梨PbCDPK29基因克隆及对MeJA的响应  

Cloning of PbCDPK29 Gene from Pear and Its Response to MeJA

作　　者：刘韶华 刘慧芳 李六林[1] LIU Shaohua;LIU Huifang;LI Liulin(College of Horticulture,Shanxi Agricultural University,Taigu 030801,China)

机构地区：[1]山西农业大学园艺学院,山西太谷030801

出　　处：《山西农业科学》2022年第12期1614-1621,共8页Journal of Shanxi Agricultural Sciences

基　　金：国家梨产业技术体系白梨育种岗位(CARS-29-05);国家重点研发计划项目(2018YFD0201407)。

摘　　要：钙依赖蛋白激酶(Calcium-Dependent Protein Kinases,CDPKs)是植物中普遍存在的一类丝氨酸/苏氨酸蛋白激酶,在植物生长发育和响应逆境过程中起重要的调控作用。前期研究发现,PdCDPK基因家族中,雪花梨中PbCDPK29在梨黑星病菌处理后表达下调,为明确其结构和功能,试验从雪花梨叶片中克隆了PbCDPK29基因,利用生物信息学软件对该基因的一级、二级、三级结构及亲缘关系、保守结构域、顺式作用元件进行预测,并采用荧光定量PCR对MeJA处理后不同时间点PbCDPK29基因表达进行差异显著性分析。结果表明,PbCDPK29基因完整开放阅读框1554 bp,编码517个氨基酸,分子质量为58.26 ku;系统进化树分析表明,梨PbCDPK29蛋白与苹果MdCDPK32蛋白的亲缘关系最近,序列相似性达到97%;多序列比对分析发现,PbCDPK29与其他已知CDPK蛋白都具有CDPK家族典型的4个保守结构域;启动子分析显示,PbCDPK29基因启动子序列包含茉莉酸甲酯(MeJA)响应元件;通过qRT-PCR分析发现,PbCDPK29基因的表达受到MeJA的诱导。综合发现,MeJA诱导可缓解梨黑星病感染,推测外源MeJA可能通过诱导PdCDPK29表达,进而缓解梨黑星病的侵染。Calcium-dependent protein kinases(CDPKs)are a class of serine/threonine protein kinases widely presenting in plants,which play an important regulatory role in plant growth and development and response to stress.Based on our previous research,in the PdCDPK gene family,PbCDPK29 in Xuehua pear was downregulated with the treatment of Venturia nashicola.To clarify its structure and function,in this study,the PbCDPK29 gene was cloned from the leaves of Xuehua pear.The primary,secondary,tertiary structure and genetic relationship,conservative domain,cis acting element of PbCDPK29 was predicted by bioinformatics software.Difference significance analysis of the expression of PbCDPK29 at different time points after MeJA treatment was conducted by quantitative real-time PCR.The results showed that the complete open reading frame of the PbCDPK29 gene was 1554 bp,encoding 517 amino acids,with 58.26 ku of the relative molecular weight.The phylogenetic tree analysis showed that PbCDPK29 protein in pears had the closest genetic relationship with MdCDPK32 protein in apples,the sequence homology reached 97%.Multiple sequence alignment analysis found that all of PbCDPK29 and other known CDPK proteins contained four typical conserved domains of CDPK family.Promoter analysis indicated that the promoter of PbCDPK29 gene possessed methyl jasmonate(MeJA)response element.The qRT-PCR analysis showed that the expression of the PbCDPK29 gene was induced by MeJA.Comprehensively,it was found that MeJA induction could alleviate the infection of V.nashicola,it was speculated that exogenous MeJA might alleviate the infection of V.nashicola by inducing the expression of PdCDPK29.

关 键 词：梨 PbCDPK29 基因克隆 茉莉酸甲酯 

分 类 号：S661.2[农业科学—果树学]