Indian J. Agric. Res.. 38 (3) : 157 - 163. :~004

INDEXING OF VIRUSES OF POLYHOUSE GROWN CAPSICUM(CAPSICUM ANNUUM L.) IN PUNE - CAUSING

SEVERE SYMPTOMS AND EPIPHYrOSISRaj Verma, S.J. Singh, R.K. Singh and Satya Pr~kash

IARI Regional Station. -'Agricultural College Estate. Shivajinagar. Pune . 411 005. India

ABSTRACTSurveys of polyhouse grown Capsicum in Pune for virus diseases was carried during 2000­

2001. A variety of symptoms were observed in these polyhouse grown Capsicum plants. Thecommon symptoms observed were: stunting, general yellowing of plants, leaf deformation, reducedsize of newly developed leaves, systemic mosaic mottling, vein-banding, vein clearing, blistering andfiliform leaves. Symptoms on fruit were reduction in size, discolouration, str_ and blistering.Occurrence of four viruses viz., cucumber mosaic cucumovirus (CMV), PotatOV (PVY), tobaccoetch (TEV) and pepper veinal mottle (PVMV) potyviruses were recorded on the basis of symptomdiagnosis, mechanical transmission, reaction on selected test plants and serological relationship byDAS-EUSA, The most prevalent virus was CMV followed by PVY, PVMV and TEV. The study alsorevealed that these viruses could infect Capsicum plants alone or in combination. PVMV and TEVwere detected for the first time in Capsicum from Maharashtra.

INTRODUCTION abnormal discolouration on their surface. TheOne of the major constraints to the samples collected were used subsequently to

production of pepper, which is an economically inoculate healthy seedlings of C. annuum cv.important crop in India, is infection by viruses. California Wonder. Mechanical transmissionThere are about 10 different viruses reported . was done by using chilled potassium phosphateto infect pepper in India. The virus complex buffer O.lM, pH 7.2. These were rraintainedcauses serious economic loss to the growers under insect proof glasshouse for symptomaffecting both yield and fruit quality. expressions. Further, the isolates wereIdentification of the virus is imperative, if control inoculated to a set of test plants to group themeasures are to be effective. In the present virus. The presence of virus in the samples waspaper, we report the occurrence of potato virus "s.c0 determined using the Double antibody

~dwich Enzyme Linked ImmunosorbentY (PVY), tobacco etch virus (TEV), pepper A:ssay (DAS-ELISA). Briefly, the coatingveinal mottle virus (PVMV), potyviruses and antibody were diluted 1: 100 in coating buffer~\lcumber mosaic virus (CMV) cucumovirus in and loaded on ELISA plates. Test samples

,\polyhouse grown.Capsicum in Pune. diluted 1:10 in coating buffer, pH 9.6, wereMATERIAL AND METHODS added to wells of ELISA plates and incubatedTo determine the prevalence of overnight at 4°C. Goat antirabbit IgG

Capsicum annuum (bell pepper) viruses in conjugated antibodies diluted in conjugatepolyhouse, a survey was undertaken in and buffer, was added to the plate and incubated inaround Pune during 2000. Survey was the wells for 2 hrs at 37°C. Enzyme substrateconducted during the mid-developmental stage p-nitrophenyl phosphate (Sigma) was added inof the plants. Plants were randomly evaluated 10% diethanolamine at pH 9.8. Antiserabased on symptoms thought to be caused by against CMV, TEV and PVMV were purchasedvirus infection such asthlorosis, vein-clearing, from Bio-Red Labs Pvt. Ltd., India and of PVYmosaic, mottling, leaf malformation and fruits was obtained from IARI, New Delhi. Virusthat were smaller and malformed and showed positive and negative controls were included

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in all tests. Colour reaction was noted andsample was considered virus positive, if thereaction was greater than two times of healthy.plant extract.

RESULTS AND DISCUSSIONThe survey conducted in polyhouse,

revealed the occurrence of Capsicum virusdiseases. Based on the reaction on selected testplants and identification through ELISA,presence of sap transmissible virus isolates ofPVY, TEV, PVMV and CMV was established(Table 1).

