Pigeonpea genotypes (89) were evaluated for resistance to Pigeonpea sterility mosaic virus Bangalore isolate (PPSMV-B). Of these, three genotypes, ICP 7035, MAL 14 and MAL 19, were found resistant, and two genotypes, ICP 6997 and ICP 8862, were tolerant to PPSMV-B. All the resistant lines tested negative to virus in enzyme-linked immunosorbent assay (ELISA) using PPSMV polyclonal antiserum. The resistant lines can be used in breeding programme for developing PPSMV-resistant high yielding cultivar

Chikkadevaiah, M V

Kumar, P L

Nagaraj, K M

Rangaswamy, K T

ICRISAT Open Access Repository

      www.IndianJournals.com            Members Copy, Not for Commercial Sale     Downloaded From IP - 220.225.236.59 on dated 3-Dec-2012Indian Journal of Plant Protection Vol. 34. No. 2, 2006 (216-220)				 !!" #$%&"'&(University of Agricultural Sciences, GKVK, Bangalore - 560 065, Karnataka, India.†International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Patancheru - 502 324, Andhra Pradesh, India.)*Pigeonpea genotypes (89) were evaluated for resistance to Pigeonpea sterility mosaic virus Bangalore isolate(PPSMV-B). Of these, three genotypes, ICP 7035, MAL 14 and MAL 19, were found resistant, and two genotypes,ICP 6997 and ICP 8862, were tolerant to PPSMV-B. All the resistant lines tested negative to virus in enzyme-linked immunosorbent assay (ELISA) using PPSMV polyclonal antiserum. The resistant lines can be used in breedingprogramme for developing PPSMV-resistant high yielding cultivars.%"+ Cajanus cajan, host resistance, eriophyid mite, sterility mosaic, PPSMV"Pigeonpea (Cajanus cajan (L.) Millsp.) is one of the mostimportant grain legumes predominantly grown in the semi-arid tropics of India, contributing to >80% of the total worldproduction (FAO, 2005). However, the productivity ofpigeonpea in India is much lower than the potential yieldsof 2,000-2,500 kg ha-1 (Dhar, 2000). Of various causes thatlimit pigeonpea yield, sterility mosaic disease (SMD) is themost damaging disease recognized in all the pigeonpeagrowing countries of Asia (Kumar et al., 2004a). It is causedby Pigeonpea sterility mosaic virus (PPSMV) and istransmitted by an eriophyid mite, Aceria cajani (Acari:Arthropoda) (Kumar et al., 2003). SMD-affected plantsshow mosaic and mottling symptoms on leaves, withseverely reduced or no flowering (sterile). SMD symptomsdepend on the pigeonpea genotype, are categorized into threetypes: genotypes that show severe mosaic (SM) and sterility;mild mosaic (MM) with partial sterility; and chlorotic ringspots (RS) without any noticeable sterility. Susceptiblecultivars that produce SM symptoms infected early in thegrowth stage (i.e., <45 day-old plants) result in >90% yieldlosses (Jones et al., 2004).SMD management through acaricidal sprays to control thevector mite is not considered economically viable and eco-friendly. Systematic resistance breeding was initiated atInternational Crops Research Institute for the Semi AridTropics (ICRISAT), Patancheru, India during 1975 andseveral cultivars with field resistance / tolerance to SMDwere identified (Nene et al., 1981b). However, the task ofdeveloping resistant varieties has become complicated inview of the occurrence of geographical isolates of PPSMV(Kumar et al., 2004b). The pigeonpea genotypes, which werereported to be resistant to SMD at one location, were foundto be susceptible at other locations (Amin et al., 1993). Acomprehensive study over a period of four consecutive years,using a set of seven differentials, at nine different locationsin India, revealed the occurrence of five different variantsof the SMD pathogen in India (Reddy et al., 1993). Recentstudies based on bio-chemical characterization of PPSMVindicated that PPSMV isolates at Patancheru (P), Bangalore(B) and Coimbatore (C) are distinct from each other (Kumaret al., 2005a), and host resistance to PPSMV is scarce in thegermplasm. Moreover, a few genotypes that showedresistance to P isolate succumbed to infection against B andC isolates, indicating that these isolates have an ability toovercome resistance selected against P isolate (Reddy et al.,1993; Jones et al., 2004). Hence, in the present studypigeonpea genotypes were evaluated to identify sources ofresistance to PPSMV-B isolate."