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作物学报 ›› 2016, Vol. 42 ›› Issue (12): 1787-1797.doi: 10.3724/SP.J.1006.2016.01787

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

谷子转录因子SiNF-YA5通过ABA非依赖途径提高转基因拟南芥耐盐性

黄锁1,**,胡利芹1,**,徐东北1,2,李微微1,3,徐兆师1,李连城1,周永斌1,2,刁现民1,贾冠清1,马有志1,陈明1,*   

  1. 1中国农业科学院作物科学研究所/农作物基因资源与基因改良国家重大科学工程/农业部麦类生物学与遗传育种重点实验室,北京100081;2西北农林科技大学农学院/旱区作物逆境生物学国家重点实验室,陕西杨凌712100;3哈尔滨师范大学生命科学与技术学院/黑龙江省分子细胞遗传与遗传育种重点实验室,黑龙江哈尔滨150025
  • 收稿日期:2016-03-06 修回日期:2016-06-20 出版日期:2016-12-12 网络出版日期:2016-07-04
  • 通讯作者: 陈明,E-mail:chenming02@caas.cn,Tel:13683360891
  • 基金资助:

    本研究由国家转基因新品种生物培育科技重大专项(2016ZX08002-002)和中国农业科学院创新工程资助。

Transcription Factor SiNF-YA5 from Foxtail Millet (Setaria italica) Conferred Tolerance to High-salt Stress through ABA-independent Pathway in Transgenic Arabidopsis

HUANGSuo1,**,HULi-Qin1,**,XUDong-Bei1,2,LIWei-Wei1,3,XUZhao-Shi1,LILian-Cheng1,ZHOUYong-Bin1,2,DIAOXian-Min1,JIAGuan-Qing1,MAYou-Zhi1,CHENMing1,*   

  1. 1 Institute of Crop Science, Chinese Academy of Agricultural Sciences / National Key Facility For Crop Gene Resource and Genetic Improvement / Key Laboratory of Biology and Genetic Improvement of Triticeae Crop, Ministry of Agriculture, Beijing 100081, China; 2 College of Agronomy, Northwest A＆F University / State Key Laboratory of Arid Region Crop Adversity Biology, Yangling 712100, China; 3 College of Life Science and Technology, Harbin Normal University / Key Laboratory of Molecular Cytogenetics and Genetic Breeding of Heilongjiang Province, Harbin 150025, China
  • Received:2016-03-06 Revised:2016-06-20 Published:2016-12-12 Published online:2016-07-04
  • Contact: Chan Ming,E-mail:chenming02@caas.cn,Tel:13683360891
  • Supported by:

    ThisworkwasfundedbytheNationalKeyProjectforResearchonTransgenicBiology(2014ZX08002-002)

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摘要：

核转录因子Y (nuclear transcription factor Y，NF-Y)类转录因子在植物生长发育和非生物胁迫响应基因表达调控中发挥重要的作用，NF-Y由3种亚基(NF-YA、NF-YB、NF-YC)组成。本研究从抗逆性强的谷子品种龙谷25中克隆1个新的NF-Y类转录因子基因SiNF-YA5。该基因序列为924 bp，编码307个氨基酸，分子量为33.76 kD，等电点为9.19。SiNF-YA5在149~210位氨基酸之间含有CBF保守结构域。亚细胞定位分析表明，SiNF-YA5定位于细胞膜和细胞核。基因功能分析显示，在不同浓度高盐处理下，和野生型拟南芥(WT)相比SiNF-YA5转基因拟南芥种子萌发率更高；苗期SiNF-YA5转基因拟南芥根表面积和植株鲜重显著高于WT，证明过表达SiNF-YA5基因可以显著提高植物耐盐性。基因表达分析结果显示，在SiNF-YA5转基因拟南芥中参与盐胁迫响应的基因NHX1和LEA7的表达量明显高于WT。另一方面，SiNF-YA5转基因拟南芥与WT相比对于ABA的敏感性差异不显著，以上结果证明SiNF-YA5主要通过ABA非依赖途径提高转基因植物对高盐胁迫的耐性。

关键词: 谷子, NF-Y类转录因子, 高盐胁迫, ABA非依赖途径

Abstract:

