苹果bZIP转录因子基因MdAREB2功能的初步研究
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摘要
对‘嘎拉’苹果中的1个bZIP转录因子基因MdAREB2(序列号:MDP0000248567)进行功能初步研究。蛋白进化树分析表明,苹果MdAREB2与拟南芥AtAREB2具有很高的同源性。利用PlantCare数据库进行启动子顺式作用元件的预测分析,MdAREB2启动子序列中存在脱落酸响应元件ABRE。实时定量PCR检测表明,MdAREB2明显受ABA诱导。拟南芥种子萌发阶段经过0.5和2μmol·L~(-1)ABA处理后发现突变体abi5中过量表达MdAREB2能够恢复对ABA的敏感性。拟南芥幼苗生长阶段,经过20μmol·L~(-1) ABA处理后发现,突变体abi5中过量表达MdAREB2能够部分恢复对ABA的敏感性。
A bZIP transcription factor gene MdAREB2(MDP0000248567)was cloned from‘Gala’apple(Malus × domestica Borkh.). The evolutionary tree analysis indicated that MdAREB2 had the very high homology with AtAREB2. The promoter of Md AREB2 was analyzed and predicted by Plant Care databases,and the results showed that the promoter sequence contained abscisic acid response element ABRE. Quantitative real-time PCR showed that MdAREB2 was significantly induced by ABA. This study found that with 0.5 μmol · L~(-1) ABA and 2 μmol · L~(-1) ABA treatment,excessive expression of MdAREB2 in mutant abi5 could restore its sensitivity to ABA in seed germination stage. In the seedling growth stage,with 20 μmol · L~(-1) ABA treatment,excessive expression of MdAREB2 in mutant abi5 could partially restore its sensitivity to ABA.
A bZIP transcription factor gene MdAREB2(MDP0000248567)was cloned from‘Gala’apple(Malus × domestica Borkh.). The evolutionary tree analysis indicated that MdAREB2 had the very high homology with AtAREB2. The promoter of Md AREB2 was analyzed and predicted by Plant Care databases,and the results showed that the promoter sequence contained abscisic acid response element ABRE. Quantitative real-time PCR showed that MdAREB2 was significantly induced by ABA. This study found that with 0.5 μmol · L~(-1) ABA and 2 μmol · L~(-1) ABA treatment,excessive expression of MdAREB2 in mutant abi5 could restore its sensitivity to ABA in seed germination stage. In the seedling growth stage,with 20 μmol · L~(-1) ABA treatment,excessive expression of MdAREB2 in mutant abi5 could partially restore its sensitivity to ABA.
引文
An J P,Wang X N,Yao J F,Ren Y R,You C X,Wang X F,Hao Y J.2017.Apple Md MYC2 reduces aluminum stress tolerance by directly regulating Md ERF3 gene.Plant and Soil,418:255-266.
Bensmihen S,Rippa S,Lambert G,Jublot D,Pautot V,Granier F,Parcy F.2002.The homologous ABI5 and EEL transcription factors function antagonistically to fine-tune gene expression during late embryogenesis.The Plant Cell,14(6):1391-1403.
Finkelstein R R,Lynch T J.2000.The Arabidopsis abscisic acid response gene ABI5 encodes a basic leucine zipper transcription factor.The Plant Cell,12(4):599-609.
Fujita Y,Fujita M,Satoh R,Maruyama K,Parvez M M,Seki M,Yamaguchi-Shinozaki K.2005.AREB1 is a transcription activator of novel ABRE-dependent ABA signaling that enhances drought stress tolerance in Arabidopsis.The Plant Cell,17(12):3470-3488.
Fujita Y,Yoshida T,Yamaguchi-Shinozaki K.2013.Pivotal role of the AREB/ABF-Sn RK2 pathway in ABRE-mediated transcription in response to osmotic stress in plants.Physiologia plantarum,147(1):15-27.
Hong Lan,Liu Xiu,Li Ling.2011.AREB/ABF transcription factors and their involvement in ABA signal transduction.Plant Physiology Journal,47(3):211-217.(in Chinese)洪岚,刘旭,李玲.2011.植物AREB/ABF转录因子及其参与的ABA信号转导.植物生理学报,47(3):211-217.
Hossain M A,Cho J I,Han M,Ahn C H,Jeon J S,An G,Park P B.2010.The ABRE-binding b ZIP transcription factor Os ABF2 is a positive regulator of abiotic stress and ABA signaling in rice.Journal of Plant Physiology,167(17):1512-1520.
Huang X S,Liu J H,Chen X J.2010.Overexpression of Ptr ABF gene,a b ZIP transcription factor isolated from Poncirus trifoliata,enhances dehydration and drought tolerance in tobacco via scavenging ROS and modulating expression of stress-responsive genes.BMC Plant Biology,10(1):230.
