i o no ft h ef u m i g a n ti n I nt h e田 c i l, i n f . 配t e dcucumberl e a v e swereb u r i e da tt h ed i s t a n c eo f30and60cmfrom 也en 但 z l ef o rf u m i g a t i o n ,buttheotherconditionssuchastheburyingdepth , s d < 瑚 . g e s ,andtimeo ff u m i g a t i o nweret h e回 mea st h ef o r m e rex 戸r i m e n t . As ,CG 恥品1Vi n回 i lwasi n a c t i v a t e da tt h eminimumd偶 a g eo f showni nT a b l e12 ,r e g a r d l e s so ft h eb u r y i n gs 戸t sandd e p t h s . 18g/m' 回 4 . Mor ρh%gyandA n t i g e n i c i t y01CGMMVP a r t i c /e sT r e a t e dw i t h M e t h y /Bromide Fromt h em a t e r i a l st r e a t e dw i t hm e t h y lb r o m i d ef o r48h o u r si nc l o s e dp l a s t i c ∞ntainers,1/500dilutionsoftheextractswerepreparedandexaminedelectron 配 o p i 回 1 1 y . Ther a t i oo ft h es h o r tp a r t i c l 白 田e medt oi n c r 回開 a f t e rt h e佐 田t . m i c r c h o u g hc o m p l e t ec o 1 1 a戸 eo ft h ev i r u sp a r t i c l e swasn o t0 1 蹴 r v e d . Ther a t i o ment,t o ft h ev i r u s伊 丘i c l e s300: t25mμinl e n g t ht oa 1 1t h ev i r u sp a r t i d 白 m伺 s u r e dwere ,50,and36% i nm a t e r i a l st r e a t e da t55 ,110,160,and480g / r r i l ,r e . 37,57 s p e c t i v e l y ,whereぉ 67% i nt h eu n t r e a t e dm a t e r i a l s .I n a c t i v a t e dv i r u sp a r t i c l e s , a f t e rt h et 回 a t m e n tw i t hm e t h y lb r o m i d e ,didn o t1ωet h e i ra n t i g e n i c i t yandr 回 c . -CG MMV詑 rumi nt h em i c r o a g g l u t i n a t i o nt e s t . t e dw i t ha n t i V . DISCUSSION o nthebasisoftheh trangeofthevirusthati sn e a r l yl i m i t e dt oC u c u r b i . 田 ,t h es t r o n gs t a b l e n 白 s , andt h em o r p h o l o g i c a lr e s e m b l a n c et oTMVw i t h t a c e a e h ev i r u su n d e rd i s c u s s i o ni s m o d a l l e n g t ho f300mμandt h ew i d t ho f18m μ,t c o n c l u d e dt obei d e n t i 伺 1 w i t hcucumbergr 配 nm ot t 1 em o s a i cv i r u s . CGMMVwas 蹴 r i b e dbyA i n s w o r t handt h et y 戸 s t r a i no ft r u sv i r u sw酪 named錨 cucum. f i r s t d ,andthenceforth thev i r u sh a s been r e p o r t e d from many b e rv i r u s3i n1935 ∞untri,白 inEuro戸 (1,3,11,16,18,21,23,24,26,32). Otherstrainsof ,sucha scucumberaucubam o s a i cv i r u s(-cucumber t h i sv i r u sa r eknownh i t h e r t o v i r u s4,C ucumisv i r u s2A)i nEurope( 1,3,1 1,27,2 8 )and C u c u m i sv i r u s2 c andi t sv a r i a n t si nl n d i a( 1 9 ,29,3 0 ) . 6 4 T .I n o u y e .N .I n o u y e . M.A錯 匂n i& K . Mit s u h a 旬: Thev i r u sr e p o r t e di nt h i sa r t i c l e ,however,hass e v e r a lp o i n t st h a ta r ei n ∞n s i s t e n tw i t ht h en a t u r eo ft h ev i r u so r i g i n a l l yd e s c r i b e dbyA i n s w o r t ha sw e l la so f l ¥ 仏 t 1 V児 p o r t e d . T h i sv i r u si si n f e c t i v et oC u c u r b i t aμ ρoand t h es t r a i n so fCG 釦 l a n a c e o u sp l a n t si n c l u d i n gN . tabacum ,whereasAinsworth'sCV3i sn o t . ,a st h esymptomsoncucumberl e a v e si n d u c e dbyt h i sv i r u sa r ed i s F u r t h e r m o r e t i n c tfromt h ea u c u b amo 田i c ,t h i sv i r u sc a n n o tb ei d e n t i f i e da sCV4 . A l t h o u g h 回 c l e a rn e c r o t i c1 民 a ll