城市绿地中立枯丝核菌和齐整小核菌的qPCR快速检测方法  

Quantitative real-time PCR for rapid detection of Rhizoctonia solani and Sclerotium rolfsii in urban green space

作　　者：雒淑红 骆玉珍 赵莺莺 张维维 刘文 何山文 安磊 王永杰[1] 韩继刚 LUO Shuhong;LUO Yuzhen;ZHAO Yingying;ZHANG Weiwei;LIU Wen;HE Shanwen;AN Lei;WANG Yongjie;HAN Jigang(College of Food Science and Technology,Shanghai Ocean University,Shanghai 200120,China;Shanghai Academy of Landscape Architecture Science and Planning,Shanghai 200232,China;College of Resources and Civil Engineering,Northeastern University,Shenyang 110819,Liaoning,China)

机构地区：[1]上海海洋大学食品学院,上海200120 [2]上海市园林科学规划研究院,上海200232 [3]东北大学资源与土木工程学院,辽宁沈阳110819

出　　处：《浙江农林大学学报》2022年第5期1087-1095,共9页Journal of Zhejiang A&F University

基　　金：上海市绿化和市容管理局科技攻关项目(G200201);上海市科学技术委员会科研计划项目(19dz1203300)。

摘　　要：【目的】土传病原真菌立枯丝核菌Rhizoctonia solani和齐整小核菌Sclerotium rolfsii严重威胁园林绿化植物正常生长。建立针对这2种土壤病原真菌的快速定量检测方法。【方法】通过筛选2种病原菌特异性引物,优化反应条件。【结果】初步建立了2种病原菌的实时荧光定量PCR(qPCR)检测方法。引物ST-RS1/ITS4和SRITSF/SRITSR可以分别用于立枯丝核菌和齐整小核菌的qPCR检测,其灵敏度分别达24×10^(6)和22×10^(6)拷贝·L,2次重复反应的变异系数分别为3.37%~4.61%和0.66%~8.61%。对上海绿地土壤样品的检测结果表明:立枯丝核菌和齐整小核菌的检出率分别为100%和19%。【结论】建立的qPCR检测方法具有较强特异性、较高灵敏度和较强重复性,可以用于上海城市绿地土壤中立枯丝核菌和齐整小核菌的快速、有效定量检测。[Objective] This study aims to establish a rapid quantitative detection method for Rhizoctonia solani and Sclerotium rolfsii,2 soil-borne pathogenic fungi that seriously threaten the normal growth of landscape plants in Shanghai. [Method] The reaction conditions were optimized by screening 2 pathogen specific primers. [Result] A quantitative real-time PCR(qPCR) method was established for the detection of the two pathogens. The primers ST-RS1/ITS4 and SRITSF/SRITSR could be used for qPCR detection of R. solani and S. rolfsii, with sensitivity of 24×10^(6) and 22×10^(6) copies·L, respectively. The coefficients of variation of the 2repeated reactions were 3.37%-4.61% and 0.66%-8.61%, respectively. The detection results of soil samples in Shanghai green space showed that the detection rates of R. solani and S. rolfsii were 100% and 19%, respectively. [Conclusion] The established qPCR method has high specificity, sensitivity and repeatability, and can be used for rapid and effective quantitative detection of R.solani and S.rolfsii in Shanghai urban green soil.

关 键 词：城市绿地土壤 立枯丝核菌 齐整小核菌 qPCR 

分 类 号：Q949.32[生物学—植物学]