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蒙古冰草MwDREB3基因的克隆及表达分析
摘要
以蒙古冰草(Agropyron mongolicum)为研究材料,利用同源序列法克隆得到1个DREB3类转录因子基因,命名为MwDREB3,采用实时荧光定量RT-PCR技术分析MwDREB3基因的表达特征,为蒙古冰草优异抗性基因的挖掘和利用提供理论依据。结果表明:该基因全长1056bp,包含1个完整的开放阅读框,长度为774bp,编码257个氨基酸,该蛋白近5'端含有1个保守的AP2结构域。系统进化树分析表明MwDREB3编码的蛋白与小麦(Triticum aestivum)的DREB、山羊草(Aegilops columnaris)DRF1的进化关系较近。该基因在干旱、高盐、ABA诱导条件下,表达量上调;在低温条件下,表达量下调。
To explore and exploit the excellent resistance gene resources of Mongolian wheatgrass(Agropyron Mongolicum Keng),A DREB3 transcription factor gene was cloned by homologous sequences from the Mongolian wheatgrass,named MwDREB3(1056bp),and the open reading frame was 774 bp,encoding 257 amino acids,containing a conserved AP2 domain nearby the 5'end.Phylogenetic analysis revealed that MwDREB3 had a closer relationship with DREBs from wheat(Triticum aestivum) and Aegilops(Aegilops columnaris).Expression patterns of MwDREB3 gene were analyzed using real-time quantitative RT-PCR,the MwDREB3 expression was up under the conditions of drought,high salt and ABA induction,and was down under the low temperatures.
引文
[1]Yamaguchi-Shinozaki K,Kazuo Shinozaki A.Novel cis-acting element in an Arabidopsis gene is involved in responsiveness to drought,low temperature or high-salt stress[J].Plant Cell,1994.6C2):251-264[2]Stockinger E J,Gilmour S J.Thomashow M F.Arabidopsis Thaliana CBF1 encodes an AP2 domain-containing transcription Activator that binds to the C-repeat/DRE,acisacting DNA regulatory element that stimulates transcription in response to low temperature and water deficit[J].Proceedings of the National Academy of Sciences USA,1997,94(3):1035-1040[3]Gilmour S J,Zarka D G,Stockinger E J,et al.Low temperature regulation of the Arabidopsis CBF family of AP2 transcriptional activators as an Early step in cold-induced COR gene expression[J].Plant Journal,1998,16(4):433-443[4]Shinozaki K,Dennis E S.Cell signaling and gene regulation global analyses of signal transduction and gene expression profiles[J].Current Opinion in Plant Biology,2003,6(5):405-409[5]Matsukura S,Mizoi J,Yoshida D,et al.Comprehensive analysis of rice DREB2-type genes that encode transcription factors involved in the expression of abiotic stress-responsive genes[J].Molecular Genetics and Genomics,2010,283(2):185-196[6]Chen M,Xu Z,Xia L,et al.Cold-induced modulation and functional analyses of the DRE-binding transcription factorgene,GmDREB3,in soybean(Glycine max L.)[J].Journal of Experimental Botany,2009,60(1):121-135[7]Shen Y G,Zhang W K,He S J,et al.An EREBP/AP2-type protein in Triticum aestivum was a DRE-binding transcription factor induced by cold,dehydration and ABA stress[J].TAG Theoretical and Applied Genetics,2003,106(5):923-930[8]Egawa C,Kobayashi F,Ishibashi M,etal.Differential regulation of transcript accumulation and alternative splicing of a DREB2 homo-log under abiotic stress conditions in common wheat[J].Genes and Genetic Systems(Japan),2006,81(2):77-91[9]Oh S J,Kwon C W,Choi D W,et al.Expression of barley HvCBF4 enhances tolerance to abiotic stress in transgenic rice[J].Plant Bio-technology Journal,2007,5(5):646-656[10]Shan D P,Huang J G,Yang Y T,et al.Cotton GhDREB1 increases plant tolerance to low temperature and is negatively regulated by gibberellic acid[J].New Phytologist,2007,176(1):70-81[11]Qin F,Sakuma Y,Li J,etal.Cloning and functional analysis of a novelDREBl/CBF transcription factor involved in cold-responsive gene expression in Zea mays L.[J].Plant and Cell Physiology,2004,45(8):1042-1052[12]Islam M S,Wang M H.Expression of dehydration responsive element-binding protein-3(DREB3)under differentabiotic stresses in tomato[J].Biochemistry and Molecular Biology Reports,2009,42(9):611-616[13]Chen J H,Xia X L,Yin W L.Expression profiling and functional characterization ofaDREB2-type gene from Populus euphratica[J],Biochemical and Biophysical Research Communications,2009,378(3):483-487[14]Benedict C,Skinner J S,Meng R,et al.The CBFl-dependent low temperature signalling pathway,regulon and increase in freeze tolerance are conserved in Populuss pp[J].Plant Cell and Environment,2006,29(7):1259-1272[15]郭本兆.中国植物志[M].北京:科技出版社,1987,9(3):7-119[16]李立会,董玉琛.冰草属研究进展[J].遗传,1993,15(1):45-48[17]张众,师文贵,逯晓萍.蒙古冰草研究概况[C].中国草学会第六届二次会议暨国际学术研讨会,呼和浩特:2004,7[18]郑殿升.中国农业生物多样性保护与可持续利用现状调研报告[R].北京:气象出版社,2000[19]郭丹丽,王崎崎,吴晓庆,等.新疆无苞芥锌指蛋白基因的克隆及表达分析[J].西北植物学报,2014,34(2):243-250[20]石凤敏.蒙古冰革MwLEA3基因功能研究及MwRRT基因的克隆[D].呼和浩特:内蒙古农业大学,2011[21]云锦凤,赵彦,石凤敏,等.蒙古冰草肌动蛋白基因片段的克隆与组织表达分析[J].草业学报,2011(2):170-176[22]赵彦.蒙古冰草抗旱相关基因克隆、表达及RNAi载体构建[D].呼和浩特:内蒙古农业大学,2010[23]Dubouzet J G,Sakuma Y,Ito Y?et al.encode transcription acti-vators that function in drought-,high-salt-and cold-responsivegene expression[J].Plant Journal,2003,33(4):751-763[24]Meissner R C,Jin H.Cominelli E,et al.Function search in a large transcription factor gene family in Arabidopsis:assessing the potential of reverse genetics to identify insertional mutations in R2R3 MYB genes[J].Plant Cell,1999,11(10),1827-1840[25]Kranz H D,Denekamp M,Greco R,et al.Towards functional characterisation of the members of the R2R3-MYB gene family from Arabidoposis thaliana[J].Plant Journal,1998,16(2):263-276[26]Maleck K,Levine A,Eulgem T,et al.The transcriptome of Arabidopsis thaliana during systemic acquired resistance[J].Nat Genet,2000,26(4):403-410[27]Abe H,Urao T,Ito T,etal.Arabidopsis AtMYC2(bHLH)and AtMYB2(MYB)function as transcriptional activators in abscisic acid signaling[J].Plant Cell,2003,15(1):63-78[28]Shinozaki K,Yamaguchi-Shinozaki K,Seki M.Regulatory network of gene expression in the drought and cold stress responses[J].Current Opinion in Plant Biology,2003,6(5):410-417[29]Yamaguchi-Shinozaki K,Shinozaki K.Organization of cis-acting regulatory elements in osmotic-and cold-stress-responsive promoters[J].Trends in Plant Science,2005,10(2):88-94
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