生物技术通报 ›› 2016, Vol. 32 ›› Issue (7): 40-47.doi: 10.13560/j.cnki.biotech.bull.1985.2016.07.006

• 技术与方法 • 上一篇    下一篇

基于高通量测序的发芽苦荞转录组学研究

陈春旭1, 李琦2, 郭元新1, 杜传来1, 丁志刚1   

  1. 1. 安徽科技学院食品药品学院,凤阳 233100;
    2. 深圳市坪山新区环境监测站,深圳 518118
  • 收稿日期:2015-09-12 出版日期:2016-07-25 发布日期:2016-07-25
  • 作者简介:陈春旭,男,硕士,助教,研究方向:食品饮料生产工艺及品质控制;E-mail:ccx1205@126.com
  • 基金资助:
    安徽省自然科学基金项目(1308085MC32),安徽科技学院农产品加工及贮藏工程重点学科项目(AKZDXK2015B04)

Transcriptome Analysis of Germinated Tartary Buckwheat Based on High-throughput Sequencing Technology

CHEN Chun-xu1, LI Qi2, GUO Yuan-xin1, DU Chuan-lai1, DING Zhi-gang1   

  1. 1. College of Food and Drug,Anhui Science and Technology University,Fengyang 233100;
    2. Pingshan Environmental Monitoring Station,Shenzhen 518118
  • Received:2015-09-12 Published:2016-07-25 Online:2016-07-25

摘要: 采用新一代高通量测序技术Illumina SolexaHiseq 2500对发芽荞麦转录组进行测序,结合生物信息学方法开展基因表达谱研究和功能基因预测。通过测序,获得了42 953 962个序列读取片段(reads),包含了5.37 Gb碱基序列信息。对reads进行序列组装,获得45 278个单基因簇(unigenes),平均长度862 bp,序列信息达到了39 Mb。另外,从长度分布、GC含量、表达水平等方面对unigenes进行评估,数据显示测序质量好,可信度高。数据库中的序列同源性比较表明,2 127个unigenes与其他生物的己知基因具有不同程度的同源性。发芽苦荞转录组中的unigenes与细胞进程、细胞和蛋白结合相关。将unigenes与KOG数据库进行比对,根据其功能大致可分为24类。以KEGG数据库作为参考,依据代谢途径可将unigenes定位到328个代谢途径分支,包括核糖体代谢通路、碳水化合物代谢等,并且筛选出38条参与GABA合成的氧化磷酸化代谢的unigenes。SSR位点查找发现,从71 366个unigenes中共找到7 141个SSR位点。SSR不同重复基序类型中,出现频率最高的为A/T,其次是AAG/CTT和AT/AT。

关键词: 发芽苦荞, Illumina, 转录组, 高通量测序

Abstract: Illumina SolexaHiseq 2500 high-throughput sequencing technology was used to get the comprehensive transcriptome from germinated tartary buckwheat. As a result,42 953 962 sequence reads containing 5.37 Gb nucleotide sequence information were obtained. After de novo assembly by the software of Trinity,a total of 45 278 unigenes were generated,corresponding to a total of 39 Mb with an average length 862 bp. In addition,the data from the evaluation of the unigenes indicated fine sequencing quality and high reliability from the aspects of length distribution,GC content,and expression level. The comparison of sequence homology in database showed that 2 127 unigenes had various degrees of homology with other known biological genes. The unigenes in the transcriptome of germinated tartary buckwheat were correlated with cellular processes,cell and protein binding. According to KOG database,the unigenes were broadly divided into 24 categories. Referring to KEGG database,unigenes were located into 328 metabolic pathways,including ribosome,carbohydrate metabolism and so on. And 38 unigenes involved in the synthesis of GABA in oxidative phosphorylation metabolism were screened. Total 7 141 unigenes were found from 71 366 by SSR and the highest frequency was A/T,followed by AAG/CTT and AT/AT.

Key words: germinated tartary buckwheat, Illumina, transcriptome, high-throughput sequencing technology