基于GBS测序的全基因组SNP揭示贵州地方茶组植物资源的亲缘关系

Genome-wide SNP developed by genotyping-by-sequencing revealed the phylogenetic relationship of Sect. Thea(L.) Dyer resources in Guizhou

  • 摘要: 【目的】分析贵州地方茶组植物资源的亲缘关系,为明确贵州地方茶组植物资源的遗传关系及其保护、利用提供科学依据。【方法】以从贵州省内不同区域收集的41份茶组植物资源及贵州省茶叶研究所茶树种质资源圃保存的18份省内外育成茶树品种为材料,利用基于GBS(Genotyping by sequencing)的简化基因组测序技术对其基因组SNP位点进行检测,基于获得的高质量SNP位点对这些材料进行遗传特征分析。【结果】从59份茶组植物材料获得45.84 Gb高质量序列(Clean reads)数据,平均每个材料为795.6 Mb,约占改良版茶树基因组大小(2.93 Gb)的26.5%,平均比对率为72.62%,经过滤后得到248772个高质量SNP位点,其中83.98%的高质量SNP位点分布在基因间区,16.02%分布于基因区;有22614个SNP位点分布在内含子,15038个SNP位点分布在外显子区,2203个SNP位点分布在非翻译区(UTR)。59份茶组植物材料的观察杂合度(Ho)为0.016~0.081,期望杂合度(He)为0.006~0.064,F为-0.331~0.737。主成分分析结果、系统发育进化树构建情况及遗传结构分析结果均显示59份茶组植物材料可分为3个类群,其中全部茶种(Camellia sinensis)材料归在一个类群、疏齿茶(C. remotiserrata)和大厂茶(C. tachangensis)归在一个类群、9份突肋茶(C. costata)单独归在一个类群,但疏齿茶与大厂茶及两个区域的大厂茶均处于独立的亚类群,此外茶种中的阿萨姆变种(C. sinensis var. assamica,CSA)和中国变种(C. sinensis var. sinensis,CSS)也处于不同的进化分支;突肋茶与疏齿茶和大厂茶的亲缘关系较其与茶种的亲缘关系更近。茶种、疏齿茶、大厂茶和突肋茶4类茶组植物存在互相融合的遗传背景。根据地理来源和种质类型可将59份茶组植物材料分为11个种群,不同种群间具有较低的基因流,但种群内部具有较高的基因流。【结论】不同茶组植物在基因组水平上存在明显差异,经典形态学分类上合二为一的大厂茶和疏齿茶在遗传结构上也存在差异,即分为2个变种更合适;茶组植物种内亲缘关系与其地理来源直接相关,在育种实践中应尽量避免相同地理来源材料间的杂交应用。

     

    Abstract: 【Objective】To clarify the genetic relationship of local Sect. Thea(L.) Dyer resources in Guizhou, and to provide scientific basis for their protection and utilization.【Method】A total of 41 Sect. Thea(L.) Dyer resources collected from different regions of Guizhou and 18 tea plant varieties bred from Guizhou and other provinces that kept by Institute of Tea, Guizhou Academy of Agricultural Sciences were used as materials. The genotyping-by-sequencing(GBS) was used to detect the single nucleotide polymorphism(SNP) sites in their genomes, and the genetic characteristics of the 59 Sect. Thea(L.) Dyer resources were analyzed based on the obtained high-quality genomic SNP sites.【Result】A total of 45.84 Gb clean reads were obtained from the 59 Sect. Thea(L.) Dyer resources, with an average of 795.6 Mb per material, accounting for 26.5% of the improved version of tea reference genome(2.93 Gb), and the average alignment rate was 72.62%. A total of 248772 high-quality genomic SNPs were obtained, of which 83.98% and 16.02% were distributed in intergenic region and genic region, respectively. There were 22614 SNP sites distributed in introns, 15038 SNP sites distributed in exon regions, and 2203 SNP sites distributed in untranslated regions(UTR). The results showed that the observed heterozygosity(Ho) of them ranged from 0.016 to 0.081, the expected heterozygosity(He) was 0.006 to 0.064, and F value was-0.331 to 0.737. The 59 samples were divided into three groups based on principal component analysis(PCA), phylogenetic tree construction and genetic structure analysis. Among them, all of the Camellia sinensis were in one group, C. remoterrata and C. tachangensis were in one group, and the nine C. costata were in one group. However, C. remoterrata and C. tachangensis and the C. tachangensis from two different regions could be further divided into different independent subgroups, and the C. sinensis var. assamica(CSA) and C. sinensis var. sinensis(CSS) were also in the different branches of evolutionary tree. Additionally, based on the phylogenetic tree, the genetic relationships between C. costata and C. remoterrata or C. tachangensis were more closer to that between C. costata and C. sinensis. The gene structure analysis showed that there was a fusion genetic background among the four classifications of Sect. Thea(L.) Dyer resources(C. sinensis, C. remoterrata, C. tachangensis and C. costata) used in this study. According to geographical origin and germplasm type, the 59 Sect. Thea(L.) Dyer resources were divided into 11 groups, and the gene flow analysis showed that there was a low level of gene flow among different groups and a high level of gene flow within the group.【Conclusion】There are obvious differences among different Sect. Thea(L.) Dyer groups at the genomic level. Based on the gene-tic structure, C. tachangensis and C. remoterrata are more appropriate to be divided into two varieties, although they are combined in the classical morphological classification. The intraspecific genetic relationship of different Sect. Thea(L.) Dyer is directly related to their geographical origin. Therefore, the same geographical source material hybrid application should be avoided in breeding practice.

     

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