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ACTA HORTICULTURAE SINICA ›› 2016, Vol. 43 ›› Issue (12): 2473-2480.doi: 10.16420/j.issn.0513-353x.2016-0198

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Development of TaqMan Real-time PCR Assay of Banana bunchy top virus

SUN Jie,WANG Wan,RAO Xue-qin*,and LI Hua-ping*   

  1. (State Key Laboratory of Conservation and Utilization of Subtropical Agro-bioresources,Guangdong Province Key Laboratory of Microbial Signals and Disease Control,College of Agriculture,South China Agricultural University,Guangzhou 510642,China)
  • Online:2016-12-25 Published:2016-12-25

Abstract:

A TaqMan real-time PCR assay was developed by designing a set of primers and probe based on the conserved replicase gene of Banana bunchy top virus(BBTV). The results showed that the assay was more sensitive than Endpoint PCR in detecting both the standard DNA plasmid and total DNA extracted from field samples,respectively,and had no cross reaction with two other common virusesCucumber mosaic virus(CMV)and Banana streak virus(BSV)in the banana plants. Seven times of reproducibility tests were analyzed and the results showed that Ct coefficient of variation was 0.93% and the Ct value kept stable in each reaction. This TaqMan real-time PCR assay was utilized to detect BBTV concentrations in diseased micropropagated banana shoots,and the results indicated that the concentrations of BBTV in the micropropagated banana shoots of‘Yueyoukang 1increased along with tissue-cultured generations,which was higher in the top leaves than those in other leaves.

Key words: banana, Banana bunchy top virus, TaqMan real-time PCR, micropropagated banana shoot

CLC Number: