蒙古岩黄芪全长转录组测序及微卫星位点特征分析

Full-length Transcriptome Sequencing and Microsatellite Characterization in Hedysarum mongolicum

蒙古岩黄芪(Hedysarum mongolicum Turcz.)是中国西北干旱半干旱地区防风固沙、保持水土的重要物种,目前其转录组学和遗传学研究很少。本研究利用PacBio Sequel测序平台对蒙古岩黄芪在-0.5 MPa干旱胁迫下的植株转录组进行全长测序,在对转录本进行聚类纠错后利用NR、GO、KEGG、Pfam、egg NOG、Swiss-Prot 6个数据库进行功能注释,分别有8202(NR)、6290(GO)、4087(KEGG)、7182(Pfam)、8108(eggNOG)和7433(Swiss-Prot)个转录本获得功能注释。通过简单序列重复(Simple sequence repeat)分析,共生成20 G数据,并根据ROI是否含有引物和嵌合体进行分类,得到145462条转录本,其中非全长转录本的有19962(13.72%),全长转录本为125493(85.16%),平均序列长度为2322 bp;Quiver校正得到15690个Isforms,准确度大于99%的高质量一致性序列有15688条。在高质量的转录本中,共有4717个SSR位点,单核苷酸序列占31.20%,1~3个核苷酸重复序列占主导位置,占总SSR位点的91.99%。研究结果增加了对蒙古岩黄芪遗传特性的认知,为下一步开展全长基因克隆、基因表达谱分析以及功能基因的挖掘与利用等提供参考。

Hedysarum mongolicum Turcz.is an important species for wind protection,sand fixation and soil conservation in semi-arid areas of Northwest China.However,few transcriptomic and genetic studies have been conducted.In this study,Pac Bio Sequel sequencing platform was used to perform full-length sequencing of H.mongolicum transcriptome under-0.5 MPa PEG-6000 osmotic stress.After cluster correction of the transcriptome,six databases including NR,GO,KEGG,Pfam,egg NOG and Swiss-Prot were used for functional annotation.Functional annotation were obtained in 8202(NR),6290(GO),4087(KEGG),7182(Pfam),8108(egg NOG)and 7433(Swiss-Prot)transcripts,respectively.A total of 20 Gb data were generated by simple sequence repeat(SSRs)analysis.Accordingto ROI classification,145462 transcripts were obtained,of which 19962(13.72%)were non-fulllengthtranscriptsand 125493(85.16%)were full-lengthtranscripts.Theaveragesequence lengthwas 2322 bp.Quiver corrected 15690 ISForms,and 15688 high quality consistent sequences with accuracy greater than 99%.Among the high-quality transcripts,there were a total of 4717 SSR sites,of which single nucleotide sequences accounted for 31.20%,and 1~3 nucleotide repeats were dominant,accounting for 91.99%of the total SSR sites.The results of this study is increasing the understanding of the genetic characteristics of H.mongolicum,and laying a solid foundation for the further development of full-length gene cloning,gene expression profile analysis,and the mining and utilization of functional genes.