大豆长链非编码RNA lncRNA77580转基因材料的转录组分析  

Transcriptome Analysis of Soybean lncRNA77580 Transgenic Hairy Root

作　　者：牛风娟 陈向前 高飞[2] 孙现军[1] 胡正[1] 张惠媛 王丽侠[1] 姜奇彦[1] 张辉[1] NIU Feng-juan;CHEN Xiang-qian;GAO Fei;SUN Xian-jun;HU Zheng;ZHANG Hui-yuan;WANG Li-xia;JIANG Qi-yan;ZHANG Hui(Institute of Crop Sciences,Chinese Academy of Agricultural Sciences,Beijing 100081;College of Life and Environmental Sciences,Minzu University of China,Beijing 100081)

机构地区：[1]中国农业科学院作物科学研究所,北京100081 [2]中央民族大学生命与环境科学学院,北京100081

出　　处：《植物遗传资源学报》2022年第3期842-856,共15页Journal of Plant Genetic Resources

基　　金：国家自然基金(31601302);国家重点研发计划项目(2016YFD0100304);中国农业科学院科技创新工程项目;江苏省现代作物生产协同创新中心;现代作物生产省部共建协同创新中心。

摘　　要：长链非编码RNAs(lncRNAs,long non-coding RNAs)是长度在200个核苷酸以上的功能性RNA分子,在植物的生长发育及逆境应答中发挥重要调控功能。前期研究发现一个大豆盐胁迫响应lncRNA,称其为lncRNA77580,它的大片段缺失和过表达影响其邻近蛋白编码基因的表达。本研究通过lncRNA77580转基因材料的转录组分析,探究lncRNA77580可能的功能及调控网络。在大豆发状根中过表达lncRNA77580,转基因复合体表现为盐敏感;利用dual-sgRNA/Cas9成功实现lncRNA77580大于4 kb的大片段敲除,突变效率为12.5%。利用lncRNA77580转基因材料进行RNA-seq分析发现,lncRNA77580过表达和大片段缺失显著改变了转基因发状根的基因表达谱。差异表达基因(DEGs,differentially expressed genes)GO和KEGG富集分析表明,lncRNA77580过表达或敲除主要影响了大量转录因子和信号传导类基因的差异表达,这些差异表达基因与植物逆境应答及生长发育密切相关。上述结果说明lncRNA77580可能是在大豆逆境胁迫响应及生长发育过程中具有重要功能的一个长链非编码RNA。Long non-coding RNAs(lncRNAs)are functional RNA molecules with>200 nucleotides in length,and play an important regulatory role in the growth and development of plant.We previously identified a salt stress-related lncRNA in soybean,called lncRNA77580.Deletion and overexpression of lncRNA77580 were found to alter the expression of several neighboring protein-coding genes.In this study,we explored the function and regulation network of lncRNA77580 by the transcriptome analysis of lncRNA77580 transgenic hairy root.Over-expressing lncRNA77580 in soybean hairy roots showed salt sensitive.We generated the edited lines harboring>4kb fragment deletion in lncRNA77580 using dual-sgRNA/Cas9 technology(mutation efficiency:12.5%).By conducting the transcriptome sequencing on the hairy roots with lncRNA77580 overexpressing or knockout,significant changes on transcriptional profiles of the transgenic hairy roots were detected.GO enrichment and KEGG pathway analysis of differentially expressed genes(DEGs)showed that overexpressing or knockout of lncRNA77580 had modified the transcriptional profiles in a large number of transcription factors and signaling genes,which are annotated to be related with plant stress response and plant growth and development.Collectively,these results suggested that lncRNA77580 is a long non-coding RNA with important functions in stress response of soybean and plant growth and development.

关 键 词：大豆 长链非编码RNA 转录组分析 逆境胁迫 基因编辑 发状根 

分 类 号：S565.1[农业科学—作物学]