Journal of Nuclear Agricultural Sciences ›› 2019, Vol. 33 ›› Issue (7): 1318-1329.DOI: 10.11869/j.issn.100-8551.2019.07.1318

• Induced Mutations for Plant Breeding·Agricultural Biotechnology • Previous Articles     Next Articles

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

YANG Xu*, YANG Zhiling, TAN Mei, CHENG Xiaoyan   

  1. Key Laboratory of Tree Breeding of Zhejiang Province/Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Fuyang, Zhejiang 311400
  • Received:2017-09-06 Revised:2017-12-22 Online:2019-07-10 Published:2019-05-09

厚朴转录组特征分析及EST-SSR标记的开发

杨旭*, 杨志玲, 谭美, 程小燕   

  1. 中国林业科学研究院亚热带林业研究所/浙江省林木育种技术研究重点实验室,浙江 富阳 311400
  • 通讯作者: *同第一作者。
  • 作者简介:杨旭,女,助理研究员,主要从事药用植物资源保护和利用研究。E-mail:yangxu2119@126.com
  • 基金资助:
    国家自然科学基金(31270585)

Abstract: In order to investigate genomic information and develop molecular markers of Houpoëa 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.

Key words: Houpoëa officinalis;, transcriptome, Illumina paired-end sequencing, EST-SSR

摘要: 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。

关键词: 厚朴, 转录组, Illumina-Solexa测序技术, EST-SSR