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本研究基于苦荞转录组,筛选克隆一条与黄酮合成相关的MYB转录因子基因,命名为FtMYB56。生物信息学分析表明,该基因cDNA全长825 bp编码蛋白含有274个氨基酸,包括R2和R3保守结构域,属于R2R3-MYB家族。实时荧光定量PCR检测结果显示,FtMYB56基因随苦荞发育在各组织具有不同的表达趋势,其中在叶中随发育期至花期达到峰值后迅速减少,而在生殖器官随种子成熟过程急速上升。FtMYB56在拟南芥中过表达的黄酮成分分析发现,两个阳性转基因株系中花青素、芦丁和总黄酮含量极显著增加(p<0.01),分别为野生型的3.51和3.98倍、1.20和1.42倍及0.77和0.68倍基因表达量检测表明,拟南芥黄酮合成途径AtDFR、AtFLS和AtANS等晚期基因均极显著上调(p<0.01),而AtPAL、AtCHS、AtCHI和AtF3'H等早期基因表达无明显变化(p>0.05)。综上可见,苦荞转录因子FtMYB56可能通过增强黄酮合成途径晚期基因的表达,同时增加花青素和黄酮醇支路的合成代谢来提高黄酮物质的积累。
Abstract：In this study,a MYB transcription factor gene related to flavonoids synthesis,named FtMYB56,was screened and cloned from tartary buckwheat(Fagopyrum tataricum) based on its transcriptome.Bioinforma-tical analysis showed that FtMYB56 cDNA with 825 bp in full length encodes a putative protein of R2 R3-MYB type transcription factor protein with 274 amino acids.Real-time PCR analysis indicated that FtMYB56 had a different expression profiles in different tissues in the whole growing period of tartary buckwheat.Particularly,its expression level decreased rapidly in leaves after reaching the peak at flowering stage,but increased dramatically in reproductive organs with seed development.Furthermore,in two FtMYB56-overexpression Arabidopsis lines,anthocyanins,rutin and flavonoids showed a significant higher accumulation than that of WT(p <0.01),and their concentrations were 3.51 and 3.98 times,1.20 and 1.42 times,and 0.77 and 0.68 times of wild type,respectively.Moreover,A tDFR,A tFLS and AtANS,the late biosynthetic genes(EBGs) in flavonoid pathway,were expressed prominently in transgenic Arabidopsis more than that in WT(p <0.01).However,for the expression levels of early biosynthetic genes(EBGs) including A tPA L,A tCHS,A tCHI and A tF3'H,there was no obvious difference between transgenic lines and with WT(p>0.05).In conclusion,transcription factor FtMYB56 from tartary buckwheat may improve flavonoids accumulation through enhancing the metabolic flux of anthocyanin and flavonol branches due to the higher expressions of the late biosynthetic genes(EBGs) in flavonoid pathway.