e s i o n sont h el e a v 田 o fN .t a b a c u mv a r .S a msun CV4p r o d u c . l( 3 ),t h i sv i r u si n f 配 t sl o c a l l yt h e回 meh缶 tw i t h o u tany aαordingt oBrcake ta v i s i b l esymptom. Thev i r u ss 配 m st ob esomewhats i m i l a ri nt h eh慣 tr a n g et o Cucumisvin ぉ2 ci nI n d i a ,buttheformerprodu偲 sapparentsymptomsi nD a t u r astramonium,watermelon,and1 ∞fah,whichareinfectedsymptomlesslyby t h el a t t e rv i r u s . l ¥ 品 1 v Therewaso n c eac o n t r o v e r s yw i t hr e s p e c tt ot h er e l a t i o nb e t w e e nCG andTl ¥ 1 Vi nsome踊 p e c t s . Rochow( 2 0 )∞n c l u d e da sar e s u l to fc r o s sp r o t 配 t i o n t 白 t t h a tCV3waso n eo ft h es 戸c i a ls t r a i n so fTl ¥ 1 V , whereasF u l t o n( 1 0 )a n d Brcake ta l .( 3 )a s s e r t e dt h a tCV3wasav i r u sd i s t i n c tfrom ,andinde戸 ndentof, TMV. Therea r e回 mes i m i l a r i t i 同 h owever ,民t weent h etwov i r u s 回 i nt h e i r m o r p h o l o g yandp h y s i c a lp r o p e r t i e s ,andaw伺 k,b u tp c 洛i t i v e ,s e r o l o g i 回1r 回 ,c t i o n b e t w e e nt h etwov i r u 記 sw aso b s e r v e d( as t r o n gr e a c t i o nwasalωnoticed by K r i s t e n s e n( 1 8 ) ) . Fort h e s er e a s o n s ,someworkersregardedCGMMVasas t r a i n ¥ 1 V .N e v e r t h e l e s s ,Knight( 1 7 )showedt h a tCGMMVwasad i s t i n c tv i r u s o fTl 出c a u 記 CG 乱品1Vh add i f f e r e n tc h e m i c a lc o m p o n e n t sa scomparedw i t ht h 四 eo f Tl ¥ 1 V . Ther e s u l t so ft h ec r o s sp r o t e c t i o nt e s tbetweent h etwov i r u s e si nt h i s e n t i a l l yi na g r e e m e n tw i t ht h o s eo ft h ep r e v i o u sw o r k e r s ,noc l e a r s t u d y a r e白 s n gob 記 r v e d . c r o s sp r o t e c t i o ne f f e c t民 i Ast h er 白 u l t so ft h es e r o l 唱i c a lr e a c t i o n si n c l u d i n gCG l ¥ 仏 1 V , TMV ,and ORSV ,CGMMV,i ns of a ra st h er 白 u l t so fex 戸r i m e n t so b t a i n e dbyt h ea u t h o r s a r ec o n c e r n e d,民emst ob er e l a t e ds e r o l o g i c a l l yd i s 匂n t l yw i t hORSVa n dTl ¥ 1 V . F u r t h e re x p e r i m e n t sont h es e r o l o g i c a lr e l a t i o n s h i pamongCGMMV ,ORSV,and Tl ¥ 1 Va r eu n d e ri n v e s t i g a t i o n,andt h ed e t a i l e dr e s u l t sa b o u tt h ep r o b l e mw i l lb e r e p o r t e di na n o t h e rp a p e r . S i n c et h ef i r s to u t b r e a ko fCGMMVi ncucumberi nJ a p a ni nt h es p r i n go f 曲 te x c l u s i v e l yl i m i t e dt oonev a r i e t y ,F1 K urume-OchiaiH,出e 1 9 6 6wasalm s e e d b o r n eo rc o n t a m i n a t e dv i r u si nt h i sv a r i e t ywass u s 戸c t e dt ob et h ef i r s t 釦 u r c eo ft h ee p i d e m i c . Exceptt h a tt h ea f f i r m a t i o no ft h ei n f e c t i o nt h r o u g hs e e d s ,theIX溺 i b i l i t yo ft h es e e dt r a n s m i s s i o no f p r o d u c e di n1965c o u l dn o tbeo b t a i n e d 田 w ered e c t e c t e dfromt h es e e d t h ev i r u sc a n n o tb ed e n i e db e c a u s ev i r u sp a r t i c l C伺 t s .A l 回 , al i m i t e dnumbero fs e e d l i n g so u to ft h es e e d st h a thadb e e na r t i f i c i a l l y c o n t a m i n a t e dw i t ht h ev i r u sont h e i rs u r f a c e sd e v e l o p e dt h esymptomso fi n f e c t i o n . ta l .