Jakoby M,Weisshaar B,Dr?ge-Laser W,Vicente-Carbajosa J,Tiedemann J,Kroj T,Parcy F.2002.b ZIP transcription factors in Arabidopsis.Trends in Plant Science,7(3):106-111.
Kang J Y,Choi H I,Im M Y,Kim S Y.2002.Arabidopsis basic leucine zipper proteins that mediate stress-responsive abscisic acid signaling.The Plant Cell,14(2):343-357.
Lee S C,Choi H W,Hwang I S,Choi D S,Hwang B K.2006.Functional roles of the pepper pathogen-induced b ZIP transcription factor,CAb ZIP1,in enhanced resistance to pathogen infection and environmental stresses.Planta,224(5):1209-1225.
Liu Ya-jing,Ma Qi-jun,Li Hao-hao,Lu Jing,Hao Yu-jin,You Chun-xiang.2017.Molecular cloning and functional characterization of Md CBL3in apple.Acta Horticulturae Sinica,44(1):1-10.(in Chinese)刘亚静,马齐军,李浩浩,路静,郝玉金,由春香.2017.苹果Md CBL3基因的克隆与功能鉴定.园艺学报,44(1):1-10.
Ma Yan-lin,Ma Jian-zhong,Wang Yong-gang.2014.Molecular cloning,prokaryotic expression and purification of abi5 gene from Arabidopsis thaliana L.Plant Physiology Journal,50(2):185-191.(in Chinese)马燕林,马建忠,王永刚.2014.拟南芥abi5基因的分子克隆及其在原核细胞中的表达和纯化.植物生理学报,50(2):185-191.
Nijhawan A,Jain M,Tyagi A K,Khurana J P.2008.Genomic survey and gene expression analysis of the basic leucine zipper transcription factor family in rice.Plant Physiology,146(2):333-350.
Orellana S,Ya?ez M,Espinoza A,Verdugo I,González E,Ruiz-Lara S,Casaretto J A.2010.The transcription factor Sl AREB1 confers drought,salt stress tolerance and regulates biotic and abiotic stress-related genes in tomato.Plant,Cell&Environment,33(12):2191-2208.
Pang Shao-ping,Dong Cui-cui,Ma Yan-yan,Qian Chun,He Shao-lan,Xie Rang-jin,Deng Lie.2017.The roles of two Cs AREB transcription factors in the development of citrus fruit.Acta Horticulturae Sinica,44(3):441-451.(in Chinese)庞少萍,董翠翠,马岩岩,钱春,何绍兰,谢让金,邓烈.2017.Cs AREB转录因子在柑橘果实发育中的作用.园艺学报,44(3):441-451.
Rodriguez-Uribe,Laura,Mary A,O'connell.2006.A root-specific b ZIP transcription factor is responsive to water deficit stress in tepary bean(Phaseolus acutifolius)and common bean(P.vulgaris).Journal Of Experimental Botany,57(6):1391-1398.
Satoh R,Fujita Y,Nakashima K,Shinozaki K,Yamaguchi-Shinozaki K.2004.A novel subgroup of b ZIP proteins functions as transcriptional activators in hypoosmolarity-responsive expression of the Pro DH gene in Arabidopsis.Plant and Cell Physiology,45(3):309-317.
Su Ling,Wang Shu-hui,Hao Yu-jin,Wang Xiao-fei,You Chun-xiang.2016.Molecular cloning and functional characterization of the ethylene signaling related gene Md EIL1 in apple.Acta Horticulturae Sinica,43(3):409-418.(in Chinese)苏玲,王淑惠,郝玉金,王小非,由春香.2016.苹果乙烯信号转导相关基因Md EIL1的克隆及功能鉴定.园艺学报,43(3):409-418.
Sun Ming-yue,Zhou Jun,Tan Qiu-ping,Fu Xi-ling,Chen Xiu-de,Li Ling,Gao Dong-sheng.2016.Analysis of basic leucine zipper genes and their expression during bud dormancy in apple(Malus×domestica).Scientia Agricultura Sinica,49(7):1325-1345.(in Chinese)孙明岳,周君,谭秋平,付喜玲,陈修德,李玲,高东升.2016.苹果b ZIP转录因子家族生物信息学分析及其在休眠芽中的表达.中国农业科学,49(7):1325-1345.
Yang X,Yang Y N,Xue L J,Zou M J,Liu J Y,Chen F,Xue H W.2011.Rice ABI5-Like1 regulates abscisic acid and auxin responses by affecting the expression of ABRE-containing genes.Plant Physiology,156(3):1397-1409.