Mosaic, dark green vein-banding andmottling were prominent symptoms on leavesof Capsicum plants infected by PVY (Fig. 1).Young leaves show crinkling and fruit formedwere reduced in size. The virus isolate gavelocalized reaction on Chel1opodiumamaranticoJor and systemic symptoms onNicotiana tabacum, N: tabacum cv. WhiteBurley, Datura meteJ, LycopersiconescuJentum, Nicandra physaJoides, N.gJutinosa, Petunia hybrida and PhysalisfJoridana (Table 2). The virus isolate resembledin symptomatology and host range with thosedescribed by Bidari and Reddy (1990) andGreen and Kim (1998) in chilli. The symptomson the plants infected with CMV consisted ofmosaic with systemic puckering and blisteringand various types of leaf distortions mainlyexhibited by filiform leaf lamina (Fig. 2). Infectedplants exhibited marked stunting in growth andsevere reduction in size of leaves and fruits.The surface of the fruits were broken into wartyswellings and were therefore frequentlydistorted (Fig. 3). These symptoms resembledthose described by Anjaneyulu and Apparao,(1967), Lokhart and Fischer, (1976), Kaper andWaterworth (1981). The virus isolate producedchlorotic local lesions on ChenopodiumamaranticoJor and systemic mosaic onNicotiana tabacum cv. White Burley, NicotianagJutinosa, Cucumu sativus, Datura stramonium,D. meteJ, L. esculentum, N. physaJoides, N.

gJutinosa and P. fJoridana (Table 2). The hostrange of the virus isolate is closely related withthose CMV reported by Bidari and Reddy(1990).

Capsicum plants infected with PVMVshow initial vein-clearing, mosaic mottling, leafmalformation and reduction of leaf size (Fig.4). Plants were much stunted with shortenedinternodes. Fruits on infected plants werestreaked of mottled with areas of lighter greenand were sometimes distorted (Fig. 5). The hostrange of the isolate consisted of local lesionshosti.e.,c. amaranticoJar and systemic hostsas Lycopersicon escuJentum, N. gJutinosa, N.tabacum cv. White Burley and Petunia hybrids(Table 2). The isolate resembled in symptomsto those described earlier by Brunt and Kenten .(1971), Brunt etaJ. (1978), Prasada Rao (1976)and Bidari and Reddy (1990) on chilli.

The characteristic symptoms.of TEVon Capsicum plants were prominent vein­clearing on young leaves followed by mildchlorosis and cupping of leaves. Older leavesshow greenvein-banding along the veins at thebase of the lamina (Fig. 6). Fruits of infectedplants were small, showed line patterns. Thesesymptoms agrees with the strain of TEVreported earlier (Greenleaf, 1953,1956; McKeen, 1954; Bidari and Reddy, 1986) inCapsiCum. The isolate produced necrotic locallesions on C. amaranticoJorand gave systemicreactions on Datura meteJ and D. stramonium,L. esculentum, N. physaJoides, N. gJutinosa,N. tabacum and P. fJoridana (Table 2). Theisolate resembled closely with TEV as reportedby Bidari and Reddy (1990),

The incidence of symptomatic plantsvaried largely in polyhouses. Of the four virusestested, CMV and PVY were found in 80-85%and 70-72% of the plants respectively. PVMVwas detected in 54% and whereas TEV wasfound in 10% of the plants surveyed. Doublevirus infection was d~tected in 50% of allsamples that tested positive of DAS-ELISA.

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Table 1. Viruses detE!cted by DAS-ElISA in p6lyhouse grown Capsicum

S.No. Viruses Serological reactions

1. P\fY + +2. CMV + + +3. TEV + +4. PVMV + +5. CMV + PVMV + +. + +6. CMV+PVY + +. +7. PVY+PVMV +. + +8. CMV + PVMV + TEV +. + +. +

+ = Mild reaction,+ + =Strong reaction.+ + + = Very strong reaction.