( 2 8 ) andY a k o v l e v a( 3 2 ),t h er a t i oo fs 民 d A c c o r d i n gt oVanK∞te 乱仏1Vi sf a i r l yh i g hwhens e e d sa r et e s t e da f t e rt h es h o r tp e r i o d t r a n s m i s s i o no fCG o fs t o r a g e ,e .g .,w i t h i nay e a r . I nt h i ss t u d y ,however ,o n l yo n eo u to f1 0 2 C u c u m b e rG r e e nM o t t l eM ω a i cV i r 国 65 e d l i n g sgrownfromt h es e e d so b t a i n dfromt h ea r t i f i c i a l l yi n f ,民t e dcucumbers foundt obei n f 配 t e d . Thisr 田 ,u l ti sc o n t r a d i c t o r yt ot h o s eo fVanK, ∞te ta . l ando fYakovleva ,andi ti ss t i l li n c o n c l u s i v e whetherorn o tt h ed i f f e r e n c ewas 伺 u s e dbyt h ed i f f e r e n c eo ft h ev i r u ss t r a i n so ro ft h ecucumberv a r i e t i 田 u s e d . Br 伺 d b e n t ,i nh i ss e r i 白 o fr e p o r t sont h ee p i d e m i o l o g yo ftomatomo 回 i c( 5 , 6,8 ) ,showedthat TMVinfections∞curredbytheseed-transmittedvirusonly whenぬmato田 e d l i n g swe 回 t r a n s p l a n t e d,whereasnoc l e a rc o r r e l a t i o nwase s t a b l i s h e di nt h i ss t u d ybetweent h er a t i oo fs e e dt r a n s m i s s i o nandt r a n s p l a n t i n gi n CG 恥仏W. Asi nt h ec 出 eo f TMVi ntomato ,s o i lt r a n s m i s s i o ni nCGMMVi ss u s 戸c t e d t ob ea n o t h e ri m p o r t a n t回 u r c eo ft h eprimaryi n f e c t i o n ,thoughnode包 i l e dworks a s句 t h i sp o i n thaveap 戸a r e d . Ryden( 2 1 )r e 戸r t e dt h a ti r r i g a t e dwaterc o n t a m i 民 eo ft h e∞c u r r e n c e,andt h a tt h ev i r u sp a r n a t e dw i t hCGMMVc o u l dbeas o u t i c l e sc o u l dp出 st h r o u g ht h el a y e ro f回 i la tl e a s t80cm. Althoughex 戸r i m e n t s ont h e回 i lt r a n s m i s s i o nwerei n s u f f i c i e n ti nt h i ss t u d yandmanymustbedonei n t h ef u t u r e ,thefollowingfactswerenoticed. Theremainingr∞t sd e c a y e dmuch morer a p i d l yi nt h ef i e l dc o n d i t i o n si nsummert h a ni nt h esubmergedc o n d i t i o n s , andt h en回 r e rt h es u r f a c et h er ∞t swe 詑 b u r i e di n田 i l ,themorerapidlypr o c e e d e dt h ed e c a y i n g . I nw e l lt u r n e do v e r回 i ld田 om 戸s i t i o nwasf a i r l yr a p i d . N e v e r t h e l 白鳥 i twouldt a k eamuchl o n g e r戸 r i o do ft i m ef o rt h ev i r u st od i s a p 戸a r c o m p l e t e l yfromt h es o i l . Inp r a c t i c e ,ther∞treminantsburiedind関 戸r回 i l t h a nt e s t e di nt h i ss t u d ymaybecomeani m p o t a n tproblemi nt h ef u t u r e . Cucumberr ∞t s記 emt obeal i t t l eloweri ns u s c e p t i b i l i t yt oCGMMVt h a n t h ea e r i a lp a r t so ft h ep l a n t s ,becausea l lt h ep l a n t si n o c u l a t e da tr ∞t sweren o ta l waysi n f e c t e di ns p i t eo f100p e rc e n t .i n f e c t i o na si n o c u l a t e donl e a v e s ,andi t t a k 田 , moredaysf o r釦 mep l a n t st op r o d u c et h es 戸n ptomsa si n ∞u l a t e da tr ∞ ,t s 釦 t h ep l a n t si n o c u l a t e dont h el e a v 白 ( t h ed e l a yi nsymptomap 戸 町 釦. c ei sn o t t h s ol o n ga sTMVi nt o m a t o ) . Spreado ft h ed i s e a s ewasi m p l i e dt obec a u 総 db yn a t u r a lc o n t a c to fcu 叩 m b e rl e a v e sandmanualh a n d l i n g sd u r i n gcucumbergrowinga si nt h e但 s eo ftomato m o s a i c .