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1 ISSN 3?8-824 THE JOURNAT OT TURI(I$II PHYTOPATIIOLOOY PUBLISHED BY THE TURKISH PHYIOPATHOLOGICAL SOCIEry Vlume : 22 Number : 1 Jan. 1g93

2 TURKISH PHYTOPATHOLOGICAL SOCIETY { President Vice- President General Secrctary Treasurer Chief f Eclitrial Bard Dg. Dr. Hikmet SAYGILI Dr. Ayge UCKAII Dr. Emin ONAN r. Umu rlen Dr. K.Necder ONGEN The furnal f Turkish Phytpathlgy is published by Turkish Phytpathlgical Sciety and issued twice r three times a year t frm vlume. The subscriptn rate per vlume is $ 21. EGE UMVERSITESI BASIMEVI BRNvA- lzmlr - l99s

3 THE J()URNAL ()F TURKISH PHYTOPATHOLOGY TURKISH PHYTOPATHOLOGICAL SOCIETY vl.22 JAN NO. 1 CONTENTS Page Studies n Citrus Stubbrn Disease and Sesame Phylldy in Sesame and Their Related L.eafhpper Vectrs. H. BA$PINAR, S. KORKMM, N. ruetce, A. Q NAR, N. UYGUN, u KERST NG Seasnal Changes in Transmissin f Citrus Stubbrn Disease Pathgen Spirplasma g11d by Budwd Grafting s. KRKMAZ, A. Q NAR, U. KERST NG The Rle f the Phytalexins n the Resistance t Chickpea Blight (Ascchyta rabiei (Pass.) Labr.) in Chickpeas F. S. DO -AR, A. GURCAN Occurence and Distributin f Fungal Diseases n Lentil in Ankara Prvince A. KARAHAN, Y. Z. KATIRqOGLU Research n Sme Bilgic and Emerging Prperties f Black Grass (Alpecurus mysurides Hudsn) B. TA$TAN, A. ytldlrtm, A. ERCig Big - Vein Virus Disease f Lettuce in Eaurum, TURK YE T. DKEN, s. AgtKGz, E. DEMiRci Recent Recrds On Virus Diseases f Vegetables in Greenhuses u. rinx...45

4 J. Turk. Phytpath., Yl.22, N. 1-8-, 1993 rssn 378-8?A studies n citrus Stubbrn Disease and sesame phylldy in Sesame and Their Related kafhper Vectis Hiiseyin BASPINAR, SavaE KORKMAZ Niiket ONtrlGn, Ahmet QINA& Nedim tl"fgun Department f Plant Prtectin Faculty f Agriculture University f eukurva TR-133-ADANA, Tiirkiye Ulrich KERSTING Instute f Phytpathlgy University f Bnn NuBalle 9 D-53 BONN 1, FRG ABSTRACT Th3 -c9ryence f (afhnners and the ppuratin dynamic f circurifer tucmarceps (M. & R.) and rsius ricnralh (Matsumura) lhmpteia, Cicadeiliade) were sytdied n sping and summer sesune by D-VAC cltectii in Adana frm May t September in 191. Furthermre, the incidence f spirplasma cilri sagli et al.'and sesarne phylldy in infected sesame plants were bserved in fietd. Bti diseases were vectred by c.lwzmuceps and. rienralk, respectively. The infectin f c. haematceps and sesarne by s- citri was determined by ELISA in labrairy. - Leafhppers were mre n sping sesame -dminant cmpared t summer crp, hwever, the number f c. haemu"eps ani.7*nal* in simnter sesene utnum_ lere_l the amunt f btly spe-cies in spring sesame. The fint c. haematceps cllected in May were all hibemating females indiere haftin{ s. citri, ss detennined by by ELISA. The first s. citri infected sesame platns were detected in the mid,f i[;, which prbably infected just after germinafih in.<pril. - wth increasing ppulatin f c. Iucmtceps the number f s. citri - infected plnts raised up 125 plants in summer sesame dfu mia f August."Mst f the,lr, plants were cncunent Wcted with S. citri plus,rri, pni,aay MLCi, thus symp_ tms psitively attibutable t S. citri were rarely bserved. This study cnclusively demnstrated the irnprtance f sesame, especially f sum_ mer sesame, and the assciated c. hucmnrceps pputaiin fr the-epidemnigt y S. cilrt. INTRODUCTION Stubbrn disease, caused by spirplasma crln sagli et al (Mycplasmatales: Spirplasmatacea) is ne f the mst serius and destrutive diseases f citrus in the Eastren Mediterranean Regin f Tiirkiye and the Near East (BvE 19g6,

