酪氨酸促进三色堇花斑形成的转录组解析
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摘要
为揭示酪氨酸对三色堇花斑形成的分子机制及转录组特征,探究酪氨酸影响三色堇花青素的合成途径,运用转录组学的方法,以清水为对照,采用3 mmol/L酪氨酸处理三色堇非色斑区,并进行转录组测序,获得19 438条表达显著差异基因,其中13 037条上调,6 401条下调。在花青素相关合成途径中,有9个基因差异表达,与花色相关的转录因子中,有1个基因差异表达。结果分析表明酪氨酸处理可以促进三色堇花青素合成基因和调控基因上调,从而促进花色素的形成。为今后三色堇的花色育种以及遗传信息的改良提供了新的思路。
To reveal the molecular mechanism and transcriptome characteristics of pansy floral blotcheseffected by tyrosine and to exploring the effect of tyrosine on anthocyanins biosynthesis of pansy, transcriptome methods,transcriptome sequencing of Viola×wittrockiana non-blotches area with treatments of H2 O or 3 mmol/L tyrosine were performed. 19 438 different expressing genes were obtained by de novo assembly method. 13 037 Unigenes were up-regulated and 6 401 Unigenes were down-regulated. 9 genes differentially expressed in anthocyanin biosynthesis, and 1 anthocyanin-related transcription factor genes differentially expressed. The results showed that tyrosine treatment could promote the up-regulation of anthocyanin biosynthesis genes and regulatory genes thus accelerating anthocyanin formation. provideing a new idea and theoretical basis for pansy to improve color breeding and genetic information in the future.
To reveal the molecular mechanism and transcriptome characteristics of pansy floral blotcheseffected by tyrosine and to exploring the effect of tyrosine on anthocyanins biosynthesis of pansy, transcriptome methods,transcriptome sequencing of Viola×wittrockiana non-blotches area with treatments of H2 O or 3 mmol/L tyrosine were performed. 19 438 different expressing genes were obtained by de novo assembly method. 13 037 Unigenes were up-regulated and 6 401 Unigenes were down-regulated. 9 genes differentially expressed in anthocyanin biosynthesis, and 1 anthocyanin-related transcription factor genes differentially expressed. The results showed that tyrosine treatment could promote the up-regulation of anthocyanin biosynthesis genes and regulatory genes thus accelerating anthocyanin formation. provideing a new idea and theoretical basis for pansy to improve color breeding and genetic information in the future.
引文
Baudry A.,Caboche M.,and Lepiniec L.,2006,TT8 controls its own expression in a feedback regulation involving TTG1and homologous MYB and bHLH factors,allowing a strong and cell-specific accumulation of flavonoids in Arabidopsis thaliana,Plant J.,46(5):768-779
Conesa A.,Gotz S.,Garcia-Gomez J.M.,Terol J.,Talon M.,and Robles M.,2005,Blast2GO:a universal tool for annotation,visualization and analysis in functional genomics research,Bioinformatics,21(18):3674-3676
de Vetten N.,Quattrocchio F.,Mol J.,and Koes R.,1997,The an11 locus controlling flower pigmentation in petunia encodes a novel WD-repeat protein conserved in yeast,Plants and animals,Genes&Development,11(11):1422-1434
Grabherr M.G.,Haas B.J.,Yassour M.,Levin J.Z.,Thompson D.A.,Amit I.,Adiconis X.,Fan L.,Raychowdhury R.,Zeng Q.D.,Chen Z.H.,Mauceli E.,Hacohen N.,Gnirke A.,Rhind N.,Palma F.D.,Birren B.W.,Nusbaum C.,Lindblad-Toh K.,Friedman N.,and Regev A.,2011,Full-length transcriptome assembly from RNA-Seq data without a reference genome,Nat.Biotechnol.,29(7):644-652
