谷子逆境应答相关的钙依赖蛋白激酶基因SiCDPK1的克隆与表达

CDPK是一类重要的钙信号感受蛋白和响应蛋白,在植物非生物胁迫应答方面具有重要的作用。为探究耐旱作物谷子CDPK在抗逆胁迫中的应答机制,利用RT-PCR技术从谷子幼苗cDNA中克隆到一个与逆境胁迫相关的CDPK基因,命名为SiCDPK1(GenBank登录号为KC249975.1)。以拟南芥CDPK基因序列为查询序列,预测谷子基因组含有28个CDPK基因。其系统发育分析表明,谷子CDPK基因家族由4个亚类组成,其中SiCDPK1属于第II亚类,其全长1596 bp,编码531个氨基酸,预测蛋白分子量为59.5 kD,等电点pI为5.94,含有典型CDPK的保守结构。启动子调控区含有与多种逆境胁迫相关的调控元件。实时定量结果显示,SiCDPK1基因受PEG、ABA、高盐、自然干旱胁迫诱导表达。本试验为谷子抗逆应答机制的深入研究奠定了良好的理论基础。

Cloning and Expression Analysis of a Calcium-Dependent Protein Kinase Gene SiCDPK1 in Setaria italica

CDPK (calcium-dependent protein kinase) is a kind of essential calcium sensors and calcium responders, which plays important roles in response to various abiotic stresses in plants. In this research, we cloned the abiotic stress response related CDPK gene with RT-PCR in Setaria italica, designated as SiCDPK1 (GenBank accession number KC249975.1). We predicted 28 CDPKs in the Setaria italica genome by using the known Arabidopsis CDPK sequences as query sequences. The phylogenetic analysis showed the SiCDPK gene family was divided into four subgroups, in which SiCDPK1 belongs to subgroup II. The ORF of SiCDPK1 contains 1596 bp, which encodes 531 amino acids. The predicted protein molecular weight is 59.5 kD and pI is 5.94. SiCDPK1 has conserved protein domains of CDPK. The regulatory element analysis of promoter in SiCDPK1 showed a lot of cis-acting elements associated with different abiotic stresses. The RT-PCR results showed that SiCDPK1 was induced by PEG, ABA, salinity and drought. The research on SiCDPK1 paves a way for unraveling the mechanism of abiotic stresses in plants.