5 CITRUS STUBBORN DISEASE AND SESAME PHYLLODY qegfevan 1987, QINAR et al 1993). The citrus stubbrn disase pathgen (CSD) is transmitted by the leafhpper Circulifer hacmtceps (M. & R. ) (Hmptera: Cicadelliade) (FOS ei ul L986, KERSTING and gengonca 1992). Prbable due t this vectr transmissin up t 5 V f. zll Navel range trees are infected by the CSD pathgen in the Qukurva regin, where abut 85 V f the ttal Turkish citrus prductin is cncentrated (GULLU 1989). A number f nn-citrus hsts f,s. citri are knwn in the Qukurva regin (UY- GUR er al I99L, KERSTING et ul 1992, BASPINAR et al 1993'). flwever, a recent study shwed that nly sesame (sesamum indicum (L)). serves as an excellent hst, bth fr the vectr C. lucmatceps and the pathgen S. citri (KERSTING et al t993). Mrever sesame is frequently infected by sesame phylldy MLO (mycplasma-like rganism) in Asia, a disease knwn t be transmitted by the leafhppers Orsins rientalh (Matsumura) and, Orsins celluhsw (Lindberg) (VASUDI- VA and SAHAMBI L959, CHOOPAI{YA 1973). This disease was previusly reprted frm Ttirkiye (TURKOdLU and FiDeN 1985) and bserved 6y the uuihrs n nearly all sesame fields alng the suth cast f TiiLrkiye and in Suth East Anatlia in L99. Fr the reasn that sesame is an imprtant crp in the Eastern Mediterranean Regin, cvering abut 1. ha/year and because f its significance fr the epidemilgy f the CSD pathgen, the ppulatin changes f the mst imprtant leafhpper sepecies and the incidence f S. cilri and sesa-e phylldy MLO were studied n sesame in the qukurva regin f Tiirkiye in MATERIALS AND METHODS Leafhppers were cllected bth in spring and summer sesame in Adana thrughut the vegetatin perid frm May t September in The sesame field cnsisted f fur varieties, namely, Gijlnarmara, Muganll 6zberk-82, and a lcal variety. The spring sesame was swed n April, 15th and the summer sesame n June, 25 th. The ttal experimental area included 112 rws f 5 plants each fr bth the spring and su'nmer date. Leafhpper samplings were cnducted three times a mnth by a mechanical insect cllectr (D-VAC) and were standardized by sucking a singls plant with 1 repetitins fr three secnds. S. citrt was detected in sesa-e and in leafhppers fllwing the technique described by BOVE ct al (1987) and MARKHAM u al (t983). Fr this purpse, sesame plants shwing either yellwing, stunting r phylldy and the leafhpper assciated with these plants were analyzed fr the presence f S. citri. In additin, surveys fr mllicute infected sesame and leafhppers were cnducted in Kz^n and Silifte an imprtant grwing area f ses:rme in 1991.

6 H. BA$PINAR, S. KORKMAZ, N. ONELGE, A QINAR, N. UYGUN, U. KERSTING REST'LTS During the ne year study ttaly L5 lealhpper species were determined n spring and s '-mer sese'''e. As it is shwn in table!, Asymnutrasca decedens (pali), Empasca decipiera Pali mrphlgical indistinguishable frm each ther - cicdulinn bipurctella (Matsrrmura), Macrsteles quadripututulzrzs (Kbm.), c. hacmatrrceps,. ricntalh and Pssmnntettix prvincinlis (L). were cmmn and ccurred in cmparatively high number. Acnurella prlixa (Lethrerry), Anceralagaltia ri- Darai (ss.)., A. hcvis (Ribaut), Austragallia sinuta (Ribaut), Balchaha lube (Kirkaldy) as well as B. pwrctara (Fabricius), Batracmrphus glaber Haupt, Eucelis atsins Ribaut and Exitianrc cupich (stdl) were rare, and bserved, in general, nly in spring sesame. The number f lealhpper species n the lust crp was as duble hrgh (15 species) than in sunmer sesame (7 species). Much the sa-e is t say fr T the abundance f leafhppers with the exceptin f C. lracmatceps and O. ricntalis, since mre individuals were encuntered in 1[s srmmer crp. Fr the reasn that C. hacmtceps and O. ricntslis are imprtant vectr species, their ppulatn fluctuatins n the spring and summer ses:rme crp as *ell as the number f S. citri and sesame phylldy infected plants are described in detail. At the fust sampling date, 26 c. lucmatceps were cllected, being all females and again 11 females *"1s 5ampled ten days later. The fust males were bserved end f May. The c. lucmntceps ppulatin reached a small peak beginning f June. Frm this date n, the number f. C. humuceps decreased until harvesting time end f August. n the spring sesame. ricmulis never reached high numbers starting with trv individuals in May and nly up t five individuals were sa-- pled end f August (Fig. 1). The fust plant shwing yellwing symptms were discvered mid f May. Later in the seasn plants with virescence r phylldy were discvered. S. citri was detecded in all plants by ELISA, shwing either yellwing, stunting r symptms attributable t sesame phylldy. The number f infected plants increased slightly in spring sesame valuing 25 plants end f August (Fig. 1) On the summer crp f sesame, L9 C. lucmuceps were cllected beginning f August. The ppulatin strngly increased and reached its maximum level end f August with abut 46 individuals/per D-vac cllectin. Later the ppulatin severerly decreased and nly 8 individuals were sampled mid f september. The first. rientalis were bserved end f July (1 individuals) and then he ppulatin grew gradually up t 27 individuals/per D-vac cllectin begin"ing f september (rig. z). The firs plants shwing yellwing r phylldy symptms in the secnd crp were bserved beginning f August. Later the number f infected plants increased dramatically and at end f sesame seasn up t L25 plants were fund t be infected (Fig.2).

