COBRA基因编码COBRA蛋白,是一种细胞壁可扩展性蛋白.COBRA蛋白通过参与细胞壁的扩展性调节调控植物细胞伸长生长等生理过程.为探究COBRA基因在大豆发育过程中的潜在功能,文章利用生物信息学方法对大豆COBRA家族基因(GmCOBRAs)进行全基因组水平鉴定.通过对COBRA蛋白结构域进行BLASTP搜索,确定GmCOBRAs;以生物信息学方法分析其进化关系、基因结构、保守基序和共线性关系;利用转录组数据分析其不同组织部位表达模式;利用再生能力强的大豆品种东农50和再生能力弱的Keburi为材料进行器官发生试验;利用实时荧光定量PCR检测GmCOBRAs基因在丛生芽诱导和丛生芽伸长阶段的表达模式.结果表明,本研究共鉴定出大豆基因组中的24个GmCOBRAs,约为拟南芥COBRA基因数量2倍.系统发育分析表明,GmCOBRAs可分为2个亚族,分布在14条染色体上.亚细胞定位预测表明24个GmCOBRAs蛋白主要位于细胞膜.在大豆全基因组复制事件中,片段重复对GmCOBRAs扩大发挥重要作用.组织表达模式分析显示,GmCOBRAs基因在大豆不同器官和组织中表达差异较大,其中GmCOBRA12、GmCOBRA22和GmCOBRA14的表达水平较高,表明这些基因可能参与整个大豆发育过程中细胞壁扩张的调节.对6个在种子中高表达的GmCOBRAs进行qRT-PCR试验发现,GmCOBRA21在丛生芽诱导和伸长阶段中显著表达,表明该基因在大豆再生过程中可能扮演关键角色.亚细胞定位表明GmC0BRA21定位于细胞膜.GmC0BRAs在大豆再生过程中发挥重要作用,其中GmC0BRA21可能为该过程中的核心基因之一.
COBRA gene encodes COBRA protein,which was a cell wall expandable protein.COBRA protein regulated physiological processes such as plant cell elongation and growth by participating in the regulation of cell wall expansibility.In order to explore the potential functions of COBRA gene family during soybean development,this paper used bioinformatics methods to identify soybean COBRA family genes(GmCOBRAs)at the whole genome level.In this study,the BLASTP search of the domain of COBRA protein was used to determine GmCOBRA s,and its evolutionary relationship,gene structure,conservative motif and collinearity analysis were analyzed by bioinformatics,and the expression patterns of different tissue parts were analyzed by transcriptome data.Using soybean Dongnong 50 with strong regeneration ability and Keburi with weak regeneration ability as materials for organogenesis experiment,real-time fluorescence quantitative PCR was used to detect the expression pattern of GmCOBRA s in tufted bud induction and tufted bud elongation stage.In this study,24 GmCOBRA s in soybean genome were identified,more than twice the number of COBRA genes in Arabidopsis thaliana.Phylogenetic analysis showed that GmCOBRA s could be divided into two subfamilies and distributed in 14 chromosomes in soybean.The subcellular localization prediction results showed that 24 GmCOBRAs genes were mainly located in the cell membrane.Fragment repetition played an important role in the expansion of GmCOBRA family genes in the whole genome replication event of soybean.The analysis of tissue expression pattern showed that there were significant differences in the expression of GmCOBRA family genes in different organs and tissues of soybean,in which the expression levels of GmCOBRA 12,GmCOBRA 22 and GmCOBRA 14 were higher,and these genes may be involved in the regulation of cell wall expansion during soybean development.Among the six GmCOBRAs highly expressed in seeds,GmCOBRA21 was significantly expressed in the induction and elongation of clustered buds after qRT-PCR experiments,indicating that the gene may play a key role in soybean regeneration.Through subcellular localization,it was found that GmCOBRA 21 was located in the cell membrane.It can be speculated that GmCOBRAs may play an important role in soybean regeneration,and GmCOBRA21 may be one of the core gene in this process.