1 Guangxi Subtropical Crops Research Institute, Nanning 530002, China; 2 Guangxi Subtropical Fruits Processing Research Center of Engineering Technology, Nanning 530002, China;
摘要茶多酚是茶树(Camelliasinensis)的主要次级代谢产物,与茶树的新陈代谢、生长发育及品质非常密切。茶树酚类物质的合成涉及多种酶。目前针对茶树酚类合成途径特定酶开发的 SSR 分子标记引物及应用还较少。本研究基于茶树全基因组序列,以茶树酚类物质合成途径12个关键酶的基因及其侧翼序列开发获得了 158 对 SSR 分子标记引物,其中类黄酮 3'-羟化酶开发获得的 SSR 引物最多,有 29 对,肉 桂酸羟化酶最少,只有1对。所开发的158对SSR引物,位于基因间隔区的最多,依次是内含子区、上游区及外显子区,位于5′端上游非编码区的最少。这些引物总计有122个重复单元,其中二核苷酸最多,之后依次为三核苷酸、四核苷酸及五核苷酸,最少的是六核苷酸。从158对SSR引物中挑选68对进行了引物多态性筛选,将获得的13对多态性好的引物开展72份茶树种质资源遗传多样性的研究,共获得105种带型,同一样品的重复样可聚在一起,表明所筛选的引物能很好地区分种质资源,具有较好的准确性。72份茶树种质资源的遗传相似系数为0.8120~0.9849。本研究为茶树基于主要内含物酚类物质开展核心种质构建、新品种选育及品种鉴定等提供依据,同时也可为针对特定基因开发SSR分子标记提供参考方法。
Abstract：Tea polyphenols are the main secondary metabolites of tea plant (Camellia sinensis). They are closely related to the metabolism, growth and quality of tea plant. The synthesis of tea tree phenolic involves a variety of enzymes. At present, the development and application of SSR molecular marker primers for specific enzymes in the tea plant synthesis pathway are still rare. This study was based on the whole genome sequence of tea plant; 158 pairs of SSR molecular marker primers were developed based on 12 key enzyme genes and their flanking sequences of tea plant phenolic synthesis pathway. Among them, the most SSR primers were obtained from flavonoid 3'-hydroxylase gene, with 29 pairs, and the least was cinnamate 4-hydroxylasegene, with only 1 pair. Among the 158 SSR primers, the most were located in the intergenic region, followed by the intron region, upstream region and exon region, and the least were located in the 5' untranslated region. These primers had 122 repeat units, among which dinucleotide was the most, followed by trinucleotide, tetranucleotide and pentanucleotide, and hexanucleotide was the least. 68 pairs of SSR primers were selected from 158 pairs of primers for primer polymorphism screening. The genetic diversity of 72 tea germplasm resources was studied by using 13 pairs of primers with good polymorphism. A total of 105 banding type were obtained, and the duplicate samples of the same sample can be gathered together. The genetic similarity coefficient (GS) of 72 tea germplasm resources ranged from 0.812 0 to 0.984 9. The results showed that the selected primers could distinguish the germplasm resources well and had good accuracy. This study can provide a basis for core germplasm construction, new variety selection and variety identification based on phenolic substances in tea plant, and also provide a reference method for developing SSR molecular markers for specific genes.
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