Iseli C.,Jongeneel C.V.,and Bucher P.,1999,Estscan:a program for detecting,evaluating,and reconstructing potential coding regions in EST sequences,Proc.Int.Conf.Intell.Syst.Mol.Biol.,99:138-148
Li J.,2014,Transcriptome sequencing of Viola×wittrockiana Gams.and searcing,expression validation for protential genes related to petal pigment biosythesis,Thesis for M.S.,Hainan University,Supervisor:Wang J.,pp.29-38(李婧,2014,三色堇(Viola×wittrockiana Gams.)转录组测序及花色相关基因的发掘与表达验证,硕士学位论文,海南大学,导师:王健,pp.29-38)
Li Q.,2013,Cloning and expression analysis of anthocyanin synthase genes in pansy,Thes is for M.S.,Hainan University,Supervisor:Wang J.,pp.9-28(李琴,2013,三色堇花色素合成结构基因的克隆与表达差异分析,硕士学位论文,海南大学,导师:王健,pp.9-28)
Li R.Q.,Yu C.,Li Y.R.,Lam T.W.,Yiu S.M.,Kristiansen K.,and Wang J.,2009,Soap2:an improved ultrafast tool for short read alignment,Bioinformatics,25(15):1966-1967
Ma H.,Pooler M.,and Griesbach R.,2009,Anthocyanin regulatory/structural gene expression in Phalaenopsis,J.AM.Soc.Hortic.Sci.,134(1):88-96
Ma Q.P.,Li H.,Zou Z.W.,Arkorful E.,Lv Q.R.,Zhou Q.Q.,Chen X.,Sun K.,and Li X.H.,2018,Transcriptomic analyses identify albino-associated genes of a novel albino tea germplasm'Huabai 1',Hortic.Res.,5:54
Martins T.R.,Berg J.J.,Blinka S.,Rausher M.D.,and Baum D.A.,2013,Precise spatialtemporal regulation of the anthocyanin biosynthetic pathway leads to petal spot formation in Clarkia gracilis(Onagraceae),New Phytol.,197(3):958-969
Matsubara K.,Kei S.B.,Koizumi M.,Kodama H.C.,and Ando T.,2012,RNA silencing in white petunia flowers creates pigmentation patterns invisible to the human eye,J.Plant Physiol.,169(9):920-923
Mortazavi A.,Williams B.A.,McCue K.,Schaeffer L.,and Wold B.,2008,Mapping and quantifying mammalian transcriptomes by RNA-Seq,Nat.Methods,5(7):621-628
Petroni K.,and Tonelli C.,2011,Recent advances on the regulation of anthocyanin synthesis in reproductive organs,Plant Sci.,181(3):219-229
Polturak G.,Heinig U.,Grossman N.,Battat M.,Leshkowitz D.,Malitsky S.,Rogachev I.,and Aharoni A.,2018,Transcriptome and metabolic profiling provides insights into betalain biosynthesis and evolution in Mirabilis jalapa,Mol.Plant,11(1):189-204
Quattrocchio F.,Wing J.F.,van der Woude K.,Mol J.N.,and Koes R.,1998,Analysis of bHLH and MYB domain proteins:Species specific regulatory differences are caused by divergent evolutionof target anthocyanin genes,Plant J.,13(4):475-488
Rosler J.,Krekel F.,Amrhein N.,and Schmid J.,1997,Maize phenylalanine ammonia-lyase has tyrosine ammonia-lyase activity,Plant Physiol.,113(1):175-179
Shang Y.,Venail J.,Mackay S.,Bailey P.C.,Schwinn K.E.,Jameson P.E.,Martin C.R.,and Davies K.M.,2011,The molecular basis for venation patterning of pigmentation and its effect on pollinator attraction in flowers of Antirrhinum,New Phytol.,189(2):602-615
Spelt C.,Quattrocchio F.,Mol J.,and Koes R.,2002,ANTHO-CYANIN1 of Petunia controls pigment synthesis,vacuolar pH,and seed coat development by genetically distinct mechanisms,Plant Cell,14(19):2121-2135
Stracke R.,Ishihara H.,Huep G.,Barsch A.,Mehrtens F.,Niehaus K.,and Weisshaar B.,2007,Differential regulation of closely related R2R3-MYB transcription factors controls flavonol accumulation in different parts of the Arabidopsis thaliana seedling,Plant J.,50(4):660-677