Yoshida T,Fujita Y,Maruyama K,Mogami J,Todaka D,Shinozaki K,Yamaguchi Shinozaki,Kazuko.2015.Four Arabidopsis AREB/ABFtranscription factors function predominantly in gene expression downstream of Sn RK2 kinases in abscisic acid signalling in response to osmotic stress.Plant,Cell&Environment,38(1):35-49.
Yu Y,Wang J,Shi H,Gu J,Dong J,Deng X W,Huang R.2016.Salt stress and ethylene antagonistically regulate nucleocytoplasmic partitioning of COP1 to control seed germination.Plant Physiology,170(4):2340-2350.
Zhang Ji-yu,Qu Shen-chun,Gu Zhong-ren,Du Xiao-li,Du Bei-bei,Zhang Zhen.2011.Biology function of b ZIP transcription factors in plants.Acta Botanica Boreali-Occidentalia,31(5):1066-1075.(in Chinese)张计育,渠慎春,郭忠仁,杜小丽,都贝贝,章镇.2011.植物b ZIP转录因子的生物学功能.西北植物学报,31(5):1066-1075.
Zhang Y,Zhang G,Xia N,Wang X J,Huang L L,Kang Z S.2008.Cloning and characterization of a b ZIP transcription factor gene in wheat and its expression in response to stripe rust pathogen infection and abiotic stresses.Physiological and Molecular Plant Pathology,73(4):88-94.
Bensmihen S,Rippa S,Lambert G,Jublot D,Pautot V,Granier F,Parcy F.2002.The homologous ABI5 and EEL transcription factors function antagonistically to fine-tune gene expression during late embryogenesis.The Plant Cell,14(6):1391-1403.
Finkelstein R R,Lynch T J.2000.The Arabidopsis abscisic acid response gene ABI5 encodes a basic leucine zipper transcription factor.The Plant Cell,12(4):599-609.
Fujita Y,Fujita M,Satoh R,Maruyama K,Parvez M M,Seki M,Yamaguchi-Shinozaki K.2005.AREB1 is a transcription activator of novel ABRE-dependent ABA signaling that enhances drought stress tolerance in Arabidopsis.The Plant Cell,17(12):3470-3488.
Fujita Y,Yoshida T,Yamaguchi-Shinozaki K.2013.Pivotal role of the AREB/ABF-Sn RK2 pathway in ABRE-mediated transcription in response to osmotic stress in plants.Physiologia plantarum,147(1):15-27.
Hong Lan,Liu Xiu,Li Ling.2011.AREB/ABF transcription factors and their involvement in ABA signal transduction.Plant Physiology Journal,47(3):211-217.(in Chinese)洪岚,刘旭,李玲.2011.植物AREB/ABF转录因子及其参与的ABA信号转导.植物生理学报,47(3):211-217.
Hossain M A,Cho J I,Han M,Ahn C H,Jeon J S,An G,Park P B.2010.The ABRE-binding b ZIP transcription factor Os ABF2 is a positive regulator of abiotic stress and ABA signaling in rice.Journal of Plant Physiology,167(17):1512-1520.
Huang X S,Liu J H,Chen X J.2010.Overexpression of Ptr ABF gene,a b ZIP transcription factor isolated from Poncirus trifoliata,enhances dehydration and drought tolerance in tobacco via scavenging ROS and modulating expression of stress-responsive genes.BMC Plant Biology,10(1):230.
Jakoby M,Weisshaar B,Dr?ge-Laser W,Vicente-Carbajosa J,Tiedemann J,Kroj T,Parcy F.2002.b ZIP transcription factors in Arabidopsis.Trends in Plant Science,7(3):106-111.
Kang J Y,Choi H I,Im M Y,Kim S Y.2002.Arabidopsis basic leucine zipper proteins that mediate stress-responsive abscisic acid signaling.The Plant Cell,14(2):343-357.
Lee S C,Choi H W,Hwang I S,Choi D S,Hwang B K.2006.Functional roles of the pepper pathogen-induced b ZIP transcription factor,CAb ZIP1,in enhanced resistance to pathogen infection and environmental stresses.Planta,224(5):1209-1225.
Liu Ya-jing,Ma Qi-jun,Li Hao-hao,Lu Jing,Hao Yu-jin,You Chun-xiang.2017.Molecular cloning and functional characterization of Md CBL3in apple.Acta Horticulturae Sinica,44(1):1-10.(in Chinese)刘亚静,马齐军,李浩浩,路静,郝玉金,由春香.2017.苹果Md CBL3基因的克隆与功能鉴定.园艺学报,44(1):1-10.