& "Seeds of 89 pigeonpea genotypes, obtained from ICRISAT,Patancheru and Indian Institute of Pulse Research (IIPR),Kanpur, were evaluated against PPSMV–B isolate byplanting in SMD screening nursery during Kharif (rainyseason) 2001. Pigeonpea cultivars, TTB 7 and ICP 8863were used as susceptible controls. Each genotype wasplanted in two replicated rows of five-meter length, with 25plants per row. The susceptible check ICP 8863 was plantedafter every two-test entries. As PPSMV-B is nottransmissible by mechanical inoculation, viruliferous mites,-.      www.IndianJournals.com            Members Copy, Not for Commercial Sale     Downloaded From IP - 220.225.236.59 on dated 3-Dec-2012were used for inoculation of all the test plants at two-leafstage (14-20 days after germination) (Nene and Reddy,1976). In this, mite-infested leaflets obtained from SMD-affected plants (maintained at University of AgricultureSciences, GKVK, Bangalore) were stapled to the leaves oftest plants. Mites from the stapled leaf migrate onto the testseedling resulting in virus transmission.The test entries were graded as resistant, tolerant orsusceptible based on per cent disease incidence and symptomtype at 75 DAS as per the rating scale given by Nene et al.(1981a) and Gupta et al. (1988) with minor modifications(Table 1). Selected test entries were evaluated for PPSMV-B by double antibody sandwich-enzyme-linkedimmunosorbent assay (DAS-ELISA) using the antibodiesproduced to PPSMV Patancheru isolate that can detect allthe PPSMV isolates, as described by Kumar et al. (2004b).Briefly, young leaflets from symptomatic and apparentlyhealthy plants were extracted in phosphate-buffered saline(1:10 w/v), and 100 µl of this was loaded into wells of ELISAplates pre-coated with PPSMV polyclonal antibodies at1:10,000 dilution. Penicillinase (PNC)-labelled PPSMVIgGs was used at 1:1,500 dilution to detect trapped antigen.Sodium penicillin G was used at 0.05 mg ml-1 in 0.015%(w/v) bromothymol blue buffer, pH 7.4. Absorbance valuesat 620nm (A620) were measured in an ELISA plate reader(Multiskan, Labsystems) and readings were considered tobe virus positive if the absorbance values of a sample werethree-folds or more than those given by the virus-free controlsamples. Observations on symptom type and severity wererecorded at 15 days intervals upto 75 DAS, by which time100% incidence was recorded in susceptible controls andall the plants showed severe symptoms.""The test genotypes were classified as resistant, moderatelyresistant, tolerant, moderately susceptible and susceptiblebased on the criteria given in Table 1. Recent developmentsfollowing the identification of SMD causal agent have paveda way for accurate monitoring of PPSMV incidence andprecise identification of resistant sources (Jones et al., 2004).Susceptible controls, ICP 8863, and TTB-7 showed 100%infection and they developed typical SMD symptoms in 14-20 days post inoculation (dpi) confirming the reliability ofvirus inoculation method (Table 2).Majority of the test entries developed severe mosaicsymptoms within 25-30 dpi, with 40-100% incidence (Table2). Only ICP 7035, MAL 14 and MAL 19, did not showany symptoms and were negative for virus in