烟草冷杉醇合成关键基因NtCPS2的SNP功能标记开发与应用  

Development and Application of SNP Functional Markers of NtCPS2, a Key Gene for Cis-abienol Synthesis in Tobacco

作　　者：王国平[1,2] 刘旦 李洋洋[1,3] 屈亚芳 李依婷 罗成刚[1] 冯全福[1] 常爱霞[1] Wang Guoping;Liu Dan;Li Yangyang;Qu Yafang;Li Yiting;Luo Chenggang;Feng Quanfu;Chang Aixia(Tobacco Research Institute of Chinese Academy of Agricltural Sciences,Qingdao,266100;Yuxi Zhongyan Tobacco Seed CO.,Ltd,Yuxi,653100;Central South Agricultural Experimental Station of China Tobacco,Changsha,410004)

机构地区：[1]中国农业科学院烟草研究所,青岛266100 [2]玉溪中烟种子有限责任公司,玉溪653100 [3]中国烟草中南农业试验站,长沙410004

出　　处：《分子植物育种》2020年第24期8178-8186,共9页Molecular Plant Breeding

基　　金：农业科技创新计划(ASTIP-TRIC01);中国烟草总公司湖南省公司重点项目(18_20Aa02)共同资助。

摘　　要：冷杉醇是一些香料烟和部分雪茄烟中主要的赖百当类物质,对烟草的香气风格起着重要作用,然而烤烟中基本不含有。开发分子标记是将这种香气特征转入烤烟的有效途径。本研究基于等位基因特异性PCR (AS-PCR)技术,设计了一系列引物,并最终筛选到引物对1F/R1与其互补引物对8F/R1具有良好的多态性,这两对引物在6个亲本和F1群体也得到了验证。利用两个引物对47份烤烟品种和98份‘中烟100’的高香气突变体进行基因分型,同时利用GC-MS检测表型,最终获得材料‘大白筋599’、‘革新三号’、‘NC2326’和‘TBT6’突变株含有冷杉醇。结果表明,这两对标记可用于烟草育种分子标记辅助选择,含冷杉醇的烤烟种质资源可作为烟草育种的重要亲本材料。Cis-abienol, an important compound responsible for flavor and aroma characteristics of tobacco, is a major labdanoid diterpene secreted from trichomes of most oriental tobacco and some cigar tobacco varieties, but is seldom present in flue-cured tobacco. Developing molecular markers is one of the most effective ways for transferring this aroma trait to flue-cured tobacco. Based on the technique of allele-specific PCR(AS-PCR), in the present study, we designed a series of primers. Finally, the primer 1F/R1 and its complementary primer 8F/R1 showed good polymorphism, and they were well validated in six parents and the F1 population. Genotyping and GC-MS were used to identify the 47 flue-cured tobacco plants and 98 high aroma mutants of ’Zhongyan100’ with the two primer pairs. At last, varieties ’Dabaijin599’, ’Gexin3’, ’NC2326’, and one mutant line TBT6 were confirmed to contain cis-abienol. The results indicated that these primers can be used for molecular markerassisted selection in tobacco breeding. The flue-cured germplasms containing cis-abienol can be useful as parents in tobacco breeding.

关 键 词：AS-PCR 冷杉醇 烟草 NtCPS2 SNP标记 

分 类 号：S572[农业科学—烟草工业]