太子参SnRK2I基因的克隆及在水分胁迫条件下的响应

Cloning of SnRK2I Gene from Pseudostellaria heterophylla and the Response Under the Condition of Water Stress

目的:获取太子参蔗糖非酵解型蛋白激酶2(SnRK2)基因,分析该基因在水分胁迫条件下的响应情况。方法:基于对SnRK2基因的同源性及太子参转录组数据库的分析,以不同水分胁迫处理的太子参鲜品为材料,利用基因克隆技术克隆得到太子参SnRK2基因序列,采用实时荧光定量PCR技术分析该基因与水分胁迫的相关性。结果:从太子参中克隆得到1条PhSnRK2I基因,cDNA全长为1 261 bp,编码305个氨基酸,分子量为47.27 kDa。该蛋白具有与SnRK2家族相似的ATP结合位点和Ser/Thr激酶活化环。实时荧光定量PCR分析结果表明,PhSnRK2I在土壤相对含水量为86%和44%时表达量较高。结论:成功克隆得到太子参PhSnRK2I基因序列,分析结果表明PhSnRK2I在中度水分胁迫下响应最积极,在太子参水分胁迫信号转导中发挥着重要作用。该研究可为进一步研究太子参PhSnRK2I基因在水分胁迫响应过程中的调控机理研究提供实验基础。

Objective:To obtain the sucrose non-fermenting-1-related protein kinase 2(SnRK2)gene of Pseudostellaria heterophylla,and to analyze its response under water stress.Methods:Based on the analysis of homology of SnRK2 gene and transcriptome database of Pseudostellaria heterophylla,fresh Pseudostellaria heterophylla under different water stress was used as material,the SnRK2 gene sequence of Pseudostellaria heterophylla was cloned by using gene cloning technology.The correlation between the gene and water stress was analyzed by using quantitative real-time PCR.Results:One PhSnRK2I gene from Pseudostellaria heterophylla was obtained by cloning,cDNA had a total length of 1 261 bp,encoded 305 amino acids,and the protein molecular weight was 47.27 kDa.The ATP binding site and Ser/Thr kinase activation rings of PhSnRK2I protein were similar to SnRK2 family.RT-qPCR results showed that PhSnRK2I had the highest expression when the relative water content of soil were 86% and 44%.Conclusion:The PhSnRK2I gene is cloned successfully from Pseudostellaria heterophylla,the analysis results suggest that PhSnRK2I of Pseudostellaria heterophylla has the most positive response under moderate water stress.It plays an important role in transmitting signal of water stress in Pseudostellaria heterophylla.This study provides an experimental basis for further studies on the regulation mechanism in the response process of PhSnRK2I gene of Pseudostellaria heterophylla under water stress.