芒果MinMYB10基因的克隆及表达分析载体的构建
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摘要
MYB(myeloblastosis)转录因子主要参与调控植物类黄酮代谢、植物生长发育等。为了研究在芒果果实中类黄酮代谢调控机制,本研究从金煌果肉中克隆得到一个MYB基因,将其命名为MinMYB10,其全长cDNA序列为1021 bp,开放阅读框为837bp。该基因编码的蛋白有278个氨基酸残基,分子重量为31.60ku,等电点为5.89。通过系统发育分析发现MinMYB10与拟南芥是亲缘关系较近的蛋白。本研究还构建了基因沉默载体pTRV2-MinMYB10和过表达载体pGreenII 62-SK-MinMYB10,以期在后续研究中探明MinMYB10基因在芒果果实花青素代谢中的作用。
The MYB(myeloblastosis) transcription factors are mainly involved in the regulation of flavonoid metabolism, growth and development in plants. In order to study how the flavonoid metabolic pathways are regulated in mango fruit, a MYB gene, designated as MinMYB10, was cloned from the flesh of Jinhuang mango fruit. The full-length cDNA sequence of MinMYB10 was 1021 bp long, containing an open reading frame of 837 bp which would encode a peptide of278 amino acid residues with a molecular weight of 31.60 ku and an isoelectric point of 5.89. Phylogenetic analysis showed that the encoded protein sequence was closely related to its homolog from Arabidopsis thaliana. We have also constructed a silencing vector, pTRV2-MinMYB10, and an over expression vector, pGreenII 62-SK-MinMYB10, aiming to decipher the role of MinMYB10 in the metabolism of anthocyanin in mango fruit in our upcoming researches.
The MYB(myeloblastosis) transcription factors are mainly involved in the regulation of flavonoid metabolism, growth and development in plants. In order to study how the flavonoid metabolic pathways are regulated in mango fruit, a MYB gene, designated as MinMYB10, was cloned from the flesh of Jinhuang mango fruit. The full-length cDNA sequence of MinMYB10 was 1021 bp long, containing an open reading frame of 837 bp which would encode a peptide of278 amino acid residues with a molecular weight of 31.60 ku and an isoelectric point of 5.89. Phylogenetic analysis showed that the encoded protein sequence was closely related to its homolog from Arabidopsis thaliana. We have also constructed a silencing vector, pTRV2-MinMYB10, and an over expression vector, pGreenII 62-SK-MinMYB10, aiming to decipher the role of MinMYB10 in the metabolism of anthocyanin in mango fruit in our upcoming researches.
引文
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[4]Legay S,Lacombe E,Goicoechea M,et al.Molecular characterization of EgMYBl,a putative transcriptional repressor of the lignin biosynthetic pathway[J].Plant Science,2007,173(5):542-549.
[5]FornaléS,Lopez E,Salazar-Henao J E,et al.AtMYB7:a new player in the regulation of UV-sunscreens in Arabidopsis thaliana[J].Plant Cell Physiology,2014,55(3):507-516.
[6]Li M,Dong X J,Peng J Q,et al.De novo transcriptome sequencing and gene expression analysis reveal potential mechanisms of seed abortion in dove tree(Davidia involucrata Baill.)[J].BMC Plant Biology,2016,16(82):1-21.
[7]Schwinn K E,Boase M R,Bradley J M,et al.MYB and b HLH transcription factor transgenes increase anthocyanin pigmentation in petunia and lisianthus plants,and the petunia phenotypes are strongly enhanced under field conditions[J].Frontiers in Plant Science,2014,5:603.
[8]罗学兵.芒果的营养价值、保健功能及食用方法[J].中国食物与营养,2011,17(7):77-79.
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[12]Song S,Qi T,Huang H,et al.The Jasmonate-ZIM domain proteins interact with the R2R3-MYB transcription factors MYB21 and MYB24 to affect Jasmonate-regulated stamen development in Arabidopsis[J].The Plant Cell,2011,23(3):1000-1013.
[13]Mccarthy R L,Zhong R,Fowler S,et al.The poplar MYBtranscription factors,PtrMYB3 and PtrMYB20,are involved in the regulation of secondary wall biosynthesis[J].Plant&Cell Physiology,2010,51(6):1084-1090.
[14]Ma Q H,Wang C,Zhu H H.TaMYB4 cloned from wheat regulates lignin biosynthesis through negatively controlling the transcripts of both cinnamyl alcohol dehydrogenase and cinnamoyl-CoA reductase genes[J].Biochimie,2011,93(7):1179-1186.
[15]Zhong R,Lee C,Mc Carthy R L,et al.Transcriptional activation of secondary wall biosynthesis by rice and maize NAC and MYB transcription factors[J].Plant&Cell Physiology,2011,52(10):1856-1871.
[16]Stracke R,Werber M,Weisshaar B.The R2R3-MYB gene family in Arabidopsis thaliana[J].Current Opinion in Plant Biology,2001,4(5):447-456.
[17]Lin-Wang K,Micheletti D,Palmer J,et al.High temperature reduces apple fruit colour via modulation of the anthocyanin regulatory complex[J].Plant,Cell&Environment,2011,34:1176-1190.
[18]Shen X,Zhao K,Liu L,et al.A role for Pac MYBA in ABA-regulated anthocyanin biosynthesis in red-colored sweet cherry cv.Hong Deng(Prunus avium L.)[J].Plant&Cell Physiology,2014,55(5):862-880.
