基于RNA-Seq数据筛选的银叶真藓耐干相关基因表达模式研究

Expression patterns of desiccation tolerance-associated genes of Bryum argenteum screened based on RNA-Seq data

本文以前期获得的银叶真藓在干旱-复水过程中的转录组数据为基础,选择了9个在干旱-复水条件下基因表达差异明显的相关耐干响应基因,并利用荧光定量PCR技术研究其在干旱-复水、ABA诱导胁迫及盐胁迫下的表达模式。研究结果表明,RT-q PCR实验获得的9个基因在干旱-复水下的表达趋势与RNA-Seq基因表达谱中基因表达规律基本一致,因此较好地验证了RNA-Seq数据的可靠性。在干旱-复水过程中,9个基因均不同程度受到干旱和复水诱导表达,但复水阶段基因诱导表达更为强烈,且大多数基因表达量在复水2 h达到高峰,如ERF、Dehydrin(DHN)和HSF3基因等。在ABA和盐胁迫处理下,9个耐干相关基因中有8个同时受ABA和盐诱导上调表达,其中ERF、DHN和LEA基因受ABA和盐胁迫上调表达最为显著。综上,ERF、DHN和LEA等基因同时受干旱-复水,ABA和高盐强烈诱导表达,可作为将来重点研究的抗逆基因。

Based on our previous dehydration-rehydration transcriptome data of Bryum argenteum, nine desic- cation tolerance associated genes were selected and their gene expression patterns under dehydration-rehydra- tion, salt and ABA treatments were investigated using real-time quantitative PCR analysis. The main results were as follows： RT-qPCR results well validated the RNA-seq data, most of genes had similar gene expression trends compared with RNA-seq data under desiccation-rehydration treatment. Nine genes were induced by the dehydration-rehydration treatment at different degrees, and most of genes significantly highly induced at rehy- dration stage, and their abundance were reached maxima at 2 h, such as ERF,, Dehydrin and HSF3 genes. The eight out of nine genes were induced by both ABA and salt treatments, especially the relative expression levels of ERE DHN and LEA genes were dramatically increased under ABA and salt stresses. In conclusion, genes like ERE, DHN and LEA were intensively induced by dehydration-rehydration, salt and ABA treatments, which should be potential stress-tolerant candidate genes for further gene cloning.