利用RNA-seq技术分析淹水胁迫下转BnERF拟南芥差异表达基因

RNA-sequencing Analysis of Differentially Expressed Genes in Wild-type and BnERF-transgenic Arabidopsis Under Submergence Treatment

为探究淹水胁迫下BnERF调节的耐淹防御相关途径,应用RNA-seq技术,对淹水6小时后的拟南芥(Arabidopsis thaliana)野生型(WT)和转BnERF株系(E33)幼苗进行基因表达分析。结果表明,淹水3天后,E33表现出较强的耐淹性,地上部生长状况和根系发育均明显强于野生型。E33幼苗未淹水处理时相对于野生型单独上调的基因有9个,4个为膜结合蛋白,其中2个参与MAPK级联途径,其它5个参与氧化胁迫及水分调节途径;与未淹水野生型相比,无论是未淹水处理还是淹水6小时后的E33幼苗中缺氧响应、抗氧化防护及细胞、器官发育相关基因的表达量均上调。另外,淹水6小时后E33的差异基因并未完全覆盖淹水6小时后野生型的差异基因;E33幼苗中缺氧响应、氧化胁迫响应、能量的产生与转变、乙醇代谢途径中的基因以及乙烯响应因子基因的表达量都明显高于野生型。上述结果表明,BnERF直接或间接调节植物的淹水胁迫相关生理代谢途径,参与淹水胁迫的防御过程。

RNA-sequencing （RNA-seq） was used to analyze differential gene expression in the wild-type （WT） and BnERF-transgenic Arabidopsis （E33） after 6 h submergence treatment to find pathways involved in defense against submergence treatment regulated by BnERF. E33 showed more tolerance with stronger shoot and root growth than the WT under 3 d submergence. Nine genes were specifically upregulated in E33 without submergence treatment as com- pared with the WT. Four genes encoded membrane-bound proteins and two of these participated in the mitogen-activated protein kinase cascade; the other five were involved in oxidative stress and water regulatory pathways. Genes responsive to hypoxia, antioxidant defense and plant development were upregulated in E33 under the control condition or 6 h sub- mergence as compared with the WT without submergence treatment. As compared with the WT, differently expressed genes in E33 did not significantly overlap with the WT when both were treated for 6 h submergence. Furthermore, genes responsive to hypoxia, oxidative stress, production and transformation of energy, and ethanol metabolic pathways as well as ethylene response factor genes showed higher expression level in E33 than that in the WT, so these pathways are regulated by BnERF directly or indirectly to participate in defense against submergence stress.