双重PCR区分检测花生黑腐病菌和花生基腐病菌  

Establishment of a duplex PCR for differentially diagnosing of Cylindrocladium parasiticum and Neocosmospora vasinfecta

作　　者：潘斐 敖莉丝 潘汝谦[1] 纪春艳 PAN Fei;AO Li-si;PAN Ru-qian;JI Chun-yan(College of Plant Protection,South China Agricultural University,Guangzhou 510642,China)

机构地区：[1]华南农业大学植物保护学院,广东广州510642

出　　处：《中国油料作物学报》2024年第4期913-919,共7页Chinese Journal of Oil Crop Sciences

摘　　要：黑腐病和基腐病是威胁花生生产的重要病害。两种病害在田间发病症状十分相似,凭症状区分诊断困难。本研究利用calmodulin(cmdA)基因序列设计特异性引物,通过优化反应体系和反应条件,针对花生黑腐病菌和基腐病菌进行了双重PCR的区分检测。结果表明,在反应体系浓度为1.8×、引物浓度为0.2μmol/L、退火温度为62℃、反应循环数为37的条件下,可特异性地分别扩增出两条目的条带(274 bp和409 bp),检测灵敏限度为100 pg/μL。成功建立了可准确、快速区分检测花生黑腐病菌和花生基腐病菌的双重PCR,可为准确诊断和及时防控花生两种易混淆的病害提供科学的参考。Cylindrocladium black rot and Neocosmospora foot rot are important diseases that seriously threaten healthy peanuts production.It is difficult to distinguish these two diseases due to their similar symptoms in the fields.In this study,duplex PCR detection was developed for the differential diagnosis of Cylindrocladium parasiti⁃cum and Neocosmospora vasinfecta based on calmodulin(cmdA)sequences.Two specific target bands(274 bp and 409 bp)were successfully amplified under the optimum conditions:reaction concentration of 1.8×,primer concentration of 0.2μmol/L,annealing temperature of 62℃,and number of reaction cycles of 37.The detection limit of the duplex PCR system was 100 pg/μL The established duplex PCR assay was specific and efficient,could be used to distinguish accurately and control timely of the two confusing peanut diseases.

关 键 词：花生 黑腐病菌 基腐病菌 双重PCR 

分 类 号：S432.1[农业科学—植物病理学]