( 4 ,7) Toreducethechancesoftouchingtheplan包 withcontaminated handso ri m p l e m e n t s ,trisodiumorthophosphateprovedt obea seH 配 t i v ea g a i n s t CGMMV田 a g a i n s tTMV( 4 ),w h i l eTeepold i dn o tg i v ec o n s i s t e n tr e s u l t s . The h i g hv a l u 田 o fpHo f飴ops o l u t i o ns 田 mt obet h eimmediate伺 U関 o ft h ei n a c t i v a t i n ge f f e c t ,andt h i sc h e m i c a lmayber ' 配 o mmendeda sad i s i n f e c t a n ta g a i n s tCG ・ MMV. A c c o r d i n gt oBroadbente ta . l( 9) ,TMVi ntomatob u r i e dinωilwasn o t t( i n a c t i v a t e dbymethylbromidea t2l b / 1 0 0f t 但 ・ 97g/m')f o r120h o u r sunder g l a s s h o u s ec o n d i t i o n si nw i n t e r . However ,Wiggse ta l . (31)g e t戸別t i v er 回 u l t s B andshowedt h a tf u m i g a t i o na t3l b / 1 o of to f田 i lf o r48h o u r swass u f f i c i e n tt o i n a c t i v a t eTMVb u r i e di ns o i li nc l 倒 dc o n t a i n e r s . Also ,Johnson. e ta l .( 1 5 ) r e p o 巾 dt h a tfrom3t o4l bo fmethylbromide戸 r100s q .f twerer e q u i r e df o r t h ec o m p l e t ei n a c t i v a t i o no fTMVon48h o u r st r e a t m e n t . 記 W箇 6 6 ,N .I n o u y e ,M .A s a t a n i& K .M i 飽曲a t a: T .I n o u y e 笛 u l t so b t a i n e dbyt h ea u t h o r swere也a tf u m i g a t i o na t640g /mS ( c a .4 Ther a u r i e di n駒 i li nc l o 総d l b / 1 0 0f t)f o r48h o u r swasn関 dedωinactivateTMVb t a i n e r s . T h i si si na g r 田 m entwi 出t h o 田 o fWiggse ta . l andJ o h n s o n p l a s t i c∞n l . Atc ∞l e rtem 戸r a t u r 白,舗 i nautumn ,h owever , l a r g e rd o s a g 田 o ft h e e ta c h e m i c a lwerer e q u i r e df o rt h ei n a c t i v a t i o no fTMV ,whicha l s oc o i n c i d e sw i t h , 田u l to fWiggse ta l . t h er F u r t h e r m o r e ,i tbecamee v i d e n tfromt h e' r 白u l t so b t a i n e di nt h i ss e r i e so f t e s t st h a tCGMMVc o u l dbemore回.si l yi n a c t i v a t e dw i t hm e t h y lb r o m i d et h a n TMV. Fori n s t a n c e ,methylbromidei s国 edi nJ a 戸 na t回 . 33gI ml a g a i n s t l ・ hou 民, a nda tc a .18g/m a g a i n s tt h esame P h y t o p h t h o r ar o to fcucumberi nv i n y l d i 記a 舘 i nt h ef i e l d . I fu艶 da th i g ht e m p e r a t u r 田 部 i nsummer ,t h e r emayb e 釦 mep r o s p e c 匂 o ft h ep r a c t i c a la p p l i c a t i o no fm e t h y lb r o m i d ea g a i n s tCGMMV , f o r ,accordingt ot h ea u t h o r ぜr e s u l t s ,18g/mlf o r48h o u r si n a c t i v a t e dt h ev i r u s i nt h ef i e l dt 田t si nsummer. However ,a sf u m i g a t i o n sweremadeo n l yf o rt h ei n f 配 t e dc u c u m b e rt i s s u e s landi ti sn o tc l e a rw h e t h e ro rn o tm e t h y l b u r i e da tt h emaximum30cmi n回 i b r o m i d e伺 nbea l 回 e f f e c t i v et oi n a c t i v a t eCGMMVi ni n f e c t e dr o o t sr e m a i n e d d白 戸ri n田 i l ,andasthem配 hanismsofthevirustransmissioni n回 i landt h e 戸r m e a b i l i t yo fm e t h y lb r o m i d et h r o u g h回 i la r es t i l l四 回l v e d ,i tw i l lb et ∞ e a r l yt o阻 yt h a tm e t h y lb r o m i d e回 nbeu民 di np r a c t i c e . Atl e a s ti m p l e m e n t si n v i n y l h o u 艶 a nde q u i p m e n tf o rg r a v e lc u l t u r emaybef u m i g a t e deH 配 t i v e l yw i t h t h i sc h e m i c a lu n d e rh i g htem 戸r a t u r e∞n d i t i o n s . a i t oe ta l .