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Development of specific real-time PCR primers for detecting Pseudomonas syringae pv. Actinidiae in China
Abstract
Pseudomonas syringae pv. Actinidae (Psa) is the most devastating disease of kiwifruit cultivation. In this study, to detect Psa in China high specifically and sensitively, a method of real-time PCR was developed. The specific primers YF32/YR32 was set for the 590 bp hrpW gene fragment, with amplicon of 248bp in length. The standard curves of real-time PCR clearly showed a suitable condition of real-time PCR and an excellent linear of the data. The specificity and sensitivity assay showed this method could specifically discriminate between Psa and Psa-related pseudomonads and sensitivity threshold was 100fg/μL. In the actual effect validation experiments, the results of all of 5 twig samples with symptoms were positive, and 3 of 5 symptomless samples were positive with minimum DNA concentration 8.45×10‐5ng/μL. The methods of this study could make an important contribution for the prevention and diagnosis of the bacterial canker disease of kiwifruit in China