Nuclear transcription factor Y (NF-Y) consisting of three subunits, NF-YA, NF-YB, and NF-YC, plays an essential role in many biologic processes, including growth, development, and abiotic stress response. In this study, a NF-Y like transcription factor gene SiNF-YA5 was isolated from foxtail millet variety Longgu 25. The full-length sequence of SiNF-YA5 gene is 924 bp, encoding 307 amino acids. Molecular weight and isoelectric point of SiNF-YA5 protein are 33.76 kD and 9.19, respectively. There is a conserved CBF domain from the 149th to the 210th amino acids of SiNF-YA5. According to the subcellular localization analysis, SiNF-YA5 was mainly localized and expressed on the plasma membrane and nucleus in plant cell. Gene functional analysis showed that under different NaCl concentration treatments, the germination rate of SiNF-YA5 transgenic Arabidopsis was significantly higher than that of wild-type (WT) Arabidopsis during seed germination stage; root surface area and fresh weight of SiNF-YA5 transgenic Arabidopsis remarkably increased compared with WT during seedling stage. Those results indicated that the overexpression of SiNF-YA5 in transgenic plants could enhance tolerance to high salt. Gene expression analysis showed that the expressions of two salt stress related genes, namely NHX1 and LEA7, increased significantly in SiNF-YA5 transgenic plants. On the other hand, there was no obvious difference in sensitivity to ABA between SiNF-YA5 transgenic Arabidopsis and WT showed during seed germination and seedling stages indicating that SiNF-YA5 could enhance salt tolerance through ABA-independent pathway in transgenic plants.

Key words: Foxtail millet (Setaria italic), NF-Y like transcription factor, High salt stress, ABA independent signaling pathway

