蕹菜溃疡病菌分子检测方法的建立及田间监测  

Detection and Field Monitoring of Water Spinach Canker

作　　者：陈思妍 薛洋 韩海亚 林敏怡 黎旭宇[4] 尹德明 陶杰 李世茂 彭李亚 席卓君 周佳暖 CHEN Siyan;XUE Yang;HAN Haiya;LIN Minyi;LI Xuyu;YIN Deming;TAO Jie;LI Shimao;PENG Liya;XI Zhuojun;ZHOU Jianuan(National Key Laboratory of Green Pesticide/Guangdong Province Key Laboratory of Microbial Signals and Disease Control/Integrative Microbiology Research Center,South China Agricultural University,Guangzhou,Guangdong 510642,China;Dongguan Agricultural Technology Extension Administration office,Dongguan,Guangdong 523010,China;Agriculture and Forestry Irrigation Bureau of Xiegang Town,Dongguan,Guangdong 523590,China;Agricultural Technology Service Center of Dalingshan Town,Dongguan,Guangdong 523820,China;Agricultural Technology Service Center of Gaobu Town,Dongguan,Guangdong 523270,China;Agricultural Technology Service Center of Mayong Town,Dongguan,Guangdong 523133,China;Agricultural Technology Service Center of Daojiao Town,Dongguan,Guangdong 523170,China;Shenzhen Agricultural Science and Technology Promotion Center,Shenzhen,Guangdong 518101,China)

机构地区：[1]绿色农药国家重点实验室/广东省微生物信号与病害防控重点实验室/华南农业大学群体微生物研究中心,广东广州510642 [2]东莞市农业技术推广管理办公室,广东东莞523010 [3]东莞市谢岗镇农林水务局,广东东莞523590 [4]东莞市大岭山镇农业技术服务中心,广东东莞523820 [5]东莞市高埗镇农业技术服务中心,广东东莞523270 [6]东莞市麻涌镇农业技术服务中心,广东东莞523133 [7]东莞市道滘镇农业技术服务中心,广东东莞523170 [8]深圳市农业科技促进中心,广东深圳518101

出　　处：《福建农业学报》2023年第3期339-345,共7页Fujian Journal of Agricultural Sciences

基　　金：广东省重点领域研发计划项目(2018B020205003)。

摘　　要：【目的】蕹菜是我国南方重要的经济作物,近年来随着蕹菜种植面积不断扩大,由穿孔黄单胞菌(Xanthomonas perforans)引起的蕹菜溃疡病日趋严重,对生产造成了巨大损失。建立高效快捷的蕹菜溃疡病菌检测技术,可为有效防控病害奠定基础。【方法】采用普通PCR检测技术,针对蕹菜溃疡病菌X. perforans基因组中特有序列TC2-1_002562(编码噬菌体终止酶大亚基家族蛋白)和TC2-1_002580(编码噬菌体家族蛋白)设计两对特异性检测引物,在同一个PCR体系中达到了快速检测蕹菜溃疡病菌的目的。【结果】建立的PCR检测方法可操作性强,对蕹菜溃疡病菌具有良好的特异性。利用特异性引物对来自广东多个地区的蕹菜植株、土壤和水体进行病原菌分子检测,发现第一批样本仅在5份来自东莞的蕹菜叶片上检测到病原菌,第二批样本分别在土壤、水体和植株上检测到病原菌。【结论】所建立的PCR检测方法能快速检测穿孔黄单胞菌,可用于对蕹菜溃疡病的早期诊断。该病在东莞、深圳等地呈普遍发生态势,田间检测结果表明,病害发展迅速,留种蕹菜种苗或种子很可能是该病的初侵染源,种植环境的水、土壤也是病害循环的重要环节,这对于今后进一步研究病害循环规律、制定针对性防控策略具有重要意义。【Objective】An efficient and rapid detection method was developed to monitor the spread of a newly discovered bacterial canker disease infected by Xanthomonas perforans that seriously impacted the production of water spinach(Ipomoea aquatica),a recently introduced and increasingly important cash crop in southern China.【Method】Two pairs of primers were designed to detect X.perforans based on the specific sequences in the genomes including TC2-1_002562 and TC2-1_002580,which encode a phage terminator large subunit family protein and a phage family protein,respectively.The applicability of the proposed methodology with a single PCR reaction was verified.【Result】The newly developed PCR method exhibited a high specificity for detecting X.perforans.In the first sampling batch of water spinach plants,planting soils,and water collected from regions in Guangdong Province,the pathogen was only detected in 5 specimens of water spinach leaves from Dongguan.However,during the subsequent second sampling 10 days later,the pathogen was found in numerous specimens from all sources.【Conclusion】The established PCR method could rapidly detect X.perforans for early disease diagnosis on water spinach.As the field monitoring indicated,the endemic was prevalent in Dongguan and Shenzhen,possibly some other localities,but spreading fast.The origin of the infection might come from the stored water spinach seedlings and/or seeds.The area water and soil could also link to the spread of the disease.Urgent study to timely develop effective prevention and control measures is in order.

关 键 词：蕹菜 溃疡病 穿孔黄单胞菌 双引物PCR检测 

分 类 号：S436.36[农业科学—农业昆虫与害虫防治]