Optimization and validation of designed primer pairs in tagging virus-host factors of abaca bunchy top virus (ABTV) and gene-specific markers in abaca (Musa textilis Nee)

Date

7-2015

Degree

Bachelor of Science in Agricultural Biotechnology

College

College of Agriculture and Food Science (CAFS)

Adviser/Committee Chair

Antonio G. Lalusin

Abstract

Six newly designed markers were subjected to optimization of polymerase chain reaction (PCR) conditions. Three of these primers were designed for tagging virus-host factors of abaca bunchy top virus (ABTV) and three of these were designed as gene-specific primers for abaca. Only three out of the six primers were successfully optimized ? the Proliferating Cell Nuclear Antigen (PCNA), RAD54 and 348bI2. It was identified that the optimum annealing temperatures for each of the markers were 57 °C, 61 °C and 63.8 °C, respectively. The effectiveness of the optimized primers was validated through screening across 38 abaca samples consisting of 20 mutants, 15 backcrosses, and three parentals -Abuab, Pacol and Inosa. Results showed that the primers PCNA and RAD54 are effective in detecting samples susceptible to abaca bunchy top virus (ABTV) and the primer 348bI2 is effective in detecting abaca samples with good fiber quality. Among the 38 samples, TPA 16 and BC2 C were found to be resistant to ABTV while the samples TPA 2, TPA 20, TPA 9, BM2 3, AM2 46 and MB 20 were found to be resistant to ABTV at the same time having good fiber quality.

Language

English

Location

UPLB Main Library Special Collections Section (USCS)

Call Number

LG 993.5 2015 A127 /B36

Notes

Major in Crop Biotechnology

Document Type

Thesis

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