基于RNA-seq的番茄响应根结线虫MiPDCD6蛋白的SA信号通路基因鉴定与表达分析  

Identification and Expression Analysis of Tomato SA Signaling Pathway Genes in Response to Root Knot Nematode MiPDCD6 Protein Based on RNA-seq

作　　者：邓小大 袁永强 蔡书静 郑礼军 徐春玲[1] 王新荣[1] DENG Xiaoda;YUAN Yongqiang;CAI Shujing;ZHENG Lijun;XU Chunling;WANG Xinrong(College of Plant Protection,South China Agricultural University,Guangdong Province Key Laboratory of Microbial Signals and Disease Control,Guangzhou 510642,China)

机构地区：[1]华南农业大学植物保护学院,广东省微生物信号与作物病害防控重点实验室,广东广州510642

出　　处：《华北农学报》2023年第1期184-195,共12页Acta Agriculturae Boreali-Sinica

基　　金：广东省基础与应用基础研究基金(2019A1515012080);国家自然科学基金项目(31171825,30771409)。

摘　　要：为了揭示南方根结线虫MiPDCD6蛋白抑制番茄PTI免疫的分子机理,以番茄品种新金丰1号MiPDCD6超表达苗为试验材料,以番茄品种新金丰1号组培苗为对照,通过转录组测序技术分别对番茄MiPDCD6超表达苗和对照苗进行转录组测序。以番茄栽培品种Heinz 1706基因组作为参考基因组进行对比,利用FPKM法计算基因表达量,设定参数(|log_(2)FC|>1且P<0.05)筛选差异表达基因。利用Gene Ontology(GO)数据库进行差异表达基因GO功能富集分析,统计每个GO term中的差异表达基因数目,计算出基因富集的显著性,找出富集显著的功能条目。利用KEGG数据库对差异表达基因进行Pathway富集分析,超几何分布检验方法计算每个Pathway中差异表达基因富集的显著性。以FDR和基因数量来衡量KEGG的富集程度。结合基因差异表达分析和功能富集分析,研究MiPDCD6蛋白对番茄PTI免疫相关通路基因的影响。结果发现:在超表达MiPDCD6番茄植株中,与野生型番茄相比,MiPDCD6过表达番茄植株中有2366个差异表达基因(DEGs),其中1354个上调基因,1012个下调基因。这些DEGs中,通过GO和KEGG注释,植物激素信号转导(sly04075)、植物-病原互作(sly04626)、植物MAPK信号通路(sly04016)和丙环素生物合成(sly00940)等KEGG通路中富集到大量的差异表达基因。SA生物合成途径包括ICS和PAL,MiPDCD6过表达番茄植株中SA合成通路PAL1和PAL-like基因以及SA信号转导途径TGA9、TGA10-like和PR1a2基因均显著下调,表明MiPDCD6基因可能抑制SA合成,从而抑制植物PTI免疫。In order to reveal the molecular mechanism of MiPDCD6 protein suppressing tomato PTI immunity,the MiPDCD6 overexpression seedlings of tomato variety Xinjinfeng 1 were used as experimental materials,and the tissue culture seedlings of tomato variety Xinjinfeng 1 were used as control.Transcriptome sequencing was performed on tomato MiPDCD6 overexpressing seedlings and control seedlings,respectively.With tomato cultivars Heinz 1706,comparing the genome as a reference genome,FPKM method was used to calculate quantity of gene expression,set parameters(|log_(2)FC|>1 and P<0.05)in screening the differentially expressed genes(DEGs).Gene ontology(GO)database was used to analyze the GO functional enrichment of DEGs,count the number of DEGs in each GO term,calculate the significance of gene enrichment,and find out the functional term with significant enrichment.KEGG database was used for Pathway enrichment analysis of DEGs,and hypergeometric distribution test was used to calculate the significance of enrichment of DEGs in each Pathway.The enrichment degree of KEGG was measured by FDR and gene number.Combined with gene differential expression analysis and functional enrichment analysis,the effect of MiPDCD6 protein on tomato PTI immune-related pathway genes was studied.The results showed that there were 2366 DEGs in MiPDCD6 overexpressed tomato plants compared with wild-type tomato,including 1354 up-regulated genes and 1012 down-regulated genes.In these DEGs,a large number of differentially expressed genes were concentrated in KEGG pathways such as plant hormone signal transduction(sly4075),plant-pathogen interaction(sly04626),plant MAPK signal pathway(sly04016)and procycloid biosynthesis(sly00940)through GO and KEGG annotation.SA biosynthesis pathways included ICS and PAL.In the MiPDCD6 overexpressed tomato plants,PAL1 and PAL-like genes in SA synthesis pathways and TGA9,TGA10-like and PR1a2 genes in SA signal transduction pathways were significantly down-regulated,suggesting that MiPDCD6 may inhibit SA synthesis thus inhib

关 键 词：番茄 根结线虫 转录组 差异表达基因 SA信号通路 MiPDCD6 

分 类 号：Q78[生物学—分子生物学] S641.2[农业科学—蔬菜学]