白魔芋热激转录因子AaHSFA2a和AaHSFA2c的分离、亚细胞定位及表达分析
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摘要
以白魔芋(Amorphophallus albus)为材料,分离得到AaHSFA2a和AaHSFA2c基因的全长cDNA,序列比对分析发现,AaHSFA2a基因的开放阅读框(ORF)为1 023 bp,可编码340个氨基酸残基的多肽;AaHSFA2c基因含有一个内含子,开放阅读框为1 026 bp,可编码341个氨基酸残基的多肽。亚细胞定位结果显示,AaHSFA2a和AaHSFA2c蛋白均定位在细胞核中。实时荧光定量结果表明,AaHSFA2a和AaHSFA2c基因在白魔芋叶中的表达量明显高于根和球茎,均在热处理1 h时达到表达顶峰,并且AaHSFA2a基因的相对表达量明显高于AaHSFA2c基因。因此,推测AaHSFA2a和AaHSFA2c参与了魔芋热应激响应过程。
The full length c DNAs of AaHSF A 2a and AaHSFA2 c were isolated from Amorphophallus albus.Sequence alignment analysis showed that the open reading frame(ORF) of AaHSFA2 a gene was 1 023 bp, encoding a polypeptide of 340 amino acids residues. AaHSFA2 c gene contained one intron, with the ORF of 1 026 bp, and it encoded a polypeptide of 341 amino acid residues. Subcellular localization indicated that both AaHSFA2 a and AaHSFA2 c proteins were located in the nucleus. qRT-PCR analysis indicated that the expression level of AaHSFA2 a and AaHSFA2 c in leaves of Amorphophallus albus was significantly higher than that in roots and corms, and both of them reached the peak at 1 h after heat treatment, but the relative expression level of AaHSFA2 a was significantly higher than that of AaHSFA2 c. These results predicted that AaHSFA2 a and AaHSFA2 c might be involved in the heat stress response of Amorphophallus albus.
The full length c DNAs of AaHSF A 2a and AaHSFA2 c were isolated from Amorphophallus albus.Sequence alignment analysis showed that the open reading frame(ORF) of AaHSFA2 a gene was 1 023 bp, encoding a polypeptide of 340 amino acids residues. AaHSFA2 c gene contained one intron, with the ORF of 1 026 bp, and it encoded a polypeptide of 341 amino acid residues. Subcellular localization indicated that both AaHSFA2 a and AaHSFA2 c proteins were located in the nucleus. qRT-PCR analysis indicated that the expression level of AaHSFA2 a and AaHSFA2 c in leaves of Amorphophallus albus was significantly higher than that in roots and corms, and both of them reached the peak at 1 h after heat treatment, but the relative expression level of AaHSFA2 a was significantly higher than that of AaHSFA2 c. These results predicted that AaHSFA2 a and AaHSFA2 c might be involved in the heat stress response of Amorphophallus albus.
引文
Baniwal S.K.,Bharti K.,Chan K.Y.,Fauth M.,Ganguli A.,Kotak S.,Mishra S.K.,Nover L.,Port M.,Scharf K.D.,Tripp J.,Weber C.,Zielinski D.,and Von Koskull-Doring P.,2004,Heat stress response in plants:a complex game with chaperones and more than twenty heat stress transcription factors,J.Biosciences,29(4):471-487
Banti V.,Mafessoni F.,Loreti E.,Alpi A.,and Perata P.,2010,The heat-inducible transcription factor HsfA2 enhances anox ia tolerance in Arabidopsis,Plant Physiol.,152(3):1471-1483
Chan-Schaminet K.Y.,Baniwal S.K.,Bublak D.,Nover L.,and Scharf K.D.,2009,Specific interaction between tomato HsfA1 and HsfA2 creates heter o-oligomeric superactivator complexes for synergistic activation of heat stress gene ex pression,J.Biol.Chem.,284(31):20848-20857
Charng Y.Y.,Liu H.C.,Liu N.Y.,He W.T.,Wang C.N.,Chang S.H.,and Wang T.T.,2007,A heat-inducible transcription factor,HsfA2,is required for extension of acquired thermotolerance in Arabidopsis,Plant Physiol.,143(1):251-262
Fragkostefanakis S.,R觟th S.,Schleiff E.,and Scharf K.D.,2015,Prospects of engineering thermotolerance in crops through modulation of heat stress transcription factor and heat shock protein networks,Plant Cell Environ.,38(9):1881-1895
