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1、作物学报ACTAAGRONOMICASINICA2014,40(7):1304−1310http://zwxb.chinacrops.org/ISSN0496-3490;CODENTSHPA9E-mail:xbzw@chinajournal.net.cnDOI:10.3724/SP.J.1006.2014.01304甘蓝型油菜2个GPAT6同源基因的克隆与表达分析11111,21,2,*刘聪肖旦望胡学芳邬克彬官春云熊兴华12湖南农业大学作物基因工程湖南省重点实验室,湖南长沙410128;湖南农业大学油料作物研究所,湖南长沙
2、410128摘要:甘油-3-磷酸酰基转移酶(sn-glycerol-3-phosphateacyltransferase,GPAT)是三酰甘油生物合成的关键酶,催化三酰甘油合成的起始步骤,在植物中参与调节生长发育、脂质的合成和逆境应答等过程。本试验通过RT-PCR从甘蓝型油菜品种湘油15中克隆得到GPAT6基因的2个拷贝,分别命名为BnGPAT6-1(GenBank登录号为KJ000117)和BnGPAT6-2(KC106728)。它们的编码区序列(codingDNAsequence,CDS)均为1506bp,编码501个
3、氨基酸。预测其蛋白结构N端含有一个类卤酸脱卤酶(haloaciddehalogenase-likehydrolase,HAD-like)活性结构域,C端含有一个溶血磷脂酰基转移酶(lysophospholipidacyltransferase,LPLAT)功能域。蛋白序列比对和进化树分析表明,甘蓝型油菜与白菜(B.rapa)、甘蓝(B.oleracea)、拟南芥(Arabidopsisthaliana)和琴叶拟南芥(A.lyrata)中的GPAT6序列相似性最高。组织表达结果表明BnGPAT6基因在花中的表达量最高,在未成
4、熟的胚中表达量呈先升高后降低的趋势。BnGPAT6的表达受ABA的抑制;在干旱和6-BA胁迫下几乎同时升高;在盐胁迫下,短时间内升高,达到峰值后降低;在水渍条件下没有明显变化。关键词:甘蓝型油菜;甘油-3-磷酸酰基转移酶;基因克隆;表达分析;逆境胁迫CloningandExpressionAnalysisofTwoHomologousGenesCodingsn-Glycerol-3-PhosphateAcyltransferase6inBrassicanapus11111,21,2,*LIUCong,XIAODan-Wan
5、g,HUXue-Fang,WUKe-Bin,GUANChun-Yun,andXIONGXing-Hua12CropGeneEngineeringKeyLaboratoryofHunanProvince,HunanAgriculturalUniversity,Changsha410128,China;OilseedCropsInstitute,HunanAgriculturalUniversity,Changsha410128,ChinaAbstract:Akeyenzymesn-glycerol-3-phosphateac
6、yltransferase(GPAT)catalyzestheinitialstepofTAGbiosyntheticpathway,andisinvolvedinmanyprocessesincludingplantgrowth,developmentandresponsetoabioticstresses.Inthisstudy,twoB.napusGPAT6homologswereclonedfromtheleafofthecultivarxiangyou15usingRT-PCR,anddesignatedasBn
7、GPAT6-1andBnGPAT6-2,respectively.ThecodingDNAsequences(CDS)oftwoBnGPAT6genesare1506bpinlength,encodingtwodiffer-entpolypeptidesof501aminoacidresidues.Theproteinswerepredictedtoconsistofahaloaciddehalogenase-likehydrolasedomainandalysophospholipidacyltransferasedom
8、ain.MultiplesequencealignmentsandphylogeneticanalysisofGPATproteinsshowedthatBnGPAT6-1andBnGPAT6-2sharehighsimilaritywithGPAT6proteinsfromB.rapa,B.olera
