苎麻镉响应转录因子BnMYB1的克隆和表达分析
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摘要
V-MYB禽成髓细胞性白血病病毒癌基因同源物(v-myb avian myeloblastosis viral oncogene homolog,MYB)类转录因子在植物生长发育以及应答逆境胁迫中发挥重要作用。本研究在苎麻(Boehmeria nivea)镉胁迫表达谱的基础上,采用RACE(rapid-amplification of cDNA ends)方法从苎麻品种中苎一号中获得了BnMYB1转录因子的全长cDNA序列,该基因(Gen Bank No.:MF741318)包含一个1 029 bp的开放读码框,编码342个氨基酸。BnMYB1蛋白包含一个保守的SANT(switching-defective protein 3(Swi3),adaptor 2(Ada2),nuclear receptor co-repressor(N-Co R),transcription factor(TFIIB))家族结构域,为典型的1R型MYB转录因子,与啤酒花(Humulus lupulus,CBY88798.1)、甜橙(Citrus sinensis,XP_006477144.1)、绿豆(Vigna radiata,XP_014509497.1)、梅(Armeniaca mume,XP_008238436.1)、鹰嘴豆(Cicer arietinum,XP_004508939.1)SANT/MYB蛋白的相似性分别为86%、77%、79%、80%、76%。系统进化树分析表明,BnMYB1蛋白与啤酒花HlMYB4亲缘关系最近。qRT-PCR分析表明,BnMYB1为组成型表达基因,但在叶中的表达量显著高于根(P<0.05),且该基因受镉诱导,表达量随时间和镉处理浓度的增加而发生变化。这些结果表明BnMYB1是一个镉应答因子,并且可能在植物对镉胁迫的适应中发挥重要作用,为苎麻耐镉分子机制研究提供理论依据。
MYB(v-myb avian myeloblastosis viral oncogene homolog) transcription factor plays an important role in plant development and their response to various environmental stresses. In this paper, a MYB gene named BnMYB1 was isolated from ramie(Boehmeria nivea) by rapid amplification of cDNA ends(RACE) based on the analysis of expression profiling of cadmium response genes in ramie. The full length of BnMYB1(Gen Bank No.: MF741318) coding sequence was 1 029 bp, which encoded 342 amino acid.BnMYB1 protein belonged to 1 RMYB subfamily which had only one conserved SANT(switching-defective protein 3(Swi3), adaptor 2(Ada2), nuclear receptor co-repressor(N-Co R), transcription factor(TFIIB))domain. The similarity comparison revealed that BnMYB1 shared 86%, 77%, 79%, 80% and 76% ofsimilarity with SANT/MYB proteins of Humulus lupulus(CBY88798.1), citrus sinensis(XP_006477144.1),Vigna radiate(XP_014509497.1), Prunus mume(XP_008238436.1) and Cicer arietinum(XP_004508939.1).Phylogenetic analysis revealed that BnMYB1 was closely to Hl MYB4 of Humulus lupulus. q RT-PCR analysis indicated that BnMYB1 was a constitutive gene, but the expression of BnMYB1 in leaf was significantly higher than that of in root(P<0.05). Furthermore, BnMYB1 gene was up-regulated by Cd stress, and the expression of BnMYB1 changed along with the stress time and Cd concentration. Collectively, our data suggest that BnMYB1 is a cadmium-responsive factor and may play potential roles in the plant adaption to cadmium stress.