7 CITRUS STUBBORN DISEASE AND SESAME PHYLLODY Table 1 : Occurence and number f leafhppers in spring and summer sesame in Adana in l.cafhpper species Leafhpper in 17 D-vac samplings n sesame (spring crp) Occurence Number Icafhpper in 8 D-vac samplings n sesame (summer crp) Occurence Number Acnurella prlixa Anaceratagallia laevis Anaceratagallia ribauti As5mmetrasca decedens * Empasca deciepiens Austragallia sinuata Balclutha hebe Balclutha punctata Batracmrphus glaber Cicadulina bipunctella Circulifer pacipennis Euscelis alsius Exitianus capicla Macrsteles quadripunctulatus Orsius rientalis Psammtettix prvincialis 1 I 2 L7 1 3 ) I t L ) L L

8 H. BA$PINAR, S. KORKMAZ, N. ONELGE, A QINAR, N. UYGUN, U. KENSTING 6 ZO a brs q c -E 1 l * i z ),. 1 """'n May May,25 I I lr I I I ftu I I lr I t iis May Jun Jun Jun Jul Jul Jul Aug Aug Aug q 6 e 6! Ef z Figure I : Ppulatin changes f Circulifer haematceps and Orsius rientalis as well as number f Spirlasma citri and sesame phylldy infected sesame plants in spring sesame in Adana in s45 e E fr4 a Fss Es Ezs z ' ;15 3t E zs 3. * C, haematceps "-." O. rientalis -.- Sesame 16. Aug 12 6ts r 3 E a Eg 6e s 4 -E E f 2z 17. Sep Figure 2 : Ppulatin changes f Circuliferr pacipennis and rsius rientalis as well as number f Spirlasma citri and sesame phylldy infected sesame plants in summer sesame in Adana in 1991.

9 CITRUS STUBBORN DISEASE AND SESAME PHYLLODY S. citri infected C. hacmaarceps and sesame was als cmmn arund Kzan and Silifte in DISCUSSION The fust C. hacmnceps cllected n spring sesame beginning f May were all females. This result cnfirms the bservatin f LODOS (1986), sayrng that C. lucnutceps is hibernating as adult. NIEISON (1975) stated that the related species C. terclhts (Baker) hibernates as mated females in U.S.A. and that females ccurred frm May n. As determined by ELISA, thse individuals cllected in May were harbring the citrus stubbrn disease pathgen. This strngly indicate that hibernating C. twcmtceps females are f majr significance fr the epidemilgy f S. citri. Mst likely the frst infectin ccurred by hibernating females in April r May and these infected plants serve as a pathgen reservir frm which S. citri will be spread t ther plants, e.g. citrus, by.the ff spring f these females. In cincidence with the ccurrence f S. citri infected C. hacmdceps, the flrst plants shwing symptms attributable t the CSD pathgen were bserved mid f May and the infectin was prved by ELISA. These sesame plants were prbably infected just after germinatin in Mid f April, befre the lust leafhpper samples were dne. The number f C. hazmuceps, vectring S. cilri was much higher in summer sesame than in spring sesame. Much the same is t say fr the number f S. cllri - and seseme phylldy MlO-infected plants, which was abut five times higher in the late swed sesams cmpared t the early swing date. The reasn is that nt nly O. ricntulh is able t transmit the MLO, but recently, als C. lwemilceps was fund t be a vectr f a sesame phylldy MLO h Iran (SAHLEHI and IZADPANAH 1992). Fr the reasn that in a cncurrent infected plant (S. citri plus sesame phylldy MLO) the S. cdn symptms are masked by MLO's the CSD - infectin is nt detectable symptmatlgically. Even symptms attributable t sesame phylldy might nt ccur r are untypical. Thus, fr any wrk n epidemilgy f S. citrf r sesrme phylldy MLO supplementary detectin methds, like ELISA, culture and micrscpt has t be used. 6znr Susamda Turunggil Stubbrn ve Susam Filldi Hastatklan ys f,rrnlann Cicadellid Vektiirleri Uzerinde Qahgmalar Bu gahgma Mays - Eyliil 1991 tarihleri arasmda ytiriitiillmiie ve D-vac b6cek tplamn aleti ile tlrneklemeler yaprlarak Circulifer Incmatceps (M & R.) ve Orsius ricfiulis (Matsumura)'in birinci ve ikinci iiriin susamda bulunma z manlarr ve ppulasyn dalgalanmalan incelenmig ve aylca. Spirpltsma citri Sagli et al

