›› 2018, Vol. 30 ›› Issue (3): 426-431.DOI: 10.3969/j.issn.1004-1524.2018.03.11

• Plant Protection • Previous Articles     Next Articles

Duplex PCR detection of Burkholderia gladioli pv. alliicola and Erwinia chrysanthemi

YANG Wanfeng1, LIU Yan2, LU Chenchen1, SHAO Peize1, CHEN Yunqing1, ZHAO Wenjun3   

  1. 1. Lianyungang Entry-Exit Inspection and Quarantine Bureau, Lianyungang 222042, China;
    2. Lianyungang Academy of Agricultural Sciences, Lianyungang 222001, China;
    3. Chinese Academy of Inspection and Quarantine, Beijing 100029, China
  • Received:2017-07-19 Online:2018-03-20 Published:2018-03-21

Abstract: The objective of this study is to establish a duplex PCR technology for simultaneously detecting Burkholderia gladioli pv. alliicola and Erwinia chrysanthemi. Based on the 16S-23S rRNA intergenic transcribed spacer sequence of B. gladioli pv. alliicola in GenBank, the primers B3/B6 were designed. The duplex PCR assay had been developed using the combining primers B3/B6 and ADE1/ADE2 which were specific primers to detect E. chrysanthemi. The reaction conditions were optimized and the specificity and sensitivity of the duplex PCR were tested. The expected DNA fragment could be specifically amplified from the genomic DNA of B. gladioli pv. alliicola and E. chrysanthemi. Specificity was confirmed using the duplex PCR assay to detect B. gladioli pv. alliicola and E. chrysanthemi in the artificially inoculated tulip leaf samples. The duplex PCR developed in this study can be used in detecting the two pathogens for imported tulip bulbs quarantine.

Key words: Burkholderia gladioli pv. alliicola, Erwinia chrysanthemi, duplex PCR, detection

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