MYB(v-myb avian myeloblastosis viral oncogene homolog) transcription factor plays an important role in plant development and their response to various environmental stresses. In this paper, a MYB gene named BnMYB1 was isolated from ramie(Boehmeria nivea) by rapid amplification of cDNA ends(RACE) based on the analysis of expression profiling of cadmium response genes in ramie. The full length of BnMYB1(Gen Bank No.: MF741318) coding sequence was 1 029 bp, which encoded 342 amino acid.BnMYB1 protein belonged to 1 RMYB subfamily which had only one conserved SANT(switching-defective protein 3(Swi3), adaptor 2(Ada2), nuclear receptor co-repressor(N-Co R), transcription factor(TFIIB))domain. The similarity comparison revealed that BnMYB1 shared 86%, 77%, 79%, 80% and 76% ofsimilarity with SANT/MYB proteins of Humulus lupulus(CBY88798.1), citrus sinensis(XP_006477144.1),Vigna radiate(XP_014509497.1), Prunus mume(XP_008238436.1) and Cicer arietinum(XP_004508939.1).Phylogenetic analysis revealed that BnMYB1 was closely to Hl MYB4 of Humulus lupulus. q RT-PCR analysis indicated that BnMYB1 was a constitutive gene, but the expression of BnMYB1 in leaf was significantly higher than that of in root(P<0.05). Furthermore, BnMYB1 gene was up-regulated by Cd stress, and the expression of BnMYB1 changed along with the stress time and Cd concentration. Collectively, our data suggest that BnMYB1 is a cadmium-responsive factor and may play potential roles in the plant adaption to cadmium stress.
引文
黄益宗,郝晓伟,雷鸣,等.2013.重金属污染土壤修复技术及其修复实践[J].农业环境科学学报,32(3):409-417.(Huang Y Z,Hao X W,Lei M,et al.2013.The remediation technology and remediation practice of heavy metals-contaminated soil[J].Journal of Agro-Environment Science,32(3):409-417.)
雷梅,岳庆玲,陈同斌,等.2005.湖南柿竹园矿区土壤重金属含量及植物吸收特征[J].生态学报,25(5):1146-1151.(Lei M,Yue Q L,Chen T B,et al.2005.Heavy metal concentrations in soils and plants around Shizhuyuan mining area of Hunan province[J].Acta Ecologica Sinica,25(5):1146-1151.)
李宗道.1980.麻作的理论与技术[M].上海:上海科学技术出版社,pp.124-256.(Li Z D.1980.Theory and Technology of Bast Fiber Crops[M].Shanghai:Shanghai Scientific and Technical Publishers,pp.124-256.)
王芸,张建辉,赵晓军.2007.污灌农田土壤镉污染状况及分布特征研究[J].中国环境监测,23(5):71-74.(Wang Y,Zhang J H,Zhao X J.2007.The study of the pollution situation and distribution characteristics of cadmium in farmland soil irrigated by waste water[J].Environmental Monitoring in China,23(5):71-74.)
佘玮,揭雨成,邢虎成,等.2010.湖南冷水江锑矿区苎麻对重金属的吸收和富集特性[J].农业环境科学学报,29(1):91-96.(She W,Jie Y C,Xing H C,et al.2010.Uptake and accumulation of heavy metal by ramie(Boehmeria nivea)growing on antimony mining area in Lengshuijiang City of Hunan Province[J].Journal of Agro-Environment Science,29(1):91-96.)
袁长春,莫健新,袁柳娇,等.2017.重金属胁迫下白骨壤数字基因表达谱分析[J].林业科学研究,30(2):206-213.(Yuan C C,Mo J X,Yuan L J,et al.2017.Digital gene expression profiling analysis of Avicennia marina under heavy metal stress treatment[J].Forest Research,30(2):206-213.)
袁柳娇,傅梦妮,余如凤,等.2017.秋茄(Kandelia candel L.)响应重金属胁迫的数字基因表达谱[J].东北林业大学学报,45(02):22-28.(Yuan L J,Fu M N,Yu R F,et al.2017.Digital gene expression profiling analysis of Kandelia candel L.in response to artificial heavy metal stress[J].Journal of Northeast Forestry University,45(2):22-28.)
朱守晶,余伟林,石朝燕,等.2015.苎麻谷胱甘肽还原酶基因(Bn GR1)的克隆和表达分析[J].农业生物技术学报,23(10):1318-1326.(Zhu S J,Yu W L,Shi Z Y,et al.2015.Cloning and expression analysis of glutathione reductase gene(Bn GR1)from ramie(Boehmeria nivea L.)[J].Journal of Agricultural Biotechnology,23(10):1318-1326.
Ambawat S,Sharma P,Yadav N R,et al.2013.MYB transcription factor genes as regulators for plant responses:An overview[J].Plant Molecular Biology,19(3):307-321.
