Development of a polymerase chain reaction-based technique for the detection of Philippine banana strains of Ralstonia solanacearum (Smith) Yabuuchi et al.

Abstract

Based on the DNA sequence of a repetitive element previously cloned and characterized from the bugtok banana strain of Ralstonia solanacearum, different primers were designed and used in polymerase chain reaction (PCR). Outward primers produced several amplicons and did not distinguish banana from non-banana strains. In contrast, the 21-mer oligonucleotides (M114F and M114R primer pair) annealed at 65 C amplified a unique 2.28 kb PCR fragment that distinguished the banana strains from the rest. The M114 primers, however, when used together with 759/760 primers specific for R. solanacearum in multiplex PCR reactions, did not produce the desired 2.28 kb fragment. This study primarily reports the development of a PCR-based technique that is specific to R. solanacearum strains infecting Philippine banana by establishing specific primers based on a repetitive element and by establishing the most stringent annealing temperature of the PCR process.

Source or Periodical Title

Philippine Agricultural Scientist

ISSN

317454

Page

385-393

Document Type

Article

Subject

Amplification, Detection, PCR, Primers, Ralstonia solanacearum

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