薄壳山核桃和山核桃CDPK基因家族的鉴定及表达分析  

Identification and Expression Analysis of CDPK Gene Family in Pecan(Carya illinoinensis)and Chinese Hickory(Carya cathayensis)

作　　者：赵娟 朱凯凯 范平桦 谭鹏鹏[1,2] 彭方仁 李永荣[3] ZHAO Juan;ZHU Kai-Kai;FAN Ping-Hua;TAN Peng-Peng;PENG Fang-Ren;LI Yong-Rong(Co-Innovation Center for Sustainable Forestry in Southern China,Nanjing Forestry University,Nanjing 210037,China;College of Forestry,Nanjing Forestry University,Nanjing 210037,China;Nanjing Green Universe Pecan Science and Technology Co.Ltd.,Nanjing 211500,China)

机构地区：[1]南京林业大学南方现代林业协同创新中心,南京210037 [2]南京林业大学林学院,南京210037 [3]南京绿宙薄壳山核桃科技有限公司,南京211500

出　　处：《农业生物技术学报》2022年第3期442-456,共15页Journal of Agricultural Biotechnology

基　　金：江苏省自然科学基金(BK20190749);国家自然科学基金(31870672)。

摘　　要：钙依赖蛋白激酶(calcium-dependent protein kinase,CDPK)是植物特有的一类丝氨酸/苏氨酸型蛋白激酶,在植物生长发育和逆境信号转导过程中具有十分重要的作用。为了探讨CDPK在薄壳山核桃(Carya illinoinensis)和山核桃(Carya cathayensis)生长发育过程中的作用,本研究利用生物信息学手段从薄壳山核桃和山核桃基因组中鉴定CDPK基因家族,利用MEGA7.0软件进行多序列比对和系统进化树构建,并进行基因结构和保守基序分析。结果显示,本研究成功鉴定了28个薄壳山核桃CDPK基因家族成员(CILCDPK1~28)和25个山核桃CDPK基因家族成员(CCACDPK1~25);进一步构建薄壳山核桃、山核桃、拟南芥(Arabidopsis thaliana)以及水稻(Oryza sativa)的CDPK成员系统进化树,聚类分析结果显示CDPK蛋白分为4个亚组;顺式作用元件分析结果表明,所有CDPK基因均含有激素响应元件,其中数目最多的是脱落酸响应元件。薄壳山核桃和山核桃的转录组数据分析显示,3个CILCDPKs和2个CCACDPKs可能在胚胎发育的3个关键阶段发挥重要作用;薄壳山核桃嫁接愈合过程中,3个CDPK家族基因高表达,可能在薄壳山核桃愈伤组织和维管组织形成时期发挥重要作用。qPCR分析结果显示,CILCDPK1/3/6/11/13/19/25在干旱胁迫下基因表达上调。本研究为探讨CDPK基因家族在植物生长发育及干旱胁迫响应中的作用机制提供参考。Calcium-dependent protein kinase(CDPK)is a specific type of Ser/Thr protein kinase to plants,which plays an important role in plant growth,development and stress signal transduction.In order to explore the function of CDPK genes in the growth and development of pecan(Carya illinoinensis)and Chinese hickory(Carya cathayensis),bioinformatics methods were applied to identify the members of CDPK gene family from pecan and Chinese hickory genome.MEGA7.0 software was used to carry on the multiple sequence alignment,classification and phylogenetic tree construction.The gene structures and conserved motifs of CDPK family were also analyzed.The results showed that 28(CILCDPK1~28)and 25(CCACDPK1~25)CDPK members were successfully identified in pecan and Chinese hickory,respectively.In order to explore the homologous evolutionary relationship between the two plants and other species,a phylogenetic tree was constructed using CDPKs in pecan,Chinese hickory,Arabidopsis and rice(Oryza sativa).The phylogenetic tree showed that all CDPK proteins could be divided into 4 subgroups.Cis-acting element analysis showed that all CDPK genes contained hormone-response elements,and abscisic acid-response elements were the most abundant.Transcriptome data analysis of pecan and Chinese hickory showed that 3 CILCDPKs and 2 CCACDPKs might play an important role in the 3 key stages of embryonic development.In addition,3 CDPK genes showed high expression during pecan grafting and healing,which might play important roles in callus and vascular tissue formation of pecan.qPCR analysis showed that CILCDPK1/3/6/11/13/19/25 were up-regulated under drought stress.The present study provides a reference basis for exploring the mechanism of CDPK gene family in plant growth and development and drought stress response.

关 键 词：薄壳山核桃 山核桃 钙依赖蛋白激酶(CDPK) 基因家族 基因表达 

分 类 号：S-3[农业科学]