蕙兰CyfaSTK基因的克隆与表达分析  

Cloning and expression analysis of the CyfaSTK gene from Cymbidium faberi

作　　者：费越 夏胜应 熊海燕 刘志雄[1] Fei Yue;Xia Sheng-Ying;Xiong Hai-Yan;Liu Zhi-Xiong(College of Horticulture and Gardening, Yangtze University , Jingzhou, Hubei 434025, China;Jiangling County Housing and Urban-Rural Development Bureau , Jiangling, Hubei 434100, China)

机构地区：[1]长江大学园艺园林学院,湖北荆州434025 [2]江陵县住房和城乡建设局,湖北江陵434100

出　　处：《植物科学学报》2019年第5期602-609,共8页Plant Science Journal

基　　金：国家自然科学基金项目(31101202);长江大学优秀硕士学位论文培育计划(YS2018049)~~

摘　　要：采用同源克隆的方法,从蕙兰( Cymbidium faberi Rolfe)花芽中克隆获得 CyfaSTK 基因的cDNA序列,并对其进行生物信息学分析及基因表达分析。结果显示,该基因全长843 bp,其中开放阅读框(ORF)长 705 bp ,共编码234个氨基酸和1个终止密码子。同源蛋白序列比对及分子系统发育分析结果表明,CyfaSTK蛋白属于D类MADS-box转录因子STK-like进化系,含有MADS、I、K和C等4个结构域,其C末端转录激活区含有2个保守的基元:AG motif Ⅰ和AG motif Ⅱ,此外,还具有一个在天门冬目植物中相对保守的基元MD motif。基因表达的组织特异性分析结果显示:蕙兰 CyfaSTK 基因在花萼、花瓣、唇瓣、药帽、子房中均有表达,但在叶片中不表达,其中在子房中的表达量与其他组织相比,差异达到极显著水平;CyfaSTK 在花芽经过休眠后的萌动期表达量最高,且在开花当天该基因表达量有上升趋势。研究结果表明 CyfaSTK 基因不仅参与调控蕙兰花器官的发育过程,且对子房及合蕊柱的正常发育具有重要作用。The cDNA sequence of the CyfaSTK gene (GenBank accession number: MH917915. 1) was cloned from the flower bud of Cymbidium faberi Rolfe by homologous cloning. Results showed that the gene was 843 bp in length and the open reading frame (ORF) was 705 bp long, encoding a total of 234 amino acids and a stop codon. Molecular phylogenetic analysis and homologous protein comparison showed that the CyfaSTK protein belonged to the transcription factor STK -like evolution line of the AG subfamily of the D-class MADS-box gene family, which contained four distinct domains of MADS, I, K, and C terminal. The C terminal transcriptional activation region contained two conserved motifs: i. e., AG motif Ⅰ and AG motif Ⅱ. In addition, there was also a relatively conserved MD motif in Asparagales plants. Tissue specificity analysis showed that the C. faberi CyfaSTK gene was expressed in sepals, petals, lips, anther caps, gynostemia, and ovaries, but not in juvenile leaves. Expression of the C. faberi CyfaSTK gene in the ovary was significantly higher than that in other tissues. Among the dynamic changes in the flower bud at different developmental stages, the expression of CyfaSTK was the highest at the germination stage after dormancy and increased on the day of flowering. These results suggest that the D-class CyfaSTK gene not only regulates organ development in C. faberi , but also plays an important role in the normal development of the gynostemium and ovary.

关 键 词：蕙兰 花发育 MADS-BOX基因 CyfaSTK基因 

分 类 号：Q943.2[生物学—植物学]