拟南芥CPK10/CPK30双突变体的构建及表型分析

Construction and Phenotype Analysis of CPK10/CPK30 Double Mutantin Arabidopsis

CDPKs是植物细胞内一类重要的钙感受器,拟南芥CPK10属于CDPK家族成员。为研究CPK10及与其同源性较高的CPK30是否共同参与逆境响应。首先构建了cpk10×cpk30双突变体,然后进行多种逆境下的生理表现检测,并利用RT-PCR方法分析2个基因的表达情况。结果显示,模拟干旱、盐、ABA处理拟南芥幼苗后,双突变体与野生型无差异;成苗期双突变体与单突变体对干旱敏感程度类似。转录水平检测到,在干旱胁迫时双突变体中RD29A的表达与野生型及单突变体趋势相反,呈下降趋势;响应ABA的OST1在双突变体中受干旱诱导后0.5 h明显表达上调。成功获得了cpk10×cpk30双突变体,由生理表型和表达分析结果推测CPK10与CPK30可能共同参与了依赖ABA的干旱逆境信号转导过程,且二者之间存在功能冗余。

Calcium-dependent protein kinases（ CDPKs） function as Ca2 ＋sensor in various plant physiological processes. CPK10 was one member of CDPK family. In order to investigate the molecular mechanisms of CPK10 and it＇s high homology CPK30 in response to stress,cpk10 × cpk30 double mutant was firstly constructed. The phenotypes in response to various stress were detected,and the RT-PCR method was used to analyzed the two genes expression. The results showed that there was no difference in the seed germination stage between double mutant and wild-type plants in the treatment of drought,salt and ABA. In the mature stage,double mutant exhibited no difference to drought stress compared with wild-type and single mutant. Nevertheless,the expression levels of RD29 A was decreased in the double mutant and showed opposite trend in wild-type and single mutant during drought stress. The expression of ABA-responsive gene OST1 was enhanced obviously in double mutant after drought stress 0. 5 h. The cpk10 × cpk30 double mutant was successfully obtained. Our data indicated that CPK10 and CPK30 might be involved in ABA-dependent signal transduction pathway,and showed redundant functions.