Guo M.,Yin Y.X.,Ji J.J.,Ma B.P.,Lu M.H.,and Gong Z.H.,2014,Cloning and expression analysis of heat-shock transcription factor gene CaHsfA2 from pepper(Capsicum annuum L.),Genet.Mol.Res.,13(1):1865-1875
Guo M.,Lu J.P.,Zhai Y.F.,Chai W.G.,Gong Z.H.,and Lu M.H.2015,Genome-wide analysis,expression profile of hea shock factor gene family(CaHsfs)and characterisation of CaHsfA2 in pepper(Capsicum annuum L.),BMC Plant Biology,15(1):151
Kotak S.,Port M.,Ganguli A.,Bicker F.,and Von Koskull-D觟ring P.,2004,Characterization of C-terminal domains of Arabidopsis heat stress transcription factors(Hsfs)and identification of a new signature combination of plant class A Hsfs with AHA and NES motifs essential for activator function and intracellular localization,Plant J.,39(1):98-112
Liu A.L.,Zou J.,Zhang X.W.,Zhou X.Y.,Wang W.F.,Xiong XY.,Chen L.Y.,and Chen X.B.,2010,Expression profiles of class a rice heat shock transcription factor genes under abiotic stresses,J.Plant Biol.,53(2):142-149
Liu H.C.,and Charng Y.Y.,2013,Common and distinct functions of Arabidopsis class A1 and A2 heat shock factors in diverse abiotic stress responses and development,Plant Physiol.,163(1):276-290
Liu J.G.,Yao Q.H.,Zhang Z.,Peng R.H.,Xiong A.S.,Xu F.,and Zhu H.,2005,Isolation and characterization of a cDNA encoding two novel heat-shock factor OsHSF6 and OsHSF12in Oryza Sativa L.,J.Biochem.Mol.Biol.,38(5):602-608
Nishizawa A.,Nosaka R.,Hayashi H.,Tainaka H.,Maruta T.Tamoi M.,Ikeda M.,Ohme-Takagi M.,Yoshimura K.Yabuta Y.,and Shigeoka S.,2011,Hsf A1d and Hsf A1e involved in the transcriptional regulation of Hsf A2 function as key regulators for the Hsf signaling network in response to environmental stress,Plant Cell Physiol.,52(5):933-945
Nishizawa A.,Yabuta Y.,Yoshida E.,Maruta T.,Yoshimura K.and Shigeoka S.,2006,Arabidopsis heat shock transcription factor A2 as a key regulator in response to several types of environmental stress,Plant J.,48(4):535-547
Nover L.,Bharti K.,D觟ring P.,Mishra S.K.,Ganguli A.,and Scharf K.D.,2001,Arabidopsis and the heat stress transcription factor world:how many heat stress transcription factors do we need?Cell Stress and Chaperones,6(3):177-189
Ogawa D.,Yamaguchi K.,and Nishiuchi T.,2007,High-leve overexpression of the Arabidopsis HsfA2 gene confers no only increased themotolerance but also salt/osmotic stress tolerance and enhanced callus growth,J.Exp.Bot.,58(12)3373-3383
Scharf K.D.,Berberich T.,Ebersberger I.,and Nover L.,2012The plant heat stress transcription factor(Hsf)family:structure,function and evolution,Biochim.Biophys.Acta Gene Regul.Mech.,1819(2):104-119
Schramm F.,Ganguli A.,Kiehlmann E.,Englich G.,Walch D.,and Von Koskull-D觟ring P.,2006,The heat stress transcription factor HsfA2 serves as a regulatory amplifier of a subset of genes in the heat stress response in Arabidopsis,Plant Mol.Biol.,60(5):759-772
Sheng Y.T.,Wang Y.L.,Meijer H.J.G.,Yang X.Y.,Hua C.,Ye W.W.,Tao K.,Liu X.Y.,Govers F.,and Wang Y.C.,2015,The heat shock transcription factor Ps HSF1 of Phytophthorasojae is required for oxidative stress tolerance and detoxifying the plant oxidative burst,Environ.Microbiol.,17(4):1351-1364
Verceles A.C.,Diaz-Abad M.,Geiger-Brown J.,and Scharf S.M.,2012,Testing the prognostic value of the rapid shallow breathing index in predicting successful weaning in patients requiring prolonged mechanical ventilation,Heart&Lung The Journal of Acute and Critical Care,41(6):546-552
Wang X.Y.,Huang W.L.,Liu J.,Yang Z.M.,and Huang B.R.,2017,Molecular regulation and physiological functions of a novel FaHsfA2c cloned from tall fescue conferring plant tolerance to heat stress,Plant Biotechnol.J.,15(2):237-248
