转录组学分析外源H2S调控黄瓜响应盐胁迫的机理

Transcriptomics Analysis of the Mechanism of Exogenous H2S Regulating Cucumber in Response to Salt Stress

为了探明硫化氢调控植物响应盐胁迫的机理,以黄瓜幼苗为试材,通过浇灌Hoagland营养液(CK)、200 mmol/L NaCl(T)和200 mmol/L NaCl+15μmol/L NaHS(H2S供体试剂,S)处理,利用Illumina HiSeq高通量测序对黄瓜根系进行转录组测序及差异表达基因分析。结果发现,9个样本获得高质量序列共365.35 Mb,与参考基因组进行比对,比例为86.27%~88.64%的序列总数为315.74 Mb。差异表达基因分析显示:T组较CK组上调基因有1168个,下调基因有1076个;S组较T组上调基因有435个,下调基因有218个。GO富集分析显示:差异表达基因主要富集在分子功能大类中的结合和催化活性类,生物过程大类中的代谢过程、细胞过程和单一生物过程类及细胞组分大类中的膜和膜组分类中。KEGG富集分析将3个比较组的差异表达基因分别归为139,75,127个代谢通路,T-S比较组的差异表达基因主要富集在植物激素信号转导、内质网中的蛋白质加工和光合作用通路中。筛选10个可能与H2S调控黄瓜响应盐胁迫相关的基因:CSC1样蛋白ERD4基因、NADH型硝酸还原酶样基因、蛋白质TIFY 10B样基因、丙二烯氧化物环化酶基因、BTB/POZ和TAZ结构域蛋白1样基因、麦冬蛋白-3样基因、丝裂原激活蛋白激酶激酶3样基因、转录因子bHLH18样基因、IAA-氨基酸水解酶ILR1样4基因和钾通道AKT1基因。研究H2S调控黄瓜响应盐胁迫的机理,为提高植物耐盐性提供了理论数据和参考依据。

In order to verify the mechanism of hydrogen sulfide to regulate the plant response to salt stress,cucumber seedlings were used as materials and treated with Hoagland(CK),200 mmol/L NaCl(T)and 200 mmol/L NaCl+15 mmol/L NaHS(the donor of H2S,S).The expression of transcriptome genes of cucumber roots were studied by using Illumina HiSeq high-throughput sequencing technology.A total of 365.35 Mb of Clean reads were obtained from 9 samples.Mapped reads was 315.74 Mb,and the percentage was 86.27%-88.64%.Analysis of differentially expressed genes revealed that T group had 1168 genes up-regulated and 1076 genes down-regulated compared with CK group,and S group had 435 genes up-regulated and 218 genes down-regulated compared with T group.The Gene Ontology(GO)enrichment analysis revealed that the differentially expressed genes were mainly concentrated in binding and catalytic activity in molecular functional classes;in biological process classes,they were mainly enriched in metabolic processes and cellular processes and single-organism process;Among cell components,the membrane and membrane part classes were mainly concentrated.KEGG enrichment analysis of the differentially expressed genes could be divided into 139,75 and 127 pathway of 3 comparison groups,respectively.The differentially expressed genes in S group compared to the T group were mainly abundant during plant hormone signal transduction,protein processing in endoplasmic reticulum and photosynthesis pathway.Screening of 10 genes that might be related to H2S regulating cucumber response to salt stress:CSC1-like protein ERD4,nitrate reductase(NADH)-like,protein TIFY 10B-like,allene oxide cyclase,BTB/POZ and TAZ domain-containing protein 1-like,tricalbin-3-like,mitogen-activated protein kinase kinase 3-like,transcription factor bHLH18-like,IAA-amino acid hydrolase ILR1-like 4 and potassium channel AKT1.Studying the mechanism of H2S regulating cucumber response to salt stress provides theoretical data and reference basis for improving plant salt tolerance.