摘要
本文分别设计和验证了烟草黑胫病菌和根黑腐病菌的特异性检测引物,结合镰刀菌属通用引物,进行了烟草三类根腐类病致病菌-烟草黑胫病菌、根黑腐病菌和镰刀菌的多重PCR检测,并对影响多重PCR的退火温度、引物用量、d NTP用量、模板用量分别进行了分析。结果显示,退火温度在43?61℃范围内,烟草黑胫病菌、根黑腐病菌和镰刀菌的引物用量在0.2:0.32?0.48:0.2μmol·L^(-1),d NTP用量在0.1?0.2 mmol·L^(-1)μL/25μL反应体系中,可以经济且高效的实现对烟草黑胫病菌、根黑腐病菌和镰刀菌的特异性同步检测。
Specific primers for Phytophthora parasitica var. nicotianae and Thielaviopsis basicola were designed andverified. Based on these results and reported specific primer for Fusarium spp., a detection system using multiplex PCR wasdeveloped and optimized. Annealing temperature, the concentrations of primers and dNTPs were examined. An economicand efficient protocol were established and applied to detect the infected tobacco. In this protocol, annealing temperature was43 to 61 ℃, primers for Phytophthora parasitica var. nicotianae, Thielaviopsis basicola and Fusarium spp. were0.2：0.32-0.48：0.2 μmol·L-1, respectively and dNTP was 0.1 to 0.2 mmol·L-1.
出处
《山东农业大学学报（自然科学版）》
CSCD
2016年第4期520-524,共5页
Journal of Shandong Agricultural University：Natural Science Edition
基金
中国烟草总公司山东省公司面上项目:烟草细菌性叶斑病快速鉴定技术研究与应用
关键词
烟草
根腐类病害
多重PCR
同步检测
Tobacco
root rot diseases
multiplex PCR
simultaneous detection
