菜用大豆CIPK基因对逆境胁迫及激素的响应特征

Expression Character of Vegetable Soybean CIPKs in Response to Abiotic Stresses and Hormones

CIPK(calcineurin B-like-interacting protein kinase)是一类丝氨酸/苏氨酸蛋白激酶,在植物响应逆境胁迫和激素信号转导中发挥重要作用。本研究利用大豆基因组数据库,在全基因组水平鉴定获得52个CIPK蛋白激酶。蛋白比对分析发现所有Gm CIPK含有高度保守特征性的N端激酶区、连接区和C端调控区。系统进化树分析发现大豆Gm CIPK与拟南芥、水稻CIPK分类一致,分为4个亚家族,且每个亚家族含有3个不同物种的成员,表明Gm CIPK基因的分化早于物种的分化。启动子分析表明,多数Gm CIPK基因的启动子区含有逆境和激素应答元件。组织表达分析发现,Gm CIPK基因呈现多样化的组织表达特性。进一步选取组织表达量相对较高的14个Gm CIPK进行荧光定量PCR分析,结果表明这些菜用大豆CIPK基因在不同程度上均受高温、干旱、高盐胁迫以及ABA、ACC、SA、Me JA激素的诱导表达。采用蛋白同源比对和蛋白互作在线数据库对拟南芥及大豆同源CIPK蛋白激酶与其他蛋白的互作关系进行了预测分析,发现17对同源CIPK与其他蛋白(激酶、磷酸酶、转录因子等)存在互作。本研究为菜用大豆CIPK基因的功能研究与利用奠定了基础。

Calcineurin B-like-interacting protein kinase（CIPKs） was one kind of serine/threonine kinases, pla- ying crucial roles in abiotic stresses and hormones signal transduction pathways. Based on soybean genome data- base, a total of 52 CIPK genes were identified from soybean. Multiple alignments showed that all soybean CIPK pro- teins had the conserved regions which were composed of an N-terminal kinase domain, a junction region and a C-ter- minal regulatory domain. Phylogenetic analysis of CIPKs from soybean ,Arabidopsis and rice revealed that all CIPKs were classified into four subfamilies and each subfamily was composed of homologous CIPKs from these three spe- cies. The classification of CIPKs from soybean was consistent with Arabidopsis and rice, which implied that the dif- ferentiation of CIPK genes occurred before the evolution of plant species. Promoter sequences analysis suggested that most GmCIPKs had stress and hormone related cis-acting elements in their promoter regions. Tissues expression anal- ysis revealed that each GmCIPK had different expression pattern in different tissues. Furthermore, 14 GmCIPKs with relatively higher transcript level in different tissues were selected to investigate their transcriptional response to vari- ous stimuli by real-time PCR. The results showed the comprehensive responses of the tested GmCIPKs to heat, drought,sah stressors and ABA,ACC,SA, MeJA signaling. By BLASTP analysis, 17 orthologous GmCIPKs involved in CIPK interaction networks were identified between Arabidopsis and soybean based on the online database of pro- tein interaction. These candidate CIPK-interacting proteins included kinase, phosphatase and transcription factor and so on. The findings of this study will provide a foundation for the functional characterization of the GmCIPK genes in vegetable soybean.