Molecular detection and characterization of banana bunchy top virus and somaclonal variants in tissue culture banana plants
KrishiKosh
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Title |
Molecular detection and characterization of banana bunchy top virus and somaclonal variants in tissue culture banana plants
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Creator |
W. A. R. T, Wickramaarachchi
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Contributor |
K. T, Rangaswamy
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Subject |
bananas, dna, planting, pcr, biological phenomena, viruses, proteins, diseases, tissue culture, productivity
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Description |
In the recent years, banana viral diseases are effectively managed by using virus free tissue culture planting materials. Virus indexing of large number of tissue culture planting materials indicated the presence of viruses in very low percentage. Field survey of banana plantations for prevalence of viruses revealed the low incidence of viral diseases in banana crops grown using tissue culture plants compared to sucker grown plantations. However, the incidence of off types ranging from 1.3 to 24.0 per cent was observed in banana crop established using tissue culture plants. Dwarf off type was the most frequent off type observed and contributed to 70 per cent of the total off types recorded. RAPD primer OPJ-04 which supported the amplification of an approximate product of 1.5 kb from DNA extracted from normal plants but not from dwarf plants was found suitable for detecting dwarf off types from micropropagated banana cv. Grand Naine. Multiplex PCR technique with a SCAR marker and an internal control was developed to increase the reliability of the detection of dwarf off types. The specific polyclonal antibodies produced against bacterially expressed coat protein of BBTV showed strong reaction with crude sap from infected plant tissues in western blot, dot blot immunoassay and DAC-ELISA. The antiserum had a titre of 1:2000. Multiplex PCR technique was developed for parallel detection of both BBTV and BSV using virus specific primers. A Novel, rapid and cost effective technique was developed for mass indexing of tissue culture banana samples for BBTV with a relatively less number of PCR reactions using combinatorial screening format. A PCR technique for detection of BBTV in viruliferous aphids was standardized using the template DNA extracted from minimum of five aphids with “Jalali” DNA extraction method. Analysis of sequences of major genomic components of Bangalore and Sri Lankan isolates of BBTV showed clustering of both isolates together with other Indian isolates in South Pacific group. High degree of genetic homology of Sri Lankan isolate with Bangalore isolate and with other isolates from India suggested a close genetic relationship of Sri Lankan and Indian BBTV isolates. |
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Date |
2016-05-26T10:41:16Z
2016-05-26T10:41:16Z 2011-11-18 |
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Type |
Thesis
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Identifier |
Th-9939
http://krishikosh.egranth.ac.in/handle/1/66332 |
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Language |
en
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Format |
application/pdf
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Publisher |
University of Agricultural Sciences, Bengaluru
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