Teng S.,Keurentjes J.,Bentsink L.,Koornneef M.,and Smeekens S.,2005,Sucrose-specific induction of anthocyanin biosynthesis in Arabidopsis requires the MYB75/PAP1 gene,Plant Physiol.,139(4):1840-1852
Wang Z.J.,Wei X.,Liang J.W.,Wang C.S.,and Kang Y.,2017,Effect of fucoidan on b16 murine melanoma cell melanin formation and apoptosis:African Journal of Traditional Complementary and Alternative Medicines,14(4):149-155
Yamagishi M.,2010,Oriental hybrid lily Sorbonne homologue of LhMYB12 regulates anthocyanin biosyntheses in flower tepals and tepal spots,Mol.Breeding,28(3):381-389
Yang W.H.,Peng T.,Li T.G.,Cen J.R.,and Wang J.,2018,Tyramine and tyrosine decarboxylase gene contributes to the formation of cyanic blotches in the petals of pansy(Viola×wittrockiana),Plant Physiol.Biochem.,127:269-275
Ye J.,Fang L.,Zheng H.K.,Zhang Y.,Chen J.,Zhang Z.J.,Wang J.,Li S.T.,Li R.Q.,Bolund L.,and Wang J.,2006,WEGO:a web tool for plotting GO annotations,Nucleic Acids Res.,34(W):293-297
Zhou Z.,2014,Tyrosine induces anthocyanin biosynthesis in Ara bidopsis thaliana,Thesis for M.S.,Hunan Agricultural University,Supervisor:Ren C.M.,pp.9-34(周舟,2014,酪氨酸诱导拟南芥花青素生物合成的研究,硕士学位论文,湖南农业大学,导师:任春梅,pp.9-34)
Conesa A.,Gotz S.,Garcia-Gomez J.M.,Terol J.,Talon M.,and Robles M.,2005,Blast2GO:a universal tool for annotation,visualization and analysis in functional genomics research,Bioinformatics,21(18):3674-3676
de Vetten N.,Quattrocchio F.,Mol J.,and Koes R.,1997,The an11 locus controlling flower pigmentation in petunia encodes a novel WD-repeat protein conserved in yeast,Plants and animals,Genes&Development,11(11):1422-1434
Grabherr M.G.,Haas B.J.,Yassour M.,Levin J.Z.,Thompson D.A.,Amit I.,Adiconis X.,Fan L.,Raychowdhury R.,Zeng Q.D.,Chen Z.H.,Mauceli E.,Hacohen N.,Gnirke A.,Rhind N.,Palma F.D.,Birren B.W.,Nusbaum C.,Lindblad-Toh K.,Friedman N.,and Regev A.,2011,Full-length transcriptome assembly from RNA-Seq data without a reference genome,Nat.Biotechnol.,29(7):644-652
Iseli C.,Jongeneel C.V.,and Bucher P.,1999,Estscan:a program for detecting,evaluating,and reconstructing potential coding regions in EST sequences,Proc.Int.Conf.Intell.Syst.Mol.Biol.,99:138-148
Li J.,2014,Transcriptome sequencing of Viola×wittrockiana Gams.and searcing,expression validation for protential genes related to petal pigment biosythesis,Thesis for M.S.,Hainan University,Supervisor:Wang J.,pp.29-38(李婧,2014,三色堇(Viola×wittrockiana Gams.)转录组测序及花色相关基因的发掘与表达验证,硕士学位论文,海南大学,导师:王健,pp.29-38)
Li Q.,2013,Cloning and expression analysis of anthocyanin synthase genes in pansy,Thes is for M.S.,Hainan University,Supervisor:Wang J.,pp.9-28(李琴,2013,三色堇花色素合成结构基因的克隆与表达差异分析,硕士学位论文,海南大学,导师:王健,pp.9-28)
Li R.Q.,Yu C.,Li Y.R.,Lam T.W.,Yiu S.M.,Kristiansen K.,and Wang J.,2009,Soap2:an improved ultrafast tool for short read alignment,Bioinformatics,25(15):1966-1967
Ma H.,Pooler M.,and Griesbach R.,2009,Anthocyanin regulatory/structural gene expression in Phalaenopsis,J.AM.Soc.Hortic.Sci.,134(1):88-96
Ma Q.P.,Li H.,Zou Z.W.,Arkorful E.,Lv Q.R.,Zhou Q.Q.,Chen X.,Sun K.,and Li X.H.,2018,Transcriptomic analyses identify albino-associated genes of a novel albino tea germplasm'Huabai 1',Hortic.Res.,5:54
