大豆GmCBL10基因的克隆与功能分析

Cloning and Function Analysis of GmCBL10 Gene in Soybean

为了剖析大豆GmCBL10在盐胁迫下的表达模式,本研究采用PCR技术从‘东农50’中克隆了GmCBL10基因全长序列,由792个碱基组成,共编码263个氨基酸。生物信息学分析发现Gm CBL10不含信号肽,有一个跨膜结构域和多个磷酸化位点,PSORT预测其定位于细胞质中。在The Bio-Analytic Resource for Plant Biology (Bar)数据库中分析了GmCBL10的拟南芥同源基因AtCBL10在不同非生物胁迫下的表达模式。对GmCBL10的启动子序列进行分析,发现Gm CBL10启动子区域含有多种与非生物胁迫应答及生理功能相关的顺式作用元件。对GmCBL10进行系统进化树分析,发现大豆GmCBL10与同为豆科的菜豆、木豆、红豆等的相似性很高。对大豆幼苗进行盐胁迫处理,荧光定量PCR结果显示Gm CBL10在盐诱导条件下上调表达,表明Gm CBL10可能作为正向调控因子参与大豆的盐胁迫响应。本研究分析了大豆GmCBL10在盐胁迫下的表达差异,为揭示大豆的抗盐机制提供了初步理论依据。

To analyze the expression of GmCBL10 in soybean under NaCl treatment, the full length sequence of GmCBL10 gene was cloned from soybean ’DN50’ by PCR technology, which consisted of 792 bases and encoded263 amino acids. Bioinformatics analysis showed that GmCBL10 had no signal peptide, but a transmembrane domain and several phosphorylation sites. PSORT analysis predicted that GmCBL10 was localized in cytoplasm.We analyzed the expression pattern of the homologous gene of Gm CBL10 in Arabidopsis thaliana under different abiotic treatments in the Bio-Analytic Resource for Plant Biology(Bar) database. The analysis of Gm CBL10 promoter sequence found that GmCBL10 promoter contained a variety of cis-elements related to abiotic stress response and physiological function. Phylogenetic tree analysis found that the genetic relationship between GmCBL10 and several leguminous plants, such as PvCBL10, CcCBL10 and VaCBL10, was closer. The qRT-PCR results showed that the GmCBL10 expression level was significantly increased after salt treatment, indicating that GmCBL10 could be used as a positive regulator to participate in the salt stress response. This study analyzed the difference in the expression of soybean GmCBL10 under salt stress, which could provide preliminary theoretical basis for the salt resistance mechanism of soybean.