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

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.