Martins T.R.,Berg J.J.,Blinka S.,Rausher M.D.,and Baum D.A.,2013,Precise spatialtemporal regulation of the anthocyanin biosynthetic pathway leads to petal spot formation in Clarkia gracilis(Onagraceae),New Phytol.,197(3):958-969
Matsubara K.,Kei S.B.,Koizumi M.,Kodama H.C.,and Ando T.,2012,RNA silencing in white petunia flowers creates pigmentation patterns invisible to the human eye,J.Plant Physiol.,169(9):920-923
Mortazavi A.,Williams B.A.,McCue K.,Schaeffer L.,and Wold B.,2008,Mapping and quantifying mammalian transcriptomes by RNA-Seq,Nat.Methods,5(7):621-628
Petroni K.,and Tonelli C.,2011,Recent advances on the regulation of anthocyanin synthesis in reproductive organs,Plant Sci.,181(3):219-229
Polturak G.,Heinig U.,Grossman N.,Battat M.,Leshkowitz D.,Malitsky S.,Rogachev I.,and Aharoni A.,2018,Transcriptome and metabolic profiling provides insights into betalain biosynthesis and evolution in Mirabilis jalapa,Mol.Plant,11(1):189-204
Quattrocchio F.,Wing J.F.,van der Woude K.,Mol J.N.,and Koes R.,1998,Analysis of bHLH and MYB domain proteins:Species specific regulatory differences are caused by divergent evolutionof target anthocyanin genes,Plant J.,13(4):475-488
Rosler J.,Krekel F.,Amrhein N.,and Schmid J.,1997,Maize phenylalanine ammonia-lyase has tyrosine ammonia-lyase activity,Plant Physiol.,113(1):175-179
Shang Y.,Venail J.,Mackay S.,Bailey P.C.,Schwinn K.E.,Jameson P.E.,Martin C.R.,and Davies K.M.,2011,The molecular basis for venation patterning of pigmentation and its effect on pollinator attraction in flowers of Antirrhinum,New Phytol.,189(2):602-615
Spelt C.,Quattrocchio F.,Mol J.,and Koes R.,2002,ANTHO-CYANIN1 of Petunia controls pigment synthesis,vacuolar pH,and seed coat development by genetically distinct mechanisms,Plant Cell,14(19):2121-2135
Stracke R.,Ishihara H.,Huep G.,Barsch A.,Mehrtens F.,Niehaus K.,and Weisshaar B.,2007,Differential regulation of closely related R2R3-MYB transcription factors controls flavonol accumulation in different parts of the Arabidopsis thaliana seedling,Plant J.,50(4):660-677
Teng S.,Keurentjes J.,Bentsink L.,Koornneef M.,and Smeekens S.,2005,Sucrose-specific induction of anthocyanin biosynthesis in Arabidopsis requires the MYB75/PAP1 gene,Plant Physiol.,139(4):1840-1852
Wang Z.J.,Wei X.,Liang J.W.,Wang C.S.,and Kang Y.,2017,Effect of fucoidan on b16 murine melanoma cell melanin formation and apoptosis:African Journal of Traditional Complementary and Alternative Medicines,14(4):149-155
Yamagishi M.,2010,Oriental hybrid lily Sorbonne homologue of LhMYB12 regulates anthocyanin biosyntheses in flower tepals and tepal spots,Mol.Breeding,28(3):381-389
Yang W.H.,Peng T.,Li T.G.,Cen J.R.,and Wang J.,2018,Tyramine and tyrosine decarboxylase gene contributes to the formation of cyanic blotches in the petals of pansy(Viola×wittrockiana),Plant Physiol.Biochem.,127:269-275
Ye J.,Fang L.,Zheng H.K.,Zhang Y.,Chen J.,Zhang Z.J.,Wang J.,Li S.T.,Li R.Q.,Bolund L.,and Wang J.,2006,WEGO:a web tool for plotting GO annotations,Nucleic Acids Res.,34(W):293-297
Zhou Z.,2014,Tyrosine induces anthocyanin biosynthesis in Ara bidopsis thaliana,Thesis for M.S.,Hunan Agricultural University,Supervisor:Ren C.M.,pp.9-34(周舟,2014,酪氨酸诱导拟南芥花青素生物合成的研究,硕士学位论文,湖南农业大学,导师:任春梅,pp.9-34)
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