( 2 2 ),p u r i f i e ds o ・ 1 ibomewheatm儲 a i c吋r u s戸 r t i c l e s Aαordingt oS c o l l a 戸e dwhent r e a t e dw i t hm e t h y lb r o m i d e . N e v e r t h e l e s s ,CGlV仏N p a r t i c l 白 r e t a i n e dl a r g e rp o r t i o n so ft h e i rm o r p h o l o g ya f t e rt 民 a t m e n t ,ex 伺 p t出a tc u t t i n g o ft h ev i r u sp a r t i c l 田 s eemedt o∞c u randt h enumbero f田 g m e n t so ft h ep a r t i c l 白 wereo b s e r v e dt oi n c 配 a s es o m e w h a t . An t i g e n i c i t yo ft h ei n a c t i v a t e dCG l ¥ 品N r e m a i n e du n a f f e c t e da f t e rt r e a t m e n t . Tog i v ea田 l u t i o nt ot h em e c h a n i s m so fi n a c t i v a t i o nbym e t h y lb r o m i d eo ft h e v i r u s ,andt of i n dar 白 s o nwhyTl¥Ni ss t a b l e ra g a i n s tm e t h y lb r o m i d e出anCG,arethefutureproblemst ob e田 I v e d . MMV SUMMARY' I nt h es p r i n go f1 9 6 6 , av i r u sd i 田: a s eo fcucumber∞cuηedw i d e l yandc a u s e d as e r i o u sdamagei nv i n y l h o u s 白 a tmanyl o c a l i t i e si nt h ew e s tp a r to fJ a p a n . i cv i r u s(CGMThec a u s a lv i r u swasi d e n t i f i e dw i t hc u c u m b e rg r e e nm o t t l em曲 a l ¥ N )newt oJ a p a ni nt h i sp a p e r . V e i n b a n d i n gandd i s t i n c tg r 田 nm o t t l i n go f l e a v ,民 r e v e r s i b l ew i l t i n go fi n f e c t e dp l a n t s , a sw e l la s民 v e r em o t t l i n go fd e f o r m e d f r u i t sa r et h ec h a r a c t e r i s t i c symptomsi nc u c u m b e r . Thec a u 回 1 v i r u si n f ,配t s v a r i o u ss 戸c i e so fc u c u r b i tp l a n t ss y s t e m i c a l l ya n dp r o d u c e dd i s t i n c tsymptoms i n many o ft h es u s c e p t i b l ep l a n t s . I tf o r m sl o c a ll e s i o n si np e t u n i a and Daturas t r a m o n i u m . I nSamsunandWhi t eB u r l e yt o b a ∞0,inf,配tioni s1 民a l w i t h o u tv i s i b l es y m p t o m s . Thev i r u si ni n f 配 t e dp l a n tj u i 伺 i si n a c t i v a t e da t80 ー C u c u m b e rG r 開 1M o t t l eM ω a i cV i r u s 67 8 900Cf o rt e nminut 白 e x p o s u r e . I tw i t h s t a n d sd i l u t i o nt o1 0 anda g i n gf o r8 o 四 o ft h ev i r u sap 戸訂部 T MV-1ikes t r a i g h tr α l s300 monthsa t20C . P a r t i c l 8mμind i a m e t e r . Noa p p a r e n tc r o s sp r o t e c t i o nwaso b s e r v e d mμin1 e n g t hand1 e s t e di nS a msunt o b a c ∞ . Purifiedviruswas b e t w e e nt h ev i r u sandT島W whent 配 t e dcucumber1 e a v 田, c l a r i f i e dbyc h 1 o r o f o r m b u t a n o 1m i x o b t a i n e dfromt h ei n f 回 a ndf o l l o w i n gd i f f e r e n t i a 1c e n t r i f u g a t i o n s . Antiserumw i t ht i t e ro f1:2, 048 t u W舗 p r e p a r e dfromr a b b i t s ,i n 炉t i n g出epur 証i e dv i r u si n t r a v e n o u s 1 yandi n t r a m u s ,白t ,CGMMVr e a c t e dw i t h anti-TMV田 r u m . c u l a r 1 y . I nt h em i c r o p r e c i p i t i nt o ntheotherhand,anti.CGお仏N 時 四m w錨 observedtoreactwithORSV,but n o tw i t hTMV. 