白桦BpTCP7基因启动子的克隆及表达分析
|
摘要
【目的】研究Bp TCP7启动子的表达模式,为深入分析Bp TCP7基因功能提供一定参考。【方法】克隆了Bp TCP7基因上游1 791 bp启动子序列,通过PLACE和Plant CARE网址对顺式作用元件进行分析,然后转入拟南芥,并分析Bp TCP7启动子的组织表达特性及盐、旱胁迫应答反应。【结果】Bp TCP7启动子序列中含有与细胞周期、开花、叶片发育、激素及胁迫响应等相关的顺式作用元件。该启动子在拟南芥中的表达模式呈现为营养生长阶段和生殖生长阶段均有表达,且在叶片中表达明显,与此同时,Bp TCP7启动子在幼嫩、成熟叶片的边缘均有表达。与对照相比,Na Cl、PEG胁迫诱导处理后Bp TCP7基因表达量有升高的现象。【结论】Bp TCP7基因参与白桦的叶片发育、激素应答、胁迫响应(盐、干旱)等生物学过程,对营养、生殖生长阶段各组织器官的发育也有一定的调控作用。
【Objective】Bp TCP7 gene,a member of the CIN branch of TCP family,plays an important role in plant growth and development. In order to characterize the function of Bp TCP7. 【Method】A 1 791 bp flanking sequence upstream of the translation initiation codon was cloned,and putative cis-regulatory elements were deciphered from the promoter sequence of Bp TCP7 using PLACE and Plant CARE web tools,and transferred into Arabidopsis thaliana. Then,the expression patterns were investigated during salt and drought stress. 【Result】The results showed that cis-regulatory elements involved in cell cycle,flowering,leaf development,multiple hormone-responsiveness,and stress response were included in the promoter region. The Gus reporter gene that was drived the Bp TCP7 promoter was detected in all tissues and organs during the reproductive and vegetative phases. Notably,the Bp TCP7 promoter was expressed in the leaf margin of young and mature leaves. The expression level of Bp TCP7 gene was increased in transgenic A. thaliana after Na Cl and polyethylene glycol( PEG) stresses compared with the control.【Conclusion】In conclusion,Bp TCP7 participates in biological processes of leaf development,hormone response,drought and salt stress response,and to some extent regulates the development of all tissues and organs during the reproductive and vegetative growth phases.
【Objective】Bp TCP7 gene,a member of the CIN branch of TCP family,plays an important role in plant growth and development. In order to characterize the function of Bp TCP7. 【Method】A 1 791 bp flanking sequence upstream of the translation initiation codon was cloned,and putative cis-regulatory elements were deciphered from the promoter sequence of Bp TCP7 using PLACE and Plant CARE web tools,and transferred into Arabidopsis thaliana. Then,the expression patterns were investigated during salt and drought stress. 【Result】The results showed that cis-regulatory elements involved in cell cycle,flowering,leaf development,multiple hormone-responsiveness,and stress response were included in the promoter region. The Gus reporter gene that was drived the Bp TCP7 promoter was detected in all tissues and organs during the reproductive and vegetative phases. Notably,the Bp TCP7 promoter was expressed in the leaf margin of young and mature leaves. The expression level of Bp TCP7 gene was increased in transgenic A. thaliana after Na Cl and polyethylene glycol( PEG) stresses compared with the control.【Conclusion】In conclusion,Bp TCP7 participates in biological processes of leaf development,hormone response,drought and salt stress response,and to some extent regulates the development of all tissues and organs during the reproductive and vegetative growth phases.
引文
[1]DOEBLEY J,STEC A,HUBBARD L.The evolution of apical in maize[J].Nature,1997,386(6624):485-488.DOI:10.1038/386485a0.
[2]LUO D,CARPENTER R,VINCENT C,et al.Origin of floral asymmetry in Antirrhinum[J].Nature,1996,383(6603):794-799.DOI:10.1038/383794a0.
[3]KOSUGI S,OHASHI Y.PCF1 and PCF2 specifically bind to cis elements in the rice PROLIFERATING CELL NUCLEARANTIGEN gene[J].Plant Cell,1997,9(9):1607-1619.DOI:10.1105/tpc.9.9.1607.
[4]CUBAS P,LAUTER N,DOEBLEY J,et al.The TCP domain:a motif found in proteins regulating plant growth and development[J].The Plant Journal,1999,18(2):215-222.
[5]KOSUGI S,OHASHI Y.DNA binding and dimerization specificity and potential targets for the TCP protein family[J].The Plant Journal,2002,30(3):337-348.
[6]LI C,POTUSCHAK T,COLóN-CARMONA A,et al.Arabidopsis TCP20 links regulation of growth and cell division control pathways[J].Proceedings of the National Academy of Sciences of the United States of America,2005,102(36):12978-12983.DOI:10.1073/pnas.0504039102.