[1]BoyerJS.Plantproductivityandenvironment.Science,1982,218:443–448 [2]XiongLM,SchumakerKS,ZhuJK.Cellsignalingduringcold,drought,andsaltstress.PlantCell,2002,14:S165–S183 [3]ZhuJK.Saltanddroughtstresssignaltransductioninplants.AnnuRevPlantBiol,2002,53:247–273 [4]TheArabidopsisGenomeInitiative.AnalysisofthegenomesequenceofthefloweringplantArabidopsisthaliana.Nature,2000,408:796–815 [5]RiechmannJL,RatcliffeOJ.Agenomicperspectiveonplanttranscriptionfactors.CurrOpinPlantBiol,2000,3:423–434 [6]KasugaM,LiuQ,MiuraS,Yamaguchi-ShinozakiK,ShinozakiK.Improvingplantdrought,saltandfreezingtolerancebygenetransferofasinglestress-inducibletranscriptionfactor.NatBiotechnol,1999,17:287–291 [7]Yamaguchi-ShinozakiK,ShinozakiK.Transcriptionalregulatorynetworksincellularresponsesandtolerancetodehydrationandcoldstresses.AnnuRevPlantBiol,2006,57:781–803 [8]JiangC,IuB,SinghJ.RequirementofaCCGACcis-actingelementforcoldinductionoftheBN115genefromwinterBrassicanapus.PlantMolBiol,1996,30:679–684 [9]UnoY,FurihataT,AbeH,YoshidaR,ShinozakiK,Yamaguchi-ShinozakiK.Arabidopsisbasicleucinezippertranscriptionfactorsinvolvedinanabscisicacid-dependentsignaltransductionpathwayunderdroughtandhigh-salinityconditions.ProcNatlAcadSciUSA,2000,97:11632–11637 [10]AbeH,UraoT,ItoT,SekiM,ShinozakiK,Yamaguchi-ShinozakiK.ArabidopsisAtMYC2(bHLH)andAtMYB2(MYB)functionastranscriptionalactivatorsinabscisicacidsignaling.PlantCell,2003,15:63–78 [11]MantovaniR,LiXY,PessaraU,HooftvanHuisjduijnenR,BenoistC,MathisD.DominantnegativeanalogsofNF-YA.JBiologChem,1994,32:20340–20346 [12]FrontiniM,ImbrianoC,ManniI,MantovaniR.CellcycleregulationofNF-YCnuclearlocalization.CellCycle,2004,3:217–222 [13]KahleJ,BaakeM,DoeneckeD,AlbigW.SubunitsoftheheterotrimerictranscriptionfactorNF-Yareimportedintothenucleusbydistinctpathwaysinvolvingimportinbetaandimportin13.MolCellBiol,2005,25:5339–5354 [14]SteidlS,TuncherA,GodaH,GuderC,PapadopoulouN,KobayashiT,TsukagoshiN,KatoM,BrakhageA.AsinglesubunitofaheterotrimericCCAAT-bindingcomplexcarriesanuclearlocalizationsignal:piggybacktransportofthepre-assembledcomplextothenucleus.JMolBiol,2004,342:515–524 [15]Ceribelli,M,Dolfini,D,MericoD,GattaR,ViganoAM.,PavesiG,MantovaniR.Thehistone-likeNF-Yisabifunctionaltranscriptionfactor.MolCellBiol,2008,28:2047–2058 [16]MaitySN,deCrombruggheB.RoleoftheCCAAT-bindingproteinCBF/NF-Yintranscription.TrendsBiochemSci,1998,23:174–178 [17]SiefersN,DangKK,KumimotoRW,BynumWE,TayroseG,HoltBF.Tissue-specificexpressionpatternsofArabidopsisNF-Ytranscriptionfactorssuggestpotentialforextensivecombinatorialcomplexity.PlantPhysiol,2009,149:625–641 [18]ThirumuruganT,ItoY,KuboT,SerizawaA,KurataN.Identification,characterizationandinteractionofHAPfamilygenesinrice.MolGenetGenomics,2008,279:279–289 [19]NelsonDE,RepettiPP,AdamsTR,CreelmanRA,WuJ,WarnerDC,AnstromDC,BensenRJ,CastiglioniPP,DonnarummoMG,HincheyBS,KumimotoRW,MaszleDR,CanalesRD,KrolikowskiKA,DotsonSB,GuttersonN,RatcliffeOJ,HeardJE.PlantnuclearfactorY(NF-Y)Bsubunitsconferdroughttoleranceandleadtoimprovedcornyieldsonwater-limitedacres.ProcNatlAcadSciUSA,2007,104:16450–16455 [20]Ni,ZY,HuZ,JiangQY,ZhangH.GmNFYA3,atargetgeneofmiR169,isapositiveregulatorofplanttolerancetodroughtstress.PlantMolBiol,2013,82:113–129 [21]QuB,HeX,WangJ,ZhaoY,TengW,ShaoA,ZhaoX,MaW,LiB,LiZ,TongY.AwheatCCAATbox-bindingtranscriptionfactorincreasesthegrainyieldofwheatwithlessfertilizerinput.PlantPhysiol,2014;167:411–423 [22]StephensonTJ,McIntyreCL,ColletC,XueGP.Genome-wideidentificationandexpressionanalysisoftheNF-YfamilyoftranscriptionfactorsinTriticumaestivum.PlantMolBiol,2007,65:77–92 [23]SunXC,LingS,LuZH,OuyangYD,LiuSS,YaoJ.OsNF-YB1,ariceendosperm-specificgene,isessentialforcellproliferationinendospermdevelopment.Gene,2014,551:214–221 [24]MiyoshiK,ItoY,SerizawaA,KurataN.OsHAP3genesregulatechloroplastbiogenesisinrice.PlantJ,2003,36:532–540 [25]MuJ,TanH,HongS,LiangY,ZuoJ.ArabidopsistranscriptionfactorgenesNF-YA1,5,6,and9playredundantrolesinmalegametogenesis,embryogenesis,andseeddevelopment.MolPlant,2013,6:188–201 [26]AlamMM,TanakaT,NakamuraH,IchikawaH,KobayashiK,YaenoT,YamaokaN,ShimomotoK,TakayamaK,NishinaH,NishiguchiM.Overexpressionofaricehemeactivatorproteingene(OsHAP2E)confersresistancetopathogens,salinityanddrought,andincreasesphotosynthesisandtillernumber.PlantBiotechnolJ,2015,13:85–96 [27]StephensonTJ.,McIntyreCL,ColletC,XueGP.TaNF-YB3isinvolvedintheregulationofphotosynthesisgenesinTriticumaestivum.FunctIntegrGenomics,2011,11:327–340 [28]CombierJP,FrugierF,deBillyF,BoualemA,El-YahyaouiF,MoreauS,VernieT,OttT,GamasP,CrespiM,NiebelA.MtHAP2-1isakeytranscriptionalregulatorofsymbioticnoduledevelopmentregulatedbymicroRNA169inMedicagotruncatula.GenesDev,2006,20:3084–3088 [29]ZanettiME,BlancoFA,BekerMP,BattagliaM,AguilarOM.ACsubunitoftheplantnuclearfactorNF-Yrequiredforrhizobialinfectionandnoduledevelopmentaffectspartnerselectioninthecommonbean-Rhizobiumetlisymbiosis.PlantCell,2010,22:4142–4157 [30]HackenbergD,KeetmanU,GrimmB.HomologousNF-YC2subunitfromArabidopsisandtobaccoisactivatedbyphotooxidativestressandinducesflowering.IntJMolSci,2012,13:3458–3477 [31]Leyva-GonzalezMA,Ibarra-LacletteE,Cruz-RamirezA,HerreraEstrellaL.FunctionalandtranscriptomeanalysisrevealsanacclimatizationstrategyforabioticstresstolerancemediatedbyArabidopsisNF-YAfamilymembers.2012,PLoSOne,7:e48138 [32]LiWX,OonoY,ZhuJ,HeXJ,WuJM,IidaK,LuXY,CuiXP,JinHL,ZhuJK.TheArabidopsisNFYA5transcriptionfactorisregulatedtranscriptionallyandposttranscriptionallytopromotedroughtresistance.PlantCell,2008,20:2238–2251 [33]WarpehaKM,UpadhyayS,YehJ,AdamiakJ,HawkinsSI,LapikYR,AndersonMB,KaufmanLS.TheGCR1,GPA1,PRN1,NFYsignalchainmediatesbothbluelightandabscisicacidresponsesinArabidopsis.PlantPhysiol,2007,143:1590–1600 [34]刘敬科,刁现民.我国谷子产业现状与加工发展方向.农业工程技术:农产品加工业,2013,(12):15–17 LiuJK,DiaoXM.Foxtailmilletprocessingindustrystatusanddevelopmenttrendinourcountry.AgricEngTechnol(AgricProdProcess),2013,(12):15–17(inChinese) [35]YooSD,ChoYH,SheenJ.Arabidopsismesophyllprotoplasts:aversatilecellsystemfortransientgeneexpressionanalysis.NatProtoc,2007,2:1565–1572 [36]BeehtoldN,EllisJ,PelletierG.InplantAgrobacteriummediatedgenetransferbyinfiltrationofadultArabidopsisthalianaplants.LifeSci,1993,316:1194–1199 [37]FengZJ,HeGH,ZhengWJ,LuPP,ChenM,GongYM,MaYZ,XuZS.FoxtailmilletNF-Yfamilies:genome-widesurveyandevolutionanalysesidentifiedtwofunctionalgenesimportantinabioticstresses.FrontPlantSci,2015,6:1142 [38]LiYJ,FangY,FuYR,HuangJG,WuCA,ZhengCC.NFYA1isinvolvedinregulationofpostgerminationgrowtharrestundersaltstressinArabidopsis.PLoSOne,2013,8(4):e61289 [39]SiriwardanaCL,KumimotoRW,JonesDS,HoltBF.GeneFamilyAnalysisoftheArabidopsisNF-YAtranscriptionfactorsrevealsopposingabscisicacidresponsesduringseedgermination.PlantMolBiolRep,2014,32:971–986 [40]KumimotoRW,SiriwardanaCL,GaylerKK,RisingerJR,SiefersN,HoltBF.NUCLEARFACTORYtranscriptionfactorshavebothopposingandadditiverolesinABA-mediatedseedgermination,PLoSOne,2013,8:e59481 [41]GaxiolaRA,RaoR,ShermanA,GrisafiP,AlperSL,FinkGR.TheArabidopsisthalianaprotontransporters,AtNHX1andAvpl,canfunctionincationdetoxificationinyeast.ProcNatlAcadSciUSA,1999,96:1480–1485 [42]谢崇波,金谷雷,徐海明,朱军.拟南芥在盐胁迫环境下SOS转录调控网络的构建及分析.遗传,2010,6:639–646 XieCB,JinGL,XuHM,ZhuJ.ConstructionandanalysisofSOSpathway-relatedtranscriptionaregulatorynetworkunderlyingsaltstressresponseinArabidopsis.Hereditas,2010,6:639–646(inChinesewithEnglishabstract) [43]WuYY,ChenQJ,ChenM,ChenJ,WangXC.Salt-toleranttransgenicperennialryegrass(LoliumperenneL.)obtainedbyAgrobacteriumtumefaciens-mediatedtransformationofthevacuolarNa+/H+antiportergene.PlantSci,2005,169:65–73 [44]BabuRC,ZhangJX,BlumA,HoTHD,WuR,NguyenHT.HVA1,aLEAgenefrombarleyconfersdehydrationtoleranceintransgenicrice(OryzasativaL.)viacellmembraneprotection.PlantSci,2004,166:855–862
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