田 d e t e c t e di nt h ee x t r a c to fs 田 dc o a t so fF1 Kurume-Oc h i a iH CGMMVw 町 田e d sp r o d u c e dc o m m e r c i a l l yi n1965 ,whichmightsugg白 tthepossibi・ cucuml 1 i t i < 田o f配 edt r a n s m i s s i o no ft h ev i r u s . However ,nos e e d 1 i n gi n f e c t i o nw踊 o b t a i n e d 田 d l i n g sr a i s e dfromt h e組 meb a t c ho ft h es 民 d so ft h ev a r i e t y . o u to fa b o u t400s S e e d 1 i n gi n f , 配t i o nwasc o n f i r m e dt h r o u g hs e e d swhichhadb e e na r t i f i c i a l l yc o n t a m 出 t h evi r u s , immersingcucumbers e 吋 si n t ot h ei n f 配 t e dl e a fj u i c ej u s t i n a t e dwi i n g . On 1 yonei n s t a n c eo fs e e dt r a n s m i s s i o nwaso b s e r v e do u to f b e f o r et h e回 w 1 0 2鋭 剖l i n g sgrownfromt h es e e d so b t a i n e dfromt h ei n f e c t e dcucumberp 1 a n t s . CGMMV a 鉛 即i a t e dw i t hr ,∞td e b r i so fi n f ,配t e dcucumber wasr ,配o v e 民 d 伺 s t4m o n t h s 'b u r y i n gi n釦 i 1u n d e rt h ef i e 1 d∞n d i t i o n sd u r i n gt h e戸 r i α ,l a f t e ra t1 1 t h o u g ha∞n s i d e r a b 1 edeαeaseo fv i r u sp a r t i c l e sand o fsummerandautumn,a 白 w eren o t i c e di ni n f e c t e ds o i l田 mpl 田 b u r i e dn 回 rt h es u r f a c eo ft h e i n f e c t i v i t i 田 eo f1 ∞ 配n eds o i . l Undert h esubmergedc o n d i t i o n s ,onthe g r o u n dandi nt h ag r e a td e a lo fv i r u sw回 r 配 o v e r e dw i t h o u ta p p r e c i a b 1 ed e c o m p o s i t i o n . o t h e rhand, 田 d l i n gi n f e c t i o nt h r o u g hs o i l∞ c u r r e dwhend i 田a 田 dp l a n tj u i c e As m a l lr a t i oo fs 戸 di n t ot h e田 i li nwhichcucumbers e e d 1 i n g sweregrown, anda l s owhen wasd r i p t a i n i n gr ∞t so fi n f , 配t e d c u c u r r i b e rs e e d l i n g sweret r a n s p 1 a n t e di n t ot h es o i l∞n 也 S of 紅 白 t 白 t e d ,however,noseedling i n f 配 t i o nw酪 o b s e r v e dwhen p l a n cucumber田 e d swere田 wni nt h es o i lc o n t a i n i n gt h er ,∞t so fi n f e c t e dp l a n 包 CGMMVw白 e a s i l yt r a n s m i t t e dbyi n t e r p 1 a n t∞n t a c ta tt h e i ra e r i a l戸 市 andv i r u sc o n t a m i n a t 吋 t ∞l sandhan d s . Fort h ep u r p c ぉeo fd i s i n f e c t i o no fv i r u s ,cleanw田 hingwith田 ap,dippinginT白 po1orNaaPO, s o l u c o n t a m i n a t e dhands 配 t i v e . Abovea l l , Naa P < 入 wasd e m o n s t r a t e dt obev e . 可 t i o nwerep r o v e dt obeeH e f f e c t i v ei nb r e a k i n gdownt h ev i r u sp a r t i c l e s , whichwasp r o b a b l yduet ot h eh i g h 田 o fpHo ft h e田 l u t i o n .I nt h et , 田t so fi n 悶 t t r a n s m i s s i o n ,bothA . ρh i sg o s y v a 1 u p i i(?)andc u c u r b i tl e a fb e e t l ed i dn o tt r a n s m i tt h ev i r u s . 恥1MV i n M e t h y lb r o m i d e wasc o n f i r m e dt ob ee f f e c t i v et oi n a c t i v a t eCG 田 na ndd r i e dp l a n tt i s s u 田 o fi n f 民t e dc u c u m b e r . Theminimumd 伺 a g ' 白 o ft h e gr p 1 e t ei n a c t i v a t i o no fCG 島 問V i np l a n tt i s s u 白 b u r i 吋1D釦 c h e m i c a lf o rt h e∞m l i nt i g h t l ys e a l e d20/c o n t a i n e r swe 問 f o u n dt obe55g/m f o r48h r .and1 1 0g / B 8 o r24h r .o ff u m i g a t i o ni nsummer ,althoughi twas320g/m f o r48h r .i nc ∞l m f 記 a 