藜钙依赖磷酸激酶基因CaCPK的克隆及胁迫表达

以耐盐植物藜(Chenopodium album)为材料,利用同源克隆技术获得了7个CDPK基因核心序列,并将其命名为CaCPK1–7。随后通过RACE技术成功获得CaCPK1–3的开放阅读框(ORF)序列,其ORF分别包含长度为1 632、1 704和1 590 bp的核苷酸序列。CaCPK1–3分别编码由543、567和529个氨基酸残基组成的钙依赖型蛋白激酶。定量PCR实验显示,CaCPK1–4受盐胁迫诱导明显上调表达,随胁迫时间增加不同基因呈现各异的表达规律。对CaCPK1–3在其它非生物胁迫下的表达分析显示,CaCPK1、CaCPK2和CaCPK3的表达均受外源ABA和H2O2的调控,H2O2合成抑制剂DPI和ABA合成抑制剂Na2WO4显著抑制300 mmol·L–1NaCl处理下CaCPK1、CaCPK2和CaCPK3的表达。研究结果为揭示藜在盐胁迫信号转导过程中CDPK基因家族的功能提供了理论依据。

Cloning and Expression Analysis of Calcium-dependent Phosphokinase Genes in Chenopodium album Under Stresses

By homology cloning, we obtained the core sequences of 7 calcium-dependent protein kinase （CDPK） genes, CaCPK1 to CaCPK7, from salt-tolerant Chenopodium a/bum. Quantitative real-time PCR revealed that CaCPK1, CaCPK2, CaCPK3 and CaCPK4 were significantly induced by salt stress with different patterns. The expression pattern of CaCPK1, CaCPK2 and CaCPK3 was also changed in C. a/bum under both H202 and abscisic acid （ABA） stresses. Di- phenylene iodonium （DPI, inhibitor of H202 synthesis） and sodium tungstate （Na2WO4, inhibitor of ABA synthesis） sig- nificantly inhibited the expression of CaCPKI, CaCPK2 and CaCPK3 on exposure to 300 mmoI-L-1NaCI. We obtained the full-length sequences of 3 CDPKs--CaCPK1, CaCPK2 and CaCPK3--by 3＇- and 5＇-RACE, which contain open reading frames of 1 632, 1 704, and 1 590 bp, respectively. The production of ABA and H202 induced by salt may be involved in induction of CaCPK1, CaCPK2 and CaCPK3, facilitating the analysis of the function of CDPK gene family in C. a/bum.