PMA-PCR检测方法的优化及瓜类种子细菌性果斑病的检测应用OA
Optimization of PMA-PCR Detection Method and Its Application in Detecting Bacterial Fruit Blotch in Cucurbit Seeds
西瓜细菌性果斑病是严重危害葫芦科作物的重要种传细菌病害.利用叠氮溴化丙锭(propidium monoazide,PMA)与PCR技术相结合,建立了一种可特异性检测西瓜细菌性果斑病菌活细胞的PMA-PCR方法.结果表明,当PMA浓度为10 μmol·L-1,避光孵育10 min后曝光20 min,可有效抑制死菌DNA的扩增,且不影响活菌检出;活菌的最低检测下限为1×104 CFU·mL-1.应用所建立的PMA-PCR检测方法对西瓜、葫芦和甜瓜种子进行检测,结果显示其能够快速稳定区分带菌与灭活种子,准确鉴定活菌存在.该检测方法为西瓜细菌性果斑病的种子健康检测与早期防控提供了可靠的技术依据.
Bacterial fruit blotch of watermelon is an important seed-borne bacterial disease that severely affects cucurbit crops.A PMA-PCR method was developed for the specific detection of live cells of the bacterial fruit blotch pathogen of watermelon by combining propidium monoazide(PMA)with PCR technology.The results showed that when the PMA concentration was 10 μmol·L-1,followed by incubation in the dark for 10 min and exposure for 20 min,the amplification of DNA from dead bacteria could be effectively inhibited without affecting the detection of live bacteria.The minimum detection limit for live bacteria was 1×104 CFU·mL-1.The established PMA-PCR method was applied to detect watermelon,bottle gourd,and melon seeds,and the results demonstrated that it could quickly and stably distinguish between contaminated and sterilized seeds,accurately identifying the presence of live bacteria.This detection method provides reliable technical support for seed health testing and early control of bacterial fruit blotch of watermelon.
乔琳芝;汤冰倩;周倩;何长征;唐宁;戴思慧;卜凤姣
湖南农业大学园艺学院,湖南 长沙 410128湖南农业大学园艺学院,湖南 长沙 410128||岳麓山实验室,湖南 长沙 410128湖南农业大学植物保护学院,湖南 长沙 410128湖南农业大学园艺学院,湖南 长沙 410128湖南农业大学园艺学院,湖南 长沙 410128湖南农业大学园艺学院,湖南 长沙 410128湖南农业大学园艺学院,湖南 长沙 410128||岳麓山实验室,湖南 长沙 410128
西瓜细菌性果斑病叠氮溴化丙锭活菌检测
watermelonbacterial fruit blotchpropidium monoazide(PMA)viable bacteria detection
《中国蔬菜》 2026 (8)
193-199,7
岳麓山实验室种业专项(YLS-2025-ZY02015),国家西甜瓜产业技术体系项目(CARS-25),岳麓山实验室人才专项(2025RC2058,2024RC2011)
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