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2025年4月3日 星期四
农业生物技术学报  2021, Vol. 29 Issue (7): 1260-1273    DOI: 10.3969/j.issn.1674-7968.2021.07.003
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
蓖麻RcCIPKs基因家族鉴定与冷胁迫下的表达模式分析
王晓宇1,2,3,*, 刘栩铭1, 韩孟良1, 李敏4, 吴颖5, 靳亚楠1,2, 张继星1,2,3,*
1 内蒙古民族大学 生命科学与食品学院,通辽 028000;
2 内蒙古民族大学 生命科学与食品学院 资源生物与生态研究所,通辽 028000;
3 内蒙古民族大学 科尔沁植物逆境生物学研究所,通辽 028000;
4 内蒙古民族大学 农学院,通辽 028000;
5 天津农学院,天津 300384
Identification of RcCIPKs Gene Family in Castor Bean (Ricinus communis) and Analysis of Its Expression Pattern Under Cold Stress
WANG Xiao-Yu1,2,3,*, LIU Xu-Ming1, HAN Meng-Liang1, LI Min4, WU Yin4, JIN Ya-Nan1,2, ZHANG Ji-Xing1,2,3,*
1 College of Life Science and Food, Inner Mongolia University for Nationalities, Tongliao 028000, China;
2 Institute of Resource Biology and Ecology, College of Life Science and Food, Inner Mongolia University for Nationalities, Tongliao 028000, China;
3 Horqin Plant Stress Biology Research Institute, Inner Mongolia University for Nationalities, Tongliao 028000, China;
4 Agricultural College, Inner Mongolia University for Nationalities, Tongliao 028000, China;
5 Tianjin Agricultural University, Tianjin 300384, China
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摘要 植物通过启动一系列信号转导途径来应对外部环境,这些途径通常涉及多种蛋白激酶,CBL特异性结合蛋白(CBL-interacting protein kinases, CIPKs)是其中的重要成员之一。本研究利用序列比对法在蓖麻(Ricinus communis)全基因组范围内进行CIPK基因家族鉴定;利用生物信息学手段对基因家族成员序列结构、理化性质、染色体定位、启动子顺式作用元件、蛋白保守结构域、系统进化关系以及聚类方式进行分析;利用RNA-seq与qRT-PCR对RcCIPKs在冷胁迫下的表达进行分析;利用qRT-PCR分析RcCIPKs的组织特异性表达模式与脱落酸(abscisic acid, ABA)激素诱导下的CIPK表达水平;并在烟草(Nicotiana tabacum)表皮细胞中实现RcCIPK16的亚细胞定位分析。结果显示,成功鉴定到18个RcCIPKs基因家族成员;基因分析发现RcCIPKs可分为外显子富集型与外显子贫乏型两类,定位于16个染色体片段上且呈不均匀分布;大多数RcCIPKs启动子区均含有MYB、MYC与ABA响应元件(ABA response element, ABRE)和乙烯响应元件(ethylene response element, ERE),暗示其响应激素诱导与逆境胁迫。大多数RcCIPK蛋白的基序类型与排布顺序基本相同,只有RcCIPK3RcCIPK4存在不同程度的motif缺失;进化树分析将18个RcCIPKs基因聚为A、B、C、D、E 5类,推测不同类型的CIPK蛋白功能存在差异;RNA-Seq和qRT-PCR结果表明仅有RcCIPK1RcCIPK12RcCIPK16受冷胁迫诱导表达。该3个基因的组织特异性分析显示,RcCIPK1RcCIPK12仅在叶与茎中表达,RcCIPK16仅在叶中表达;且3个基因的表达水平均受ABA激素诱导。亚细胞定位分析初步将RcCIPK16定位于细胞质中。本研究在蓖麻全基因组水平对CIPK基因家族进行鉴定,并分析了其冷胁迫下的表达水平,为进一步创制蓖麻抗冷新种质提供重要的候选基因。
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关键词 蓖麻CIPK基因家族冷胁迫表达模式    
Abstract:Plants respond to the external environment by initiating a series of signaling pathways, which usually involve multiple protein kinases, such as CBL-interacting protein kinases (CIPKs). In this study, the sequence alignment method was used to identify the CIPK gene family within the whole genome of castor bean (Ricinus communis). The bioinformatics method was used to analyze the sequence structure, physical and chemical properties, chromosome positioning, cis-acting elements of promoters, conserved domains, phylogenetic relationships, and clustering methods of the encoded protein. RNA-seq and qRT-PCR were carried out to assess the expression of RcCIPKs. qRT-PCR was used to analyze the tissue-specific expression pattern of RcCIPKs and the expression level of CIPK affected by abscisic acid (ABA) hormone. The subcellular localization of RcCIPK16 were further analyzed in the epidermal cells of Nicotiana tabacum. Eighteen RcCIPKs family members were successfully identified. Gene analysis showed that RcCIPKs were divided into exon-rich and exon-poor types, which were located on 16 chromosome segments with uneven distribution. Most RcCIPKs contained MYB, MYC, ABRE and ERE elements, which indicated that these genes were responsive to plant hormone and stress. Protein analysis showed that majorities of RcCIPKs had the identical motif types and arrangement except RcCIPK3 and RcCIPK4. RcCIPKs were grouped into A, B, C, D and E categories based on phylogenetic analysis, which inferred that diverse types of CIPKs might perform various function. The results of RNA-Seq and qRT-PCR displayed that only RcCIPK1, RcCIPK12 and RcIPK16 were induced by cold stress. The tissue-specific analysis showed that RcCIPK1 and RcCIPK12 were mainly expressed in leaves and stems, and RcCIPK16 was mainly expressed in leaves; and the expression levels of the three genes were induced by ABA hormones. Subcellular localization analysis revealed that RcCIPK16 was mainly localized in the cytoplasm. This study firstly identified RcCIPKs gene family at the whole genome level and analyzed gene expression under cold stress, which will provide important candidate genes for further development of new cold tolerant castor germplasm.
Key wordsRicinus communis    CIPK gene family    Cold stress    Expression pattern
收稿日期: 2020-12-17     
ZTFLH:  S565.6  
  Q78  
基金资助:国家自然科学基金(32060493;31760399;61971312);自治区高校“青年科技英才支持计划”(NJYT-20-B25);自治区留学人员创新创业启动支持计划;国家服务特殊需求蒙药学博士点建设专项开放基金项目(MDMYBJ2019008)
通讯作者: *xiaoyuwang1987@hotmail.com;zhangjixing@imun.edu.cn   
引用本文:   
王晓宇, 刘栩铭, 韩孟良, 李敏, 吴颖, 靳亚楠, 张继星. 蓖麻RcCIPKs基因家族鉴定与冷胁迫下的表达模式分析[J]. 农业生物技术学报, 2021, 29(7): 1260-1273.
WANG Xiao-Yu, LIU Xu-Ming, HAN Meng-Liang, LI Min, WU Yin, JIN Ya-Nan, , ZHANG Ji-Xing. Identification of RcCIPKs Gene Family in Castor Bean (Ricinus communis) and Analysis of Its Expression Pattern Under Cold Stress. 农业生物技术学报, 2021, 29(7): 1260-1273.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2021.07.003     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2021/V29/I7/1260
 
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