Table 2. Symptoms of viruses detected in polyhouse grown Capsicum on diagnostic host species

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Indicator plant Viruses-------------------PVY CMV TEV PVMV

Chenopodhan amaranticoJor II Nll II IINicotiana gJutinosa MM MM. Def. MM MMNicotiana tabacum cv. White Burley MM MM VC. MM. LD CllDatura mete! MM.LD MM.LD VC.MMD. stramonium MM.Def. MM. Rt. St.Lycopersicon escuJentum MM MM. Def. mM.lD MMCucumis sativus MVigna unguiculata NllCapsicum annuum cv. California Wonder GVB.MM MM. LD. Fl " VC. mM. Cu VC. MM. lMC. frutescens L. cv. Tabasco VC.MM MM TN MM.LDNicandra physaloides Nll, MM MM.LD mMPetumia hybrida MM MM MMPhysalis floridana Nll.MM MM mM

Cll Chlorotic local lesions,Def. Deformity,MM Mosaic mottleNll Necrotic local lesions.St. StuntingVC Vein clearingFL Filiform leavesCu Cupping.LD leaf distortion.mM Mild mosaicRt Rat tailing.TN Top necrosis,GVB Green vein banding,lM leaf malformation.

Among double infected plants, 45% were Capsicum has also been observed by Alegbejoinfected with CMV and PVMV, the most (1999), George eta/. (1995), Abdalla et a/.frequently detected viruses in the samples. (1991). Triple infection involving differentDouble infection with CMVand PW and PW combinations with ail viruses detected in theseand PVMV was also observed to low surveys was found in only 8 samples. Apercentage. Double infection of viruses in significantly percentage of samples from

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Fig. 1

Fig. 1. Capsicum leaves infected with potato virus Y (PVY) showing mosaic and dark green vein-banding

Fig. 2. Capsicum twig showing filiform leaves due to cucumber mosaic virus (CMV) infection rr

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Fig. 3. Capsicum fruits infected with CMV showing deformation

fig...

Fig. 4. Pepper veinal mottle virus infected (.apsicum leaves showing mosaic and mottlmg

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Fig. 5. PVMV infected Capsicum fruits showing light green streaks

fig. 6

Fig. 6. Capsicum leaves infected with lobacco etch virus [rEV) showing green vein·banding along the veins

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symptomatic Capsicum plants analyzed did not ELISA are the low concentration of virus inreact with the antisera against any of the four the samples being tested or abiotic agentsviruses. The absence of positive reaction may causing virus-like symptoms. The presentbe due' to any other mosaic causing agent report show for the first time the identities ofwhose antisera was not available. Other factors viruses infecting polyhouse grown Capsicumthat might explain negative results on DAS- in Pune.

REFERENCESAbdalla. OA etal. (1991). Plant Dis. Reptr., 75: 1019-1023.Alegbejo. M.D. (1999). Capsicum and Eggplant Newsl. No. 18: 60-62.Anjaneyulu. A and Apparao, A (1967). Indian Phytopath., 20(4): 380.Bidari. V.B. and Reddy, H.R. (1986). Curro Sci., 55: 254-256.Bidari, v.B. and Reddy, H.R. (1990). Mysore J. Agric. Sci.. 24: 45-51.Brunt. A.A. and Kenten, R.H. (1971). Ann. Appl. Bioi.. 69: 235-243.Brunt. AA eta/. (1978). Ann. Appl. Bioi., 88: 115-119.George. T.E. et a/. (1995). Indian Cocoa. Areanut Spices J. 19(1): 1-4.Greenleaf. W.H. (1953). Phytopath.. 43: 564-570.

. Greenleaf. W.H. (1956). Phytopath., 46: 371-375.Green, SK and Kim, J.•S. (1991). AVRDC Technical Bulletin No. 18. 60 pp.Kaper. J .M. and Waterworth, H.E. (1981). In : Handbook of Plant Virus Infections - Cumparative Diagnosis. (Karstak.

E. Ed.) Elsevier, New York. Pages 257-332.Lokhart. B.E.L. and Fischer. H.U. (1976). Plant Dis. Reptr.. 60: 262-264.Mc Keen. C.D. (1954). Canad. J Bot.. 32: 95-100.Prasada Rao. R.D.v.J. (1976). Ph. D. Thesis. University of Agricultural Sciences. Bangalore. Karnataka. India.