Baldoni E,Genga A,Cominelli E.2015.Plant MYB transcription factors:Their role in drought response mechanisms[J].International Journal of Molecular Sciences,16(7):15811-15851.
Chaney R L,Reeves P G,Ryan J A,et al.2004.An improved understanding of soil Cd risk to humans and low cost methods to phytoextract Cd from contaminated soils to prevent soil Cd risks[J].Biometals,17(5):549-553.
Christensen T H.1987.Cadmium soil sorption at low concentrations:V.Evidence of competition by other heavy metals[J].Water Air&Soil Pollution,34(3):293-303.
Cominelli E,Galbiati M,Vavasseur A,et al.2005.A guard-cell-specific MYB transcription factor regulates stomatal movements and plant drought tolerance[J].Current Biology,15(13):1196-1200.
Dai X Y,Xu Y Y,Ma Q B,et al.2007.Overexpression of an R1R2R3-MYB gene,Os MYB3R-2,increases tolerance to freezing,drought and salt stress in transgenic Arabidopsis[J].Plant Physiology,143(4):1739-1751.
Dubos C,Stracke R,Grotewold E,et al.2010.MYB transcription factors in Arabidopsis[J].Trends in Plant Science,15(10):573-581.
Feller A,Machemer K,Braun E L,et al.2011.Evolutionary and comparative analysis of MYB and b HLH plant transcription factors[J].Plant Journal,66(1):94-116.
Fujita Y,Fujita M,Shinozaki K,et al.2011.ABA-mediated transcriptional regulation in response to osmotic stress in plants[J].Journal of Plant Research,124(4):509-25.
Lin C Y,Trinh N N,FU S F,et al.2013.Comparison of early transcriptome responses to copper and cadmium in rice roots[J].Plant Molecular Biology,81(4-5):507-522.
Lipsick J S.1996.One billion years of Myb[J].Oncogene,13(2):223-235.
Liu T M,Zhu S,Tang Q,et al.2015 Genome-wide transcriptomic profiling of ramie(Boehmeria nivea L.Gaud)in response to cadmium stress[J].Gene,558(1):131-137.
Matsui K,Umemura Y,Ohme M.2008.At MYBL2,a protein with a single MYB domain,acts as a negative regulator of anthocyanin biosynthesis in Arabidopsis[J].The Plant Journal,55(6):954-967.
Paz-Ares J,Ghosal D,Wienand U,et al.1987.The regulatory C1 locus of Zea mays encodes a protein with homology to MYB-related proto-oncogene products and with structural similarities to transcriptional activators[J].Embo Journal,6(12):3553-3558.
Riechmann J L,Heard J,Martin G,et al.2000.Arabidopsis transcription factors:Genome-wide comparative analysis among Eukaryotes[J].Science,290(5499):2105-2110.
Rubio V,Linhares F,Solano R,et al.2001.A conserved MYBtranscription factor involved in phosphate starvation signaling both in vascular plants and in unicellular algae[J].Genes&Development,15(16):2122-2128.
SanitàD,Toppi L,Gabbrielli R.1999.Response to cadmium in higher plants[J].Environmental and Experimental Botany,41(2):105-130.
Shah K,Nongkynrih J M.2007.Metal hyperaccumulation and bioremediation[J].Biologia Plantarum,51(4):618-634.
She W,Zhu S J,Jie Y C,et al.2015.Expression profiling of cadmium response genes in ramie(Boehmeria nivea L.)root[J].Bulletin of Environmental Contamination and Toxicology,94(4):453-459
Su C F,Wang Y C,Hsieh T H,et al.2010.A novel MYBS3-dependent pathway confers cold tolerance in rice[J].Plant Physiology,153(1):145-158.
Vannini C,Locatelli F,Bracale M,et al.2004.Overexpression of the rice Osmyb4 gene increases chilling and freezing tolerance of Arabidopsis thaliana plants[J].The Plant Journal,37(1):115-127.
Villalobos M A,Bartels D,Iturriaga G.2004.Stress tolerante and glucose insensitive phenotypes in Arabidopsis overexpressing the Cp MYBIO transcription factor gene[J].Plant Physiology,135(1):309-324.
