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Real-Time Fluorescence PCR Identification of Alternaria alternata Causing Black Spot Disease of Fragrant Pear

WANG Feng-jun，ZHANG Xiang-lin，MA Hai-bo，FENG Xin-zhong，FENG Jun-li，HUA Peng   

  1. 1. Korla Entry-Exit Inspection and Quarantine Bureau, Korla 841000, China；
    2. Xinjiang Entry-Exit Inspection and Quarantine Bureau, Ürümqi 830000, China；
    3. College of Agriculture, Xinjiang Agricultural University, Ürümqi 830000, China；
    4. Institute of Bioengineering, Zhejiang Sci-Tech University, Hangzhou 310018, China
  • Online:2013-10-25 Published:2013-09-28

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Abstract:

A real-time fluorescence PCR method was established by using a pair of specific primers and double-fluorescence
labeling probe according to the conserved sequence of the pathogen Alternaria alternata causing fragrant pear black
spots. Totally 12 flowers, 30 fruits and 94 slides from fragrant pears were detected in this study. The real-time fluorescence
PCR was used to detect four other plant disease pathogens on pears. Results showed that only in the pathogen causing black
spots, fluorescence signal could be collected with the specific primers and probe. The assay for specific detection was more
sensitive than conventional PCR, and could detect the concentration of DNA template as low as 1.9 pg/μL without pathogen
separation and purification. This method is a reliable, sensitive, quick and easy-handling method suitable for rapid diagnosis
and dynamic monitoring of the plant pathogen.

Key words: fragrant pear, Alternaria alternata, real-time fluorescence polymerase chain reaction, TaqMan probe, rapid detection