[19]Fischer T C,Mirbeth B,Rentsch J,et al.Premature and ectopic anthocyanin formation by silencing of anthocyanidin reductase in strawberry(Fragaria ananassa)[J].New Phytologist Trust,2014,201(2):440-451.
[20]Fournier-Level A,Lacombe T,Le Cunff L,et al.Evolution of the Vv MybA gene family,the major determinant of berry colour in cultivated grapevine(Vitis vinifera L.)[J].Heredity,2010,104(4):351-362.
[21]Vimolmangkang S,Han Y,Wei G,et al.An apple MYBtranscription factor Md MYB3 is involved in regulation of anthocyanin biosynthesis and flower development[J].BMCPlant Biology,2013,13:176.
[22]赵佳,刘荣,杨帆,等.月季花青素苷相关R2R3-MYB蛋白基因的克隆和表达分析[J].中国农业科学,2015,48(7):1392-1404.
[2]Liu L,White M J,Macrae T H.Transcription factors and their genes in higher plants[J].European Journal of Biochemistry,1999,262(2):247.
[3]左然,徐美玲,柴国华,等.植物MYB转录因子功能及调控机制研究进展[J].生命科学,2012,24(10):1133-1140.
[4]Legay S,Lacombe E,Goicoechea M,et al.Molecular characterization of EgMYBl,a putative transcriptional repressor of the lignin biosynthetic pathway[J].Plant Science,2007,173(5):542-549.
[5]FornaléS,Lopez E,Salazar-Henao J E,et al.AtMYB7:a new player in the regulation of UV-sunscreens in Arabidopsis thaliana[J].Plant Cell Physiology,2014,55(3):507-516.
[6]Li M,Dong X J,Peng J Q,et al.De novo transcriptome sequencing and gene expression analysis reveal potential mechanisms of seed abortion in dove tree(Davidia involucrata Baill.)[J].BMC Plant Biology,2016,16(82):1-21.
[7]Schwinn K E,Boase M R,Bradley J M,et al.MYB and b HLH transcription factor transgenes increase anthocyanin pigmentation in petunia and lisianthus plants,and the petunia phenotypes are strongly enhanced under field conditions[J].Frontiers in Plant Science,2014,5:603.
[8]罗学兵.芒果的营养价值、保健功能及食用方法[J].中国食物与营养,2011,17(7):77-79.
[9]罗小杰,韦虎.芒果酒的研制[J].中国酿造,2007(5):89-90.
[10]Dubos C,Stracke R,Grotewold E,et al.MYB transcription factors in Arabidopsis[J].Trends in Plant Science,2010,15(10):573-581.
[11]Ma L G,Sun N,Liu X G,et al.Organ-specific expression of Arabidopsis genome during development[J].Plant Physiology,2005,138(1):80-91.
[12]Song S,Qi T,Huang H,et al.The Jasmonate-ZIM domain proteins interact with the R2R3-MYB transcription factors MYB21 and MYB24 to affect Jasmonate-regulated stamen development in Arabidopsis[J].The Plant Cell,2011,23(3):1000-1013.
[13]Mccarthy R L,Zhong R,Fowler S,et al.The poplar MYBtranscription factors,PtrMYB3 and PtrMYB20,are involved in the regulation of secondary wall biosynthesis[J].Plant&Cell Physiology,2010,51(6):1084-1090.
[14]Ma Q H,Wang C,Zhu H H.TaMYB4 cloned from wheat regulates lignin biosynthesis through negatively controlling the transcripts of both cinnamyl alcohol dehydrogenase and cinnamoyl-CoA reductase genes[J].Biochimie,2011,93(7):1179-1186.
[15]Zhong R,Lee C,Mc Carthy R L,et al.Transcriptional activation of secondary wall biosynthesis by rice and maize NAC and MYB transcription factors[J].Plant&Cell Physiology,2011,52(10):1856-1871.
[16]Stracke R,Werber M,Weisshaar B.The R2R3-MYB gene family in Arabidopsis thaliana[J].Current Opinion in Plant Biology,2001,4(5):447-456.
[17]Lin-Wang K,Micheletti D,Palmer J,et al.High temperature reduces apple fruit colour via modulation of the anthocyanin regulatory complex[J].Plant,Cell&Environment,2011,34:1176-1190.
[18]Shen X,Zhao K,Liu L,et al.A role for Pac MYBA in ABA-regulated anthocyanin biosynthesis in red-colored sweet cherry cv.Hong Deng(Prunus avium L.)[J].Plant&Cell Physiology,2014,55(5):862-880.
[19]Fischer T C,Mirbeth B,Rentsch J,et al.Premature and ectopic anthocyanin formation by silencing of anthocyanidin reductase in strawberry(Fragaria ananassa)[J].New Phytologist Trust,2014,201(2):440-451.
[20]Fournier-Level A,Lacombe T,Le Cunff L,et al.Evolution of the Vv MybA gene family,the major determinant of berry colour in cultivated grapevine(Vitis vinifera L.)[J].Heredity,2010,104(4):351-362.
[21]Vimolmangkang S,Han Y,Wei G,et al.An apple MYBtranscription factor Md MYB3 is involved in regulation of anthocyanin biosynthesis and flower development[J].BMCPlant Biology,2013,13:176.
[22]赵佳,刘荣,杨帆,等.月季花青素苷相关R2R3-MYB蛋白基因的克隆和表达分析[J].中国农业科学,2015,48(7):1392-1404.