Cloning and detection of rice tungro bacilliform virus in plants and leafhoppers by nucleic acid hybridization
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Authors
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G. RADHAKRISHNAN and F.R. NIAZI*
Division of Plant Pathology, Indian Agricultural Research Institute, New Delhi 110 012
Keywords:
Rice tungro bacilliform virus, cloning, detection, nucleic acid spot hybridizationAbstract
Rice tungro spherical virus (RTSV) and rice tungro bacilliform virus (RTBV) were purified from tungro infected Oryza sativa cv. TN1. RTBV-DNA was isolated and cloned into pUC18 vector. The viral nature of the two clones having an insert of ? kb, designated as pRTBV-E6, and of ? kb, designated as pRTBV-E2, was confirmed by Southern hybridization using radiolabelled pRTBV201 probe. Specificity of pRTBV-E6 and pRTBV-E2 probes was confirmed by their hybridization with the DNA of RTBV infected TN1 leaves and lack of hybridization with the DNA of citrus mosaic virus (a badnavirus). Radiolabelled pRTBV-E6 and pRTBV-E2 probes detected RTBV infection in roots, stem, symptomatic and youngest leaves of variety TN1. Nucleic acid spot hybridization (NASH) using pRTBV-E2 probe detected RTBV from viruliferous adults as well as third instar nymphs of the leafhopper vector.
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