基于RNA-Seq技术的茶树响应高温胁迫转录组差异性分析  

Difference Analysis of Heat Stress-responsive Transcriptome of Camellia sinensis Based on RNA-Seq Technology

作　　者：翟秀明[1] 唐敏[1] 李解 陈世春[1] 胡方洁 侯渝嘉[1] Zhai Xiuming;Tang Min;Li Jie;Chen Shichun;Hu Fangjie;Hou Yujia(Chongqing Engineering Technology Research Center of Tea Industry,Tea Research Institute,Chongqing Acedemy of Agricultural Sciences,Yongchuan,402160)

机构地区：[1]重庆市农业科学院茶叶研究所,茶叶工程技术研究中心,永川402160

出　　处：《分子植物育种》2020年第17期5629-5637,共9页Molecular Plant Breeding

基　　金：重庆市基础科学与前沿技术研究专项项目(cstc2016jcyjA0060);重庆市农业发展资金项目(NKY-2018AC003);重庆科技攻关计划项目(cstc2012ggB80002);重庆市农业发展资金项目(NKY-2017AB011)共同资助。

摘　　要：为探索茶树响应高温胁迫的分子生理学机制,本研究通过转录组和数字基因表达谱技术,对高温胁迫前后的‘福鼎大白茶’、‘早白尖5号’两组茶树的差异表达进行分析,并基于转录组数据筛选鉴定茶树中与适应高温胁迫相关的保护基因和调控基因。结果显示,项目共测序获得53.7 Gb数据,组装并去冗余后获得的各样本Unigene均超过60976条,总长度,平均长度,N50以及GC含量分别大于53974945 nt,840 nt,1411 bp和41.36%。使用Blast将Unigene比对到七大功能数据库进行注释,在核苷酸序列信息库(nucleotide collection,NT)中被注释的Unigene序列最多,有109581(63.53%)条;高温胁迫后,‘福鼎大白茶’高温胁迫(fuHT)vs‘福鼎大白茶’常温对照(fuCK)和‘早白尖5号’高温胁迫(zaoHT)vs‘早白尖5号’常温对照(zaoCK)两个比较组分别筛选获得5256和5765个差异性表达基因,其中‘福鼎大白茶’上调基因4379个,下调877个,‘早白尖5号’上调基因4997个,下调768个。将差异基因按GO功能分类,‘福鼎大白茶’、‘早白尖5号’高温胁迫后差异基因均富集到41个类别中,分属于GO功能分类体系的生物学过程、细胞组分和分子功能。将差异基因进行KEGG功能分析,‘福鼎大白茶’、‘早白尖5号’两个对比组显著富集的代谢通路(pathway)前3名均为代谢途径、内质网中的蛋白质处理、次级代谢物的生物合成。选取17个在高温胁迫后被诱导的基因,利用实时荧光定量PCR(qRT-PCR)对高通量测序数据进行验证,证明测序结果真实可靠。本研究筛选出大量与高温胁迫相关的响应基因,为下一步耐热茶树新品种的选育提供了更多的理论参考。In order to explore the molecular physiology mechanism of Camellia sinensis in response to high temperature stress,transcriptome and digital gene expression profiling techniques were used to analysis the different expression of'Fudingdabaicha'and'Zaobaijian5'(Camellia sinensis L.)before and after high temperature stress.Based on transcriptome data,protective genes and regulatory genes were identified.The results showed that this project obtained 53.7 Gb data.Each samples obtained more than 60976 Unigene samples after assembling and de-redundant.The total length,average length,N50 and GC content were greater than 53974445 nt,840 nt,1411 bp and 41.36%.Using Blast to annotate Unigene to the seven functional databases,Unigene sequences annotated in the NT database is the most with 109581(63.53%).After high temperature stress,5256 and 5765 differentially expressed genes were obtained from the two comparison groups of'Fudingdabaicha'high temperature stress(fuHT)vs'Fudingdabaicha'room temperature control(fuCK)and'Zaobaijian5'high temperature stress(zaoHT)vs'Zaobaijian5'room temperature control(zaoCK),among which 4379 were up-regulated in'Fudingdabaicha'group,877 were down-regulated,and 4997 were up-regulated in'Zaobaijian5'group,768 were down-regulated.The differential genes of'Fudingdabaicha'and'Zaobaijian5'before and after high temperature stress were enriched into 41 categories according to GO function,which belonged to the biological process,cellular components and molecular functions of the GO functional classification system.The differential gene was analyzed by KEGG function.The top three pathways of differential genes in'Fudingdaba icha'and'Zaobaijian5'that significantly enriched were metabolic pathways,protein processing in the endoplasmic reticulum,and biosynthesis of secondary metabolites.Seventeen genes induced after high temperature stress were selected to verify high-throughput sequencing data by real-time fluorescent quantitative PCR,which proved that the sequencing results were true and reliable.This stu

关 键 词：茶树(Camellia sinensis) 高温胁迫 转录组测序 差异表达基因 QRT-PCR 

分 类 号：S571.1[农业科学—茶叶生产加工]