过表达NtH202烟草叶片响应烟蚜侵袭转录组分析  

Transcriptome Analysis of Overexpressing NtH202 Nicotiana tabacum Leaves in Response to Tobacco Aphid Invasion

作　　者：孙计平[1] 王亚乐 李雪君[1] 孙焕[1] 平文丽[1] SUN Jiping;WANG Yale;LI Xuejun;SUN Huan;PING Wenli(Tobacco Research Institute,He'nan Academy of Agricultural Sciences/Key Laboratory for Green Preservation&Control of Tobacco DiseaseSand PestSin Huanghuai Growing Area,Xuchang He'nan 461000,China)

机构地区：[1]河南省农业科学院烟草研究所/黄淮烟区烟草病虫害绿色防控重点实验室,河南许昌461000

出　　处：《种子》2024年第6期44-54,共11页Seed

基　　金：河南省科技攻关项目“NTH202基因的抗蚜功能分析和验证”(212102110306);河南省农业科学院自主创新专项基金项目“烟草抗蚜虫种质资源筛选和抗虫机理的研究”(2020ZC17)。

摘　　要：为了筛选烟草受到蚜虫胁迫的差异表达基因,对野生型和过表达NtH202株系烟草幼苗接蚜虫,并在接虫前和接虫6 h采集叶片,提取总RNA后,进行转录组测序,对差异表达基因进行GO和KEGG分析,挖掘相关转录因子,并利用qRT-PCR方法验证。结果表明,受到烟蚜侵袭后,WT共有8694个基因表达量变化,OE共有6795个基因表达量变化,WT上调和下调差异基因数目更多,二者共同变化的差异基因有2663个。GO分类表明,烟蚜侵袭后OE富集到免疫系统进程(GO:0002376)的基因数目明显高于WT。OE和WT富集差异基因数量最多的均是植物-病原体互作途径,其次是植物激素信号转导、淀粉和蔗糖代谢通路和MAPK信号通路-植物通路。与WT相比,NtH202过表达株系共有1342个基因上调在4倍以上;GO分析表明,差异表达基因的功能主要集中在生物学过程、细胞组分和分子功能;KEGG分类显示,与WT相比较,烟蚜侵袭前后,OE均是参与植物病原体互作途径的基因最多。蚜虫侵袭后,WT和OE共同变化的基因中有9个WRKY基因上调表达、5个下调表达,NtWRKY6、NtWRKY11、NtWRKY51、NtWRKY68、NtWRKY70基因RNA测序结果与qRT-PCR结果表现一致,均为上调表达,预测WRKY转录因子在抗蚜反应中起着重要作用。受到蚜虫侵袭后,WT和OE共同变化的基因中有8个MYB上调表达、8个下调表达,MYB转录因子ODO1基因RNA测序结果与qRT-PCR结果表现一致,预测转录因子ODO1在烟草抗蚜反应中起着重要作用。In order to screen the differentially expressed genes of Nicotiana tabacum under aphid stress,Nicotiana tabacum seedlings of wild type and overexpressed NtH202 strain were exposed to aphid,and the leaves were collected before and after 6 hourSof exposure,total RNA wasextracted,and transcriptome sequencing wasperformed.The differentially expressed genes were analyzed by GO and KEGG,the relevant transcription factors were mined,and verified by qRT-PCR.The results showed that there were 8694 gene expression changes in WT and 6795 gene expression changes in OE,and the number of up-regulated and down-regulated WT differential genes washigher,and 2663 differential genes were changed by both.The GO classification showed that the number of genes enriched by OE into immune system process (GO:0002376)wassignificantly higher than that of WT.The plant-pathogen interaction pathway had the largest number of OE and WT differentially enriched genes,followed by plant hormone signal transduction,starch and sucrose metabolism pathway,and MAPK signal pathway-plant pathway.Compared with WT,a total of 1342 genes of NtH202 overexpressed strain were upregulated more than 4 times.GO analysis showed that the functionSof differentially expressed genes were mainly concentrated in biological processes,cell componentSand molecular functions.KEGG classification showed that compared with WT,OE wasthe most involved gene in plant pathogen interaction pathway before and after aphid invasion.After aphid invasion,9 genes of WT and OE were up-regulated and 5 were down-regulated.RNA sequencing results of NtWRKY6,NtWRKY11,NtWRKY51,NtWRKY68 and NtWRKY70 genes were consistent with qRT-PCR results .It waspredicted that WRKY transcription factor played an important role in anti-aphid response.After being invaded by aphids,8 MYB genes were up-regulated and 8 MYB genes were down-regulated in both WT and OE genes.The results of RNA sequencing of MYB transcription factor ODO1 gene were consistent with the results of qRT-PCR,which predicted that transcription f

关 键 词：NtH202基因 转录组 蚜虫 抗蚜性 烟草 

分 类 号：S572[农业科学—烟草工业]