盐胁迫下转DcCIPK24拟南芥的转录组比较分析

Comparative Transcriptome Analysis of DcCIPK24 Transgenic Arabidopsis Under Salt Stress

【目的】探究铁皮石斛(Dendrobium catenatum Lindl.)DcCIPK24的下游响应基因,为研究DcCIPK24提高耐盐性的分子调控途径提供指导。【方法】利用Illumina NovaSeq 6000测序平台对200 mmol/L NaCl处理和对照组的转DcCIPK24拟南芥、野生型拟南芥进行测序,分析差异表达基因的富集通路,并对通路中的差异基因进行荧光定量PCR验证。【结果】盐胁迫下,从2个转基因拟南芥株系与野生型拟南芥的比较组合中共鉴定出96个差异表达基因;GO注释分析发现96个差异基因主要被富集在分子功能相关通路;KEGG富集分析发现差异基因主要被注释在氨基糖和核苷酸糖代谢、植物MAPK信号通路和甘油酯代谢通路中,选取上调表达的差异基因AtGPAT5、AtSIRK、AtMEE25、AtUGE5和下调表达基因AtAMT1-2、AtATTI3、AtCYP71A25、AtLHCB4.3进行实时荧光定量PCR验证,结果表明盐胁迫下8个差异基因表达趋势与转录组数据基本一致。【结论】盐胁迫下,DcCIPK24主要通过氨基糖和核苷酸糖代谢途径、植物MAPK信号途径和甘油酯代谢通路响应耐盐。

【Objective】This study aimed to explore the downstream response genes of Dendrobium catenatum Lindl.DcCIPK24,and provided guidance for studying the molecular regulatory pathway of DcCIPK24 to improve salt tolerance.【Method】The DcCIPK24 transgenic Arabidopsis thaliana and wild type plants under 200 mmol/L NaCl treatment and normal conditions were used for RNA-seq on the illumina NovaSeq 6000 sequencing platform.The enrichment pathway of differentially expressed genes(DEGs)was analyzed,and their expression patterns were verified by quantitative real-time PCR.【Result】A total of 96 DEGs were identified from transgenic A.thaliana and wild type plants under salt stress.GO annotation analysis showed that 96 DEGs were mainly enriched in molecular function pathways.KEGG enrichment analysis found that DEGs enriched in amino sugar and nucleotide sugar metabolism,plant MAPK signaling pathway and glycerolipid metabolism.The DEGs including four upregulated genes(AtGPAT5,AtSIRK,AtMEE25,and AtUGE5)and four downregulated genes(AtAMT1-2,AtATTI3,AtCYP71A25,and AtLHCB4.3)were selected for qRT-PCR validation.The results showed that the trends of 8 DEGs’expression were basically consistent with the RNA-seq data.【Conclusion】DcCIPK24 responded to salt tolerance mainly through amino sugar and nucleotide sugar metabolism,plant MAPK signaling pathway,and glycerolipid metabolism.