Wang Z Y,Tobin E M.1998.Constitutive expression of the CIRCADIAN CLOCK ASSOCIATED 1(CCA1)gene disrupts circadian rhythms and suppresses its own expression[J].Cell,93:1207-1217.
Weber M,Trampczynska A,Clemens S.2006.Comparative transcriptome analysis of toxic metal responses in Arabidopsis thaliana and the Cd2+-hypertolerant facultative metallophyte Arabidopsis halleri[J].Plant Cell and Environment,29(5):950-963.
Weng X,Wang L,Wang J,et al.2014.Grain number,plant height,and heading date7 is a central regulator of growth,development,and stress response[J].Plant Physiology,164(2):735-747.
雷梅,岳庆玲,陈同斌,等.2005.湖南柿竹园矿区土壤重金属含量及植物吸收特征[J].生态学报,25(5):1146-1151.(Lei M,Yue Q L,Chen T B,et al.2005.Heavy metal concentrations in soils and plants around Shizhuyuan mining area of Hunan province[J].Acta Ecologica Sinica,25(5):1146-1151.)
李宗道.1980.麻作的理论与技术[M].上海:上海科学技术出版社,pp.124-256.(Li Z D.1980.Theory and Technology of Bast Fiber Crops[M].Shanghai:Shanghai Scientific and Technical Publishers,pp.124-256.)
王芸,张建辉,赵晓军.2007.污灌农田土壤镉污染状况及分布特征研究[J].中国环境监测,23(5):71-74.(Wang Y,Zhang J H,Zhao X J.2007.The study of the pollution situation and distribution characteristics of cadmium in farmland soil irrigated by waste water[J].Environmental Monitoring in China,23(5):71-74.)
佘玮,揭雨成,邢虎成,等.2010.湖南冷水江锑矿区苎麻对重金属的吸收和富集特性[J].农业环境科学学报,29(1):91-96.(She W,Jie Y C,Xing H C,et al.2010.Uptake and accumulation of heavy metal by ramie(Boehmeria nivea)growing on antimony mining area in Lengshuijiang City of Hunan Province[J].Journal of Agro-Environment Science,29(1):91-96.)
袁长春,莫健新,袁柳娇,等.2017.重金属胁迫下白骨壤数字基因表达谱分析[J].林业科学研究,30(2):206-213.(Yuan C C,Mo J X,Yuan L J,et al.2017.Digital gene expression profiling analysis of Avicennia marina under heavy metal stress treatment[J].Forest Research,30(2):206-213.)
袁柳娇,傅梦妮,余如凤,等.2017.秋茄(Kandelia candel L.)响应重金属胁迫的数字基因表达谱[J].东北林业大学学报,45(02):22-28.(Yuan L J,Fu M N,Yu R F,et al.2017.Digital gene expression profiling analysis of Kandelia candel L.in response to artificial heavy metal stress[J].Journal of Northeast Forestry University,45(2):22-28.)
朱守晶,余伟林,石朝燕,等.2015.苎麻谷胱甘肽还原酶基因(Bn GR1)的克隆和表达分析[J].农业生物技术学报,23(10):1318-1326.(Zhu S J,Yu W L,Shi Z Y,et al.2015.Cloning and expression analysis of glutathione reductase gene(Bn GR1)from ramie(Boehmeria nivea L.)[J].Journal of Agricultural Biotechnology,23(10):1318-1326.
Ambawat S,Sharma P,Yadav N R,et al.2013.MYB transcription factor genes as regulators for plant responses:An overview[J].Plant Molecular Biology,19(3):307-321.
Baldoni E,Genga A,Cominelli E.2015.Plant MYB transcription factors:Their role in drought response mechanisms[J].International Journal of Molecular Sciences,16(7):15811-15851.
Chaney R L,Reeves P G,Ryan J A,et al.2004.An improved understanding of soil Cd risk to humans and low cost methods to phytoextract Cd from contaminated soils to prevent soil Cd risks[J].Biometals,17(5):549-553.
Christensen T H.1987.Cadmium soil sorption at low concentrations:V.Evidence of competition by other heavy metals[J].Water Air&Soil Pollution,34(3):293-303.