10 H. BA$PINAB, S. KORKMAZ, N. ONELGE, A. QINAR, N. UYGUN, U. KENSTING ve Susam filld hastahermn tarla igerisindeki yayhgr izlenmiqtir. Bu hastahklar srrasryla C. heerytpps ve O. ricntalrs tarafindan tagrnmaktadt. C. hacmalceps ve susamda S. cirrfdn varhp,r labratuvarda ELISA testi ile saptanmrgtr. Cicadellid[erin birinci iirii,n susamda tiir saysr bakrmrndan daha zengin ldu$u, ancak C. hwceps ve O. ricntalls'in di[er tiirlere giire hem birinci ve hem de ikinci iiriin susrnda daha yiiksek diizeyde ppulasynlar lugturduklan belirlenmigtir. Kry ;agiren diei bireylerin biinyelerindeki S. citn ELISA testi ile rtaya knmu$tur. Terla iginde infekteli ilk bitki mays rtalarmda saptanmrg lup, bu bitkinin ginlenmden hemen snra vektirrlerin hastah[r tagrmasryla infekteli hale gelebilece$ diisiirtilmektedir. C. lucmatw.qs ppulasynunun artmasryla birlikte S. citri ile infekteli bitki sayrsrnda da bir artrs giidenmig lup, apusts rtalannda 125 adet infekteli bitki ile bu sayr en yuksek diieeye ulagmrgtu. Bu bitkilerin biiyiilk g[unlu[u hem S. citri ve hem de Filldi etmeni ile infekteli fulrrnmug sadece S. citri simptmu gbsteren bitki saysr sn derece diisii& saptanmrgtu. Bu gafrgmayla susam bitkisinin iizellikle ikinci tiriin susamrn gerek S. citri lle n- fektili gk sayrda bitkiyi ve gerekse ytiksek ppulasyn diizeylerinde c. lwcmalceps'i birlikte bulundurmasr nedeniyle S. citri'nin epidemiyljisinde gk iinemli bir yeri ldu[u rtaya knulmugtur. ACKNOWLEDGEMENT Dr. Ulrich Kersting gratefully acknwledges his schlarship frm the Alexander vn Humbldt Fundatin, Fedr - Lynen - prgrarnme, FRG. LITERATURE CITED BA$PINAR, H., S.UYGUR, F.N. UYGUR, N. UYGUN and U. KERSTING, D$t Akdeniz Bblgesi turunggil bahgelerinde zararh lan Stubbrn hastahs $plrcpksrw crrn Sagli et al.)'nin epidemiyljisinde yabancr tlann rlii. TiiLrkiye I. Herblji Kngresi, Adana, 3-5. gubat BOVE, J.M., Stubbrn disease and its natural transmissin in the Mediterranean area and the Near East. FAO Plant Prt. Bull., 34,L5-24. BOVE, J.M., J.G. VIGNAULT and C. SAILLARD,1987. Spirnlasma citri detectin by enz5anelinked imm,unsrbent assay (ELISA), culture and dt hybridisatin. Isr. J. Med. Sci., 23, chpatiyas L.N Mycplasma-like bdies assciated with sesame phylldy in Thailand. Phytpathlgy,6.j, L536 - L537. QAGLAYAN, K., Turunggil yediverenleeme (Stubbrn : palamutlagma) hastalr$ etmenispirplasrna citri'nin izlasynu, tamsr ve patjen-vektiir iligkilerinin

11 CITRUS STUBBORN DISEASE AND SESAME PHYLLODY aragturlm351. Dktra Tezi, Qukurva Universitesi, Adana, 133 s. qinar, A., U.KERSTING, N. NELGE, S. KORKMAZ and G. $A$, Virus and virus-like diseases f citrus in the Eastern Mediterranean regin f Turkey: the citrus budwd imprvement prgramme and its prblems. In: Prc. 1,2 th IOCV Cnference,IOCV Riverside, USA. (in press). FOS, A., J.M. BOVE, J. LALLEMAND, C. SAILLARD, J.C. VIGNAULT, Y. ALI, P. BRUN and R. VOGEL, The leafhpper Nealilurus haematceps is a vectr f Spirplasma citri in the Mediterranenan arear. Ann. Inst. Pasteur..Micrbil., 137a, GULLU, M,, DSu Akdeiz Birlgesi Navel grubu prtakal ve Satsuma mandarin a$aglannda yaygrn viriis ve viriis benzeri hastahklarrn stirveyi ve indekslenmesi iizerinde gahgmalar. Dktra Tezi, Qukurva Universitesi, Adana, 266 s. KERSTING, U. and q. SENGONCAL99Z. Detectin f insect vectrs f the citrus stubbrn disease pathgen, Spirpluma crrrd Sagli et al.,in the citrus grwing area f Suth Turkey. J. Appl. Ent., 113, KERSTING, U. Q. SENGONCA and A. QINAR, lggz.detectin f Spirplasma citri n nncitrus hst plants and their assciated leaafhpper vectrs in Suthern Turkey. FAO Plant Prt. Bull., 4 (3). (in press). KERSTING, U., H,.BA$PINAR, A. QINAR, Q. SENGONCA and N. UYGUN, New findings n the epidemilgy f Spirplumt citri in the Eastren Mediteranean regin f Turkey. In: Prc. 12 th IOCV, Riverside, USA. (in press). LODOS, N., Tilrkiye entmljisi. Genel, uygulamah ve faunistik, cilt II: Ege Universitesi Ziraat Faktiltesi Yayrnlan N.: 429. Ege Universitesi Basrrnevi Brnva - iztr,tir. Ss s. MARKHAM, J.M., T.B. CLARK and R.F. WHITCOMB, Culture techniques fr spirplasmas frm arthrpds. p. 26l In: Methds in Mycplasmlgy, Vl. II. Academic Press, New Yrk. NIELSON, M.W., The leafhpper vectrs f phytpathgenic viruses (Hmptera, Cicadelliade). Taxnmy, bilgy, and virus transmmissin. Technical Bulletin N. L382. Agricultural Reseacrh Servrce, USDA, Washingtn, D.C.,386 pp. SAHLEHI, M. and IZADPANAH, Etilgy and transmissinf sesame phylldy in lran. J. Phytpathl, 135, TURKOGLU, T and U. fioan, Ege Biilgesinde ikinci iirtin larak ekilen susamlarda virtis hastahklan _ ytiniinden incelemeler. IV. Ttrkiye Fitpatlji Kngresi,8-1L Ekim 1985.izmir, Ttirkiye. UYGUR, S., F.N. UYGUR ve A. QINAR,I99l. Qukurva Bblgesi'nde Spirplasma cilrt Sagli et al.'nin knukgusu lan yabancr t tiirlerin saptanmasr. VL Tiirkiye Fitpatlji Kngresi, 7 - L1 Ekim l99l,izrnt.3il - 3I4. VASUDM, A.R.S. and H.S. SAHAMBI, Phylldy in sesamum (Sesamum ricntulis L.) Indian Phytpathl, 8, I29.

