烟草根黑腐病抗性位点RBRR1的InDel分子标记开发与分析  

Development and haplotype analysis of insertion–deletion(InDel)molecular markers for resistance locus RBRR1 to black root rot of tobacco(Nicotiana tabacum)

作　　者：冯智宇 陈学军[1] 盖晓彤 姜宁 焦芳婵[1] 吴兴富[1] 童治军[1] 刘天彦 杨林明 许美玲[1] 李永平[1] FENG Zhiyu;CHEN Xuejun;GAI Xiaotong;JIANG Ning;JIAO Fangchan;WU Xingfu;TONG Zhijun;LIU Tianyan;YANG Linming;XU Meiling;LI Yongping(Key Laboratory of Tobacco Biotechnological Breeding,National Tobacco Genetic Engineering Research Center,Yunnan Academy of Tobacco Agricultural Sciences,Kunming 650021,Yunnan,China;Qujing Tobacco Company of Yunnan Province,Qujing 655099,Yunnan,China)

机构地区：[1]云南省烟草农业科学研究院,云南省昆明市,650021 [2]云南省烟草公司曲靖市公司,云南省曲靖市,655099

出　　处：《中国烟草学报》2022年第4期67-76,共10页Acta Tabacaria Sinica

基　　金：中国烟草总公司云南省烟草公司科技项目“优质抗病烤烟新品种选育”(2019530000241001);河北省自然科学基金项目“甜瓜根系分泌物中化感物质与镰孢枯萎菌的互作关系研究”(C2019402430)。

摘　　要：【背景和目的】RBRR1是通过远缘杂交与回交导入栽培烟草的一个单显性广谱型根黑腐病抗病位点,为了开发基于该位点的简单高效稳定且成本低的紧密连锁分子标记。【方法】在已有酶切扩增多态性序列(Cleaved amplified polymorphism sequences,CAPS)标记相关序列基础上,利用云晒1号基因组与抗、感根黑腐病材料重测序信息,基于其序列插入/缺失(Insertion/Deletion,InDel)多态性设计抗病相关分子标记,并开展遗传效应和遗传变异分析。【结果】筛选到4对具有良好多态性的引物,在目标单株的分子标记辅助选择中具备便捷、高效且呈共显性的特点。遗传效应分析表明,通过分子标记辅助选择导入RBRR1位点能够极显著提高栽培烟草根黑腐病抗性。遗传变异分析结果显示,在烟草核心种质资源中仅有白肋烟TN90和SoTa2携带有RBRR1抗病基因,并发现4对引物在普通烟草自然群体中呈现2种基因型。【结论】RBRR1抗病位点在我国尚未进行广泛利用,具有较高的利用价值;而开发的4个InDel标记可用于该抗性位点的分子标记辅助选择育种和后续抗病基因的精细定位与克隆。[Background and Objective]RBRR1 is a single dominant broad-spectrum locus coffering resistance to black root rot that was introduced into tobacco(Nicotiana tabacum)by remote hybridization and backcross,and the development of efficient,stable,low-cost and user-friendly tightly linked molecular markers is an effective approach for resistance breeding by marker-assisted selection and for location of functional gene within the locus.[Methods]In this study,we anchored the region of RBRR1 locus based on the primer sequences of published CAPS markers,and acquired four polymorphism markers targeting insertion-deletion(InDel)within the region of RBRR1 using the genome sequence of Yunshai#1 as reference and the resequencing data of a pair of resistant and susceptible root black rot materials.These markers were convenient,efficient and codominant in molecular marker-assisted selection of target individuals.[Results]Genetic effect analysis showed that the introduction of RBRR1 through marker-assisted selection could significantly improve the resistance to black root rot in cultivated tobacco.Haplotype analysis showed that only TN90 and SoTa2 carried RBRR1 in the tobacco core germplasm resources,and four pairs of primers presented two genotypes in the natural population of common tobacco.[Conclusion]Collectively,our data indicated that the four InDel markers have high utilization value in marker-assisted selection and subsequent mapping of the functional gene.

关 键 词：烟草 根黑腐 根串珠霉菌 分子标记辅助选择 INDEL标记 

分 类 号：S435.72[农业科学—农业昆虫与害虫防治]