厚朴转录组特征分析及EST-SSR标记的开发

Characteristics Analysis of Houpoea officinalis Transcription and Development of EST-SSR Markers

为探明厚朴的基因组信息,开发合适的生物学工具以促进厚朴的遗传育种,本研究利用Illumina-Solexa测序技术对厚朴根、茎皮、叶不同组织的9个样本进行转录组分析,获得大量的序列信息,并以转录组序列为基础进行厚朴SSR分子标记的大规模发掘。结果表明,共获得了109 526条厚朴转录组序列,平均序列长度为1 023 bp,通过与蛋白数据库NR、Swiss-Prot、GO、KOG和KEGG五个数据库比对,分别有6 0361、39 942、65 535、51 351和27 310条厚朴转录组的Unigene被注释;厚朴转录组表达信息进行大通量SSR位点的发掘,发现了含SSR位点的序列25 925条,共27 721个SSR位点,SSR出现的频率为23.67%。厚朴转录组共发现了374种碱基重复模式,且CT/GA和AG/TC是主要的双碱基重复序列,AGA/TCT和AAG/TTC是主要的三碱基重复序列;随机挑选的180对SSR引物中有104对产生清晰可重复的条带,21对引物具有多态性,厚朴SSR引物多态性较高。本研究结果为厚朴及近缘种的遗传多样性分析、遗传图谱构建及比较基因组研究奠定了一定的理论基础。

In order to investigate genomic information and develop molecular markers of Houpo?a officinalis,Illumina-Solexa Genome sequencing platform was used to analyze different tissues of velamen,stem bark and leaf of 9 individuals to get a large number of Unigenes,and to develop abundant SSR markers on the basis of the transcription sequencing data.The results showed that a total of 109 526 Unigenes with an average length of 1 023 bp were produced,among which 60 361,39 942,65 535,51 351 and 27 310 were annotated in NR,Swiss-Prot,GO,KEGG and KOG database,respectively.A total of 25 949 sequences with 27 721 SSRs were detected through transcription of H.officinalis,SSR sites occurred with a frequency of 23.67%.Among the total of 374 identified EST-SSRs,CT/GA(31.8%)and AG/TC(27.90%)were the dominant di-nucleotide repeat motifs,AGA/TCT and AAG/TTC were the dominant tri-nucleotide repeat motifs.A total of 180 SSR markers were randomly selected for validation,among these,104 produced reproducible amplicons and 21 were polymorphic which showed that EST-SSRs of H.officinalis were polymorphic.The results of this study laid a good foundation for genetic analysis,map construction and comparative genome of H.officinalis and related species.