12 J.TurkPhytpalh.,Vl.22, N. l, 9-16, 1993 ISSN Seasnal Changes in Transmissin f Citrus Stubbrn Disease Pathgen, Spirplasma citri, by Budwd Grafting Savag KORKMAZ' Ahmet QINAR Department f Plant Prtectin Ulrich Kf,RSTING Institute f Phytpathlry University f Bnn Faculty f Agriculture University f Qukurva Nu3allee 9 T-133 ADANA Ttirkiye W-53 BONN l, FRG ABSTRACT The Mediterranean regin is the mst imprtant citrus grwing area f Ttirkiye and abut 85 % f the ttal prductin is cncentrated in this regin. Almst all citrus trees are infectedwith at least ne virus-like disease, the mst imprtant being citrus stubbrn disease (CSD), caused by Spirplasma citrisagli et al The bjective f the present study was the evaluatin f the rate f transmissin f S.citri by budwds with regard t the seasn and the perid f string at cl temperature. Budwds were cllected in February, May, August, and Nvember frm S.citri infectedatwcd navel ranges. One third f the budwdswas grafted s sur range the same day, the restwas stred in a refrigeratrat 4 Cfr ne and tw mnth, respectively. The rate f transmissin f the CSD pathgen was evaluated symptmatlgically, by bilgical indexing, and by ELISA. Out f 26 plants 15 shwed symptms attributable t S.citri, I I plants were tested psitive by bilgical indexing and 17 plants were harbring the csd pathgen as determined by EUSA. Over all I9 citrus plants were ir{ected with S.citri accrding t bilgical indexing and ELISA, hwever nly 4 plants were psitive tested symptmatlgically, by bilgical indexing, and by ELISA. As a result f this study it is strngly suggested that in any budwd imprvement prgram, bilgical indexing as the preferred methd shuld be cmbined with ELISA r culture f the CSD pathgen in the detectin f S.cilri infected trees. INTRODUCTION Citrus is f ever increasing imprtance fr Tiirkiye and the prductin reached abut 1.5 millin tns in 1991 (ANNYMus, 1991). citrus plantatins have mainly been established alng the suth cast ftiirkiye, where abut 85 % fthe ttal citrus is prduced. Unfrtunately, many citrus rchards in the Eastern Mediterranean regin are infected with several virus and virus-like diseases. One f the mst imprtant is citrus

13 SPIROPLASMA CITRI stubbrn disease (CSD), caused by Spirplasma citri Sagli et al. (Mycplasmatales: Spirplasmataceae) (SALIBE 1986, Qnar et al. 1993). Citrus stubbrn disease was first bserved in Califrnia in the beginning f the 2 th century GAWCETT A al. 1944) and later n, the disease spread widely in many Mediterranean and Middle East cuntries (CALAVAN and BOVE 1989). The presence f a stubbrn-like disease in Ttrkiye was reprted fr the first time in the 195's (CHAPOT, 1959), reaching epidemic levels in the 197's. S.citri was intrduced int Tiirkiye prbably thrugh infected yung citrus trees r budwds. In the fllwing years the disease spread rapidly and the cmmn planting f navel ranges which are highly sensitive t,s. cifri S. citri is vectred by leaftrppers, especially by species fthe genus Circulifer. ln Tiirkiye, Circuliftr haematceps cmplex (M. & R. ) was fund t be nly vectr f the stubbrn disease pathgen (KERSTING and $ENGONCAL992). Besides by leafhppers, the disease is transmitted by budwds frm infected trees r prpagatins. Hweer, it is reprted that the transmissin rate is quite variable and ranged frm t 1 % depending n variety, tissue and seasn (CALAVAN 4 al 1969, GUMPF A al 1986). The purpse f this study was t determine the transmissin rate f S. cilri by using budwds frm stubbrn infected field trees with regard t seasn and the peid fstring at cl temperature. MATERIALS AND METHODS During this study, five stubbrn infected Atwd Navel range trees (Cifras sinensis (L.) Osb.) at the Mersin - Alata Hrticultural Research Institute had been chsen as budwd surce. In May, August, Nvember 1989 and in February 199 at least 9 buds were cllected at each date. Each fthe 9 buds were separated int three grups. The first grup was immediately grafted n sur range and the ther tw grups were grafted fllwing a ne r tw mnth strage in a refrigeratr at 4C. Fr each di-fferent grup 3 virus and stubbrn free budwds were grafted as cntrl. The budwds were grafted t ne year ld sur range nucellars and wrapped with plyethylene tape fr three weeks. Thereafter the sur ranges were pruned l cm. abve the graft. The scins were frced t grw as single sht fr six mnth at 32l27C (dayl night) temperalure,t + 5V relative humidity and l6 h light in an insect-free greenhuse. T determine the transmissin rate f,s. crlrd by budwds, the citrus stubbrn disease pathgen was detected in all test plants symptmatlgicalyy, by bilgical indexing and enryme-linked imunsrbent assay (ELISA). Bilgical indexing was dne accrding a mdified technigque frm CALAVAN and CHRISTIANSEN (1965) by side-grafting a 5 cm lng Madame Vinus sht with three buds t each test plant. The shts were cvered l

