RT-PCR and CP Gene Based Molecular Characterization of A Cucumber Mosaic Ccumovirus from Aligarh, U.P., India

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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. Pu- rified sample in EM showed spherical particles c.28 nm in diameter. DAC-ELISA [1] was per- formed with crude sap, specific polyclonal anti- serum (PVAS 242a, ATCC, USA) and alkaline phosphatase-linked secondary antibodies (Deuts- che Sammlung von Mikroorganismen und Zell- kulturen 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 re- corded four-times more than the value of nega- tive controls with values that ranged between 0.289 ± 0.005 and 0.325 ± 0.003. RT-PCR was per- formed using total RNA as templates and CMV Coat Protein (CP) gene specific reverse and for- ward 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 cu- cumovirus belonging to the family Bromoviridae.

Shahid Ali
Masood Akhtar
Kangabam S. Singh
Qamar A. Naqvi
Agricultural Sciences