加工番茄耐盐突变体耐盐相关基因的转录组分析  

Transcriptome Analysis of Salt-tolerance-related Genes in Processing Tomato Salt-Tolerant Mutants

作　　者：张国儒[1] 庞胜群[1] 郭晓珊 单淑玲 Zhang Guoru;Pang Shengqun;Guo Xiaoshan;Shan Shuling(Key Laboratory of Applied Physiology and Germplasm Utilization of Fruits and Vegetables,Department of Horticulture,College of Agronomy,Shihezi University,Shihezi,83200)

机构地区：[1]石河子大学农学院园艺系特色果蔬栽培生理与种质资源利用兵团重点实验室,石河子832003

出　　处：《分子植物育种》2018年第18期5884-5896,共13页Molecular Plant Breeding

基　　金：新疆建设兵团科技支疆计划(2014AB004);石河子大学动植物育种专项(gxjs2015-yz01)共同资助

摘　　要：为筛选鉴定番茄叶片响应盐胁迫应答基因,本研究通过甲基磺酸乙酯(methyl ethyl sulfonate, EMS)诱变处理加工番茄‘JW9’获得的16株耐盐突变植株。经单独收种后,利用(RNA sequencing, RNA-Sep)技术对获得的耐盐突变体加工番茄幼苗叶片进行转录组测序分析。提取样品总RNA进行转录组cDNA文库制备后进行Illumina HiSeq/MiSeq测序。对转录组测序结果进行分析共获得了102 Gb大约810 974 186个高质量的Clean阅读序列,Cufflinks软件重构转录本后得到38 190个已知转录本,1 647个未知转录本。在测序样本中共筛选出4 367个基因在样品间显著差异表达,其中上调表达基因2 606个,下调表达基因1 761个。经qRT-PCR检测表明,10个随机选择的差异表达基因(differentially expressed genes, DEGs)表达量变化与转录组测序结果一致。通过GO和KEGG通路富集分析,从具有耐盐功能的丝氨酸/苏氨酸蛋白激酶和ATPase酶合成相关的基因中,找到5个显著差异表达基因Solyc08g066270.1和Solyc03g083420.2以及Solyc10g018530.1、Solyc09g082890.1和Solyc02g014190.2可能为耐盐相关基因。本研究从耐盐突变体与原始亲本的差异表达基因中筛选出耐盐候选基因,为进一步鉴定和克隆出番茄的耐盐基因培育耐盐品种番茄提供理论参考。In order to screen and identify tomato leaves in response to salt stress response genes, we studied 16 salt-tolerant mutant plants obtained from processing tomato ‘JW9＇were mutagenized by methyl ethyl sulfonate（EMS）. After separate seed harvesting, RNA Sequencing（RNA-Seq） was used to analyze the transcriptome of the obtained seedling leaves of processing tomato salt-tolerant mutant. Total RNA samples were extracted for transcriptome cDNA library preparation followed by Illumina HiSeq/MiSeq sequencing analysis. Analysis of transcriptome sequencing results showed that 102 Gb contained approximately 810 974 186 high quality Clean reading sequence, Cufflinks software reconstructed transcripts obtained 38 190 known transcripts and 1 647 unknown transcripts. In the sequencing samples, a total of 4 367 genes were screened out significantly differentially expressed among samples, of which 2 606 genes were up-regulated and 1 761 genes were down-regulated. The results of qRT-PCR showed that the expression of 10 randomly selected differentially expressed genes was consistent with the results of transcriptome sequencing. Through GO and KEGG pathway enrichment analysis, we found five significantly differentially expressed genes from the genes involved in the synthesis of serine/threonine protein kinases and ATPase with salt tolerance function, including Solyc08 g066270.1, Solyc03 g083420.2, Solyc10 g018530.1,Solyc09 g082890.1 and Solyc02 g014190.2 might be salt-tolerance related gene. In this study, salt-tolerant candidate genes were screened from the differential expression genes of salt-tolerant mutants and original parents, for providing theoretical references to the further identification and cloning of salt-tolerant tomato genes for tolerance to salt-tolerant tomato.

关 键 词：加工番茄 EMS RNA-SEQ 耐盐基因 QRT-PCR 

分 类 号：S641.2[农业科学—蔬菜学]