拟南芥CPK10/CPK30双突变体的构建及表型分析

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。

Construction and Phenotype Analysis of CPK10/CPK30 Double Mutantin Arabidopsis

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 pheno-types in response to various stress were detected,and the RT-PCR method was used to analyzed the two genes ex-pression.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 differ-ence to drought stress compared with wild-type and single mutant.Nevertheless,the expression levels of RD29A 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 in-volved in ABA-dependent signal transduction pathway,and showed redundant functions.