Cominelli E,Galbiati M,Vavasseur A,et al.2005.A guard-cell-specific MYB transcription factor regulates stomatal movements and plant drought tolerance[J].Current Biology,15(13):1196-1200.
Dai X Y,Xu Y Y,Ma Q B,et al.2007.Overexpression of an R1R2R3-MYB gene,Os MYB3R-2,increases tolerance to freezing,drought and salt stress in transgenic Arabidopsis[J].Plant Physiology,143(4):1739-1751.
Dubos C,Stracke R,Grotewold E,et al.2010.MYB transcription factors in Arabidopsis[J].Trends in Plant Science,15(10):573-581.
Feller A,Machemer K,Braun E L,et al.2011.Evolutionary and comparative analysis of MYB and b HLH plant transcription factors[J].Plant Journal,66(1):94-116.
Fujita Y,Fujita M,Shinozaki K,et al.2011.ABA-mediated transcriptional regulation in response to osmotic stress in plants[J].Journal of Plant Research,124(4):509-25.
Lin C Y,Trinh N N,FU S F,et al.2013.Comparison of early transcriptome responses to copper and cadmium in rice roots[J].Plant Molecular Biology,81(4-5):507-522.
Lipsick J S.1996.One billion years of Myb[J].Oncogene,13(2):223-235.
Liu T M,Zhu S,Tang Q,et al.2015 Genome-wide transcriptomic profiling of ramie(Boehmeria nivea L.Gaud)in response to cadmium stress[J].Gene,558(1):131-137.
Matsui K,Umemura Y,Ohme M.2008.At MYBL2,a protein with a single MYB domain,acts as a negative regulator of anthocyanin biosynthesis in Arabidopsis[J].The Plant Journal,55(6):954-967.
Paz-Ares J,Ghosal D,Wienand U,et al.1987.The regulatory C1 locus of Zea mays encodes a protein with homology to MYB-related proto-oncogene products and with structural similarities to transcriptional activators[J].Embo Journal,6(12):3553-3558.
Riechmann J L,Heard J,Martin G,et al.2000.Arabidopsis transcription factors:Genome-wide comparative analysis among Eukaryotes[J].Science,290(5499):2105-2110.
Rubio V,Linhares F,Solano R,et al.2001.A conserved MYBtranscription factor involved in phosphate starvation signaling both in vascular plants and in unicellular algae[J].Genes&Development,15(16):2122-2128.
SanitàD,Toppi L,Gabbrielli R.1999.Response to cadmium in higher plants[J].Environmental and Experimental Botany,41(2):105-130.
Shah K,Nongkynrih J M.2007.Metal hyperaccumulation and bioremediation[J].Biologia Plantarum,51(4):618-634.
She W,Zhu S J,Jie Y C,et al.2015.Expression profiling of cadmium response genes in ramie(Boehmeria nivea L.)root[J].Bulletin of Environmental Contamination and Toxicology,94(4):453-459
Su C F,Wang Y C,Hsieh T H,et al.2010.A novel MYBS3-dependent pathway confers cold tolerance in rice[J].Plant Physiology,153(1):145-158.
Vannini C,Locatelli F,Bracale M,et al.2004.Overexpression of the rice Osmyb4 gene increases chilling and freezing tolerance of Arabidopsis thaliana plants[J].The Plant Journal,37(1):115-127.
Villalobos M A,Bartels D,Iturriaga G.2004.Stress tolerante and glucose insensitive phenotypes in Arabidopsis overexpressing the Cp MYBIO transcription factor gene[J].Plant Physiology,135(1):309-324.
Wang Z Y,Tobin E M.1998.Constitutive expression of the CIRCADIAN CLOCK ASSOCIATED 1(CCA1)gene disrupts circadian rhythms and suppresses its own expression[J].Cell,93:1207-1217.
Weber M,Trampczynska A,Clemens S.2006.Comparative transcriptome analysis of toxic metal responses in Arabidopsis thaliana and the Cd2+-hypertolerant facultative metallophyte Arabidopsis halleri[J].Plant Cell and Environment,29(5):950-963.
Weng X,Wang L,Wang J,et al.2014.Grain number,plant height,and heading date7 is a central regulator of growth,development,and stress response[J].Plant Physiology,164(2):735-747.