[7]HOWARTH D G,DONOGHUE M J.Phylogenetic analysis of the"ECE"(CYC/TB1)clade reveals duplications predating the core eudicots[J].Proceedings of the National Academy of Sciences,2006,103(24):9101-9106.DOI:10.1073/pnas.0602827103.
[8]NAVAUD O,DADOS P,CARNUS E,et al.TCP transcription factors predate the emergence of land plants[J].Journal of Molecular Evolution,2007,65(1):23-33.DOI:10.1007/s00239-006-0174-z.
[9]DAMERVAL C,LE GUILLOUX M,JAGER M,et al.Diversity and evolution of CYCLOIDEA-like TCP genes in relation to flower development in Papaveraceae[J].Plant Physiology,2007,143(2):759-772.DOI:10.1104/pp.106.090324.
[10]GAO Y,ZHANG D,LI J.TCP1 modulates DWF4 expression via directly interacting with the GGNCCC motifs in the promoter rejion of Arabidopsis thaliana[J].Journal of Genetics and Genomics,2015,42(7):383-392.DOI:10.1016/j.jgg.2015.04.009.
[11]WELCHEN E,GONZALEZ D H.Overrepresentation of elements recognized by TCP-domain transcription factors in the upstream regions of nuclear genes encoding components of the mitochondrial oxidative phosphorylation Machinery[J].Plant Physiology,2006,141(2):540-545.DOI:10.1104/pp.105.075366.
[12]AGUILAR-MARTíNEZ J A,POZA-CARRIóN C,CUBAS P.Arabidopsis BRANCHED1 acts as an integrator of branching signals within axillary buds[J].The Plant Cell Online,2007,19(2):458-472.DOI:10.1105/tpc.106.048934.
[13]DAVIèRE J M,WILD M,REGNAULT T,et al.Class I TCP-DELLA interactions in inflorescence shoot apex determine plant height[J].Current Biology Cb,2014,24(16):1923-1928.DOI:10.1016/j.cub.2014.07.012.
[14]CRAWFORD B C,NATH U,CARPENTER R,et al.CINCINNATA controls both cell differentiation and growth in petal lobes and leaves of Antirrhinum[J].Plant Physiol,2004,135(1):244-253.DOI:10.1104/pp.103.036368.
[15]GIRAUD E,NG S,CARRIE C,et al.TCP transcription factors link the regulation of genes encoding mitochondrial proteins with the circadian clock in Arabidopsis thaliana[J].Plant Cell,2010,22(12):3921-3924.DOI:10.1105/tpc.110.074518.
[16]TATEMATSU K,NAKABAYASHI K,KAMIYA Y,et al.Transcription factor At TCP14 regulates embryonic growth potential during seed germination in Arabidopsis thaliana[J].Plant J,2008,53(1):42-52.DOI:10.1111/j.1365-313X.2007.03308.x.
[17]NATH U,CRAWFORD B C W,CARPENTER R,et al.Genetic control of surface curvature[J].Science,2003,299(5611):1404-1407.DOI:10.1126/science.1079354.
[18]MASUDA H P,CABRAL L M,VEYLDER L D,et al.ABAP1 is a novel plant Armadillo BTB protein involved in DNA replication and transcription[J].Embo Journal,2014,27(20):2746-2756.DOI:10.1038/emboj.2008.191.
[19]SCHOMMER C,PALATNIK J F,AGGARWAL P,et al.Control of jasmonate biosynthesis and senescence bymiR319 targets[J].PLoS Biol,2008,6(9):e230.DOI:10.1371/journal.pbio.0060230.
[20]KOYAMA T,FURUTANI M,TASAKA M,et al.TCP transcription factors control the morphology of shoot lateral organs via negative regulation of the expression of boundary-specific genes in Arabidopsis[J].Plant Cell,2007,19(2):473-484.DOI:10.1105/tpc.106.044792.
[21]ORI N,COHEN A R,ETZIONI A,et al.Regulation of LANCEO-LATE by miR319 is required for compound-leaf development in tomato[J].Nature Genetics,2007,39(6):787-791.DOI:10.1038/ng2036.
[22]NATH U,AGGRAWAL P,CHALLA K R.Activation of YUCCA5by the transcription factor TCP4 integrates developmental and environmental signals to promote hypocotyl elongation in Arabidopsis[J].Plant Cell,2016,28:2117-2130.DOI:10.1105/tpc.16.00360.
[23]DANISMAN S,VAN DER WAL F,DHONDT S,et al.Arabidopsis class I and class II TCP transcription factors regulate jasmonic acid metabolism and leaf development antagonistically[J].Plant Physiol,2012,159(4):1511-1523.DOI:10.1104/pp.112.200303.