田n . t h eo t h e rhand ,TMVwashard1yi n a c t i v a t e dbym e t h y lb r o m i d ea t 8 田 geo f640g/mf o r48h r .o ff u m i g a t i o ni nsummer. C h 1 o r o p i c r i nshowed t h edo n o n 6 8 T .I n o u y e,N. I n o u y e ,M. A s a t a n i &K . Mi t s u h a t a : 田. I nthef i e l dt 田t sconductedinmid. noe f f e c tonthei n a c t i v a t i o nofthev i r u s 恥仏1v b uried i n回 i la tthedepthsof 1 ー 3 0cmfromthe ground summer,CG 8gfm'for l e v e lwasi n a c t i v a t e dcompletelybymethylbromidea tthedosageof 1 48h r .offumigation. Accompanyingal i t t 1 ei n c r e a 民 i nther a t i oofs h o r t戸 r t i c l 民 modallengthofthep a r t i c l e so fCG 乱仏1v i n a c t i v a t e dbymethylbromidefumiga. 戸r edwitht h euntreatedv i r u s ,andthestrongantigenicity t i o nd i dnota l t e r e dcom oft h ei n a c t i v a t e dv i r u swasa l s odemonstrated. Acknowledgement τbe a u t h o r se x p r ,臨 t h e i rs i n c e 隠 伽n k st oM r .N . 匂s a k i,M r . n dD r .H .I s h i io fT o k u s h i m a T.YamamoωandM r .Y .K a s h i w a g io fT o k u s h i m aA g r .Ex p .S t a .,a A g r . High剖 1 ∞1f o rt h e∞H回 i o no fp l a n ts a m p l 四 a n dt h es u r v e yo ft h ed i 記 a s e α: c u π e n c e, a n da l 鉛 t oD r .Y .Komuroo fI n s t i t u t ef o rP l a n tV i r u sR白 e a r c h,a n dM r .K .T o r n a r uo fHat a n o Toba ∞。Exp .S t a .f o rt h ev a r u a b l ea d v i c e so nv i r u si d e n t i f i α t i o na n dv i r u st r a n s m i 鑓i o n . L lTERA TURECITED 1 .A i n s w o 目h ,G. C . 1 9 3 5 . Mc 回a i cd i 民 a s e so ft h ec u c u m b e r . A n n .a p p . lB i o l .2 2 : 55-67. 2 .A n n u a lM e e t i n go fP l a n tQ U a r a n t i n ei nC h u g o k u S h i k o k uR e g i o ni n1 9 6 6 . 3 .B r c a k, J .,U l r y c h o v a,M.a n dゐ h, M. 1 9 6 2 . I n f , 回i o no ft o b a c r oa n dsome Che ∞ 阿ium S戸 c i e sbyt h eCucumberV i r u s4( 3 )a n dbyi 包 n u c l e i ca c i d . V i r o l o g y16: 105-114. 4 .B r o a d b e n t, L 1 9 6 3 . Thee p i d e r n i o l o g yo ft o m a t omωai c .I I I .C I 鎚 n i n gv i r u sf r o mh a n d sa n d , ∞I s . Ann.a p p l .B i o l . 52: 225-232. t r o a d b e n t, L 1 9 65 a . Thee p i d e m i o l o g yo ft o m a t om ω a i c .V I I I .V i r u si n f e c t i o nt h r o u g h回 n a t o 5 .B r ∞t s . A n n .a p p l .B i o . l 55: 57-66. 6 .B r o a d b e n t ,L 1965b . Theepidemiol,句~ o f tornaωm 咽 i ιXI. S副 t r a n s m i s s i o no fτ'MV. A n n .a p p l .B i o . l 56: 177-205. 7 .B r o a d b e n t, L .a n dF l e t c h e r ,J .T .1 9 6 3 . Thee p i d e m i o l o g yo ft o m a t om幽 i c .I V .P e r s i s t e n c e 国 s t r u c t u r e . Ann .a p p l .B i o l5 2 : 233-241 . o fv i r u si nc l o t h i n ga n dg l a S S h O I 8 . B氏 岨d b e n t,L. a n dF l e t c h e r, J .T .1 9 6 6 . τbee p i d e m i o l 句 yo ft o r n a ぬ m ωai c .X I I . So u r c 溺 o f m e r c i a lt o m a t oc r o 戸 u n d e rg l a 鎚. A n n .a p p l .B i o l . 57: 113-120. TMVin∞m 9 .B r o a d 凶1 t,L . , R回 ,d,W.H .a n dLa s , tF .T . 1 9 6 5 . Th ee p i d e m i o l o g yo ft o m a t omωaiιX. ・ i n f e c t e dd e b r i sinωil, a n dt b ee f f ,舵包 o fs o i lp a 民泊1s t e r i l i z a t i o n . An n . P e r s i s t e n c eo fTMV a 即 1 .B i o l . 