基于SLAF-seq技术的金花茶SNP标记开发及遗传分析

为了解金花茶种质的遗传关系,利用SLAF-seq测序技术对在不同地区收集的25份金花茶种质与山茶科其他5份种质进行测序。以猕猴桃基因组为参照,对金花茶基因组DNA酶切构建SLAF-seq文库,利用SLAFseq测序技术对其进行高通量测序、SNP标记开发及其进化关系分析,在多态性SLAF标签上开发特异性SNP位点,最后根据SNP位点构建25份金花茶种质与山茶科其他5份种质的进化树。最终共开发1471113个SLAF标签,其中多态性SLAF标签有69597个;共得到443819个群体SNP位点,其中高一致性的群体SNP位点119452个。遗传分析结果表明,金花茶种群的遗传多样性水平较高,可分成2个大的类群,其中第2类群包含了全部收集的金花茶种质,其又可分为3个亚族,这3个亚族的分化有明显的地域特征。第1亚族主要是来源于越南的金花茶品系,第2亚族由2个越南的金花茶品系和5个中国的金花茶品系组成,第3亚族全部由来源于中国的金花茶品系组成。研究结果从基因组水平揭示不同地区金花茶种质之间的遗传关系,所开发的SNP位点可进一步用于金花茶种群的进化分析、重要性状的关联分析等。

Development and genetic analysis on SNP sites from Camellia nitidssima based on SLAF-seq technology

In order to learn genetic relationship among Camellia nitidssima germplasms, we collected 25 Camellia nitidssima germplasms from different regions and other five Theaceae germplasms, and sequenced the varieties by SLAFseq technology. Using Actinidia chinensis genome as reference, we constructed SLAF-seq library by specific size of DNA. And then, we carried on high-throughput sequencing, specific SNP sites developing, and evolutionary relationship analysis by using SLAF-seq technology. Finally, we developed specific SNP sites in SLAFs with polymorphism, and constructed evolutionary tree of 25 C. nitidssima and other five Theaceae varieties based on SNP sites. A total of 1 471 113 SLAFs were produced, of which 69 597 SLAFs had polymorphisms. 443 819 SNP sites were developed, of which 119 452 SNP sites had high consistency. The results of genetic analysis showed that the C. nitidssima germplasms exhibited genetic diversity at a higher level and clustered into two groups. The second group contained all of the collected C. nitidssima germplasms, and it could be divided into three subgroups. Differentiation of the three subgroups had obvious regional characteristics. The varieties in the first subgroup were mainly derived from Vietnam, ones in the second subgroup were constituted with two C. nitidssima germplasms from Vietnam and five germplasms from China, while ones in the third Dualsubgroup was entirely composed of germplasms from China. The paper revealed genetic relationship of C. nitidssima germplasms from different provenances at genomic level, and developed SNP sites could be further used for population evolution and correlation analysis of important characteristics in C. nitidssima.