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2025年5月13日 星期二
农业生物技术学报  2021, Vol. 29 Issue (6): 1215-1221    DOI: 10.3969/j.issn.1674-7968.2021.06.019
  研究资源与技术改进 本期目录 | 过刊浏览 | 高级检索 |
草莓枯萎病和炭疽病的双重LAMP快速病原鉴定
史芳芳, 王雷*
新疆生产建设兵团第十二师农业科学研究所,乌鲁木齐 830088
Rapid Pathogen Identification of Fusarium Wilt and Anthracnose in Strawberry (Fragaria ananassa) by Double LAMP
SHI Fang-Fang, WANG Lei*
The Institute of Agricultural Sciences of the 12th Division of Xinjiang Production and Construction Corps, Urumqi 830088, China
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摘要 枯萎病和炭疽病是草莓(Fragaria ananassa)最为常见的主要病害,病原分别为尖孢镰刀菌(Fusarium oxysporum)和炭疽菌(Colletotrichum spp.),目前的检测手段主要采用传统的病原菌培养法和常规PCR法,而环介导等温扩增(loop-mediated isothermal amplification, LAMP)技术可用于草莓枯萎病和炭疽病的病原菌快速检测。本研究针对尖孢镰刀菌rDNA间隔区(intergenic spacer, IGS)的保守区域设计了1套LAMP扩增引物,并设计了炭疽菌内转录间隔区(internal transcribed spacer, ITS)特异序列的LAMP引物,利用LAMP技术快速、准确地将草莓尖孢镰刀菌和炭疽菌进行分辨和检测。结果表明,建立的LAMP法能够特异性检测尖孢镰刀菌和炭疽菌,检测灵敏度为10×10-5 ng/μL,是普通PCR检测灵敏度的1 000倍。本研究建立的草莓枯萎病和炭疽病快速LAMP检测方法具有灵敏度高、快速及简便的优点,是可以向基层单位推广的检测手段,对于草莓病原菌实时监测和前期预警具有重要作用。
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史芳芳
王雷
关键词 环介导等温扩增(LAMP)枯萎病炭疽病尖孢镰刀菌炭疽菌    
Abstract:Fusarium wilt and Anthracnose are the most common and major diseases in the strawberry (Fragaria ananassa), and the pathogens are Fusarium oxysporum and Colletotrichum spp., respectively. At present, the main detection methods are traditional pathogen culture method and conventional PCR method, but loop-mediated isothermal amplification (LAMP) can be used for rapid detection of Fusarium wilt and Anthracnose in strawberry. In this study, a set of LAMP amplification primers was designed for the conserved region of intergenic spacer (IGS) in rDNA of Fusarium oxysporum, and the LAMP specific primers were designed to detect the specific sequence of internal transcribed spacer (ITS) in Colletotrichum spp. The LAMP technique was used to identify and detect strawberry Fusarium oxysporum and Colletotrichum spp. rapidly and accurately. The results showed that the established LAMP method was able to specifically detect F. oxysporum and Colletotrichum spp., and the detection sensitivity was 10×10-5 ng/uL, which was 1 000 times that of conventional PCR. As a result, the rapid LAMP detection method of Fusarium wilt and Anthracnose in strawberry developed in this study had the advantages of high sensitivity, rapidity and simplicity, which could be popularized to grass-roots units and play an important role in real-time monitoring and early warning of strawberry pathogens.
Key wordsLoop-mediated isothermal amplification (LAMP)    Fusarium wilt    Anthracnose    Fusarium oxysporum    Colletotrichum spp.
收稿日期: 2020-09-17     
ZTFLH:  S432.1  
基金资助:兵团中青年科技创新领军人才计划项目(2018CB030)
通讯作者: *gzx120731@sina.com   
引用本文:   
史芳芳, 王雷. 草莓枯萎病和炭疽病的双重LAMP快速病原鉴定[J]. 农业生物技术学报, 2021, 29(6): 1215-1221.
SHI Fang-Fang, WANG Lei. Rapid Pathogen Identification of Fusarium Wilt and Anthracnose in Strawberry (Fragaria ananassa) by Double LAMP. 农业生物技术学报, 2021, 29(6): 1215-1221.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2021.06.019     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2021/V29/I6/1215
 
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