By P. Narayanasamy
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Extra info for Molecular Biology in Plant Pathogeneses and Disease Management Disease Development
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When purified virus proteins were subjected to SDS-PAGE and silver staining, similar results were obtained (Fig. 3) [Appendix 7] As there was no reaction with antisera raised against potyviruses Bean Yellow mosaic virus, Clover yellow vein virus, Cowpea aphidborne mosaic virus, Pea seedborne mosaic virus, Bean common mosaic virus or Bean common mosaic necrosis virus, the CYMV was considered as a distinct member of Potyviridae (Larsen et al. 2003). coli. In Western blot analysis with total protein extracts from BYDV-infected plants antisera effectively recognized the 99 kDa fusion protein expressed from ORF1 and 2 (Fomitcheva et al.
2005a) have been detected efficiently by using PABs or MABs in their respective hosts, some of them carrying the pathogen (s) without showing any visible symptoms. The antisera may be prepared against antigens present on the surface of mycelium or spores. In some cases, antisera have been prepared against specific protein (s) extracted from the test fungal pathogens and separated by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE). Purified mycelial proteins were used to generate PABs against Verticillium dahliae and these antibodies reacted positively with 11 of 12 isolates of V.
Different hybridization methods such as colony and dot-blot hybridization have been performed for detection and qualitative differentiation to distinguish groups of plant pathogens, when a specific probe is available. By using appropriate restriction enzymes, restriction fragment length polymorphism (RFLPs) and analyes of natural variations in the genomes of different groups or strains of organisms can be assessed. The variations (polymorphisms) in fragment sizes can be generated by loss or gain of restriction endonuclease recognition sites, or by other events that change fragment sizes such as deletions or insertions in the DNA sequences.