| 摘要: |
| G2/有丝分裂特异性细胞周期蛋白 2(G2/mitotic-specific cyclin-2,Msc2)作为高等植物应对逆境胁迫的关键调控蛋白,参与多个抗逆境胁迫的应答。为探究RcMsc2基因的功能,该研究从蓖麻叶片组织中成功克隆了RcMsc2,并利用生物信息学分析RcMsc2蛋白的结构和潜在功能,同时借助qRT-PCR方法分析RcMsc2基因的组织表达特性和非生物胁迫表达特性。结果表明:(1)RcMsc2基因位于蓖麻第5号染色体长臂,该基因的CDS(coding sequence)区是1 299 bp,编码432个氨基酸。(2)RcMsc2蛋白拥有细胞周期(cyclin)家族特征结构域,是一个不稳定酸性亲水蛋白,无跨膜域和信号肽,相对分子量为49.38 kD。(3)RcMsc2蛋白质的二级、三级结构以α-螺旋和无规则卷曲为主。(4)RcMsc2蛋白与麻风树和巴西橡胶树的CYCB2蛋白的序列同源性最高,且同被聚为Group Ⅱ。(5)35S-RcMsc2-GFP融合蛋白定位于细胞核。(6)RcMsc2基因在蓖麻的所有组织中均有表达且主要在根和茎中发挥作用; 非生物胁迫分析表明RcMsc2基因可以被脱落酸(abscisic acid, ABA)、盐、干旱和低温处理诱导表达,并且RcMsc2基因对低温胁迫的响应最敏感。综上表明,该研究较全面地分析了RcMsc2基因的结构特征、系统进化和表达模式,为揭示RcMsc2基因在蓖麻的生长发育和应答冷胁迫过程中的功能提供了理论参考。 |
| 关键词: 蓖麻, RcMsc2, 基因克隆, 表达特性, 冷胁迫 |
| DOI：10.11931/guihaia.gxzw202207020 |
| 分类号:Q943 |
| 文章编号:1000-3142(2023)02-0336-11 |
| Fund project:国家自然科学基金(31860389,32060492); 内蒙古自治区自然科学基金(2022MS03057,2020LH03006)。 |
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| Functional analysis of RcMsc2 gene in castor(Ricinus communis) |
|
GENG Liuting1, LI Yanxiao1, ZHANG Chunlan2, YU Zhongyong3,
WANG Guiling4, XIANG Dianjun1, LIU Peng1*
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1. College of Agriculture, Inner Mongolia Minzu University, Tongliao 028000, Inner Mongolia, China;2. College of Life Sciences and Food
Engineering, Inner Mongolia Minzu University, Tongliao 028000, Inner Mongolia, China;3. Inner Mongolia Hongbo Seed
Industry Technology Limited Company, Tongliao 028000, Inner Mongolia, China;4. Zhalantun
Green Industry Development Center, Zhalantun 162650, Inner Mongolia, China
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| Abstract: |
| G2/mitotic-specific cyclin-2(Msc2), as a key regulatory protein in response to stress in higher plants, is involved in multiple responses to stresses. In order to explore the function of RcMsc2 gene which was successfully cloned from castor leaf tissue, the structure and potential function of RcMsc2 protein were analyzed by bioinformatics, and the expression characteristics of RcMsc2 gene in tissue and abiotic stress were analyzed by qRT-PCR. The results were as follows:(1)RcMsc2 gene was located in the long arm of Chromosome 5 in castor, and its CDS region was 1 299 bp, encoding 432 amino acids.(2)RcMsc2 protein has the characteristic domain of cyclin family, which was an unstable acidic hydrophilic protein without transmembrane domain and signal peptide, and its relative molecular weight was 49.38 kD.(3)The secondary and tertiary structures of RcMsc2 protein were mainly α-helix and random coil.(4)RcMsc2 protein had the highest sequence homology with CYCB2 protein of Jatropha curcas and Hevea brasiliensis, and was clustered into Group Ⅱ.(5)35S-RcMsc2-GFP fusion protein was localized to the nucleus.(6)RcMsc2 gene was expressed in all tissues of castor, and mainly played a role in roots and stems; abiotic stress analysis showed that RcMsc2 gene could be induced by abscisic acid( ABA ), salt, drought and low temperature treatment, and the response of RcMsc2 gene to low temperature stress was the most sensitive. In summary, this study comprehensively analyzed the structural characteristics, phylogenetic evolution and expression patterns of RcMsc2 gene, and provides a theoretical reference for revealing the function of RcMsc2 gene in castor growth and development and response to cold stress. |
| Key words: castor, RcMsc2, gene cloning, expression characteristics, cold stress |