Ma Yan-lin,Ma Jian-zhong,Wang Yong-gang.2014.Molecular cloning,prokaryotic expression and purification of abi5 gene from Arabidopsis thaliana L.Plant Physiology Journal,50(2):185-191.(in Chinese)马燕林,马建忠,王永刚.2014.拟南芥abi5基因的分子克隆及其在原核细胞中的表达和纯化.植物生理学报,50(2):185-191.
Nijhawan A,Jain M,Tyagi A K,Khurana J P.2008.Genomic survey and gene expression analysis of the basic leucine zipper transcription factor family in rice.Plant Physiology,146(2):333-350.
Orellana S,Ya?ez M,Espinoza A,Verdugo I,González E,Ruiz-Lara S,Casaretto J A.2010.The transcription factor Sl AREB1 confers drought,salt stress tolerance and regulates biotic and abiotic stress-related genes in tomato.Plant,Cell&Environment,33(12):2191-2208.
Pang Shao-ping,Dong Cui-cui,Ma Yan-yan,Qian Chun,He Shao-lan,Xie Rang-jin,Deng Lie.2017.The roles of two Cs AREB transcription factors in the development of citrus fruit.Acta Horticulturae Sinica,44(3):441-451.(in Chinese)庞少萍,董翠翠,马岩岩,钱春,何绍兰,谢让金,邓烈.2017.Cs AREB转录因子在柑橘果实发育中的作用.园艺学报,44(3):441-451.
Rodriguez-Uribe,Laura,Mary A,O'connell.2006.A root-specific b ZIP transcription factor is responsive to water deficit stress in tepary bean(Phaseolus acutifolius)and common bean(P.vulgaris).Journal Of Experimental Botany,57(6):1391-1398.
Satoh R,Fujita Y,Nakashima K,Shinozaki K,Yamaguchi-Shinozaki K.2004.A novel subgroup of b ZIP proteins functions as transcriptional activators in hypoosmolarity-responsive expression of the Pro DH gene in Arabidopsis.Plant and Cell Physiology,45(3):309-317.
Su Ling,Wang Shu-hui,Hao Yu-jin,Wang Xiao-fei,You Chun-xiang.2016.Molecular cloning and functional characterization of the ethylene signaling related gene Md EIL1 in apple.Acta Horticulturae Sinica,43(3):409-418.(in Chinese)苏玲,王淑惠,郝玉金,王小非,由春香.2016.苹果乙烯信号转导相关基因Md EIL1的克隆及功能鉴定.园艺学报,43(3):409-418.
Sun Ming-yue,Zhou Jun,Tan Qiu-ping,Fu Xi-ling,Chen Xiu-de,Li Ling,Gao Dong-sheng.2016.Analysis of basic leucine zipper genes and their expression during bud dormancy in apple(Malus×domestica).Scientia Agricultura Sinica,49(7):1325-1345.(in Chinese)孙明岳,周君,谭秋平,付喜玲,陈修德,李玲,高东升.2016.苹果b ZIP转录因子家族生物信息学分析及其在休眠芽中的表达.中国农业科学,49(7):1325-1345.
Yang X,Yang Y N,Xue L J,Zou M J,Liu J Y,Chen F,Xue H W.2011.Rice ABI5-Like1 regulates abscisic acid and auxin responses by affecting the expression of ABRE-containing genes.Plant Physiology,156(3):1397-1409.
Yoshida T,Fujita Y,Maruyama K,Mogami J,Todaka D,Shinozaki K,Yamaguchi Shinozaki,Kazuko.2015.Four Arabidopsis AREB/ABFtranscription factors function predominantly in gene expression downstream of Sn RK2 kinases in abscisic acid signalling in response to osmotic stress.Plant,Cell&Environment,38(1):35-49.
Yu Y,Wang J,Shi H,Gu J,Dong J,Deng X W,Huang R.2016.Salt stress and ethylene antagonistically regulate nucleocytoplasmic partitioning of COP1 to control seed germination.Plant Physiology,170(4):2340-2350.
Zhang Ji-yu,Qu Shen-chun,Gu Zhong-ren,Du Xiao-li,Du Bei-bei,Zhang Zhen.2011.Biology function of b ZIP transcription factors in plants.Acta Botanica Boreali-Occidentalia,31(5):1066-1075.(in Chinese)张计育,渠慎春,郭忠仁,杜小丽,都贝贝,章镇.2011.植物b ZIP转录因子的生物学功能.西北植物学报,31(5):1066-1075.
Zhang Y,Zhang G,Xia N,Wang X J,Huang L L,Kang Z S.2008.Cloning and characterization of a b ZIP transcription factor gene in wheat and its expression in response to stripe rust pathogen infection and abiotic stresses.Physiological and Molecular Plant Pathology,73(4):88-94.