甜瓜CmCIPK家族全基因组鉴定和逆境条件下的表达分析

Genome-wide identification of CmCIPK family and its expression analysis under abiotic stress in melon

为探究甜瓜CIPK(CBL-interacting protein kinase)基因家族成员的生物学功能,以拟南芥中26个CIPKs的氨基酸序列为参照,用BLASTP方法鉴定了18个甜瓜CIPK家族成员,并对其理化性质、染色体分布、系统进化、基因结构、蛋白保守基序、顺式元件,以及基因的表达模式进行了分析。结果显示,CIPK基因家族成员的基因长度为1499~8499 bp,它们不均匀地分布在甜瓜的9条染色体上;根据进化关系可将其分为5个亚家族,分别包含了26、10、25、26和8个成员,并且其中有4个CmCIPK基因对发生了片段复制。基因结构分析表明,10个基因为内含子缺失型,8个基因为内含子富集型。蛋白保守基序分析发现,CmCIPK家族保守性较好,所有成员均含有CIPK家族的典型特征:N端激酶域中的激活环和C端调节域中的NAF/FISL结构域。在基因上游的2000 bp序列中存在多个与植物激素和逆境相关的顺式元件,显示可能有多种转录调控。转录组分析发现,CmCIPKs的组织表达量整体表现为叶>根>雄花>雌花>果。选择CmCIPK1-like和CmCIPK12-like基因验证其组织表达模式,显示分别在叶和雄花中的表达量最高。对其进行不同逆境处理,结果表明,这2个CmCIPK基因受NaCl和脱落酸(ABA)诱导表达;在干旱处理中,这2个基因经历短时间的上调或下调后恢复至最初表达水平。以上结果说明,CmCIPK1-like和CmCIPK12-like基因可能在ABA和非生物胁迫中发挥着重要作用,可为以后CmCIPK的功能研究提供参考。

In order to explore the biological functions of CIPK(CBL-interacting protein kinase)gene family in melon,18 melon CIPK family members were identified by the BLASTP based on the amino acid sequence of CIPK gene family of Arabidopsis thaliana,and their physico-chemical properties,chromosomal distribution,systematic evolution,gene structure,protein conserved motifs,cis-elements and gene expression patterns were analyzed.The results showed that gene length of CIPK family members was 1499-8499 bp,and they were unevenly distributed on 9 chromosomes of melon;According to the evolutionary relationship,they could be divided into five sub-families,which contain 26,10,25,26 and 8 members,respectively.Among them,four pairs of CmCIPKs had fragmented duplication.Gene structure analysis showed that ten genes were intron-poor and eight genes were intron-rich.Protein conservation motif analysis found that the CmCIPK family was well-conserved,and all members contained typical features of the CIPK family:the activation loop in the N-terminal kinase domain and the NAF/FISL domain in the C-terminal regulatory domain.The presence of multiple cis-elements related to plant hormones and stress in the 2000 bp upstream sequence of their genes suggested that there might be multiple transcriptional regulation.Transcriptome analysis found that the tissue expression levels of CmCIPKs from high to low were:leaf,root,male flower,female flower and fruit.The CmCIPK1-like and CmCIPK12-like genes were selected to verify their tissue expression patterns,which showed the highest expression levels in leaves and male flowers,respectively.After different adversity treatments,the results showed that the two CmCIPK genes were induced by NaCl and abscisic acid(ABA),and they returned to the initial expression level after a short period of up-regulation or down-regulation in drought treatment.The results indicated that CmCIPK1-like and CmCIPK12-like might play important roles in ABA and abiotic stress,which could provide references for future CmCIPK functional studies.