14 S.KORKMAZ, A.QINAR and U.KERSTING by a plyethylene bag fr three weeks and later frced t grw as a single sht. All test plants were kept at 32127'C (day/night) temperature, '7 + 5 y relative humidity and 16 light in an insect-free greenhuse. The evalutin f symptms were started three mnth after grafting (ROISTACIIER 1991). All plants were tested serlgically fr the presence f,s.cilri by a DAS-ELISA (CLARK and ADAMS 1977), using a plyclnal antibdy against SPA-strain Israel. The results were evaluated in a Titertek Multiscan ELISA Reader at 45 nm; the plants were tested psitive, if the difference between sample and healty cntrl was higher than,1 OD (SAILLARD and BOVE 1983).. Rf,SULTS AND DISCUSSION In general, the survival rate f budwds cllected in May and August was cmpartively lw. Only 43.3 V and 47.8 y f the grafts survived, while 98,9 % f thse buds cllected in Nvember and February cntinued t grw. All buds taken frm healthy, greenhuse-grwn, Navelranges survived. Budwdscllected innvemberrfebruary and kept fr ne and tw mnths at 4 C shwed the same viability as thse grafted at the same day. Ntable mre buds died after string fr ne r tw mnth, if cllected in May r August (Tab. 1). After a six mnths bservatin perid nly 15 f 26 trees shwed symptms attributable t stubbrn disease (5,8 y). The symptms bserved varied frm very severe t slight. The fur plants revealing severe symptms were small with shrtened interndes and the leaves, have had symptms f zinc deficienry. The eleven plants with slight characteristis were stunted, but n leaves symptms were bserved. Using bilgical indexing eleven plants (4,2 7) were tested psitive fr CSD pathgen and by ELISA in ttal l7 plants (6,5 %) were harbring S. citri. Over all 19 citrus plants (7,3 7) were infected with the stubbrn disease pathgen accrding t the ELISA and bilgical indexing results (Tab. 1). There was nlya slight effectf seasn n the transmissin rate fs. cilribybudwds. Accrding t the ELISA and bilgical indexing results, the transmissin rate ranged frm5,6 /t 1 %ifthebudwdswerecllectedinfebruaryandmay, respectively. Over all the transmissin rate was as lw as 7,3 %. CALAVAN A al Q969) bserved a very lw percentage f transmmissin frm budwds grafted in August, Octber and January as well as a lwer transmissin rate when using buds instead f side grafts. These results clearly indicate that the CSD pathgen is distributed irregularly in many trees and that pathgen is inhibited r lst in mst budwds during much f the year. Five plants revealing n t'?ical characteristics f a,s. crlri infectin were tested psitive by bilgical indexing and ELISA. Althugh l 5 plants shwed typical symptms f an,s. cilri infectin, these bservatins were cnfirmed nly fr six plants by bilgical indexing r by ELISA. Out f eleven plants tested psitive by bilgical indexing, nine reacted psitive and tw negative in the serlgical test. Furthermre, seven citrus trees which did nt shw any symptms in bilgical indexing were tested psitive by ELISA. Slely fur plants fund t be infected symptmatlgically, by bilgical indexing, and by ELISA (Tab. 2). ll

