PMA-PCR方法进行水稻细菌性条斑病菌细胞活性检测的建立与初步应用  

Establishment and preliminary application of a PMA-PCR method for selective detection of viable Xanthomonas oryzae pv.oryzicola cells

作　　者：于贵戌 赵玉强[3] 姜培 田艳丽[1,2] 胡白石 YU Guixu;ZHAO Yuqiang;JIANG Pei;TIAN Yanli;HU Baishi(College of Plant Protection,Nanjing Agricultural University,Nanjing 210095,China;Key Laboratory of Monitoring and Control Plant Quarantine Pests,Ministry of Agriculture and Rural Affairs,Nanjing 210095,China;Institute of Botany,Jiangsu Province and Chinese Academy of Sciences(Nanjing Botanical Garden Mem.Sun Yat-Sen),Nanjing 210014,China;National Agricultural Technology Extension Service Center,Beijing 100125,China)

机构地区：[1]南京农业大学植物保护学院,南京210095 [2]农业农村部植物检疫性有害生物监测防控重点实验室,南京210095 [3]江苏省中国科学院植物研究所(南京中山植物园),南京210014 [4]全国农业技术推广服务中心,北京100125

出　　处：《植物病理学报》2023年第5期944-949,共6页Acta Phytopathologica Sinica

摘　　要：水稻细菌性条斑病是危害水稻的重要细菌病害。本文利用叠氮溴化丙锭(propidium monoazide, PMA)与PCR技术相结合,建立了一种快速有效区分水稻细菌性条斑病菌死活细胞的分子检测方法。结果表明,当样品中PMA质量浓度为5μg·mL-1,避光条件下,孵育5 min,曝光20 min, PMA可有效抑制死亡菌体细胞中的DNA扩增;且对活菌DNA的扩增无影响;最低的检测浓度为2×104CFU·mL-1。本试验建立的PMA-PCR方法能够有效的区分水稻细菌性条斑病菌死活细胞,避免了假阳性结果的产生。Bacterial leaf streak is an important rice disease caused by Xanthomonas oryzae pv.oryzicola(Xoo).In this paper,we developed a rapid and effective method using PCR combined with propidium monoazide(PMA)to distinguish dead and living Xoo cells.The results showed that PMA at 5μg·mL^(-1)could effectively inhibit the DNA amplification from dead cells after an incubation period of 5 min and light exposuring for 20 min.The minimum detectable concentration of living Xoo cells is 2×10^(4) CFU·mL^(-1).The PMA-PCR method established in this study can effectively distinguish the dead and living cells of Xoo and avoid false-positive results.

关 键 词：水稻细菌性条斑病 叠氮溴化丙锭 活菌 

分 类 号：S432.42[农业科学—植物病理学]