芒果炭疽病病原亚洲炭疽菌(Colletotrichum asianum)特异性PCR检测
Specific PCR detection of Colletotrichum asianum causing mango anthracnose
鲁萌萌 1唐利华 2黄穗萍 2陈小林 2郭堂勋 2马立安 3陆建勋 4李其利2
作者信息
- 1. 广西壮族自治区农业科学院植物保护研究所/农业农村部华南果蔬绿色防控重点实验室/广西作物病虫害生物学重点实验室,南宁 530007;长江大学生命科学学院,荆州 434025
- 2. 广西壮族自治区农业科学院植物保护研究所/农业农村部华南果蔬绿色防控重点实验室/广西作物病虫害生物学重点实验室,南宁 530007
- 3. 长江大学生命科学学院,荆州 434025
- 4. 广西壮族自治区农业科学院花卉研究所,南宁 530007
- 折叠
摘要
亚洲炭疽菌(Colletotrichum asianum)是一种能引发芒果炭疽病的优势病原菌.为建立一种能快速诊断该病原菌所引起的芒果炭疽病的PCR检测方法,本研究通过比较不同种类炭疽菌的全基因组DNA序列,获得了 1条C.asianum特异性序列,设计并筛选得到 1 对特异性引物:caf1(5'-CCGTCAGACGGAATTATCAGC-3')和 car1(5'-CCGATCCTGTCTTT-GAAATGG-3').结果表明,引物caf1/car1可以从C.asianum中扩增出长度为419 bp的特异性目标条带,灵敏度可达到1 pg·μL-1.利用该引物能够从人工接种C.asianum YN55-1的芒果叶片以及田间发生炭疽病的芒果叶片样品中快速稳定地检测出长度为419 bp的目标条带.本研究建立的特异性PCR检测方法为C.asianum导致的芒果炭疽病的田间监测和快速诊断提供了技术支持.
Abstract
Colletotrichum asianum is one of the most important pathogens that cause anthracnose on mango plants.To establish a method for rapid detection of C.asianum,a C.asianum-specific sequence was obtained by comparing genomic sequences of different Colletotrichum species,and a pair of specific primers:caf1(5'-CCGTCAGACGGAATTATCAGC-3')and car1(5'-CCGATCCTGTCTTTGAAATGG-3')was designed accor-dingly.Specificity test results showed that a 419-bp target amplicon could only be obtained from C.asianum rather than other Colletotrichum species and non-Colletotrichum fungal and bacterial species that we tested in this study using the primer pair caf1/car1,with a detection sensitivity of 1 pg-μL-1.The 419-bp amplicon could be detected from both mango leaves artificially inoculated with C.asianum strain YN55-1 and from naturally infec-ted mango leaves with anthracnose symptoms.The specific PCR system developed in this study provides a tech-nical support for monitoring and rapid diagnosis of mango anthracnose caused by C.asianum in the field.
关键词
芒果炭疽病/PCR检测/亚洲炭疽菌Key words
mango anthracnose/PCR detection/Colletotrichum asianum引用本文复制引用
基金项目
财政部和农业农村部:国家现代农业产业技术体系资助(CARS-31)
广西作物病虫害生物学重点实验室基金(20-065-30-KF-05)
出版年
2024
NETL
NSTL
万方数据