白桦2个bZIP基因的克隆及表达分析
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摘要
在本研究中,以白桦总cDNA为模板,扩增出2个bZIP基因并成功构建到T载体上;通过对白桦的根、茎、叶进行不同时间的盐胁迫处理,利用实时荧光定量PCR技术分析2个bZIP基因在胁迫下的表达情况,结果表明:2个bZIP基因在不同组织以及不同时间盐胁迫下表达量呈现不同的变化趋势,说明这2个bZIP基因能够被逆境胁迫胁迫诱导表达。本实验为后续研究bZIP基因在白桦中的功能提供前期数据参考,并为进一步的白桦遗传改良工作提供了帮助。
In this study, first of all, the total cDNA from Betula platyphylla was used as template for PCR, two bZIP genes was amplified, and successfully was built into pMD-18 T carrier, respectively. The roots, stems and leaves of B. platyphylla were treated at different time under salts. The expressions of two bZIP genes were analyzed at different time under stress by real-time fluorescence quantitative PCR. The results showed that the expression of two bZIP genes were different changes tends in the different tissues at different time, illustrating that the expressions of two bZIP genes can be induced by stress. This study provides early data reference for the function of bZIP gene in B. platyphylla, and provides help for further study on the genetic modification of B. platyphylla.
In this study, first of all, the total cDNA from Betula platyphylla was used as template for PCR, two bZIP genes was amplified, and successfully was built into pMD-18 T carrier, respectively. The roots, stems and leaves of B. platyphylla were treated at different time under salts. The expressions of two bZIP genes were analyzed at different time under stress by real-time fluorescence quantitative PCR. The results showed that the expression of two bZIP genes were different changes tends in the different tissues at different time, illustrating that the expressions of two bZIP genes can be induced by stress. This study provides early data reference for the function of bZIP gene in B. platyphylla, and provides help for further study on the genetic modification of B. platyphylla.
引文
Dou L.L.,Li G.L.,Pang C.Y.,Wei H.L.,Fan S.L.,Song M.Z.,and Yu S.X.,2016,Cloning and function analysis of Gh WRKY11 in cotton(Gossypium hirsutum),Nongye Shengwu Jishu Xuebao(Journal of Agricultural Biotechnology),24(5):625-636(窦玲玲,李光雷,庞朝友,魏恒玲,范术丽,宋美珍,喻树迅2016,棉花转录因子Gh WRKY11的克隆及功能分析,农业生物技术学报,24(5):625-636)
Fu X.Y.,2007,Cloning and functional analysis of AP2/EREBPtranscription factor in Malus robusta R.and Oryza sativa College of Forestry,Nanjing Agricultural University,Supervisor:Yao Q.H.,and Zhang Z.,pp.4-5(付晓燕,2007,八棱海棠与水稻AP2/EREBP类转录因子的克隆及功能分析,硕士学位论文,南京农业大学,导师:姚泉洪,章镇,pp.4-5)
Fujita Y.,Fujita M.,Satoh R.,Maruyama K.,Parvez M.M.,Seki M.Hiratsu K.,Ohme-Takagi M.,Shinozaki K.,and YamaguchiShinozaki K.,2005,AREB1 is a transcription activator of novel ABRE-dependent ABA signaling that enhances drough stress tolerance in Arabidopsis,Plant Cell,17(12):3470-3488
Gao S.Q.,Chen M.,Xu Z.S.,Zhao C.P.,Li L.,Xu H.J.,Tang YM.,Zhao X.,and Ma Y.Z.,2011,The soybean Gmb ZIP1transcription factor enhances multiple abiotic stress tolerances in transgenic plants,Plant Mol.Biol.,75(6):537-553
Guan L.L.,Cui Q.,Han Y.L.,Gu T.Y.,Wu Y.Y.,Hu R.G.,Du L.N.Li H.Y.,and Li X.K.,2016,Cloning and expression analysis of Ctb ZIP20 transcription factor gene from Carthamus tinctorius and construction of plant expression vector,Zhongcaoyao(Chinese Traditional and Herbal Drugs),47(8):1369-1374(官丽莉,崔琪,韩怡来,顾天瑶,武云云,胡人阁,杜林娜,李海燕,李校堃,2016,红花b ZIP20转录因子基因的克隆、表达分析及植物表达载体构建,中草药,47(8):1369-1374)
Huang X.S.,Liu J.H.,and Chen X.J.,2010,Overexpression of Ptr ABF gene,a b ZIP transcription factor isolated from Poncirus trifoliata,enhances dehydration anddrought tolerance in tobacco via scavenging ROS and modulating expression of stress-responsive genes,BMC Plant Biol.,10(1):230
