茶树转录因子CsMYB81的克隆与生物信息学分析  

Cloning and Bioinformatics Analysis of CsMYB81 Transcription Factor in Tea Plants

作　　者：罗易萍 刘赛 陈艳妮 谷美仪 张天天 刘硕谦[1,2,3] 田娜[1,2,3] Luo Yiping;Liu Sai;Chen Yanni;Gu Meiyi;Zhang Tiantian;Liu Shuoqian;Tian Na(College of Horticulture and Hardening,Hunan Agricultural University,Changsha,410128;National Research Center of Engineering Technology for Utilization of Functional Ingredients from Botanicals,College of Horticulture and Hardening,Hunan Agricultural University,Changsha,410128;Key Lab of Tea Science Ministry of Education,College of Horticulture and Hardening,Hunan Agricultural University,Changsha,410128)

机构地区：[1]湖南农业大学园艺学院,长沙410128 [2]湖南农业大学园艺学院,国家植物功能成分利用工程技术研究中心,长沙410128 [3]湖南农业大学园艺学院茶学教育部重点实验室,长沙410128

出　　处：《分子植物育种》2022年第12期3902-3912,共11页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31670689);湖南省自然科学基金项目(2020JJ4358);中央引导地方科技发展基金项目(2019XF5041)共同资助。

摘　　要：MYB转录因子广泛参与植物的生长发育、生物以及非生物胁迫的应答过程,为探究茶树Cs MYB81的生物学功能,分析其表达模式。本研究以茶树(Camellia sinensis)品种‘碧香早’为材料,克隆和分析了CsMYB81的cDNA全长序列,对其序列和其编码的氨基酸序列进行了生物信息学分析,利用实时荧光定量PCR检测喷施赤霉素后不同部位的该基因表达水平。结果表明,Cs MYB81全长为1676 bp,开放阅读框(ORF)全长1632 bp,编码543个氨基酸,蛋白分子量为59.9055 kD,等电点为4.99;具有高度保守的R2R3结构域和GAMYB特异性基序(BOX1,BOX2),二级结构预测结果表明CsMYB81蛋白含有螺旋-转角-螺旋(HTH)结构,符合MYB基因家族的结构特征。实时荧光定量PCR分析结果表明Cs MYB81在叶片和茶籽中表达较高,在茎、花和花粉中的表达量较低。对茶树喷施赤霉素(GA3),结果发现当用浓度为10 mmol/L的GA3处理叶片0.5 h时,CsMYB81呈显著表达趋势,推测CsMYB81参与茶树响应赤霉素的过程。本研究为探究茶树CsMYB81的生物学功能提供一定的科学依据,并且茶树CsMYB81在种子中高表达提示该基因可能参与了种子的萌发过程。MYB transcription factors involved in the growth and development of plants,response processes under biological and abiotic stresses.In order to explore the biological function of Cs MYB81 in tea plants,analyze its expression pattern,in this study,the full-length c DNA sequences of Cs MYB81 was cloned and analyzed using the tea plants(Camellia sinensis)cultivar’Bixiangzao’as the material,and bioinformatics analysis of its sequence and amino acid sequence was carried out.Quantitative Real-time PCR was used to analyzed the expression of CsMYB81 in different parts of tea plants after gibberellin spraying.The results indicated that the total length of CsMYB81 was 1676 bp,the open reading frame(ORF)of CsMYB81 consisted of 1632 bp and it encoded 543amino acids with a deduced molecular weight of 59.9055 kD and the theoretical isoelectric point of 4.99.The protein has a highly conserved R2R3 domain and GAMYB-specific motifs(BOX1,BOX2).The secondary structure analysis revealed that Cs MYB81 contained a unique helix-coil-helix(HTH)of MYB protein,it was consistent with the structural characteristics of MYB gene family.The results of quantitative real-time PCR analysis showed that CsMYB81 was expressed higher in tea leaves and tea seeds,and lower in tea stems,petals and pollen.Finally,the response of CsMYB81 to gibberellin was significantly expressed when tea leaves were treated with GA3at a concentration of 10 mmol/L for 0.5 h.This speculated that CsMYB81 was involved in the gibberellin response mechanism,which provides a scientific basis for further investigation of the biological functions of tea plants CsMYB81,and the high expression of CsMYB81 in seeds suggests that CsMYB81 may be involved in the germination process of seeds.

关 键 词：茶树 MYB转录因子 赤霉素 生物信息学 实时荧光定量PCR 

分 类 号：S571.1[农业科学—茶叶生产加工]