15 S.KORI{\{A2., AQINAR and U.KERSTING b 6 (! c! c.) r3 C! b 6).E q E 13 - c)! la $ q-\ t= h C c.) (! E (.) t a B rg (H (),, 5!./ =+ ze -.i c) 5 (g F q) (,) L ' Cj).c) L (J v) tt L () z Edbl cdtr < bx dc) d 5'= a JI!. ()cb B'8 EE.i (c b (n.a * 'da ) > 'trt a> (t) =s b.= c) E cq.i I.= c E l9 6g O \ \ z c.t t 6l s ci z ct v s z s ct z N c.l s z c.) ca \ \ Cg 2 F- \ \ 6l c.) '- V' (t) c\ (? rn aa tr) rf, \ av) t- nc q\ c\ b, g l Ez I E I () I I (\t GI T2 ca c; F c C; (t tr I 6 9p $ GI Fs (a ca 6l \l t* + N ae N 6l r.. d N N t-- + (\ c.) r- rf, fa = Lr C:) O F (J I 6l ca c.) c) p I z I c C) () k p ) z c I ca c\ \ \ c\ N h ( (J I ca t\ t\ r- c\ 6l \ r\ rr 6l r/) + rf, \ V' rn c\ q \ \ 6 c.l ca cl F P G tr fr I ce b(n p.) fjr c\ \ \ \ 6l! ( a I c (,) I c t.' tn v}!+ $?t)?-t N s (a \ c \ \ 6t F (t) rs \ V' F N.f l/' ln l N r-- \ N c\l F (,) a I C! a H b * (.) G}

16 SPIROPI-ASI\{A CITRI Table 2: citrus trees infected by spirptasma citri asdetermined symptmatlgically, by bilgical indexing, and by ELISA. Trees tested psitive by all three methds are idicated by bld numbers Grup N. t-a Budwd Cllectin time Grafting time Citrus plants (N.) infected with,s.cirri Symptms May l,l2 8 tsrlgical indexine ELISA 8 1-B May June 1,5,9 2,9 1,8,9 l-c 2-A July August 2I 13 2-B August September 2-C Octber 7 6,7 7 3-A Nvember 13,2,33 2,23 8,2,23 3-B Nvember December l9 l8 18,3 3-C Janurary l t6 15,16 4-A 4-B February February I,23 t6 16,23 March l6 29 L6 4-C April 23 l3

17 SPIROPLASMA CITRI These results clearly indicate that stubbrn is almst undetectable syttrpmatlically in yung citrus trees. Even plants with n Eymptms may harbr the CSD pathgen and will help t spread S. cilri if used as a budwd surce. It is bvius that even the applicatin f nly ne detectin methd might led t false negative, s that bilgical indexingshuldbe cmbinedeitherwithelisar culture fthepathgenin anybutwd imprvement prgram. 6zm TURIJNCGIL STUBBORN HASTALIGI ETMENi Sptrplasnecihi'nin A$rczu ylu irr ragrnuesindaki MEVSiMSEL DEGi$IKLIKLER Tiirkiye turunggil tanmlnrn / 85'inin yaprldrlr Akdeniz Bcilgesi'nde mevcut gk sayrdaki viriis ve viriis benzeri hastahklardan en Onemlilerinden biri de Spitplasma citri etmeninin neden lduiu Stubbrn hastalrtrdrr. Bu gahqmarun amaq hastah$n agrgdzii ile tagmmasmda agrgdderinin a[agtan ahm zamaru ve dii;iik srcakhklarda (+ 4 qc) bekletme siiresinin etkisini aragtrrmaktrr. Aqrgtideri Mayrs, Aiusts, Kastm 1989 ve $ubat 199 tarihlerinde kesilmig ve her grup iginde 3 krsma aynlmrgtrr. Aynlan krsrmlardan birincisi hemen, ikircisi bir ay ve iigtinciisti iki ay sfukta deplandrktan snra aqrlanmtgtr. Hastaltim ta$amau sipmtmljik, biyljik indeksleme ve ELISA testine gore aragtmlmr$tr. Mayrs ve A$usts'ta ahnan agrgcizlerinin tutma ranlan dtigiik bulurunu$ur. Alu aylft g6demlerden snra 26 bitkinin l5'inde zayrftan kuwetliye kadar de$gen simptmlar gddenmigtir. Biyljik indeksleme snucu ise 26 bitkiden 1l'i, ELISAtqsti gahgmalan snucu ise 17 bitki etmen ile bulagrk larak bulunmu;tur. ELISA testi ve biyljik indekslemeler snucu ise 19 bi&i bulagrk larak saptanmr$tir. Aqrgdzii alma zamamrun etkisi gk az bulunmug, en diigtik deler Alusts ayrnda alman gdzlerde belirlenmigtir. Simptmljik gtizlemler, biyljik indeksleme ve ELISA testinin her iigtine birden yanftzca 4bitki pzitif snugvermigtir. Bu nedenle aergdzii rslahprgrarnltrndabiyljik indeksleme prgramrrun ELISA veya kiiltiir testlerinden biri ile krt*tine edilmesi gerektili snucuna vanlmrqfi r. ACKNOWLEDGEMENT Dr. Ulrich KERSTING gratefully acknwledges his schlarship frun the Alexander vn Humbldt-Sffiung, Fedr-Lynen-prgramme, FRG. I4

