基于全基因组SNP分子标记分析青海云杉遗传结构

以张掖龙渠青海云杉（Picea crassifolia）无性系种子园中的106个青海云杉亲本无性系为研究材料，采用改良CTAB法，提取的青海云杉基因组DNA，构建SLAF文库并进行高通量测序，之后分析SLAF测序数据和筛选SNP位点，基于邻接法分析得到样品的聚类情况。研究得出：将测序的水稻日本晴reads与其参考基因组进行比对，显示本试验双端比对效率为95.33%，说明SLAF建库成功。本研究中所测序列的Q30较高，碱基测 序错误率低，测序质量高；本研究共开发4 058 883个SLAF标签，标签的平均测序深度为21.21×。共开发 12 275 765个青海云杉SNP标记，各青海云杉样本的SNP数量为1 890 934~4 487 841。利用已开发的高质量青海云杉SNP标记，构建了106个青海云杉的系统发育树，发现来自不同种源的青海云杉在各组中分布比较均匀，不同种源的青海云杉多聚为一类。通过SNP标记和主成分分析，这些无性系来源于同一个祖先的可能性较大，表明无性系间亲缘关系相近。为今后遗传多样性的分析、遗传图谱的构建等提供了基础数据，也为青海云杉初级种子园去劣疏伐提供依据，为高世代种子园的营建奠定基础。

Genetic Structure Analysis of Picea crassifolia Based on Genome-wide SNP Molecular Markers

A total of 106 clones of Picea crassifolia in Longqu P. crassifolia Clone Seed Orchard in Zhangye were used as materials， and the genomic DNA of P. crassifolia was extracted by improved CTAB method， the SLAF library was constructed and sequenced with high flux， and then the SLAF sequencing data were analyzed and SNP sites were screened， and the clustering of samples was obtained based on adjacency analysis respectively. The results showed that the efficiency of two terminal alignment was 95.33%， indicated that the SLAF database was successfully established. The Q30 of the sequence measured and the sequencing quality was high， but the error rate of base sequencing was low. A total of 4 058 883 SLAF tags were developed， and the average sequencing depth of the tags was 21.21×. A total of 12 275 765 P. crassifolia SNP markers were developed， and the number of SNPs in each P. crassifolia sample ranged from 1 890 934-4 487 841. Using the developed high quality SNP markers of P. crassifolia， 106 phylogenetic trees of P. crassifolia were constructed. It was found that P. crassifolia from different provenances were evenly distributed in each group， and P. crassifolia from different provenances mostly were clustered into one class. By SNP markers and principal component analysis， these clones were more likely to be derived from the same ancestor， indicated that the genetic relationship between clones was similar. It provids basic data for the analysis of genetic diversity and the construction of genetic map in the future， and also provids a basis for the elite seeds selection in P. crassifolia primary seed orchard， and lays a foundation for the construction of high generation seed orchard.