旱麦草EtAP2基因的克隆及其对干旱胁迫的响应表达
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摘要
以旱麦草(Eremopyrum triticeum)为实验材料,利用RT-PCR技术从旱麦草叶片中克隆了1个AP2/ERF家族基因,命名为EtAP2(GenBank登录号KX622583)。EtAP2基因含有1 128bp开放阅读框,编码375个氨基酸,相对分子质量40.87kD,等电点为5.36。多序列比对和进化树分析表明,该基因编码蛋白具有2个AP2保守结构域,与小麦AP2/ERF家族蛋白具有较近的亲缘关系。实时荧光定量PCR分析表明,15%PEG 6000模拟干旱胁迫可诱导EtAP2基因在根和叶中表达,且在根中对干旱胁迫的响应大于叶片。研究表明,EtAP2可能参与旱麦草对干旱逆境胁迫应答的调节。
A AP2/ERF gene was isolated fromEremopyrum triticeum by RT-PCR method,and designated as EtAP2(GenBank accession no.KX622583).EtAP2 contained an open reading frame(ORF)of 1 128 bp encoding 375 amino acids.The molecular mass of EtAP2 was 40.87 kDa with isoelectric point(PI)of5.36.Multiple sequences alignment and phylogenetic analysis indicated that EtAP2 protein contained two conserved AP2 domains,which had close genetic relationship with AP2/ERF transcription factor in Triticum aestivum.Quantitative real-time PCR analysis revealed that simulated drought stress(15% PEG6000)treatment significantly induced the expression of EtAP2,and root is more sensitive to drought stress than leaf.The results indicated than EtAP2 might involve in response to drought stress regulation in E.triticeum.
A AP2/ERF gene was isolated fromEremopyrum triticeum by RT-PCR method,and designated as EtAP2(GenBank accession no.KX622583).EtAP2 contained an open reading frame(ORF)of 1 128 bp encoding 375 amino acids.The molecular mass of EtAP2 was 40.87 kDa with isoelectric point(PI)of5.36.Multiple sequences alignment and phylogenetic analysis indicated that EtAP2 protein contained two conserved AP2 domains,which had close genetic relationship with AP2/ERF transcription factor in Triticum aestivum.Quantitative real-time PCR analysis revealed that simulated drought stress(15% PEG6000)treatment significantly induced the expression of EtAP2,and root is more sensitive to drought stress than leaf.The results indicated than EtAP2 might involve in response to drought stress regulation in E.triticeum.
引文
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[3]SAKUMA Y,LIU Q,DUBOUZET J G,et al.DNA-binding specificity of the ERF/AP2domain of Arabidopsis DREBs,transcription factors involved in dehydration-and cold-inducible gene expression[J].Biochemical and Biophysical Research Communications,2002,290:998-1 009.
[4]LIU S,WANG X,WANG H,et al.Genome-wide analysis of ZmDREBgenes and their association with natural variation in drought tolerance at seedling stage of Zea mays L[J].Plos Genetics,2013,9(9):119-129.
[5]OHTO M A,FLOYD S K,FISCHER R L,et al.Effects of APETALA2on embryo,endosperm,and seed coat development determine seed size in Arabidopsis[J].Sexual Plant Reproduction,2009,22:277-289.
[6]ZHU X L,QI L,LIU X,et al.The wheat ethylene response factor transcription factor pathogen-induced ERF1 mediates host responses to both the necrotrophic pathogen Rhizoctonia cerealis and freezing stresses[J].Plant Physiology,2014,164(3):1 499-1 514.
[7]NAKANO T,SUZUKI K,FUJIMURA T,et al.Genomewide analysis of the ERF gene family in Arabidopsis and rice[J].Plant Physiology,2006,140:411-432.
[8]DU H,HUANG M,ZHANG Z,et al.Genome-wide analysis of the AP2/ERF gene family in maize waterlogging stress response[J].Euphytica,2014,198(1):115-126.
[9]HIROSHI M,SHINJIRO Y,ATSUSHI H,et al.Dwarf and delayed-flowering 1,a novel Arabidopsis mutant deficient in gibberellin biosynthesis because of overexpression of a putative AP2transcription factor[J].The Plant Journal,2004,37(5):720-729.
[10]TAMURA K,DUDLEY J,NEI M,et al.MEGA4:molecular evolutionary genetics analysis(MEGA)software version4.0[J].Molecular Biology Evolution,2007,24(8):1596-1 599.
[11]张计育,王庆菊,郭忠仁.植物AP2/ERF类转录因子研究进展[J].遗传,2012,34(7):835-847.ZHANG J Y,WANG Q J,GUO Z R.Progresses on plant AP2/ERF transcription factors[J].Hereditas,2012,34(7):835-847.
[12]RIECHMANN J L,MEYEROWITZ E M.The AP2/EREBP family of plant transcription factors[J].Journal of Biological Chemistry,1998,379(6):633-646.
[13]NOLE-WILSON S,KRIZEK B A.DNA binding properties of the Arabidopsis floral development protein AINTEGUMENTA[J].Nucleic Acids Research,2000,28(21):4 076-4 082.
[14]HUIJUAN Z,YONGBO H,LEI H,et al.Arabidopsis AtERF014acts as a dual regulator that differentially modulates immunity against Pseudomonas syringae pv.tomato and Botrytis cinerea[J].Scientific Reports,2016,6:30 251.
[15]GOLLDACK D,LI C,MOHAN H,PROBST N.Tolerance to drought and salt stress in plants:unraveling the signaling networks[J].Frontiers in Plant Science,2014,5:151.
[16]DOSSA K,WEI X,LI D,et al.Insight into the AP2/ERF transcription factor superfamily in sesame and expression profiling of DREB subfamily under drought stress[J].BMC Plant Biology,2016,16(1):171.
[17]ZHANG G Y,CHEN M,LI L C,et al.Overexpression of the soybean GmERF3gene,an AP2/ERF type transcription factor for increased tolerances to salt,drought,and diseases in transgenic tobacco[J].Journal of Experimental Botany,2009,60(13):3 781-3 796.
[18]ZENG L,YIN Y,YOU C,et al.Evolution and protein interactions of AP2proteins in Brassicaceae:evidence linking development and environmental responses[J].Journal of Integrative Plant Biology,2016,58(6):549-563.