PJB-2019-83
Development of specific real-time PCR primers for detecting Pseudomonas syringae pv. Actinidiae in China
Chao Yu, Jin-Tian Xie, Yue-Yan Wu, Li-Meng Ma and Zhong-Hua Wang
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
To Cite this article:
Yu, C., J.T. Xie, Y.Y. Wu, L.M. Ma and Z.H. Wang. 2019. Development of specific real-time PCR primers for detecting Pseudomonas syringae pv. Actinidiae in China. Pak. J. Bot., 51(2): DOI: http://dx.doi.org/10.30848/PJB2019-2(36)
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