马铃薯抗晚疫病基因R8、RB和抗病毒病基因Rx1、Ry_(adg)的多重PCR检测  

Multiplex PCR Detection for Late Blight Resistant Genes R8,RB and Virus Resistant Genes Rx1,Ry_(adg) in Potato

作　　者：刘程 王世尧 史伟玲 宋玉浩 蒋锐 赵勇[1] 莫世春 吕典秋 王季春[1] 刘勋[1] LIU Cheng;WANG Shiyao;SHI Weiling;SONG Yuhao;JIANG Rui;ZHAO Yong;MO Shichun;LÜDianqiu;WANG Jichun;LIU Xun(Chongqing Key Laboratory of Biology and Genetic Improvement in Tuber and Root Crops,College of Agronomy and Biotechnology,Southwest University,Chongqing 400715,China;Chongqing Wuxi Agricultural and Rural Committee,Chongqing 405899,China)

机构地区：[1]西南大学农学与生物科技学院,薯类生物学与遗传育种重庆市重点实验室,重庆400715 [2]重庆市巫溪县农业农村委员会,重庆405899

出　　处：《园艺学报》2021年第2期389-396,共8页Acta Horticulturae Sinica

基　　金：重庆市技术创新与应用发展专项(cstc2019jscx-gksbX0157);国家级大学生创新创业训练计划项目(201810635026);科技部对发展中国家科技援助项目(KY201904016);国家重点研发计划项目(2018YFE0127900)。

摘　　要：在218份马铃薯材料中筛查晚疫病持久抗性基因RB和R8标记的基础上,进一步筛查了马铃薯X病毒(PVX)和马铃薯Y病毒(PVY)极端抗性基因Rx1和Ry_(adg)的标记。利用筛查出的含有Rx1和RB标记的‘德薯3号’和含有Ry_(adg)和R8标记的P182马铃薯资源材料混合DNA为模板,选择已报道的Rx1和Ry_(adg)的标记引物,并根据RB和R8序列设计特异引物,同时设计马铃薯GBSS基因特异引物为内参监控PCR反应体系有效扩增,对多重PCR延伸温度、引物组合浓度等进行优化,建立了同时检测4个抗病基因的多重PCR方法。应用该方法对选育品系样品进行筛查,鉴定到聚合上述4个抗性基因的材料4份。本研究中建立的多重PCR检测体系用于分子标记辅助选择,为马铃薯晚疫病和病毒病多抗基因聚合育种提供了简单、高效的技术方法并为多抗性基因聚合新品种选育创制新资源。Based on defined RB and R8 genes for late blight resistance in 218 germplasm resources,Rx1 and Ry_(adg) for potato virus X(PVX)and potato virus Y(PVY)extreme resistance were screened in the present study,respectively.The mixed DNA from two screened potato genotypes containing Rx1,RB and Ry_(adg),R8 was used as templates.The multiplex PCR system was developed with the reported markers for Rx1 and Ry_(adg) and designed markers for RB and R8.In addition,the amplification of the PCR reaction system was monitored by the potato GBSS gene markers.The crucial factors of multiplex PCR system including extension temperature and primers concentration were optimized.Five specific fragments were simultaneously amplified in one PCR reaction.This detection method was used to detect these R genes from advanced breeding lines.The results revealed that four breeding lines containing four above mentioned R genes were screened.In conclusion,the established multiplex PCR detection system will be used for molecular marker-assisted selection,providing a simple and efficient technical method for late blight and viral diseases multi-R genes aggregation breeding,and new resources for the new varieties selection with four R genes in potato.

关 键 词：马铃薯 晚疫病 马铃薯Y病毒 马铃薯X病毒 多重PCR 

分 类 号：S532[农业科学—作物学]