ABSTRACT A virus disease of garden sage (Salvia splendens Ker-Gawl.) was observed and characterized showing symptoms of severe mosaic, mottling and distortion of leaves being remain shortened and growth retarded. The virus was transmitted to the healthy plants of Salvia spp. as well as many other hosts by mechanical inoculation, Myzus persicae Sulzer and Aphis gossypii Glover transmit the virus in non-persistent manner. Purified sample in EM showed spherical particles c.28 nm in diameter. DAC- ELISA  was performed with crude sap, specific polyclonal anti-serum (PVAS 242a, ATCC, USA) and alkaline phosphatase-linked secondary antibodies (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH or DSMZ, Germany). The mean absorbance at 405 nm for negative and positive controls were 0.061 ± 0.008 and 0.349 ± 0.003 respectively, while infected samples were recorded four-times more than the value of negative controls with values that ranged between 0.289 ± 0.005 and 0.325 ± 0.003. RT-PCR was performed using total RNA as templates and CMV Coat Protein (CP) gene specific reverse and forward primers, gel was electrophoresed on 1% agarose, an amplification of expected size 650 bp fragment was obtained only in the infected sample which proved that the present virus is a strain of CMV, the type member of the genus cucumovirus belonging to the family Bromoviridae.
Cite this paper
Ali, S. , Akhtar, M. , Singh, K. and Naqvi, Q. (2012) RT-PCR and CP gene based molecular characterization of a cucumber mosaic cucumovirus from Aligarh, U.P., India. Agricultural Sciences, 3, 971-978. doi: 10.4236/as.2012.38118.
 Verma, N., Singh, L., Singh, A.K., Kulshreshtha, S., Raikhy, G., Hallan, V., Ram, R. and Zaidi, A.A. (2005) Ornithogalum: A new host of Cucumber Mosaic Virus (CMV) from India. Plant Pathology, 54, 256.
 Pisi, A. and Vicchi, V. (1989) Infection by Cucumber Mosaic Virus (CMV) on Salvia sclarea in Italy. Informatore fitopathologico, 39, 49-51.
 Holcomb, G.E., Buras, H., Cox, P., Owings, A.D. and Valverde, R.A. (1998) Research on disease of ornamental plants. Luisiana Agriculture, 41, 4-5.
 Bellardi, M.G., Rubies, A. and Cerato, C. (2001) Identification of an isolate of potato virus—Y tuber necrotic strain of clary sage (Salvia sclarea); Plant Dis. (St. Paul. Minn). American Phytopathological Society, 85, 1207
 van Regenmortel, M.H.V., Fauquet, C.M., Bishop, D.H.L., Carstens, E.B., Estes, M.K., Lemon, S.M., Maniloff, J., Mayo, M.A., McGeoch, D.J., Pringle, C.R. and Wickner R.B (2000) Virus taxonomy classification and nomenclature of viruses. The 7th Report of International Committee on Taxonomy of Viruses, Academic Press, California.
 Palukaitis, P., Roossinck, M., Dietzgen, R.G. and Francki, R.I.B. (1992) Cucumber mosaic virus. Advances in Virus Research, 41, 281-348.
 Suzuki, M., Kuwata, S., Kataoka, J., Masuta, C., Nitta, N. and Takanami, Y. (1991) Functional analysis of deletion mutants of Cucumber mosaic virus RNA 3 using an in vitro transcription system. Virology, 183, 106-113.
 Wahyuni, W.S., Dietzen, R.G., Handa, K. and Francki, R.I.B. (1992) Serological and biological variation between and within subgroup I and II strains of Cucumber mosaic virus. Plant Pathology, 41, 282-297.
 Hu, J.S., Li, H.P., Berry, K., Wang, M. and Jordan, R. (1995) Comparison of dot blot ELISA and RT-PCR assays for detection of two cucumber mosaic virus isolates infecting banana in Hawaii. Plant Disease, 79, 202-206.
 Ilardi, V., Mazzei, M., Loreti, S., Tomassoli, L. and Barba, M. (1995) Biomolecular and serological methods to identify strains of cucumber mosaic cucumovirus on tomato. EPPO Bulletin, 25, 321-327.