DAS-ELISAand classified as resistant. Earlier reports by Reddy and Nene(1980), Singh et al. (1989), Amin et al. (1993) andRangaswamy et al. (1997) indicated that ICP 7035 showedresistance to SMD at different locations in India, whichshows its broad-based resistance to various isolates of SMD.The genotypes ICP 6997 and ICP 8862 showed chloroticring spot symptoms, with per cent incidence of 50.9 and63.0, respectively and classified as tolerant (Table 2). InDAS-ELISA only symptomatic regions tested positive andnon-symptomatic areas were negative to the virus. Recentstudies have shown that systemic movement of the virusfrom inoculated leaves was absent in genotypes exhibitingchlorotic ringspots, and such symptoms were mostlyconfined to the site of mite inoculation indicating thatsymptoms were due to localized infection, and suchgenotypes show normal flowering pattern (P L Kumar,Personal communication). Apparent systemic symptomsobserved on such genotypes were due to multipleinoculations by the vector mites. ICP10976, MAL 10,BSMR-736 showed mild mosaic symptoms with <20%incidence and they were classified as moderately resistant;MAL 12, MAL 13, Bahar and KSMR-33 showed severemosaic symptoms but incidence was <20%, and they wereclassified as moderately susceptible; and rest of the 77Table 1. Rating scale for screening pigeonpea genotypes against sterility mosaic diseaseRating Genotype reaction to SMD CategoryNo symptoms on any plant, and no sterilitySevere mosaic symptoms on <10% plants or mild mosaic symptoms on <20% ofthe plants, without any noticeable stunting; and recovery of infected plants, withpartial sterilityRing spot symptoms on a few/all plants, and no sterilitySevere mosaic on 10-20% of the plants or mild mosaic symptoms on most plants,without any noticeable stunting, and partial sterilitySevere mosaic on >20% plants with severe stunting and near complete sterilityResistantModerately resistantTolerantModerately susceptibleSusceptible*Sterility = inhibition of flowering13579,-/Indian Journal of Plant Protection Vol. 34. No. 2, 2006 (216-220)      www.IndianJournals.com            Members Copy, Not for Commercial Sale     Downloaded From IP - 220.225.236.59 on dated 3-Dec-2012Table 2. Response of pigeonpea genotypes inoculatedwith PPSMV-B isolates using Aceria cajaniSl. No. Genotype Per cent Symptom Reactionidentity incidence type1 Bahar 11.7 SM MS2 BSMR-736 18.3 MM MR3 DPPA 85-14 90.0 SM S4 GUPH 1126-1 94.4 SM S5 GUPH 1126-9-2 97.3 SM S6 GUPH 1126-29-1 89.9 SM S7 GUPH 1126-29-2 96.9 SM S8 GUPH 1126-29-5 89.1 SM S9 GUPH 1126-47 89.7 SM S10 GUPH 1126-47-1 78.0 SM S11 GUPH 1126-47-2 92.1 SM S12 ICP 1206 67.1 SM S13 ICP 1207 22.7 SM S14 ICP 2376 87.0 SM S15 ICP 2668 81.6 SM S16 ICP 6997 50.9 RS T17 ICP 7035 0.0 NS R18 ICP 7039 94.0 SM S19 ICP 7550 94.6 SM S20 ICP 7867 95.9 SM S21 ICP 8087 93.4 SM S22 ICP 8094 74.0 SM S23 ICP 8362 81.8 SM S24 ICP 8610 100.0 SM S25 ICP 8860 85.0 SM S26 ICP 8862 63.0 RS T27 ICP 8869 81.8 SM S28 ICP 10976 13.7 MM MR29 ICP 10977 14.3 SM S30 ICP 10979 14.6 SM S31 ICP 10983 74.1 SM S32 ICP 11049 72.6 SM S33 ICP 11204 74.0 SM S34 ICP 11207 73.8 SM S35 ICP 11231 98.6 SM S36 ICP 11297 93.0 SM S37 ICP 12947 92.9 SM S38 ICP 13914 85.5 SM S39 ICP 14035 55.1 SM S40 ICP 14198 90.8 SM S41 ICP 14217 93.8 SM S42 ICP 14271 91.7 SM S43 ICP 14298 78.3 SM S44 ICP 14410 100.0 SM S45 ICP 14415 94.6 SM S46 ICP 14503 91.0 SM S47 ICP 14513 51.1 SM S48 ICP 14514 23.9 SM S49 ICP 14523 64.9 SM S50 ICP 14566 90.8 SM S51 ICP 14652 82.4 SM S52 ICP 14722 53.7 SM S53 ICP 14751 65.6 SM S54 ICP 14757 82.1 SM S55 ICP 14813 84.6 SM S56 ICP 14819 89.5 SM S57 ICP 14827 88.5 SM S58 ICP 15052 84.6 SM S59 ICP 16255 85.7 SM S60 ICP 16273 78.1 SM S61 ICP 16274 75.9 SM S62 ICP 16275 84.6 SM