| 354 | 11 | 74 |
| 下载次数 | 被引频次 | 阅读次数 |
以黄芪转录组测序获得的一条MYB基因为基础,进行克隆、生物信息学分析和荧光定量分析。克隆得到cDNA序列1 296 bp,其中包含一个长度为1 200 bp的开放阅读框,编码一个由399个氨基酸组成的蛋白质,命名为AmMYB44。预测黄芪MYB44蛋白的分子量为43.392 kD,等电点为6.39,平均亲水数是-0.459,不稳定系数为46.08,属于不稳定蛋白。实时荧光定量PCR分析显示,AmMYB44基因在干旱和低温条件下的表达均明显上调,但调节幅度存在组织差异、条件差异和处理时间的差异。在AmMYB44启动子上发现了多个与逆境和激素应答相关的顺式作用元件。推测AmMYB44基因在黄芪干旱、低温等非生物胁迫应答中具有重要的调控作用,本研究将促进黄芪的抗旱抗寒分子机制的研究。
Abstract：A MYB gene obtained from transcriptome sequencing of Astragalus membranaceus was used for cloning, bioinformatics analysis and gene expression of analysis. The 1 296 bp full length cDNA sequence was isolated by PCR, which contained an open reading frame with a length of 1 200 bp. The gene encoded a protein with 399 amino acids, and was named AmMYB44. It was predicted that the molecular weight of MYB44 protein in A. membranaceus was 43.392 kD, the theoretical isoelectric point was 6.39, the average hydrophilic number was-0.459, and the instability coefficient was 46.08, and MYB44 belonged to the unstable protein. Real-time quantitative PCR analysis showed that the expression of AmMYB44 gene was increased markedly under drought and low temperature conditions, but there were differences between the amplitude of accommodation in tissue,stress condition and treatment duration. Several cis-acting elements associated with abiotic stresses and hormone responses were found on the AmMYB44 promoter. It was conjectured that AmMYB44 gene plays an important regulatory role in the response to abiotic stresses such as drought and low temperature in A. membranaceus. The present study could advance further investigation of the molecular mechanism associated with the cold and drought stress in A. membranaceus.
Abe H.,1997,Role of Arabidopsis MYC and MYB homologs in drought-and abscisic acid-regulated gene expression,Plant Cell,9(10):1859-1868
Agarwal M.,Hao Y.,Kapoor A.,Dong C.H.,Fujii H.,Zheng X.,and Zhu J.K.,2006,A R2R3 type MYB transcription factor is involved in the cold regulation of CBF genes and in acquired freezing tolerance,J.Biol.Chem.,281(49):37636-37645
Ambawat S.,Sharma P.,Yadav N.R.,and Yadav R.C.,2013,MYB transcription factor genes as regulators for plant responses:an overview,Physiol.Mol.Biol.Plants,19(3):307-321
Baldoni E.,Genga A.,and Cominelli E.,2015,Plant MYB transcription factors:their role in drought response mechanisms,Int.J.Mol.Sci.,16(7):15811-15851
Bernsdorff F.,Doering A.C.,Gruner K.,Schuck S.,Brutigam A.,and Zeier J.,2015,Pipecolic acid orchestrates plant systemic acquired resistance and defense priming via salicylic acid-dependent and independent pathways,Plant Cell,28(1):102-129
Chen N.,Chi X.Y.,Pan L.J.,Chen M.N.,and Yu S.L.,2015,Advances in MYB transcription factors during salt-stress regulation in plants,Zhiwu Shengli Xuebao(Plant Physiology Journal),51(9):1395-1399(陈娜,迟哓元,潘丽娟,陈明娜,禹山林,2015,MYB转录因子在植物盐胁迫调控中的研究进展,植物生理学报,51(9):1395-1399)
Chen Y.H.,Yang X.Y.,He K.,Liu M.H.,Li J.L.,Gao Z.F.,Lin Z.Q.,Zhang Y.F.,Wang X.X.,Qiu X.M.,Shen Y.P.,Zhang L.,Deng X.H.,Luo J.C.,Deng X.W.,Chen Z.L.,Gu H.Y.,and Qu L.J.,2006,The MYB transcription factor superfamily of arabidopsis:expression analysis and phylogenetic comparison with the rice MYB family,Plant Mol.Biol.,60(1):107-124
