利用转录组测序分析大豆矮小突变体中差异表达基因

Transcriptome Analysis of Differentially Expressed Genes by RNA-Seq in a Dwarf Soybean Mutant

植株高度是影响农作物产量的重要性状,本实验以大豆矮小突变体为研究材料,发现GA3处理可以部分恢复其矮化表型。为了探究其至矮的分子机制,运用RNA-Seq技术测序分析了野生型和突变体的转录组差异,结果表明有多个与植物激素相关的基因差异表达,其中Glyma19g01590与植物激素GA调节有关,生物信息学分析表明差异表达基因Glyma19g01590的开放阅读框有594 bp,推测编码198个氨基酸,该蛋白具有GASA(Gibberellic Acid-Stimulated in Arabidopsis)保守区域,GASA保守区域与GA调节有关。半定量RT-PCR实验表明Glyma19g01590在突变体中的表达水平明显高于野生型,qRT-PCR实验表明Glyma19g01590在突变体的表达量大约是是野生型的24倍。根据以上结果推测Glyma19g01590基因是导致大豆矮小表型的因素之一。

Plant height is an important agronomic character for crop yield improvement.In this study a dwarf mutant could be partially rescued treated by GA3.To explore the molecular mechanism of leading to dwarf phenotype,RNA-seq was used to identify different genes between mutant and wild type,and many genes related to phytohormone were detected to be differential expressed, in which Glyma19g01590 gene expression relates to GA.Bioinformatics analysis indicated that Glyma19g01590 gene contained 594 bp in open reading frame （ORF） and presumably encoded 198 amino acids.Glyma19g01590 protein contains a GASA （Gibberellic Acid-Stimulated in Arabidopsis） domain which could be regulated by GA3.In addition,semi-quantitative RT-PCR experiment showed that the expression level of Glyma19g01590 gene in mutant was higher than that in wild type. Quantitative Real-time RT-PCR （qRT-PCR） result showed the expression quantity of Glyma19g01590 gene in Gmdwarf1 was 24 times compared with that in wild type.The results indicated that Glyma19g01590 gene was perhaps one of the genes that are responsible for dwarf phenotype.