Whiteside S.T.,and Goodbourn S.,1993,Signal transduction and nuclear targeting:regulation of transcription factor activity by subcellular localization,J.Cell Sci.,104(4):949-955
Xin H.,Zhang H.,Chen L.,Li X.X.,Lian Q.L.,Yuan X.,Hu X.Y.,Cao L.,He X.L.,and Yi M.F.,2010,Cloning and characterization of HsfA2 from Lily(Lilium longiflorum),Plant Cell Rep.,29(8):875-885
Xin H.B.,Zhang H.,Zhong X.H.,Lian Q.L.,Dong A.X.,Cao L.,Yi M.F.,and Cong R.C.,2017,Over-expression of LlHsfA2b,a lily heat shock transcription factor lacking trans-activation activity in yeast,can enhance tolerance to heat and oxidative stress in transgenic Arabidopsis seedlings,Plant Cell Tiss.Org.Cult.,130(3):617-629
Xue G.P.,Sadat S.,Drenth J.,and McIntyre C.L.,2014,The heat shock factor family from Triticum aestivum in response to heat and other major abiotic stresses and their role in regulation of heat shock protein genes,J.Exp.Bot.,65(2):539-557
Yasuda H.,Sagehashi Y.,Shimosaka E.,and Sato Y.,2013,Generation of transgenic rice expressing heat shock protein genes under cool conditions,Plant Biotechnol.,30(5):489-496
Zhang L.R.,Li Y.S.,Xing D.,and Gao C.J.,2009,Characterization of mitochondrial dynamics and subcellular localization of ROS reveal that HsfA2 alleviates oxidative damage caused by heat stress in Arabidopsis,J.Exp.Bot.,60(7):2073-2091
Banti V.,Mafessoni F.,Loreti E.,Alpi A.,and Perata P.,2010,The heat-inducible transcription factor HsfA2 enhances anox ia tolerance in Arabidopsis,Plant Physiol.,152(3):1471-1483
Chan-Schaminet K.Y.,Baniwal S.K.,Bublak D.,Nover L.,and Scharf K.D.,2009,Specific interaction between tomato HsfA1 and HsfA2 creates heter o-oligomeric superactivator complexes for synergistic activation of heat stress gene ex pression,J.Biol.Chem.,284(31):20848-20857
Charng Y.Y.,Liu H.C.,Liu N.Y.,He W.T.,Wang C.N.,Chang S.H.,and Wang T.T.,2007,A heat-inducible transcription factor,HsfA2,is required for extension of acquired thermotolerance in Arabidopsis,Plant Physiol.,143(1):251-262
Fragkostefanakis S.,R觟th S.,Schleiff E.,and Scharf K.D.,2015,Prospects of engineering thermotolerance in crops through modulation of heat stress transcription factor and heat shock protein networks,Plant Cell Environ.,38(9):1881-1895
Guo M.,Yin Y.X.,Ji J.J.,Ma B.P.,Lu M.H.,and Gong Z.H.,2014,Cloning and expression analysis of heat-shock transcription factor gene CaHsfA2 from pepper(Capsicum annuum L.),Genet.Mol.Res.,13(1):1865-1875
Guo M.,Lu J.P.,Zhai Y.F.,Chai W.G.,Gong Z.H.,and Lu M.H.2015,Genome-wide analysis,expression profile of hea shock factor gene family(CaHsfs)and characterisation of CaHsfA2 in pepper(Capsicum annuum L.),BMC Plant Biology,15(1):151
Kotak S.,Port M.,Ganguli A.,Bicker F.,and Von Koskull-D觟ring P.,2004,Characterization of C-terminal domains of Arabidopsis heat stress transcription factors(Hsfs)and identification of a new signature combination of plant class A Hsfs with AHA and NES motifs essential for activator function and intracellular localization,Plant J.,39(1):98-112
Liu A.L.,Zou J.,Zhang X.W.,Zhou X.Y.,Wang W.F.,Xiong XY.,Chen L.Y.,and Chen X.B.,2010,Expression profiles of class a rice heat shock transcription factor genes under abiotic stresses,J.Plant Biol.,53(2):142-149
Liu H.C.,and Charng Y.Y.,2013,Common and distinct functions of Arabidopsis class A1 and A2 heat shock factors in diverse abiotic stress responses and development,Plant Physiol.,163(1):276-290
Liu J.G.,Yao Q.H.,Zhang Z.,Peng R.H.,Xiong A.S.,Xu F.,and Zhu H.,2005,Isolation and characterization of a cDNA encoding two novel heat-shock factor OsHSF6 and OsHSF12in Oryza Sativa L.,J.Biochem.Mol.Biol.,38(5):602-608