Cheng S.F.,Duanmu H.Z.,Chen C.,Liu A.L.,Xiao J.L.,and Zhu Y.M.,2016,Whole genome identification of soybean MYBtranscription factors and bioinformatics analysis,Dadou Kexue(Soybean Science),35(1):52-57(成舒飞,端木慧子,陈超,刘艾林,肖佳雷,朱延明,2016,大豆MYB转录因子的全基因组鉴定及生物信息学分析,大豆科学,35(1):52-57)
Dubos C.,Gourrierec J.L.,Baudry A.,Huep G.,Lanet E.,Debeaujon I.,Routaboul J.M.,Alboresi A.,Weisshaar B.,and Lepiniec L.,2008,MYBL2 is a new regulator of flavonoid biosynthesis in Arabidopsis thaliana,Plant J.,55(6):940-953
Jin J.P.,Guo A.Y.,He K.,Zhang H.,Zhu Q.H.,Chen X.,Gao G.,and Luo J.C.,2015,Classification,prediction and database construction of plant transcription factors,Shengwu Jishu Tongbao(Biotechnology Bulletin),31(11):68-77(靳进朴,郭安源,何坤,张禾,朱其慧,陈新,高歌,罗静初,2015,植物转录因子分类,预测和数据库构建,生物技术通报,31(11):68-77)
Kranz H.D.,Denekamp M.,Greco R.,Jin H.,Leyva A.,Meissner R.C.,Petroni K.,Urzainqui A.,Bevan M.,Martin C.,Smeekens S.,Tonelli C.,Paz-Ares J.,and Weisshaar B.,1998,Towards functional characterisation of the members of the R2R3-MYB gene family from Arabidopsis thaliana,Plant J.,16(2):263-276
Lee S.B.,and Suh M.C.,2015,Cuticular wax biosynthesis is up-regulated by the MYB94 transcription factor in arabidopsis,Plant and Cell Physiol.,56(1):48-60
Li S.F.,2013,Research progress on pharmacological action of Astragalus membranaceus,Hubei Zhongyi Zazhi(Hubei Journal of TCM JUN),35(6):73-75(李淑芳,2013,中药黄芪药理作用研究进展,湖北中医杂志,35(6):73-75)
Liu J.,Osbourn A.,and Ma P.,2015,MYB transcription factors as regulators of phenylpropanoid methabolism in plants,Mol.Plant,8(5):689-708
Liu J.Q.,Yang J.H.,and Zhu G.,2011,Analysis on the life cycle of the endangered plant population of Astragalus membranaceus,Caoye Kexue(Pratacultural Science),28(2):180-184(刘建泉,杨建红,朱高,2011,濒危植物膜荚黄芪种群生活史分析,草业科学,28(2):180-184)
Niu Y.L.,Jiang X.M.,and Xu X.Y.,2016,Reaserch advances on transcription factor MYB gene family in plant,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),14(8):2050-2059(牛义岭,姜秀明,许向阳,2016,植物转录因子MYB基因家族的研究进展,分子植物育种,14(8):2050-2059)
Ogata K.,Kanei-Ishii C.,Sasaki M.,Hatanaka H.,Nagadoi A.,Enari M.,Nakamura H.,Nishimura Y.,Ishii S.,and Sarai A.,1996,The cavity in the hydrophobic core of Myb DNA-binding domain is reserved for DNA recognition and transactivation,Nat.Struct.Biol.,3(2):178-187
Paz-Ares J.,Ghosal D.,Wienand U.,Peterson P.A.,and Saedler H.,1987,The regulatory c1 locus of Zea mays encodes a protein with homology to myb proto-oncogene products and with structural similarities to transcriptional activators,EM-BO J.,6(12):3553-3558
Riechmann J.L.,Heard J.,Martin G.,Reuber L.,Jiang C.Z.,Keddie J.,Adam L.,Pineda O.,Ratcliffe O.J.,Samaha R.R.,Creelman R.,Pilgrim M.,Broun P.,Zhang J.Z.,Ghandehari D.,Sherman B.K.,and Yu G.L.,2000,Arabidopsis transcription factors:genome-wide comparative analysis among eukaryotes,Science,290(5499):2105-2110
Rosinski J.A.,and Atchley W.R.,1998.,Molecular evolution of the Myb family of transcription factors:evidence for polyphyletic origin,J.Mol.Evol.,46(1):74-83