Jakoby M.,Weisshaar B.,Droge-Laser W.,Vicente-Carbajosa J.,Tiedemann J.,Kroj T.,and Parcy F.,2002,b ZIP transcription factors in Arabidopsis,Trends Plant Sci.,7(3):106-111
Kang J.Y.,Choi H.I.,Lm M.Y.,and Kim S.Y.,2002,Arabidopsis basic leucine zipper proteins that mediate stress-responsive abscisic acid signaling,Plant Cell,14(2):343-357
Kosugi S.,and Ohashi Y.,2002,DNA binding and dimerization specificity and potential targets for the TCP protein family,Plant J.,30(3):337-348
Li D.Y.,Yuan W.X.,Zheng C.,Wang X.M.,Zhou J.,Yan C.Q.,and Chen J.P.,2017,Roles of b ZIP transcription factors in phytohormone-mediated disease resistance and stress tolerance,Zhejiang Nongye Xuebao(Acta Agriculture Zhejiangensi),29(1):168-175(李冬月,原文霞,郑超,王栩鸣,周洁,严成其,陈剑平,2017,b ZIP转录因子在植物激素介导的抗病抗逆途径中的作用,浙江农业学报,29(1):168-175)
Li Y.Y.,2013,Tab ZIP transferred white Poplus simonii×P.nigra generation and stress resistance analysis,Thesis for M.S.,College of Forestry,Northeast Forestry University,Supervisor:Jiang J.,pp.34-35(李园园,2013,转Tab ZIP基因小黑杨的获得及抗逆性分析,硕士学位论文,东北林业大学林学院,导师:姜静,pp.34-35)
Nijhawan A.,Jain M.,Tyagi A.K.,and Khurana J.P.,2008,Genomic survey and gene expression analysis of the basic leucine zipper transcription factor family in rice,Plant Physiol.,146(2):333-350
Song P.,2012,Cloning and expression analysis of wheat stressresponsive transcription fcator Ta SNAC1,Thesis for M.S.,College of Life Science,Northwest Agriculture Forestry University,Supervisor:Fan S.H.,pp.25-26(宋鹏,2012,小麦胁迫响应转录因子基因Ta SNAC1的克隆与表达分析,硕士学位论文,西北农林科技大学生命科学学院,导师:范三红,pp.25-26)
Sun Z.W.,Huang X.Q.,Li W.J.,Zhong Q.F.,Yu T.Q.,Guo Y.Q.,Li D.Q.,Yin F.Y.,and Cheng Z.Q.,2011,Molecular marker technology and its application in the gene mapping of rice,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),30(1):78-86(孙正文,黄兴奇,李维蛟,钟巧芳,余腾琼,郭怡卿,李定琴,殷富有,程在全,2011,分子标记技术及其在水稻基因定位上的应用,基因组学与应用生物学,30(1):78-86)
Wang J.C.,and Chen X.B.,2011,Advances in transcription factors related to plant abiotic stress response,Hunan Nongye Kexue(Hunan Agricultural Sciences),(17):9-12(王景晨,陈信波2011,植物非生物胁迫应答相关转录因子研究进展,湖南农业科学,(17):9-12)
Wang J.Z.,Zhou J.X.,Zhang B.L.,Vanitha J.,Ramachandran S.and Jiang S.Y.,2011,Genomewide expansion and expression divergence of the basic leucine zipper transcription factors in higher plants with an emphasis on sorghum,J.Integrative Plant Biol.,53(3):212-231
Wang Y.,Gao C.,Liang Y.,Wang C.,Yang C.,and Liu C.,2010,Anovel b ZIP gene from Tamarix hispida mediates physiological responses to salt stress in tobacco plants,J.Plant Physiol.,167(3):222-230
Xiang Y.,Tang N I.,Du H.,Ye H.Y.,and Xiong L.Z.,2008,Characterization of Osb ZIP23 as a key player of the basic leucine zipper transcription factor family for conferring abscisic acid sensitivity and salinity and drought tolerance in Rice,Plant Physiol.,148(4):1938-1952
Fu X.Y.,2007,Cloning and functional analysis of AP2/EREBPtranscription factor in Malus robusta R.and Oryza sativa College of Forestry,Nanjing Agricultural University,Supervisor:Yao Q.H.,and Zhang Z.,pp.4-5(付晓燕,2007,八棱海棠与水稻AP2/EREBP类转录因子的克隆及功能分析,硕士学位论文,南京农业大学,导师:姚泉洪,章镇,pp.4-5)
Fujita Y.,Fujita M.,Satoh R.,Maruyama K.,Parvez M.M.,Seki M.Hiratsu K.,Ohme-Takagi M.,Shinozaki K.,and YamaguchiShinozaki K.,2005,AREB1 is a transcription activator of novel ABRE-dependent ABA signaling that enhances drough stress tolerance in Arabidopsis,Plant Cell,17(12):3470-3488
Gao S.Q.,Chen M.,Xu Z.S.,Zhao C.P.,Li L.,Xu H.J.,Tang YM.,Zhao X.,and Ma Y.Z.,2011,The soybean Gmb ZIP1transcription factor enhances multiple abiotic stress tolerances in transgenic plants,Plant Mol.Biol.,75(6):537-553