Nishizawa A.,Nosaka R.,Hayashi H.,Tainaka H.,Maruta T.Tamoi M.,Ikeda M.,Ohme-Takagi M.,Yoshimura K.Yabuta Y.,and Shigeoka S.,2011,Hsf A1d and Hsf A1e involved in the transcriptional regulation of Hsf A2 function as key regulators for the Hsf signaling network in response to environmental stress,Plant Cell Physiol.,52(5):933-945
Nishizawa A.,Yabuta Y.,Yoshida E.,Maruta T.,Yoshimura K.and Shigeoka S.,2006,Arabidopsis heat shock transcription factor A2 as a key regulator in response to several types of environmental stress,Plant J.,48(4):535-547
Nover L.,Bharti K.,D觟ring P.,Mishra S.K.,Ganguli A.,and Scharf K.D.,2001,Arabidopsis and the heat stress transcription factor world:how many heat stress transcription factors do we need?Cell Stress and Chaperones,6(3):177-189
Ogawa D.,Yamaguchi K.,and Nishiuchi T.,2007,High-leve overexpression of the Arabidopsis HsfA2 gene confers no only increased themotolerance but also salt/osmotic stress tolerance and enhanced callus growth,J.Exp.Bot.,58(12)3373-3383
Scharf K.D.,Berberich T.,Ebersberger I.,and Nover L.,2012The plant heat stress transcription factor(Hsf)family:structure,function and evolution,Biochim.Biophys.Acta Gene Regul.Mech.,1819(2):104-119
Schramm F.,Ganguli A.,Kiehlmann E.,Englich G.,Walch D.,and Von Koskull-D觟ring P.,2006,The heat stress transcription factor HsfA2 serves as a regulatory amplifier of a subset of genes in the heat stress response in Arabidopsis,Plant Mol.Biol.,60(5):759-772
Sheng Y.T.,Wang Y.L.,Meijer H.J.G.,Yang X.Y.,Hua C.,Ye W.W.,Tao K.,Liu X.Y.,Govers F.,and Wang Y.C.,2015,The heat shock transcription factor Ps HSF1 of Phytophthorasojae is required for oxidative stress tolerance and detoxifying the plant oxidative burst,Environ.Microbiol.,17(4):1351-1364
Verceles A.C.,Diaz-Abad M.,Geiger-Brown J.,and Scharf S.M.,2012,Testing the prognostic value of the rapid shallow breathing index in predicting successful weaning in patients requiring prolonged mechanical ventilation,Heart&Lung The Journal of Acute and Critical Care,41(6):546-552
Wang X.Y.,Huang W.L.,Liu J.,Yang Z.M.,and Huang B.R.,2017,Molecular regulation and physiological functions of a novel FaHsfA2c cloned from tall fescue conferring plant tolerance to heat stress,Plant Biotechnol.J.,15(2):237-248
Whiteside S.T.,and Goodbourn S.,1993,Signal transduction and nuclear targeting:regulation of transcription factor activity by subcellular localization,J.Cell Sci.,104(4):949-955
Xin H.,Zhang H.,Chen L.,Li X.X.,Lian Q.L.,Yuan X.,Hu X.Y.,Cao L.,He X.L.,and Yi M.F.,2010,Cloning and characterization of HsfA2 from Lily(Lilium longiflorum),Plant Cell Rep.,29(8):875-885
Xin H.B.,Zhang H.,Zhong X.H.,Lian Q.L.,Dong A.X.,Cao L.,Yi M.F.,and Cong R.C.,2017,Over-expression of LlHsfA2b,a lily heat shock transcription factor lacking trans-activation activity in yeast,can enhance tolerance to heat and oxidative stress in transgenic Arabidopsis seedlings,Plant Cell Tiss.Org.Cult.,130(3):617-629
Xue G.P.,Sadat S.,Drenth J.,and McIntyre C.L.,2014,The heat shock factor family from Triticum aestivum in response to heat and other major abiotic stresses and their role in regulation of heat shock protein genes,J.Exp.Bot.,65(2):539-557
Yasuda H.,Sagehashi Y.,Shimosaka E.,and Sato Y.,2013,Generation of transgenic rice expressing heat shock protein genes under cool conditions,Plant Biotechnol.,30(5):489-496
Zhang L.R.,Li Y.S.,Xing D.,and Gao C.J.,2009,Characterization of mitochondrial dynamics and subcellular localization of ROS reveal that HsfA2 alleviates oxidative damage caused by heat stress in Arabidopsis,J.Exp.Bot.,60(7):2073-2091