甘蔗CBL-CIPK基因家族的鉴定和表达分析

Identification and Expression Analysis of the CBL-CIPK Gene Family in Sugarcane

【目的】甘蔗是重要的糖料作物,温度、盐碱、水分等因素是制约其生长发育的关键环境因素。类钙调磷酸酶B蛋白CBL(calcineurin B-like protein)是一类Ca^(2+)结合蛋白,通过与其特定的蛋白激酶CIPK(CBL-interacting protein kinase)作用,在Ca^(2+)信号传导通路,尤其是逆境信号传导通路中发挥重要作用。目前,甘蔗全基因组测序已完成,但其CBL-CIPK基因家族成员尚未确定,互作调控机理依然未知。本研究确定了甘蔗CBL、CIPK成员并揭示了CBL-CIPK互作关系,为研究甘蔗CBL-CIPK的互作机理提供基因资源和理论基础。【方法】以甘蔗品种‘GT58’为材料,通过RT-qPCR技术分析CBLs、CIPKs在低温、高温、NaHCO_(3)和PEG处理等4种非生物胁迫下的表达水平,利用酵母双杂交试验分析SsCBLs和SsCIPKs之间的相互作用。【结果】甘蔗全基因组中共有19个CBL基因和82个CIPK基因,分布在不同的进化分支且存在基因复制现象,基因家族成员之间理化性质差异较大,结构域与蛋白基序具有高度保守性,顺式作用元件分布多样;转录水平上,SsCBL7/SsCBL12/SsCIPK1/SsCIPK5的表达水平易受低温、高温、干旱和高盐等多种非生物胁迫的调控;蛋白水平上,SsCBL1与SsCIPK47和SsCIPK81相互作用,SsCBL8与SsCIPK47和SsCIPK81相互作用。【结论】CBL-CIPK互作网络可能在甘蔗生长发育过程中响应非生物胁迫,发挥重要作用。

【Objective】Sugarcane is an important sugar crop,and temperature,salinity,water and other factors are the key environmental factors restricting its growth and development.Calcineurin B-Like protein(CBL)family represents a group of Ca^(2+) binding proteins and plays a key role in decoding calcium signals by specifically interacting with and regulating a family of protein kinases CIPKs(for CBL-interacting protein kinases)in abiotic stress signal transduction pathway.At present,the whole genome of sugarcane has been sequenced,but its CPL-CIPK gene family members have not been identified,and the interaction regulation mechanism is still unknown.Here,the members of CBL and CIPK in sugarcane were identified and the CPL-CIPK interaction relationship was revealed,which may provide gene resources and theoretical basis for studying the mechanism of CPL-CIPK interaction in sugarcane.【Method】The expression levels of CBLs and CIPKs under low temperature,high temperature,NaHCO_(3) and PEG were analyzed by RT-qPCR from sugarcane GT58.The interactions between SsCBLs and SsCIPKs were analyzed by yeast two-hybrid assay.【Result】There were 19 CBL genes and 82 CIPK genes in the whole genome of sugarcane,which distributed in different evolutionary branches with gene replication.The physicochemical properties of gene family members were different,the structural domains and protein motifs were highly conserved,and the distribution of cis-acting elements was diverse.SsCBL7/SsCBL12/SsCIPK1/SsCIPK5 were specifically expressed under low temperature,high temperature,drought and high salt and other abiotic stress regulation.Meanwhile,SsCBL1 interacted with SsCIPK47 and SsCIPK81,and SsCBL8 interacted with SsCIPK47 and SsCIPK81.【Conclusion】The CBL-CIPK signaling system may play an important role in response to abiotic stress during the growth and development of sugarcane.