芒果炭疽病菌多重巢式PCR检测体系建立及田间快速检测

摘要

Mango anthracnose,caused by Colletotrichum gloeosporioides and C.acutatum,is a major threat to mango production.Rapid and sensitive molecular diagnosis is vital to guide the efficient control of the disease.Based on the conserved rDNA-ITS sequences,a multiple nested-PCR system was developed for the simultaneous detection of the two types of pathogens.The established multiplex nested-PCR had high specificity and sensitivity to detect pathogen DNA as low as 2 fg/μL.Combined with conventional morphological characteristics analysis,the multiplex nested-PCR method revealed that the mixed infection of the two pathogens of mango anthracnose was always existed in the field.The multiplex nested-PCR system may not only serve as a supplement of conventional morphology diagnosis,but also be used in simultaneous detection and distinguishing the types of pathogens for the guide of disease prevention and control.%以芒果炭疽病病原菌rDNA-ITS为靶标,建立并完善多重巢式PCR检测技术,结合常规形态学特征分析,对中国华南地区10个田间样品进行检测分析.结果表明:该技术特异性强、灵敏度达2fg/μL、重复性强、可操作性强,能同时检测、区分2种病原菌,能够应用于田间炭疽病害的检测工作,也是常规形态学鉴定的补充.利用该技术检测到田间芒果炭疽病存在胶孢、尖孢2种病原菌的复合侵染,该结果为后续开展有效防控措施提供理论指导和技术支撑.