Bai Y.C.,Li C.L.,Zhang J.W.,Li S.J.,Luo X.P.,Yao H.P.,Chen H.,Zhao H.X.,Park S.U.,and Wu Q.,2014,Characterization of two tartary buckwheat R2R3-MYB transcription factors and their regulation of proanthocyanidin biosynthesis,Physiol.Plant.,152(3):431-440
Bogs J.,Downey M.O.,Harvey J.S.,Ashton A.R.,Tanner G.J.,and Robinson S.P.,2005,Proanthocyanidin synthesis and expression of genes encoding leucoanthocyanidin reductase and anthocyanidin reductase in developing grape berries and grapevine leaves,Plant Physiol.,139(2):652-663
Du H.,Ran F.,Ma S.S.,Ke Y.Z.,Sun L.P.,Li J.N.,and Tang Y.X.,2016,Regulating effects of GmMYB042 gene on flavonoid biosynthesis,Zuowu Xuebao(Acta Agronomica Sinica),42(1):1-10(杜海,冉凤,马珊珊,柯蕴倬,孙丽萍,李加纳,唐益雄,2016,GmMYB042基因对类黄酮生物合成的调控作用,作物学报,42(1):1-10)
Falcone Ferreyra M.L.,Rius S.P.,and Casati P.,2012,Flavonoids:biosynthesis,biological functions,and biotechnological applications,Front.Plant Sci.,3:222
He S.M.,and Liu J.L.,2007,Study on the determination method of flavone content in tea,Fenxi Huaxue(Chinese Journal of Analytical Chemistry),35(9):1365-1368(何书美,刘敬兰,2007,茶叶中总黄酮含量测定方法的研究,分析化学,35(9):1365-1368)
Holton T.A.,Brugliera F.,and Tanaka Y.,1993,Cloning and expression of flavonol synthase from Petunia hybrida,Plant J.,4(6):1003-1010
Huang W.,Sun W.,LüH.,Xiao G.,Zeng S.,and Wang Y.,2012,Isolation and molecular characterization of thirteen R2R3-MYB transcription factors from epimedium sagittatum,Int.J.Mol.Sci.,14(1):594-610
Jaakola L.,2013,New insights into the regulation of anthocyanin biosynthesis in fruits,Trends Plant Sci.,18(9):477-483
Jin H.,and Martin C.,1999,Multifunctionality and diversity within the plant MYB-gene family,Plant Mol.Biol.,41(5):577-585
Jin W.,Wang H.,Li M.,Wang J.,Yang Y.,Zhang X.,Yan G.,Zhang H.,Liu J.,and Zhang K.,2016,The R2R3 MYB transcription factor PavMYB10.1 involves in anthocyanin biosynthesis and determines fruit skin colour in sweet cherry(Prunus avium L.),Plant Biotechnol.J.,14(11):2120-2133
Jun J.H.,Liu C.,Xiao X.,and Dixon R A.,2015,The transcriptional repressor MYB2 regulates both spatial and temporal patterns of proanthocyandin and anthocyanin pigmentation in medicago truncatula,Plant Cell,27(10):2860-2879
Jung C.,Seo J.S.,Han S.W.,Koo Y.J.,Kim C.H.,Song S.I.,Nahm B.H.,Choi Y.D.,and Cheong J.J.,2008,Overexpression of At MYB44 enhances stomatal closure to confer abiotic stress tolerance in transgenic Arabidopsis,Plant Physiol.,146(2):623-635
Liu L.,Du H.,Tang X.F.,Wu Y.M.,Huang Y.B.,and Tang Y.X.,2008,The roles of MYB transcription factors on plant defense responses and its molecular mechanism,Yichuan(Hereditas),30(10):1265-1271(刘蕾,杜海,唐晓凤,吴燕民,黄玉碧,唐益雄,2008,MYB转录因子在植物抗逆胁迫中的作用及其分子机理,遗传,30(10):1265-1271)
Lin-Wang K.,Mcghie T.K.,Wang M.,Liu Y.,Warren B.,Storey R.,Espley R.V.,and Allan A.C.,2014,Engineering the anthocyanin regulatory complex of strawberry(Fragaria vesca),Front.Plant Sci.,5:651
Luo X.,Zhao H.,Yao P.,Li Q.,Huang Y.,Li C.,Chen H.,and Wu Q.,2018,An R2R3-MYB transcription factor FtMYB15 involved in the synthesis of anthocyanin and proanthocyanidins from tartary buckwheat,J.Plant Growth Regul.,37(1):76-84
