Biological and Molecular Characterization of Two Distinct Tomato Strains of Cucumber mosaic virus Based on Complete RNA-3 Genome and Subgroup Specific Diagnosis
- 135 Downloads
Cucumber mosaic virus (CMV) is one of the most important viral pathogen infecting several plant species in India. Five isolates of CMV obtained from cucumber, muskmelon, tobacco and tomato from distinct geographical locations in India were analysed based on host-reactions and genome sequence. The majority of the isolates were very similar and only two isolates, Tfr-In and Tss-In showed distinct symptoms in tomato and high sequence diversity (77.8%) in coat protein (CP) gene. Tfr-In was isolated from tomato fruit showing grey patches in Aurangabad and Tss-In from tomato plant showing shoe-string symptoms in New Delhi. The RNA-3 genomes of Tfr-In (2,214 nt; JF279606), shared only 70.3% nucleotide sequence identity with Tss-In (2,178 nt; JF279605. The complete RNA-3 genome of Tss-In and Tfr-In were compared with that of 65 CMV isolates reported from various plants of the world, which formed four distinct subclades-IA, -IB, -IC and -II. The Tfr-In isolate clustered with the CMV subgroup-IB and Tss-In with the subgroup-II. The comparison of the RNA-3 sequence of both the isolates revealed maximum heterogeneity in the intergenic region (IR). Reverse transcriptase polymerase chain reaction (RT-PCR) based detection of CMV subgroup-I and -II was developed designing primers from flanking IR region. The specificity of the RT-PCR detection was confirmed using Tfr-In and Tss-In representing subgroup-I and -II and validated with field samples of tomato, cucurbits and chilli. This is the first report of complete RNA-3 of subgroup-IB CMV causing grey patches in tomato fruit and subgroup-II CMV causing shoe-string symptoms in tomato in India. The present and previous studies together showed that tomato in India was affected by multiple strains of CMV.
KeywordsCucumber mosaic virus Tomato CMV subgroups Shoe-string Grey-patch RT-PCR RNA-3 genome
- 1.Aglave BA, Krishnareddy M, Patil FS, Andhale MS. Molecular identification of a virus causing banana chlorosis disease from Marathwada region. Int J Biotechnol Biochem. 2007;3:13–23.Google Scholar
- 2.Akhtar KP, Ryu KH, Saleem MY, Asghar M, Jamil FF, Haq MA. Occurrence of Cucumber mosaic virus subgroup IA in tomato in Pakistan. J Plant Dis Prot. 2008;115:2–3.Google Scholar
- 5.Bhat AI, Hareesh PS, Madhubala R. Sequencing of coat protein gene of an isolate of Cucumber mosaic virus infecting black pepper (Piper nigrum L.) in India. J Plant Biochem Biotechnol. 2005;14:37–40.Google Scholar
- 8.Eck RV, Dayhoff MO. Atlas of protein sequence and structure. Silver Springs: National Biomedical Research Foundation; 1966.Google Scholar
- 11.Nei M, Kumar S. Molecular evolution and phylogenetics. New York: Oxford University Press; 2000. pp. 352.Google Scholar
- 15.Raj SK, Srivastava A, Chandra G, Singh BP. Characterisation of Cucumber mosaic virus isolate infecting Gladiolus cultivars and comparative analysis of serological and molecular methods for sensitive diagnosis. Curr Sci. 2002;83:1132–6.Google Scholar
- 21.Roossinck MJ, Bujarski J, Ding SW, Hajimorad R, Hanada K, Scott S, Tousignant M. Bromoviridae. In: van Regenmortel MHV, Fauquet CM, Bishop DHL, Carstens EB, Estes MK, Lemon SM, Maniloff J, Mayo MA, McGeoch DJ, Pringle CR, Wickner RB, editors. Virus taxonomy: classification and nomenclature of viruses. Seventh Report of the International Committee on the Taxonomy of Viruses. San Diego: Academic Press; 2000. p. 923–35.Google Scholar
- 22.Salamon P, Gajdos L, Balogh P, Solymoss E, Varro P, Milotay P, Kiss L, Salanki K. Isolation of a virulent strain of Cucumber mosaic virus from tomato (Lycopersicon esculentum Mill.) affected by fruit necrosis. Kertgazdasag Hortic. 2005;(special):155–61.Google Scholar
- 24.Sambrook J, Russell DW. Molecular cloning, a laboratory manual. 3rd ed. Cold Spring Harbor: Cold Spring Harbor Laboratory Press; 2001.Google Scholar
- 26.Singh Z, Jones RA, Jones MGK. Identification of Cucumber mosaic virus subgroup I isolates from banana plants affected by infectious chlorosis disease using RT-PCR. Plant Dis. 1995;79:716–31.Google Scholar
- 27.Srivastava A, Raj SK. High molecular similarity among Indian isolates of Cucumber mosaic virus suggests a common origin. Curr Sci. 2004;87:1126–31.Google Scholar
- 34.Wang HY, Zhang HR, Du ZY, Zeng R, Chen J. Evolutionary characterization of two Cucumber mosaic virus isolates infecting Pinellia ternata of Araceae family. 2009; http://www.paper.edu.cn/index.php/default/en_releasepaper/downPaper/200902-1424.