Cloning of the DNA 3 of the bunchy top nanavirus in abaca
2004
Villamor, D.E.V. | Galvez, L.C. | Barcial, P.M. | Bajet, N.B.,Philippines Univ. Los Banos, College, Laguna (Philippines). Dept. of Plant Pathology
Banana bunchy top nanavirus (BBTV) causes one of the most serious diseases of banana. A very similar disease called abaca bunchy top, also affects abaca and the causal virus of viruses have been assumed but not proven to be similar, if not the same viruses. The BBTV genome has six components with the open reading frame (ORF) of the DNA 3 coding for the coat protein gene. Two outwardly extending primers for the DNA 3 of BBTV (Wanichakorn et al. 2000 Arch. Virol 145:593-602) were used on a polymerase chain reaction (PCR) to amplify the putative DNA 3 of the bunchy top nanavirus in abaca (AbaBTV). A PCR product of about 1000 base-pairs (bp) was amplified from the total nucleic acid extracts of bunchy top infected banana and abaca, but none from the extracts of the correspondingly healthy banana and abaca tissues. This PCR product was cloned into a topoisomerase activated vector and three representative clones coming from each of the BBTV-infected banana and AbaBTV-infected abaca were selected. Analysis of the selected clones by colony PCR confirmed the presence of a 1000 bp insert. The result of the study provides evidence on the presence of the virus genome DNA 3 component and, in general, the similarity of the two viruses infecting the two crops at the molecular level. This is also the first report on the cloning of one particular DNA component of the bunchy top virus in abaca. Nucleotide sequence analysis of these clones is further pursued to determine the degree of similarity between the two DNA 3 components.
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