[24]LI S,ZACHGO S.TCP3 interacts with R2R3-MYB proteins,promotes flavonoid biosynthesis and negatively regulates the auxin response in Arabidopsis thaliana[J].The Plant Journal,2013,76(6):901-913.DOI:10.1111/tpj.12348.
[25]WANG S T,SUN X L,HOSHINO Y,et al.MicroRNA319positively regulates cold tolerance by targeting Os PCF6 and OsTCP21 in rice(Oryza sativa L.)[J].PLoS One,2014,9(3):e91357.DOI:10.1371/journal.pone.0091357.
[26]GUAN P,RIPOLL J J,WANG R,et al.Interacting TCP and NLPtranscription factors control plant responses to nitrate availability[J].Proc Natl Acad Sci USA,2017,114(9):2419-2424.DOI:10.1073/pnas.1615676114.
[27]HIGO K,UGAWA Y,IWAMOTO M,et al.Plant cis-acting regulatory DNA elements(PLACE)database:1999[J].Nucleic Acids Research,1999,27(1):297-300.
[28]POSTEL D,VANLEMMENS P,GODE P,et al.PlantCARE,a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences[J].Nucleic Acids Research,2002,30(1):325-327.
[29]CLOUGH S J,BENT A F.Floral dip:a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana[J].Plant Journal,2010,16(6):735-743.
[30]LI C,JIANG B,WU C,et al.The characterization of GmTIP,a root-specific gene from soybean,and the expression analysis of its promoter[J].Plant Cell Tissue&Organ Culture,2015,121(2):259-274.DOI:10.1007/s11240-014-0682-2.
[31]MUKHOPADHYAY P,TYAGI A K.Os TCP19 influences developmental and abiotic stress signaling by modulating ABI4-mediated pathways[J].Sci Rep,2015,5:9998.DOI:10.1038/srep12381.
[2]LUO D,CARPENTER R,VINCENT C,et al.Origin of floral asymmetry in Antirrhinum[J].Nature,1996,383(6603):794-799.DOI:10.1038/383794a0.
[3]KOSUGI S,OHASHI Y.PCF1 and PCF2 specifically bind to cis elements in the rice PROLIFERATING CELL NUCLEARANTIGEN gene[J].Plant Cell,1997,9(9):1607-1619.DOI:10.1105/tpc.9.9.1607.
[4]CUBAS P,LAUTER N,DOEBLEY J,et al.The TCP domain:a motif found in proteins regulating plant growth and development[J].The Plant Journal,1999,18(2):215-222.
[5]KOSUGI S,OHASHI Y.DNA binding and dimerization specificity and potential targets for the TCP protein family[J].The Plant Journal,2002,30(3):337-348.
[6]LI C,POTUSCHAK T,COLóN-CARMONA A,et al.Arabidopsis TCP20 links regulation of growth and cell division control pathways[J].Proceedings of the National Academy of Sciences of the United States of America,2005,102(36):12978-12983.DOI:10.1073/pnas.0504039102.
[7]HOWARTH D G,DONOGHUE M J.Phylogenetic analysis of the"ECE"(CYC/TB1)clade reveals duplications predating the core eudicots[J].Proceedings of the National Academy of Sciences,2006,103(24):9101-9106.DOI:10.1073/pnas.0602827103.
[8]NAVAUD O,DADOS P,CARNUS E,et al.TCP transcription factors predate the emergence of land plants[J].Journal of Molecular Evolution,2007,65(1):23-33.DOI:10.1007/s00239-006-0174-z.
[9]DAMERVAL C,LE GUILLOUX M,JAGER M,et al.Diversity and evolution of CYCLOIDEA-like TCP genes in relation to flower development in Papaveraceae[J].Plant Physiology,2007,143(2):759-772.DOI:10.1104/pp.106.090324.
[10]GAO Y,ZHANG D,LI J.TCP1 modulates DWF4 expression via directly interacting with the GGNCCC motifs in the promoter rejion of Arabidopsis thaliana[J].Journal of Genetics and Genomics,2015,42(7):383-392.DOI:10.1016/j.jgg.2015.04.009.
[11]WELCHEN E,GONZALEZ D H.Overrepresentation of elements recognized by TCP-domain transcription factors in the upstream regions of nuclear genes encoding components of the mitochondrial oxidative phosphorylation Machinery[J].Plant Physiology,2006,141(2):540-545.DOI:10.1104/pp.105.075366.
[12]AGUILAR-MARTíNEZ J A,POZA-CARRIóN C,CUBAS P.Arabidopsis BRANCHED1 acts as an integrator of branching signals within axillary buds[J].The Plant Cell Online,2007,19(2):458-472.DOI:10.1105/tpc.106.048934.