 Owens, J. and Palukaitis, P. (1988) Characterization of Cucumber mosaic virus. I. Molecular heterogeneity mapping of RNA 3 in eight CMV strains. Virology, 69, 496-502
 Owens, J., Shintaku, M., Aeschleman, P., Tahar, S.F. and Palukaitis, P. (1990) Nucleotide sequence and evolutionary relationships of Cucumber Mosaic Virus (CMV) strains. CMV RNA 3. Journal of General Virology, 71, 2243-2249.
 Szilassy, D., Sala’nki, K. and Bala’zs, E. (1999) Molecular evidence for the existence of two distinct subgroups in Cucumber mosaic cucumovirus. Virus Genes, 18, 221-227. doi:10.1023/A:1008016202128
 Rizos, H., Gunn, L.V., Pares, R.D. and Gillings, M.R. (1992) Differentiation of cucumber mosaic virus isolates using polymerase chain reaction. Journal of General Virology, 73, 2099-2103.
 Sialer, M.M., Cillo, F., Barbarossa, L. and Gallitelli, D. (1999) Differentiation of Cucumber mosaic virus sub-groups by RT-PCR, RFLP. Journal of Plant Pathology, 81, 145-148
 Palukaitis, P. and Zaitlin, M. (1997) Replicase-mediated resistance to plant virus diseases. Advances in Virus Research, 48, 349-377.
 Roossinck, M.J. (2002) Evolutionary history of Cucumber mosaic virus deduced by phylogenetic analysis. Journal of Virology, 76, 3382-3387.
 Eiras, M., Boari, A.J., Colariccio, A., Chaves, A.L.R., Briones, M.R.S., Figueira, A.R. and Harakava, R. (2004) Characterization of isolates of the cucumovirus Cucumber Mosaic Virus present in Brazil. Journal of Plant Pathology, 86, 61-69.
 Singh, D., Naqvi, Q.A. and Garg, I.D. (1999) A strain of Cucumber mosaic virus causing mosaic in marigold in India. Indian Phytopathology, 52, 114-117.
 Walkey, D.G.A. (1985) Applied plant virorogy. William Heinmann Ltd., London.
 Christie, S.R., Purciful, D.E., Grawford, W.E. and Ahmad, N.A. (1987) Electron microscopy of negatively stained clarified virus concentrates obtained from small tissue samples with appendices of negative staining techniques. Institute of Food and Agriculture Science, University of Florida Technical Bulletin, Gainesville.
 Garg, I.D. and Khurana, S.M.P. (1992) Factors influencing immune electron microscopy of flexuous potato viruses. Acta Virologica, 36, 435-442.
 Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989) Molecular coning: A laboratory manual. Cold Spring Harbor Laboratory, New York.
 Srivastava, A., Chandra, G. and Raj, S.K. (2004) Coat protein and movement protein gene based molecular characterization of Amaranthus strain of cucumber mosaic virus. Acta Virologica, 48, 229-239.
 Thompson, J.D., Higgins, D.G. and Gibson, T.J. (1994) Clustal W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighing, position-specific gap penalties and weight matrix choice. Nucleic Acids Research, 22, 4673-4680.
 Tamura, K., Dudley, J., Nei, M. and Kumar, S. (2007) Mega4: Molecular evolutionary genetics analysis (MEGA) software version 4.0. Molecular Biology and Evolution, 24, 1596-1599. doi:10.1093/molbev/msm092
 Wang, X.F., Pei, M.Y., Ma, D.F. and Tian, W.H. (1985) Studies on scarlet sage mosaic disease. Acta Phytopathologica Sinica, 15, 36-46.
 Hu, J.S., Li, H.P., Barry, K., Wang, M. and Jordan, R. (1995) Comparision of dot.blot ELISA and RT-PCR assay for detection of two cucumber mosaic isolates infecting banana in Hawaii. Plant Disease, 79, 902-906.
 Carrere, I., Tepfer, M. and Jacquemond, M. (1999) Recombinants of Cucumber Mosaic Virus (CMV): determinants of host range and symptomatology. Archives of Virology, 114, 365-379.
 Takeshita, M., Suzuki, M. and Takanami, Y. (2001) Combinations of amino acids in the 3a protein and the coat protein of cucumber mosaic virus determines symptom expression and virus spread in bottle gourd. Archives of Virology, 146, 697-711. doi:10.1007/s007050170140