Molecular virus resistance screening for abaca varietal improvement program against abaca bunchy top viruses
2023
Piamonte, R.T.
Abaca bunchy top, caused by Banana bunchy top virus (BBTV) and Abaca bunchy top virus (ABTV), posed the most serious damage to the Philippine abaca industry. The disease continues to devastate many farms in Luzon, Visayas and Mindanao regions of the country. In the abaca rehabilitation and virus resistance breeding programs, virus resistance screening is crucial and has to use a sensitive and reliable virus disease diagnostic. To fast-track the screening selection of the developed and collected abaca genotypes for resistance against bunchy top viruses, a molecular virus indexing through polymerase chain reaction (PCR) was developed and optimized. The optimized PCR condition was then used to screen the abaca germplasm collections of the National Abaca Research Center for bunchy top resistance. The method of DNA extraction and the dilution of the template DNA affect the sensitivity and reliability of BBTV detection by PCR. Virus detection was most sensitive when the template DNA was extracted by Dellaporta method. Detection level increases as DNA template concentration is reduced. The BBTV was highly detectable at 10 and 1 ng/ul template DNA concentrations using BBT1/BBT2 primers. The ABTV was detectable using SLCR/CRMB primers at 100ng/ul template DNA. The use of an internal control DNA, a Musa sequence tagged microsattelite AGMIO25/AGMI026, proved the presence of PCR inhibitors in the DNA suspension. The optimized protocol for bunchy top virus detection in abaca hastened the selection process of abaca genotypes potentially resistant to abaca bunchy top viruses.
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