[13]DAVIèRE J M,WILD M,REGNAULT T,et al.Class I TCP-DELLA interactions in inflorescence shoot apex determine plant height[J].Current Biology Cb,2014,24(16):1923-1928.DOI:10.1016/j.cub.2014.07.012.
[14]CRAWFORD B C,NATH U,CARPENTER R,et al.CINCINNATA controls both cell differentiation and growth in petal lobes and leaves of Antirrhinum[J].Plant Physiol,2004,135(1):244-253.DOI:10.1104/pp.103.036368.
[15]GIRAUD E,NG S,CARRIE C,et al.TCP transcription factors link the regulation of genes encoding mitochondrial proteins with the circadian clock in Arabidopsis thaliana[J].Plant Cell,2010,22(12):3921-3924.DOI:10.1105/tpc.110.074518.
[16]TATEMATSU K,NAKABAYASHI K,KAMIYA Y,et al.Transcription factor At TCP14 regulates embryonic growth potential during seed germination in Arabidopsis thaliana[J].Plant J,2008,53(1):42-52.DOI:10.1111/j.1365-313X.2007.03308.x.
[17]NATH U,CRAWFORD B C W,CARPENTER R,et al.Genetic control of surface curvature[J].Science,2003,299(5611):1404-1407.DOI:10.1126/science.1079354.
[18]MASUDA H P,CABRAL L M,VEYLDER L D,et al.ABAP1 is a novel plant Armadillo BTB protein involved in DNA replication and transcription[J].Embo Journal,2014,27(20):2746-2756.DOI:10.1038/emboj.2008.191.
[19]SCHOMMER C,PALATNIK J F,AGGARWAL P,et al.Control of jasmonate biosynthesis and senescence bymiR319 targets[J].PLoS Biol,2008,6(9):e230.DOI:10.1371/journal.pbio.0060230.
[20]KOYAMA T,FURUTANI M,TASAKA M,et al.TCP transcription factors control the morphology of shoot lateral organs via negative regulation of the expression of boundary-specific genes in Arabidopsis[J].Plant Cell,2007,19(2):473-484.DOI:10.1105/tpc.106.044792.
[21]ORI N,COHEN A R,ETZIONI A,et al.Regulation of LANCEO-LATE by miR319 is required for compound-leaf development in tomato[J].Nature Genetics,2007,39(6):787-791.DOI:10.1038/ng2036.
[22]NATH U,AGGRAWAL P,CHALLA K R.Activation of YUCCA5by the transcription factor TCP4 integrates developmental and environmental signals to promote hypocotyl elongation in Arabidopsis[J].Plant Cell,2016,28:2117-2130.DOI:10.1105/tpc.16.00360.
[23]DANISMAN S,VAN DER WAL F,DHONDT S,et al.Arabidopsis class I and class II TCP transcription factors regulate jasmonic acid metabolism and leaf development antagonistically[J].Plant Physiol,2012,159(4):1511-1523.DOI:10.1104/pp.112.200303.
[24]LI S,ZACHGO S.TCP3 interacts with R2R3-MYB proteins,promotes flavonoid biosynthesis and negatively regulates the auxin response in Arabidopsis thaliana[J].The Plant Journal,2013,76(6):901-913.DOI:10.1111/tpj.12348.
[25]WANG S T,SUN X L,HOSHINO Y,et al.MicroRNA319positively regulates cold tolerance by targeting Os PCF6 and OsTCP21 in rice(Oryza sativa L.)[J].PLoS One,2014,9(3):e91357.DOI:10.1371/journal.pone.0091357.
[26]GUAN P,RIPOLL J J,WANG R,et al.Interacting TCP and NLPtranscription factors control plant responses to nitrate availability[J].Proc Natl Acad Sci USA,2017,114(9):2419-2424.DOI:10.1073/pnas.1615676114.
[27]HIGO K,UGAWA Y,IWAMOTO M,et al.Plant cis-acting regulatory DNA elements(PLACE)database:1999[J].Nucleic Acids Research,1999,27(1):297-300.
[28]POSTEL D,VANLEMMENS P,GODE P,et al.PlantCARE,a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences[J].Nucleic Acids Research,2002,30(1):325-327.
[29]CLOUGH S J,BENT A F.Floral dip:a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana[J].Plant Journal,2010,16(6):735-743.
[30]LI C,JIANG B,WU C,et al.The characterization of GmTIP,a root-specific gene from soybean,and the expression analysis of its promoter[J].Plant Cell Tissue&Organ Culture,2015,121(2):259-274.DOI:10.1007/s11240-014-0682-2.
[31]MUKHOPADHYAY P,TYAGI A K.Os TCP19 influences developmental and abiotic stress signaling by modulating ABI4-mediated pathways[J].Sci Rep,2015,5:9998.DOI:10.1038/srep12381.