【摘要】 【目的】哈密瓜细菌性果斑病在中国为新发生的病害,其病原菌为燕麦食酸菌西瓜亚种(Acidovorax avenae subsp.Citrulli),危害瓜果造成品质下降。该病为典型的种传细菌性病害,因此种子检疫成为防治此病害的重要手段。【方法】通过实验,将生物学、免疫学和分子生物学检测技术有机结合,建立了一套针对哈密瓜果斑病的种子带菌检测方法Bio-IMS-Real-timePCR。【结果】经过人工模拟种子带菌检测实验,结果表明该方法可成功地检测出1000粒种子中的1粒带菌种子(带菌量约为1.04×105CFU/种子,经计算,可以检测的最初浓度为2CFU·ml-1ASCM培养液),且信号较强。【结论】该检测方法的建立,大大提高了对哈密瓜细菌性果斑病菌检测的精度,也为生产实践中哈密瓜果斑病的防治提供了重要的技术支持。更多还原

【Abstract】 【Objective】Hami melon bacterial fruit blotch (HBFB) is a new disease in China and the pathogen is Acidovorax avenae subsp. citrulli, which damages the fruits and lowers their qualities. HBFB is a typical seedborne bacterial disease, and the important prevention and cure method is the quarantine of the seeds with bacteria.【Method】In this study, the immuno-detection was combined with PCR by using the superparamagnetic beads to form Bio-IMS-Real-time PCR, which increase the precision of bacteria detection. The integrated method of bacterial detection was formed through combining the ASCM culturing, IMS and real-time PCR. 【Result】As a result, one seed with bacteria (average 1.04×105 CFU/seed) in 1 000 seeds could be detected using this integrated method. The threshold of A. avenae subsp. citrulli detection for Bio-IMS-Real Time PCR was 2 CFU·ml-1 in ASCM liquid. 【Conclusion】This method can greatly increase the precision of A. avenae subsp. citrulli detection, and provide a important technical support for the prevention and control of HBFB.更多还原