S63 ICP 16276 75.0 SM S64 ICP 87119 86.4 SM S65 ICP 93001 34.9 SM S66 ICPL 93003 73.2 SM S67 ICPL 96047 66.0 SM S68 ICPL 96053 70.0 SM S69 ICPL 96057 89.8 SM S70 ICPL 96061 62.8 SM S71 ICPL 99048 82.9 SM S72 ICPL 99051 100.0 SM S73 ICPL 99054 91.7 SM S74 ICPL 99055 96.2 SM S75 ICPX 900148-SMB 92.9 SM S76 IPH-487-75-1 73.2 SM S77 KPL-43 48.2 SM S78 KPL-272 92.3 SM S79 KSMR-33 13.5 SM MS80 MAL 10 17.7 M M MR,-0 Resistance to Sterility Mosaic Virus in Pigeonpea K M Nagaraj et al.,ContinuedTable 2. Continued      www.IndianJournals.com            Members Copy, Not for Commercial Sale     Downloaded From IP - 220.225.236.59 on dated 3-Dec-201281 MAL 12 14.5 SM MS82 MAL 13 13.7 SM MS83 MAL 14 0.0 NS R84 MAL 15 21.4 SM S85 MAL 18 23.9 SM S86 MAL 19 0.0 NS R87 PI-397430 85.3 SM S88 PR-5149 75.8 SM S89 PWS-1 89.5 SM SSusceptible controlsTTB-7 100.00 SM SICP 8863 100.00 SM SSM = Severe mosaic; RS = Ring spot; MM = Mild mosaic;NS = No symptoms; S = Susceptible; R = Resistant; MR =Moderately resistant; T = Tolerant; MS = Moderately susceptiblegenotypes showed severe mosaic symptoms with >20%incidence and were classified as susceptible (Table 2). Theresults showed that nearly 87% of the genotypes evaluatedwere susceptible to PPSMV-B isolate. Evaluation of wildCajanus species for SMD resistance indicated that fewergenotypes were resistant to PPSMV-B isolate (Kumar et al.,2005b). Genotypes, ICP 14410, ICP 8610 and ICPL 99051showed 100% incidence. In DAS-ELISA, the leaf samplescollected from genotypes, which exhibited mild mosaic(MAL 10, ICP 10976 and BSMR 736), chlorotic ring spots(ICP 6997 and ICP 8862) reacted positive to virus and allthe asymptomatic plants and also resistant genotypes (ICP7035, MAL 14 and MAL 19) tested negative. Broad-basedresistance to PPSMV in ICP7035 has been confirmed byvarious studies (Reddy et al., 1993). The genotypes MAL14 and MAL 19 needs to be further evaluated against otherPPSMV isolates in India.Owing to its resistance to PPSMV-B, superior agronomicperformance in multilocational trials, and its use forvegetable as well as grain purpose, ICP7035 has beenreleased for cultivation in Zone-5 region of Karnataka state(Rangaswamy et al., 2005). This genotype is also being usedin several breeding programmes as PPSMV-B resistancedonor. Similarly, resistant and tolerant varieties identifiedin this study can be exploited for cultivation in SMD endemicareas and also in resistance breeding programmes to mitigatelosses against SMD.)!%"&The first author is grateful to IIPR, Kanpur, India, forproviding the seed material of some genotypes evaluated inthis study. The work presented in this document is partlysupported from a project (No. R8205) funded by the CropProtection Program, Department for InternationalDevelopment (DFID), United Kingdom, for the benefit ofdeveloping countries. The views expressed are notnecessarily those of DFID.Amin, K. S., Reddy, M. V., Nene, Y. L., Raju, T. N., Shukla,P.,Zote, K. K., Arjunan, G., Bendre, J. N., Rathi, Y. P. S.,Sinha, B. K., Gupta, R. P., Anil Kumar, T. B., Chauhan, V.B., Singh, G., Jha, D. K. and Gangadharan, K. 1993.Multilocational evaluation of pigeonpea for broad basedresistance to sterility mosaic disease in India. Indian Journal ofAgriculture Sciences 63 : 542-546.Dhar, V. 2000. Seven decades of research on sterility mosaic ofpigeonpea. Indian Journal of Pulses Research 13 : 1-10.FAO 2005. FAOSTAT Data. http://www.faostat.fao.org/.Gupta, R. P., Singh, D. P. and Katiyar, R. P. 1988. Response ofpigeonpea lines/germplasm to sterility mosaic.  