延胡索转录组表达序列标签微卫星标记的开发及其功能分析

Development and function analysis of EST-SSR markers in the transcriptome of Corydalis yanhusuo

目的研究延胡索块茎和叶转录组水平上表达序列标签微卫星(EST-SSR)的分布规律,分析其差异性,并探索含EST-SSR基因的生物学功能,为延胡索遗传多样性评估和遗传育种提供方法和依据。方法利用生物信息学方法构建延胡索块茎和叶的转录组EST-SSR序列库,识别和搜索其EST-SSRs序列,利用TBtools进行GO功能分析,使用KAAS进行KEGG通路富集分析。结果延胡索的块茎和叶转录组中分别有7952个和137197个EST-SSR位点,其序列长度占其转录组长度的比例分别为0.87%、0.89%,丰度分别为475.76、458.91个·Mb^(-1)。延胡索块茎和叶转录组均为三碱基重复类型最多,其中,以AAG/CTT重复类型最多。通过对延胡索块茎含有EST-SSR的unigene进行功能富集分析表明:含EST-SSR的unigene序列的差异表达基因与化合物结合功能、有机物代谢功能、核糖体功能、氧化磷酸化等通路有关。结论延胡索块茎和叶转录组EST-SSR的比例、丰度和密度高度一致,其中,含EST-SSR的unigene具有显著的生物学功能。

OBJECTIVE To study distribution patterns of EST-SSR sequence at the transcriptomic level of tuber and leaves of Corydalis yanhusuo,and to explore the differences and biological functions of unigene containing EST-SSR,so as to provide methods and basis for genetic diversity assessment and genetic breeding of C.yanhusuo.METHODS The EST-SSR sequence library of the transcriptome of tuber and leaves of C.yanhusuo was constructed by bioinformatics method,and EST-SSR sequences were identified and searched.TBtools was used for GO function analysis,and KAAS was used for KEGG pathway enrichment analysis.RESULTS There were 7952 and 137197 EST-SSR loci in the tuber and leaf transcriptome of C.yanhusuo,respectively,and the sequence length accounted for 0.87%and 0.89%of the length of transcription,and the abundance was 475.76 and 458.91/Mb,respectively.The transcriptome of tuber and leaf of C.yanhusuo had the highest number of triple base repeats,in which,AAG/CTT is the most repetitive type.The functional enrichment analysis of unigene containing EST-SSR in tuber showed that unigene containing EST-SSR were related to compound binding function,organic metabolic function,ribosome function,oxidative phosphorylation and other pathways.CONCLUSION The ratio,abundance and density of EST-SSR in the transcripts of tubers and leaves of C.yanhusuo are highly consistent,and unigene containing EST-SSR has significant biological function.