| 城市绿地中立枯丝核菌和齐整小核菌的qPCR快速检测方法 |
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| 引用本文： | 雒淑红,骆玉珍,赵莺莺,张维维,刘文,何山文,安磊,王永杰,韩继刚.城市绿地中立枯丝核菌和齐整小核菌的qPCR快速检测方法[J].浙江农林大学学报,2022,39(5):1087-1095. |
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| 作者姓名： | 雒淑红 骆玉珍 赵莺莺 张维维 刘文 何山文 安磊 王永杰 韩继刚 |
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| 作者单位： | 1.上海海洋大学 食品学院，上海 2001202.上海市园林科学规划研究院，上海 2002323.东北大学 资源与土木工程学院，辽宁 沈阳 110819 |
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| 基金项目： | 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) |
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| 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24×106和22×106拷贝·L?1，2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。图2表5参29
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| 关 键 词： | 城市绿地土壤 立枯丝核菌 齐整小核菌 qPCR |
| 收稿时间： | 2021-12-14 |
Quantitative real-time PCR for rapid detection of Rhizoctonia solani and Sclerotium rolfsii in urban green space |
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| LUO Shuhong,LUO Yuzhen,ZHAO Yingying,ZHANG Weiwei,LIU Wen,HE Shanwen,AN Lei,WANG Yongjie,HAN Jigang.Quantitative real-time PCR for rapid detection of Rhizoctonia solani and Sclerotium rolfsii in urban green space[J].Journal of Zhejiang A&F University,2022,39(5):1087-1095. |
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| Authors: | LUO Shuhong LUO Yuzhen ZHAO Yingying ZHANG Weiwei LIU Wen HE Shanwen AN Lei WANG Yongjie HAN Jigang |
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| Institution: | 1.College of Food Science and Technology, Shanghai Ocean University, Shanghai 200120, China2.Shanghai Academy of Landscape Architecture Science and Planning, Shanghai 200232, China3.College of Resources and Civil Engineering, Northeastern University, Shenyang 110819, Liaoning, China |
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| Abstract: | 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 24×106 and 22×106 copies·L?1, respectively. The coefficients of variation of the 2 repeated 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. Ch, 2 fig. 5 tab. 29 ref.] |
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