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1、棉花學(xué)報(bào)CottonScience2017,29(2):166―176陸地棉基因40-1的克隆及表達(dá)分析司愛君袁田琴袁陳紅袁余渝淵新疆農(nóng)墾科學(xué)院棉花研究所/農(nóng)業(yè)部西北內(nèi)陸區(qū)棉花生物學(xué)與遺傳育種重點(diǎn)實(shí)驗(yàn)室/新疆兵團(tuán)棉花遺傳改良與高產(chǎn)栽培重點(diǎn)實(shí)驗(yàn)室,新疆石河子832000冤摘要院揖目的銥WRKY轉(zhuǎn)錄因子通過激活下游一系列抗逆應(yīng)答基因的表達(dá)而提高植株的綜合抗性遙本研究旨在研究WRKY轉(zhuǎn)錄因子在陸地棉中的功能遙揖方法銥通過基因芯片篩選鑒定出1個(gè)逆境誘導(dǎo)的陸地棉WRKY轉(zhuǎn)錄因子基因袁由于此基因編碼蛋白含有1個(gè)典型的WRKY結(jié)構(gòu)域袁且與擬南芥40具
2、有較高相似性袁故命名為40-1遙采用電子克隆及反轉(zhuǎn)錄-聚合酶鏈反應(yīng)技術(shù)獲得40-1基因cDNA全長(zhǎng)袁并對(duì)其進(jìn)行序列分析及結(jié)構(gòu)與功能預(yù)測(cè)遙揖結(jié)果銥40-1的開放閱讀框?yàn)?81bp袁編碼326個(gè)氨基酸袁預(yù)測(cè)蛋白相對(duì)分子質(zhì)量為35.78伊103袁等電點(diǎn)為8.39遙GhWRKY40-1在擬南芥原生質(zhì)體中瞬時(shí)表達(dá)定位于細(xì)胞核袁且具有轉(zhuǎn)錄激活活性遙RT-PCR結(jié)果表明袁40-1受甘露醇誘導(dǎo)上調(diào)表達(dá)遙揖結(jié)論銥推測(cè)40-1可能參與干旱脅迫應(yīng)答反應(yīng)袁并且在植物逆境脅迫中發(fā)揮重要作用遙上述分析為進(jìn)一步從分子水平上驗(yàn)證其抗逆生物學(xué)功能以及揭示棉花非生物脅迫抗
3、逆機(jī)制奠定了基礎(chǔ)遙關(guān)鍵詞院棉花曰WRKY曰轉(zhuǎn)錄因子曰基因克隆曰序列分析中圖分類號(hào)院S562.035文獻(xiàn)標(biāo)志碼院A文章編號(hào)院1002-7807淵2017冤02-0166-1110.11963/issn.1002-7807.201702006SiAijun,TianQin,ChenHong,YuYu(832000,)[Objective]WRKYtranscriptionfactorsenhanceplantresistancebyactivatingtheexpressionofaseriesofstress-re-sponsivedow
4、nstreamgenes.ThepurposeofourresearchwastoanalyzefunctionsofWRKYtranscriptionfactorsinuplandcot-ton.[Method]Astress-inducedupland-cottonWRKYtranscriptionfactorgenewasscreenedbycDNAmicroarrayanalysis.Thegene,whichwasfoundtocontainatypicalWRKYdomainandtoshareahighlysimilara
5、minoacidsequencewith40,wasaccordinglynamed40-1.Thefull-lengthcDNAofGhWRKY40-1wasobtainedbyelectroniccloningandreversetranscription-polymerasechainreaction(RT-PCR)technology,withproteinsequenceanalysisandstructure-functionpredictionperformedusingbioinformaticssoftware.[Re
6、sult]Accordingtoourresults,40-1containsa981-bpopenreadingframeencoding326aminoacidresidues,withapredictedrelativemolecularmassof35.78kDaandanisoelectricpointof8.39.Inprotoplasts,GhWRKY40-1waslocalizedinthenucleusandwasfoundtohavetranscriptionalactivationactivity.RT-PCRan
7、alysisshowedthat40-1wasup-regulatedundermannitolstress.[Conclusion]Wehypothesizethat40-1isinvolvedindroughtstressresponseandplaysanimportantroleinplantstressresponses.Ourcharacterizationandbioinformaticsanalyseshavelaidthefoundationforfutureconfirmationofthebiologicalfun
8、ctionsofresistanceatthemolecularlevelandelucidationofmechanismsofabioticstressresistanceincotton.cotton
