山药块茎膨大期DoCDPK1基因生物信息学及低温胁迫下的表达分析  

Bioinformatics and expression analysis of DoCDPK1 gene under low temperature stress during tuber expansion stage of yam

作　　者：高晶晶[1] 张艳芳[1] 邢丽南 葛明然 季祥 霍秀文[1] GAO Jingjing;ZHANG Yanfang;XING Li’nan;GE Mingran;JI Xiang;HUO Xiuwen(College of Horticulture and Plant Protection,Inner Mongolia Agricultural University,Hohhot 010019,Inner Mongolia,China;Inner Mongolia Agricultural and Animal Husbandry Technology Extension Center,Hohhot 010011,Inner Mongolia,China)

机构地区：[1]内蒙古农业大学园艺与植物保护学院,呼和浩特010019 [2]内蒙古自治区农牧业技术推广中心,呼和浩特010011

出　　处：《中国瓜菜》2024年第7期42-51,共10页China Cucurbits And Vegetables

基　　金：国家自然科学基金项目(32260759);内蒙古自治区自然科学基金项目(2022MS03052)。

摘　　要：钙依赖性蛋白激酶(CDPK)在植物的生长发育及逆境胁迫方面发挥着重要作用。以大和长芋和毕克齐山药为试验材料,克隆钙依赖性蛋白激酶基因DoCDPK1,并对其进行生物信息学分析,为DoCDPK1基因的功能研究奠定基础。采用生物信息学方法分析DoCDPK1基因结构,构建瞬时表达载体,对DoCDPK1蛋白进行亚细胞定位;采用qRT-PCR分析DoCDPK1基因在不同山药品种的不同发育阶段及低温胁迫处理下的表达模式。结果表明(:1)DoCDPK1基因序列长度为2023 bp,编码521个氨基酸。(2)DoCDPK1与几内亚薯蓣CDPK蛋白序列的一致性为97%,且DoCDPK1存在STKc_CAMK结构域。(3)瞬时表达分析表明,DoCDPK1蛋白定位于细胞核与细胞膜。(4)大和长芋山药中的CDPK活性更高,DoCDPK1可能参与调控山药块茎膨大后期的生长发育。(5)经4℃低温胁迫后,CDPK活性降低,DoCDPK1表达量上调,且表达水平在不同时间处理下存在差异。综上所述,DoCDPK1可能参与调控山药块茎膨大后期的生长发育及低温胁迫响应过程。Calcium-dependent protein kinase plays an important role in plant growth and osmotic stress.Using Dahechangyu and Bikeqi yam as experimental materials,the calcium dependent protein kinase gene DoCDPK1 was cloned and bioinformatic analysis was performed to lay a foundation for the functional study of DoCDPK1.The structure of DoCDPK1 was analyzed by bioinformatics and subcellular localization of DoCDPK1 protein was performed by constructing transient expression vector.qRT-PCR was used to analyze the expression patterns of DoCDPK1 at different developmental stages and under low temperature stress in different yam varieties.The results indicated:(1)The length of DoCDPK1 gene sequence was 2023 bp,encoding 521 amino acids.(2)The consistency of the protein sequence between DoCDPK1 and CDPK protein sequence of Dioscorea cayenensis subsp.was 97%,and DoCDPK1 had STKc_CAMK domain.(3)Transient expression analysis showed that DoCDPK1 protein was localized in cell nucleus and membrane.(4)The CDPK activity in Dahechangyu was higher,and DoCDPK1 might be involved in regulating the growth and development of yam tuber at the later stage of enlargement.(5)After low temperature stress at 4℃,CDPK activity decreased,and the expression of DoCDPK1 was up-regulated,with differences in expression levels observed under different time treatments.In conclusion,DoCDPK1 gene may be involved in regulating the growth and development of yam tuber and its response to low temperature stress.

关 键 词：山药 钙依赖性蛋白激酶 亚细胞定位 低温胁迫 基因表达量 

分 类 号：S632.1[农业科学—蔬菜学]