Martin C.,and Paz-Ares J.,1997,MYB transcription factors in plants,Trends in Genetics,13(2):67-73
Mehrtens F.,Kranz H.,Bednarek P.,and Weisshaar B.,2005,The Arabidopsis transcription factor MYB12 is a flavonolspecific regulator of phenylpropanoid biosynthesis,Plant Physiol.,138(2):1083-1096
Qin Y.,Wang M.,Tian Y.,He W.,Han L.,and Xia G.,2012,Over-expression of TaMYB33 encoding a novel wheat MYB transcription factor increases salt and drought tolerance in Arabidopsis,Mol.Biol.Rep.,39(6):7183-7192
Shi Q.Q.,Zhou L.,Li K.,and Wang Y.,2015,Transcriptional regulation involved in anthocyanin biosynthesis in plants,Linye Kexue Yanjiu(Forest Research),28(4):570-576(史倩倩,周琳,李奎,王雁,2015,植物花色素合成的转录调控研究进展,林业科学研究,28(4):570-576)
Thwe A.A.,Kim Y.J.,Li X.,Kim Y.B.,Park N.I.,Kim H.H.,Kim S.J.,and Sang U.P.,2014,Accumulation of phenylpropanoids and correlated gene expression in hairy roots of tartary buckwheat under light and dark conditions,Appl.Biochem.Biotechnol.,174(7):2537-2547
Witham P.K.,Yamashiro C.T.,Livak K.J.,and Batt C.A.,1996,A PCR-based assay for the detection of Escherichia coli Shiga-like toxin genes in ground beef,Appl.Environ.Microbiol.,62(4):1347-1353
Xing W.,and Jin X.L.,2015,Recent advances of MYB transcription factors involved in the regulation of flavonoid biosynthesis,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),13(3):689-696(邢文,金晓玲,2015,调控植物类黄酮生物合成的MYB转录因子研究进展,分子植物育种,13(3):689-696)
Xu F.,Ning Y.,Zhang W.,Liao Y.,Li L.,Cheng H.,and Cheng S.,2014,An R2R3-MYB transcription factor as a negative regulator of the flavonoid biosynthesis pathway in Ginkgo biloba,Funct.Integr.Genomic.,14(1):177-189
Zhang L.,Li X.,Ma B.,Gao Q.,Du H.,Han Y.,Li Y.,Cao Y.,Qi M.,Zhu Y.,Lu H.,Ma M.,Liu L.,Zhou J.,Nan C.,Qin Y.,Wang J.,Cui L.,Liu H.,Liang C.,and Qiao Z.,2017,The tartary buckwheat genome provides insights into rutin biosynthesis and abiotic stress tolerance,Mol.Plant,10(9):1224-1237
Zhang K.,Logacheva M D.,Meng Y.,Hu J.,Wan D.,Li L.,JanovskáD.,Wang Z.,Georgiev M.I.,Yu Z.,Yang F.,Yan M.,and Zhou M.,2018,Jasmonate-responsive MYB factors spatially repress rutin biosynthesis in Fagopyrum tataricum,J.Exp.Bot.,69(8):1955-1966
Zhang L.,Zhao G.,Xia C.,Jia J.,Liu X.,and Kong X.,2012,Overexpression of a wheat MYB transcription factor gene,TaMYB56-B,enhances tolerances to freezing and salt stresses in transgenic Arabidopsis,Gene,505(1):100-107
Zhang W.,Ning G.,Lv H.,Liao L.,and Bao M.,2009,Single MYB-type transcription factor At CAPRICE:a new efficient tool to engineer the production of anthocyanin in tobacco,Biochem.Bioph.Res.Co.,388(4):742-747
Zhou T.S.,2016,Characterization of flavonoid 3'-hydoxylase from the plant and its application in metabolic engineering,Dissertation for Ph.D.,Northwest Agricultural Forest Technology University,Supervisor:Yang Y.J.,pp.71-73(周天山,2016,茶树类黄酮3'-羟基化酶功能分析及其在代谢工程中的应用,博士学位论文,西北农林科技大学,导师:杨亚军,pp.71-73)
基本信息:
DOI：10.13417/j.gab.039.004664
中图分类号:S517
引用信息:
[1]余艳,廖小龙,肖玉华,等.苦荞转录因子基因FtMYB56的克隆及其对黄酮合成的调控[J].基因组学与应用生物学,2020,39(10):4664-4671.DOI:10.13417/j.gab.039.004664.
基金信息:
四川农业大学创新训练计划(编号:201710626040)资助
2019-10-25
2019-10-25
2019-10-25