烟草鸢尾丝囊霉菌的生物学特性及分子检测  

Biological Characteristics and Molecular Detection of Aphanomyces iridis Harmful to Tobacco Seedlings

作　　者：李小杰[1] 刘剑君 白静科 刘畅[1] 陈玉国[1] 苗圃[3] 邱睿[1] 陆施羽 李淑君[1] LI Xiaojie;LIU Jianjun;BAI Jingke;LIU Chang;CHEN Yuguo;MIAO Pu;QIU Rui;LU Shiyu;LI Shujun(Tobacco Research Institute of Henan Academy of Agricultural Sciences,Key Laboratory for Green Preservation&Control of Tobacco Diseases and Pests in Huanghuai Growing Area,Xuchang,Henan 461000,China;Henan Provincial Tobacco Company,Zhengzhou 450046,China;Luoyang Branch of Henan Provincial Tobacco Company,Luoyang,Henan 471000,China)

机构地区：[1]河南省农业科学院烟草研究所,烟草行业黄淮烟区烟草病虫害绿色防控重点实验室,河南许昌461000 [2]中国烟草总公司河南省公司,郑州450046 [3]河南省烟草公司洛阳市公司,河南洛阳471000

出　　处：《中国烟草科学》2022年第6期53-59,共7页Chinese Tobacco Science

基　　金：河南省农业科学院科技创新团队专项项目(2022TD26);河南省烟草公司科技项目(2022410000240018)。

摘　　要：针对近年来河南省许昌、洛阳、三门峡等部分烟区烟草漂浮苗鸢尾丝囊霉根腐病发生较重的问题,采用菌丝生长速率法初步分析了病原菌的生物学特性,并根据NCBI数据库中鸢尾丝囊霉菌代表菌株CBS 524.87的5个编码基因CDS序列,设计筛选鸢尾丝囊霉菌的特异性扩增引物并应用。生物学特性分析结果表明,在PDA培养基上,鸢尾丝囊霉菌菌丝生长的适宜温度为25~35℃,致死温度为50℃,处理10min;生长适宜pH为4.0~8.0,最适pH为6.0;连续光照条件有利于菌丝生长。筛选获得了特异性扩增鸢尾丝囊霉菌的引物对7个,对于基因组DNA扩增的灵敏度约为0.182 ng/μL;利用特异引物AiT3分别对接种育苗基质和烟苗进行分子检测,可特异性的检测出鸢尾丝囊霉菌,检测的灵敏度分别为每克基质含菌量为2.5×10-2g病原菌菌丝和0.5 ng/μL烟苗根系基因组DNA。本研究为鸢尾丝囊霉菌的快速分子检测提供了技术支撑,对苗期鸢尾丝囊霉根腐病病原的准确识别和预测预报提供了依据。In view of the serious occurrence of root rot caused by Aphanomyces iridis in tobacco seedling bed of Henan Province in recent years, this study was carried out to clarify the biological characteristics of the pathogen and fro rapid detection at early stages.The biological characteristics of the pathogen were studied with the mycelial growth rate method, and specific amplification primers for A. iridis were designed and screened for application based on the CDS sequences of five coding genes of the representative strain CBS 524.87 in NCBI database. The results showed that the suitable temperature for mycelium growth of A. iridis was 25-35 ℃ on PDA plates, and the lethal temperature was 50 ℃ for 10 min. The suitable pH for A. iridis growth was 4.0-8.0 and the optimum pH was 6.0. Continuous light was beneficial to mycelium growth for A. iridis. Seven pairs of primers were designed and screened for the specific amplification of A. iridis, and the sensitivity to genomic DNA amplification was about 0.182 ng/μL. A. iridis could be specifically detected by the specific primer pair AiT3 for molecular detection of inoculated seedling substrate and tobacco seedlings respectively, with the detection sensitivity being 2.5×10-2 g hyphae of per gram of substrate and 0.5 ng/μL tobacco seedling root genomic DNA. The results of this study provide technical support for the rapid molecular detection of A. iridis, and provide an important basis for the accurate identification and prediction of root rot caused by A. iridis at seedling stages.

关 键 词：烟草 鸢尾丝囊霉 生物学特性 特异引物 分子检测 

分 类 号：S435.72[农业科学—农业昆虫与害虫防治]