InternationalPigeonpea Newsletter 7 : 32-34.Jones, A. T., Kumar, P. L., Saxena, K. B., Kulkarni, N. K.,Muniyappa, V. and Waliyar, F. 2004. Sterility mosaic disease -the “green plague” of pigeonpea:  advances in understanding theetiology, transmission and control of a major virus disease. PlantDisease 88 : 436-445.Kumar, P. L., Jones, A. T. and Reddy, D. V. R. 2003. A novelmite-transmitted virus with a divided RNA genome closelyassociated with pigeonpea sterility mosaic disease.Phytopathology 93 : 81-91.Kumar, P. L., Jones, A. T. and Waliyar, F. 2004a. Biology, etiologyand management of pigeonpea sterility mosaic disease. AnnualReview of Plant Pathology 3 : 77-100.Kumar, P. L., Jones, A. T. and Waliyar, F. (Eds). 2004b. Serologicaland nucleic acid based methods for the detection of plant viruses:methods manual, ICRISAT,  Patancheru - 502 324, AndhraPradesh, India, 120pp.Kumar, P. L., Latha, T. K. S., Chauhan, V. B., Aparna, V. andWaliyar, F. 2005a. Studies on Pigeonpea sterility mosaic virusisolates in India. In Abstracts 2nd Asian Congress of PlantPathology 25-28 June 2005, National University of Singapore,Singapore. p 85-86.Kumar, P. L., Latha, T. K., Kulkarni, N. K., Raghavendra, K.,Saxena, K. B., Waliyar, F., Rangaswamy, K. T., Muniyappa,V., Sabitha Doriswamy and Jones, A. T. 2005b. Broad-basedresistance to pigeonpea sterility mosaic disease in wild relativesof pigeonpea (Cajanus: Phaseoleae). Annals of Applied Biology146 : 371-379.,-1Indian Journal of Plant Protection Vol. 34. No. 2, 2006 (216-220)Table 2. continued      www.IndianJournals.com            Members Copy, Not for Commercial Sale     Downloaded From IP - 220.225.236.59 on dated 3-Dec-2012Nene, Y. L. and Reddy, M. V. 1976. A new technique to screenpigeonpea for resistance to sterility mosaic. Tropical GrainLegume Bulletin 5 : 23-24.Nene, Y. L., Kannaiyan, J. and Reddy, M. V. 1981a. Pigeonpeadiseases: Resistance screening technique. Information Bulletin4: ICRISAT, Patancheru, Andhra Pradesh, India. pp. 14.Nene, Y. L., Kannaiyan, J. and Reddy, M. V. l981b. Resistanceto major pigeonpea diseases. In: Proceedings of InternationalWorkshop on Pigeonpea. Dec. 15-16, 1980. ICRISAT,Patancheru, India, pp.121-128.Rangaswamy, K. T., Sulladmath, V. V. and Byregowda, M.1997. Reaction of certain pigeonpea entries and varieties tosterility mosaic disease. In: Abst. Symposium on EconomicallyImportant Diseases of Crop Plants, IPS Southern chapter, Univ.Agric. Sci., Bangalore, Dec. 18 -20, pp. 74.Rangaswamy, K. T., Muniyappa, V., Kumar, P. L., Saxena,K. B., Byregowda, M., Raghavendra, N., Pandurangaiah,K., Kumar, R. V., Waliyar, F. and Jones, A. T. 2005. ICP7035 - A sterility mosaic resistant vegetable and grain purposepigeonpea variety for Karnataka state of India. InternationalChickpea and Pigeonpea Newsletter 12 : 42-44.Reddy, M. V. and Nene, Y. L. 1980. Estimation of yield loss due tosterility mosaic. In: Proceedings of International Workshop onPigeonpea 15-19, Dec. 1980. ICRISAT center, India Vol. 2,Patancheru, A.P. pp. 305-312.Reddy, M.V., Raju, T. N., Nene, Y. L., Ghanekar, A. M., Amin,K. S., Arjunan, G., Astaputra, J. V., Sinha, B. K.,Muniyappa, V., Reddy, S. V., Gupta, R. P. andGangadharan, K. 1993. Variability in sterility mosaic pathogenin India. Indian Phytopathology 46 : 206-212.Singh, R. P., Sharada Joshi, Shrestha, D., Karna, R. S. L. andBharati, M. P. 1989. Pigeonpea germplasm resistant to sterilitymosaic identified in Nepal. International Pigeonpea Newsletter9 : 21-22.Received and revised : 30-05-06, 17-11-06 Accepted : 23-11-06,,2 Resistance to Sterility Mosaic Virus in Pigeonpea K M Nagaraj et al.,

Evaluation of pigeonpea genotypes for resistance to pigeonpea sterility Mmsaic Vvrus - B Isolate

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Evaluation of pigeonpea genotypes for resistance to pigeonpea sterility Mmsaic Vvrus - B Isolate

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