|
|||||
|
|
| 生姜腐皮镰刀菌的分离鉴定及PCR快速检测方法构建 | |
| 引用本文： | 周洁,张玲玲,袁继荣,秦曼丽,刘续立,何杨,吴林,贾切,刘奕清.生姜腐皮镰刀菌的分离鉴定及PCR快速检测方法构建[J].植物病理学报,2022,52(4):681-690. |
| 作者姓名： | 周洁 张玲玲 袁继荣 秦曼丽 刘续立 何杨 吴林 贾切 刘奕清 |
| 作者单位： | 长江大学 园艺园林学院/香辛作物研究院,荆州 434025; 重庆市幅沅农业生物技术研究院有限公司,永川 402160 |
| 基金项目： | 湖北省重点研发项目(2020BBA037);中央引导地方专项项目(2020ZYYD020);重庆调味品产业体系重大专项项目(2021-07);重庆英才计划创新创业团队(CQYC201903201) |
| 摘 要： | 由腐皮镰刀菌(Fusarium solani)引起的生姜枯萎病是一种毁灭性真菌土传病害,其病原菌的快速高效检测,有助于枯萎病的早期诊断及预警。通过回接试验、形态学观测以及系统进化树分析,对湖北生姜产区分离的菌株进行鉴定,获得生姜腐皮镰刀菌(F. solani)。基于镰刀菌属通用引物TEF-1αF/TEF-1αR基因序列,以腐皮镰刀菌基因组DNA为模板进行扩增测序,根据序列信息设计筛选出一对腐皮镰刀菌特异性引物,构建了基于普通PCR的快速高效分子检测方法,并对接种过病菌的生姜植株和土壤进行检测验证。通过病原菌的菌落形态、孢子显微结构观察、真菌通用引物ITS1/ITS4鉴定和致病性测定,确定引起生姜枯萎病的病原菌为腐皮镰刀菌(F. solani)。用设计的特异性引物F8/R8进行PCR,特异扩增获得约381 bp的目标条带,检测灵敏度为454 pg·μL-1,利用该引物对带菌生姜幼苗和土壤进行检测验证,证实可从发病生姜幼苗和带菌土壤中特异性检测到腐皮镰刀菌(F. solani)。本研究明确了引起湖北生姜枯萎病的病原菌为腐皮镰刀菌(F. solani),并建立了PCR快速检测方法。该检测方法简便、灵敏、高效,可用于生姜枯萎病的早期诊断与预防。 |
| 关 键 词： | 生姜 枯萎病 腐皮镰刀菌 PCR检测 |
| 收稿时间： | 2021-05-06 |
Isolation and identification of Fusarium solani from ginger rhizomes and establishment of a rapid PCR detection method |
|
| ZHOU Jie,ZAHNG Lingling,YUAN Jirong,QIN Manli,LIU Xuli,HE Yang,WU Lin,JIA Qie,LIU Yiqing.Isolation and identification of Fusarium solani from ginger rhizomes and establishment of a rapid PCR detection method[J].Acta Phytopathologica Sinica,2022,52(4):681-690. | |
| Authors: | ZHOU Jie ZAHNG Lingling YUAN Jirong QIN Manli LIU Xuli HE Yang WU Lin JIA Qie LIU Yiqing |
| Institution: | College of Horticulture and Landscape Architecture/Spice Crops Research Institute, Yangtze University, Jingzhou 434025,China; Chongqing Fuyuan Agricultural Biotechnology Research Institute Co. Ltd, Yongchuan, 402160,China |
| Abstract: | Ginger wilt caused by Fusarium solani is a destructive fungal soil-borne disease, and the rapid and efficient detection of its pathogens is of great significance for the early diagnosis and early warning of wilt. Through back inoculation, morphological observation and phylogenetic tree analysis, we identified the strains isolated from Hubei ginger-producing areas, and subsequently, F. solani was obtained as one of the Fusarium wilt pathogens. Based on the reported general primer TEF-1αF/TEF-1αR gene sequence of Fusarium, using F. solani genomic DNA as a template for amplification and sequencing, a pair of F. solani specific primers was designed and screened according to the obtained sequence information, and a rapid and efficient molecular detection method based on ordinary PCR was developed, which was then tested and verified on ginger plants and soil. Based on the colony morphology, spore microstructure observation, gene analysis and pathogenicity determination; the pathogen causing ginger wilt was identified as F. solani. The designed specific primer F8/R8 was used for PCR, and the target band of about 381 bp was obtained by specific amplification, with a detection sensitivity of 454 pg·μL-1. The primers were used to detect and verify the infected ginger seedlings and soil, confirming that F. solani could be specifically detected from infected ginger seedlings and soil. In this study, the pathogen causing ginger wilt in Hubei Province was identified as F. solani, and a rapid PCR detection method was established. The detection method is simple, sensitive and efficient, which can accurately and quickly identify the pathogen, and can be used for early diagnosis and prevention of ginger wilt. |
| Keywords: | ginger fusarium wilt Fusarium solani PCR detection |
| 点击此处可从《植物病理学报》浏览原始摘要信息 | |
| 点击此处可从《植物病理学报》下载免费的PDF全文 | |