Guan L.L.,Cui Q.,Han Y.L.,Gu T.Y.,Wu Y.Y.,Hu R.G.,Du L.N.Li H.Y.,and Li X.K.,2016,Cloning and expression analysis of Ctb ZIP20 transcription factor gene from Carthamus tinctorius and construction of plant expression vector,Zhongcaoyao(Chinese Traditional and Herbal Drugs),47(8):1369-1374(官丽莉,崔琪,韩怡来,顾天瑶,武云云,胡人阁,杜林娜,李海燕,李校堃,2016,红花b ZIP20转录因子基因的克隆、表达分析及植物表达载体构建,中草药,47(8):1369-1374)
Huang X.S.,Liu J.H.,and Chen X.J.,2010,Overexpression of Ptr ABF gene,a b ZIP transcription factor isolated from Poncirus trifoliata,enhances dehydration anddrought tolerance in tobacco via scavenging ROS and modulating expression of stress-responsive genes,BMC Plant Biol.,10(1):230
Jakoby M.,Weisshaar B.,Droge-Laser W.,Vicente-Carbajosa J.,Tiedemann J.,Kroj T.,and Parcy F.,2002,b ZIP transcription factors in Arabidopsis,Trends Plant Sci.,7(3):106-111
Kang J.Y.,Choi H.I.,Lm M.Y.,and Kim S.Y.,2002,Arabidopsis basic leucine zipper proteins that mediate stress-responsive abscisic acid signaling,Plant Cell,14(2):343-357
Kosugi S.,and Ohashi Y.,2002,DNA binding and dimerization specificity and potential targets for the TCP protein family,Plant J.,30(3):337-348
Li D.Y.,Yuan W.X.,Zheng C.,Wang X.M.,Zhou J.,Yan C.Q.,and Chen J.P.,2017,Roles of b ZIP transcription factors in phytohormone-mediated disease resistance and stress tolerance,Zhejiang Nongye Xuebao(Acta Agriculture Zhejiangensi),29(1):168-175(李冬月,原文霞,郑超,王栩鸣,周洁,严成其,陈剑平,2017,b ZIP转录因子在植物激素介导的抗病抗逆途径中的作用,浙江农业学报,29(1):168-175)
Li Y.Y.,2013,Tab ZIP transferred white Poplus simonii×P.nigra generation and stress resistance analysis,Thesis for M.S.,College of Forestry,Northeast Forestry University,Supervisor:Jiang J.,pp.34-35(李园园,2013,转Tab ZIP基因小黑杨的获得及抗逆性分析,硕士学位论文,东北林业大学林学院,导师:姜静,pp.34-35)
Nijhawan A.,Jain M.,Tyagi A.K.,and Khurana J.P.,2008,Genomic survey and gene expression analysis of the basic leucine zipper transcription factor family in rice,Plant Physiol.,146(2):333-350
Song P.,2012,Cloning and expression analysis of wheat stressresponsive transcription fcator Ta SNAC1,Thesis for M.S.,College of Life Science,Northwest Agriculture Forestry University,Supervisor:Fan S.H.,pp.25-26(宋鹏,2012,小麦胁迫响应转录因子基因Ta SNAC1的克隆与表达分析,硕士学位论文,西北农林科技大学生命科学学院,导师:范三红,pp.25-26)
Sun Z.W.,Huang X.Q.,Li W.J.,Zhong Q.F.,Yu T.Q.,Guo Y.Q.,Li D.Q.,Yin F.Y.,and Cheng Z.Q.,2011,Molecular marker technology and its application in the gene mapping of rice,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),30(1):78-86(孙正文,黄兴奇,李维蛟,钟巧芳,余腾琼,郭怡卿,李定琴,殷富有,程在全,2011,分子标记技术及其在水稻基因定位上的应用,基因组学与应用生物学,30(1):78-86)
Wang J.C.,and Chen X.B.,2011,Advances in transcription factors related to plant abiotic stress response,Hunan Nongye Kexue(Hunan Agricultural Sciences),(17):9-12(王景晨,陈信波2011,植物非生物胁迫应答相关转录因子研究进展,湖南农业科学,(17):9-12)
Wang J.Z.,Zhou J.X.,Zhang B.L.,Vanitha J.,Ramachandran S.and Jiang S.Y.,2011,Genomewide expansion and expression divergence of the basic leucine zipper transcription factors in higher plants with an emphasis on sorghum,J.Integrative Plant Biol.,53(3):212-231
Wang Y.,Gao C.,Liang Y.,Wang C.,Yang C.,and Liu C.,2010,Anovel b ZIP gene from Tamarix hispida mediates physiological responses to salt stress in tobacco plants,J.Plant Physiol.,167(3):222-230
Xiang Y.,Tang N I.,Du H.,Ye H.Y.,and Xiong L.Z.,2008,Characterization of Osb ZIP23 as a key player of the basic leucine zipper transcription factor family for conferring abscisic acid sensitivity and salinity and drought tolerance in Rice,Plant Physiol.,148(4):1938-1952