55: 471-483. 1 0 .F u l t o n, R .W. 1 9 5 0 . Cr 閣 p r o 肱 t i o nt 鎚包 w i t hc u c u m b e rv i r u 措 3a n d4a n dt o b a ∞o m欄 i c 討r u s . P h y t o p a t h . 40: 219-220. 1 1 .G e s ', D .K .1 9 6 5 . M o z a i c h n y eb o l e z n io g u r 回 vz a k r y t o g o g r u n t a .D o k l .A k a d .Naukb e l o n 邸 k . SSR 9: 555-557. ( R .A.M. 4 5 :5 6,1 9 6 6 ) 1 2 .I n o u y e , T .,I n o u y e, N .,A鈍 t a n i, M. a n dM i t s u h a t a, K . 1 9 6 7 . S t u d i ぉ onωα. u n b e rg r 匂 n 抗I emωaicv i r u si nJ a 開n . 1 . I d e n t i f i α t i o no ft h ev i r u s . N句 法uK enkyu51: 175mo i nJ a p a n 白 e ) 1 8 6 .( 1 3 .I n o u y e , T .,I n o u y e , N .,A羽 t a n i, M. a n dM i t s u h a t a, K . 1 9 6 7 . S t u d i 包 o n回 目.un bergz 唖e n m o t t l em田 a i cv i r u si nJ a p a n .1 1.So me白 戸r i m e n t so nv i r u st r a n s m i 畠 i o n .NogakuKenkyu 51: 187-197. ( i nJ a p a n 醍} 1 4 .I n o u y , eT .,I n o u y , eN .,A阻 t a n i,M. 阻 dM i t s u h a t a,K .1 9 6 7 . S t u d i 包 o nc ぽ u mberg r e e n m o t t l e m鑓 a i cv i r u si nJ a p a n . I I I . I na c t i v a t i o no fCGMMV i ni n f e c t e dp l a n tt i s s u e i nJ a p a n ' 闇} b u r i e di ns o i lb ym e t h y lb r o m i d e . NogakuKenkyu 51: 199-207. ( 1 5 .J o h n 鉛 n , E .M. a n dCh 且p man , R .A. 1 6 6 3 . C h e m i c a li n a c t i v a t i o nofω凶 c ∞ m制 i cv i r u si n 国光s . P I .Dis .R e p t r . 47: 389-391 . ωmator 1 6 .K l i n k o w s k i, M. a n dU s c h d r a w e i t, H .A. 1 9 6 0 . D i eV i 民 間nd e rG e m U s e p f l a n z e n . (日l a n z l i c h e V i r o l o g i eB d .1 1 .K l i n k o w s k i,M.)A k a d e m i e V e r l a g,Ber l i n . 1 7 .K n i g h t, C .A. 1 9 5 5 .A recucum 民 rv i r u 描 3a n d4s t r a i n so ft o b a α沿 m岨 i cv i r u s ? Ar e v i e w CucumberG r e e uM o t t l eNωaicVi r u s 6 9 o ft h ep r o b l e r n . V i r o l 句y 1 :2 6 1 2 6 7 . 1 8 . Kr i s t e n 掴n ,H.R、 1 9 5 6 . V i r u s s y g d o r n r n eh c 園 A g u r k e riDanmark. H o r t i c u l t u r a1 0 :1 6 1 ー 1 7 2 .R .A.M. 3 6: 1 6 5 ,1 9 5 7 . 1 9 .R e p o r to ft h eD i v i s i o no fMy ∞logyandPlantPathology. 1954. S c i .R e p .A g r .R飽 .I n s t .N. De l h i1 9 5 1 ー1 9 5 2 : 75-87. ( R .A.肌 34: 348-350 ,1 9 5 5 ) 吋10W , W.F .1 9 5 6 . I n t e r f e r e n c ew i t ht o b a c ∞ m凶 a i cv i r u si n f e c t i o nby∞ 四rnbervirus偲 3 2 0 . R, and4 . Ph 戸 叩a 出. 4 6: 133-137. 2 1 . Ryden , K. 1 9 6 6 .I n v e s t i g a t i o n sont h es p r e a do fc u c u r n l 町 g r e e nm o t t l emωaicv i r u s( C u c u m i s v i r u s2S m i t h )byw a t e r i n g . V i r u 臨 o fP l a n 包 包d i t e dbyBee r n s t e r ,A.B .R .andDi j k s t r a ,J . 7p p .3 1 7 3 1 9 .N o r t h H o l l a n dB u b .C o . , A m s t e r d a r n . 2 2 . Sai t o , Y .,百kan ぉh i , K.,I w a t a , Y . 印 dOkamoto , H. 1 9 6 4 . S t u d i e s on t h es o i l b o r n ev i r u s d i s e a 錨 o fwh 飽 t andb a r l e y .I I I . I n f l u e n c eo fc h e r n i c a l s on t h ei n f . 回 吋 鈎i l sa n dt h e v i r u s e s . B u l l .N a t .I n s t. A g r i c . S i , . c Ser i e sC, 1 7 : 41-59. 2 3 . Smimova , MrneV .A. andS h t e l n ・ M a r g o l i n a , MmeV .A. 1 9 6 2 . 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