18 S.KORKMAZ, A.QINAR and U.KERSTING LITERATURE CITI,D AIIIONYMOUS, 199 I. Les exprtatins d'agrumes du bassin Mediterranean. Statistiques, evaluatins prepartitins - situatin Secretariat General Du C.L.A.M, Madrid. pp. 79. CALAVAN, E.C. and D. W. CHRISTIANSEN, Rapid indeing fr stubbrn disease f citrus. Phytpathlgy, 55, 153. CALAVAN, E.C. and J.M. BOVE, Eclgy f Mycplasma. p 425-4E5. In: The Mycplasmas. Vl V, WHITCOMB, R.F. and J.G. TULLY, (eds). Academic Press, New Yrk CALAVAI.I, E.C., C.N. ROISTACHER and D.W. CHRISTIANSEN, Distributin f stubbrn disease virus in trees f Cilrzs sizet?sis and C. paradki at different seasns. p In:Prc. 4th IOCV Cnference,Icv Riverside, USA. CHAPOT,H., 1959First studies n the sutbbrndisease fcitrusinsmemediterranean cuntries. p In: Citrus Virus Diseases (J.M. WALLACE ed.).univ. Calif. Div. Agr. Sci. Berkeley. qrnar, A., KERSTING, N. NELGE, S.KORKMAZ and G.$AS, Virus and viruslike diseases f citrus in the Eastern Mediterranean regin f Turkey: The citrus budwd imprvement prgramme and its prblems. In: Prc. 12 th IOCV Cnference, IOCV Riverside, USA. (in press). QLARK, M.F. and A.N. ADAMS, Characteristics f the micrplate methd f enzyme - linked imunsrbent assay fr detectin f plant viruses. J. Gen. Virl., FAWCETT, H.S., J.C. JERRY and J.C. JOHNSTON,1944. The stubbrn dtsease f citrus. Calif. Citrgraph, 29, GUMPF, D.J., E.C. CALAVAN and J. PEHRSON, Spread f stubbrn disease in Califrnia citrus. Citrgtaph, 7 I, '7. KERSTING, U. and C.$ENGONCL Detectin f insect vectrs f the citrus stubbrn disease pathgen. Spirplasma citri Sagli er al, in the citrus grwing area f Suth Turkey. J. Appl. Ent, 113, ROISTACHE& C.N., A handbk fr detectin and diagnsis f citrusvirus and virus-like diseases. FAO Publicatins, Distributin and Sales Center, FAO t5

19 SPIROPLASMACITRI SAILLARD, c.and J.M. BO\IE, Applicatin f ELISA t spirplasma detectin and classificatin. p In: Methds in Mycplasmlry vl. I, Academic Press, New Yrk. SALIBE,'aA, 1986 Aprgramme fr citrus imprvement and prtectin in Turkey,Reprt t the Gvernment f Turkey. FAO Rme Italy, p l-g4. t6

20 J. Turk. Phytpath., YL22, N ,L993 ISSN The Rle f the Phytalexins n the Resistance t Chickpea Blight (Ascchyn rabiei (Pass.) I-abr.) in Chickpeas F. Sara DOLAR AbdUIIAh GURCAN University f Ankara, Faculty f Agriculture, Department f Plant Prtectin, 611 Ankara / TURfiyg ABSTRACT After inculatin with race 1 f Ascchyta rabiei, the accumulatin f ptercarpan phytalexins in the leaves and stems f the chickpea (Cicer arictinum) cultivars ILC 1929 (susceptible) and ILC 3279 Qesistant) determined t be quantitatively by HPLC.. Medicarpin and maackiain in the leaves and stems f resistant cultivar accumulated 12 hurs after inculatin. Maackiain did nt accumulate in the stems and leaves f the susceptible plants althugh medicarpin started t accumulate 24 h after inculatin. Cntents f medicaryin (22 Blg. fr.w.) and maackiain (17. glg fr.w) in resistant cultivar was enugh fr the inhibitin f fungal develpment but it was less (9. glg fnw.) in the susceptible nes- The detectin f maackiain in nly resistant plants indicated that this cmpund has an imprtant rle in the resistunce. INTRODUCTION Chickpea (Cicer ariztinum L.) is a widespread crp plant in semiarid areas f Asian and Nrth African cuntries. The ptercarpans medicarpin and maackiain have been identilied as the main phytalexeins f this plant (Keen, L975) where they accumulate during infectin vith Helnintlnsprium carbnrm Ullstrup (Ingham 1976) r Nectrfu hacmstccca (Denny and VanEtten, L981,). The mst imprtant disease recrded n chickpea in Ascchyta blight which is caused by Ascchyta rhiei (Pass.) Labr. (Telimrph Mycsphazralla rabici Kv.) There are many reprts f serius lsses f chickpea crps caused by Ascchyta blight (Nene,1982). Singh and Reddy (199) demnstrated that in favurable envirnmental cnditins yield lsses due t the disease can reach lv. Ascchyta ra- Drci-resistant cultivars f chickpea shwed, amng ther prperties (Nene, 1982) a much higher phenlic cntent f unidentilied structures when cmpared t susceptible cultivars (Vir nad Grewal. 1974). Furthermre, inculatin f seed cavities f detached pds f Cicer arictinum with spr suspensins f. A. rabizi led t the accumulatin f an antifungal cmpund which was nt chemically identified (Kunzru and Sinha, 1.97) The present study was undertaken t cmpare A. rabici -resistant and a susceptible cultivar f chickpea with regard t the accumulatin f phytalexins during infectin with spre suspensins f a virulent race (race l) fa. rabici. L7

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