辣椒抗疮痂病基因Bs2和Bs3双重PCR体系的建立及应用  

Establishment and Application of Duplex PCR to Detection of Bacterial Spot Disease Resistance Genes Bs2 and Bs3 in Pepper

作　　者：高升华 李宁[1] 王飞[1] JUNTAWONG Niran 焦春海[3] 姚明华[1] GAO Shenghua;LI Ning;WANG Fei;JUNTAWONG Niran;JIAO Chunhai;YAO Minghua(Cash Crops Research Institute,Hubei Academy of Agricultural Sciences,Hubei Key Laboratory of Vegetable Germplasm Enhancement and Genetic Improvement,Wuhan,Hubei 430064;Kasetsart University,Bangkok,Thailand 10900;Hubei Academy of Agricultural Sciences,Wuhan,Hubei 430064)

机构地区：[1]湖北省农业科学院经济作物研究所/蔬菜种质创新与遗传改良湖北省重点实验室,湖北武汉430064 [2]泰国农业大学,泰国曼谷10900 [3]湖北省农业科学院,湖北武汉430064

出　　处：《北方园艺》2020年第13期9-15,共7页Northern Horticulture

基　　金：国家重点研发计划资助项目(2017YFD0101903,2016YFE0205500);博士后科学基金资助项目(2017M620305);湖北省技术创新专项资助项目(2017ABA147);现代农业产业技术体系建设专项资金资助项目(CARS-23-G28)。

摘　　要：以栽培辣椒和野生型辣椒为试验材料,采用双重PCR体系构建的方法,建立了辣椒抗疮痂病基因Bs2和Bs3的双重PCR反应体系,以期利用该反应体系提高辣椒抗疮痂病种质资源鉴定和育种分离世代材料分子标记辅助筛选的效率。结果表明:抗疮痂病基因Bs2和Bs3的双重PCR反应体系,以Bs2-For/Bs2-Rev和Bs3-For/Bs3-Rev为特异引物,在退火温度为52℃时,具有很好的多态性和稳定性。此外,用该体系对不同辣椒种材料进行抗病性鉴定,与接种鉴定的结果一致,进一步验证了该体系的可靠性。该双重PCR反应体系可用于辣椒抗疮痂病品种选育。Taking cultivated and wild-type peppers as materials,the construction method of duplex PCR system was used to establish a novel duplex PCR marker for detection of bacterial spot disease(BS)resistance genes Bs2 and Bs3,in order to improve the efficiency of germplasm resource identification and molecular marker-assisted screening of breeding generations.The results showed that the duplex PCR marker for detection of bacterial spot disease(BS)resistance genes Bs2 and Bs3 had rich polymorphism and high stability,when Bs2-For/Bs2-Rev and Bs3-For/Bs3-Rev were used as specific primers pairs and annealing temperature was set at 52℃.In addition,the novel duplex PCR system was used to identify the resistance of different pepper species materials to bacterial spot disease,and the results were consistent with the inoculation assay,indicating that the duplex PCR system had high reliability.The application of the duplex PCR system in Bs2 and Bs3 gene could improve breeding selection efficiency.

关 键 词：辣椒 抗辣椒疮痂病基因Bs2和Bs3 双重PCR标记 

分 类 号：S436[农业科学—农业昆虫与害虫防治]