Sun X.,Liu J.Y.,Yuan X.H.,Pan X.W.,Du W.G.,Ren H.X.,Ma Y.B.,Jun ABE.,Qiu L.J.,and Liu B.H.,2013,Cloning and expression analysis of GmMYB genes induced by abiotic stresses,Zuowu Xuebao(Acta agronomica sinica),38(2):360-368(孙霞,刘晋跃,袁晓辉,潘相文,杜维广,任海洋,马永波,Jun ABE,邱丽娟,刘宝辉,2013,非生物胁迫诱导的GmMYB基因克隆与表达分析,作物学报,38(2):360-368)
Waites R.,Selvadurai H.R.N.,Oliver I.R.,and Hudson A.,1998,The PHANTASTICA gene encodes a MYB transcription factor involved in growth and dorsoventrality of lateral organs in antirrhinum,Cell,93(5):779-789
Wang C.R.,Wang Q.J.,Li X.L.,Yao Y.X.,and Hao Y.J.,2014,Screening and expression analysis of salt-induced R2R3-MYB genes in grapevine,Yuanyi Xuebao(Acta Horticulturae Sinica),41(3):529-535(王春荣,王庆杰,李晓玲,姚玉新,郝玉金,2014,葡萄中盐诱导的R2R3-MYB基因的筛选与表达分析,园艺学报,41(3):529-535)
Wang L.F.,Lu Y.X.,Wang J.K.,Qin B.,and Zhang D.,2017,Functions of plant MYB44 transcript factor and its applicationprospects for stress resistance research in rubber tree,Redai Nongye Kexue(Chinese Journal of Tropical Agriculture),37(5):30-36(王立丰,陆燕茜,王纪坤,覃碧,张冬,2017,植物MYB44转录因子的功能及其在橡胶树抗逆研究中的应用前景,热带农业科学,37(5):30-36)
Wang Y.Q.,Zhang B.,Chen W.J.,Liu D.C.,Liu B.L.,and Zhang H.G.,2015,Cloning and functional verification of R2R3-MYB transcription factor TaMYB3-4D in wheat,Xibei Zhiwu Xuebao(Acta Botanica Boreali-Occidentalia Sinica),35(4):646-652(王延谦,张波,陈文杰,刘登才,刘宝龙,张怀刚,2015,小麦R2R3-MYB转录因子TaMYB3-4D的克隆及功能分析,西北植物学报,35(4):646-652)
Wasternack C.,and Hause B.,2013,Jasmonates:biosynthesis,perception,signal transduction and action in plant stress response,growth and development,an update to the 2007 review in annals of botany,Ann.Bot.,111(6):1021-1058
Yan Z.J.,Yang C.,Gao T.M.,and Jia Z.F.,2006,Comparison of ecological and economic value of 6 kinds of common sand fixing plants,Ganhanqu Ziyuan Yu Huanjin(Journal of Arid Land Resources and Environment),20(3):163-168(闫志坚,扬持,高天明,贾志斌,2006,6种常用固沙植物的生态经济价值比较,干旱区资源与环境,20(3):163-168)
Yang C.F.,Cao Y.,Hu S.L.,Xu G.,Huang Y.,and Long Z.J.,2015,Cloning of MYB gene based on Bambusa emeiensis transcriptome and their induced expression under stresses,Shenlin Yu Huanjin Xuebao(Journal of Forest and Environment),35(1):60-66(杨传凤,曹颖,胡尚连,徐刚,黄艳,龙治坚,2015,基于慈竹转录组MYB基因的克隆及胁迫诱导表达,森林与环境学报,35(1):60-66)
Yang W.J.,Du H.,Fang F.,Yang W.S.,Wu Y.M.,and Tang Y.X.,2008,Cloning and characterization of two new MYB transcription factor genes from soybean,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),41(4):961-970(杨文杰,杜海,方芳,杨婉身,吴燕民,唐益雄,2008,大豆两个MYB转录因子基因克隆及表达分析,中国农业科学,41(4):961-970)
Yi B.X.,Yu S.Q.,Zhang J.L.,Li Z.Q.,and Gong Q.F.,2014,Aresearch review of Astragalus membranaceus,Jiangxi Zhongyiyao Daxue Xuebao(Journal of Jiangxi University of TCM),26(2):98-100(易炳学，余书琦，张金莲，李志强，龚千锋,2014,黄芪的研究概况,江西中医药大学学报,26(2):98-100)
基本信息:
DOI：10.13417/j.gab.038.003605
中图分类号:Q943.2;S567.239
引用信息:
[1]李竹韵,焦子迅,陈阳,等.黄芪AmMYB44基因的克隆与表达模式分析[J].基因组学与应用生物学,2019,38(08):3605-3613.DOI:10.13417/j.gab.038.003605.
基金信息:
国家自然科学基金项目(31670335; 31770363);; 国家大学生创新训练计划(URTP2017110023)共同资助
2019-08-25
2019-08-25