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1、園藝學(xué)報(bào)2014,(4111):2169–2178http://www.ahs.ac.cnActaHorticulturaeSinicaE-mail:yuanyixuebao@126.com蘋(píng)果PGIP基因部分家族成員啟動(dòng)子的克隆與功能分析*符聰慧,王建平,張沖,馬鋒旺,張軍科(西北農(nóng)林科技大學(xué)園藝學(xué)院,陜西楊凌712100)摘要:根據(jù)已發(fā)表的蘋(píng)果基因組序列設(shè)計(jì)特異引物,克隆了蘋(píng)果(Malus×domesticaBorkh.)品種‘秦冠’和‘太平洋玫瑰’中PGIP家族基因PGIP1和PGIP2的啟動(dòng)子片段,序列分析結(jié)果表明PGIP1與PGIP2啟動(dòng)子序列相
2、似度為71%,‘秦冠’和‘太平洋玫瑰’的PGIP1、PGIP2啟動(dòng)子相似度都達(dá)到了99%。兩個(gè)品種PGIP1啟動(dòng)子中TATA-box和CAAT-box的數(shù)量明顯多于PGIP2,PGIP1和PGIP2啟動(dòng)子分別存在茉莉酸甲酯和水楊酸響應(yīng)元件,暗示PGIP1和PGIP2存在不同的抗病響應(yīng)途徑。構(gòu)建了啟動(dòng)子︰︰GUS融合表達(dá)載體,通過(guò)對(duì)農(nóng)桿菌介導(dǎo)法轉(zhuǎn)化煙草葉片中的GUS活性分析,比較了不同啟動(dòng)子的活性。結(jié)果表明:PGIP1啟動(dòng)子活性在品種間差異不顯著;PGIP2啟動(dòng)子活性在品種間差異顯著,‘秦冠’是‘太平洋玫瑰’的2.37倍,可能與品種抗病性差異有關(guān);同一品種中
3、PGIP1啟動(dòng)子活性顯著高于PGIP2。關(guān)鍵詞:蘋(píng)果;PGIP基因;啟動(dòng)子;順式作用元件中圖分類(lèi)號(hào):S661.1文獻(xiàn)標(biāo)志碼:A文章編號(hào):0513-353X(2014)11-2169-10CloningandFunctionAnalysisofPartofPGIPGeneFamilyMemberPromotersfromApple*FUCong-hui,WANGJian-ping,ZHANGChong,MAFeng-wang,andZHANGJun-ke(CollegeofHorticulture,NorthwestA&FUniversity,Yanglin
4、g,Shaanxi712100,China)Abstract:PrimersweredesignedaccordingtothepublishedsequenceoftheapplegenomeandtwoPGIPgenepromotersofMalus×domesticaBorkh.‘Qinguan’and‘PacificRose’werecloned.PGIP1were71%homologywithPGIP2genepromotersinbothapplecultivar;BothPGIP1andPGIP2genepromotersof‘Qinguan
5、’and‘PacificRose’sharedasequenceidentityof99%.PGIP1promotershadmoreTATA-boxandCAAT-boxthanthatofPGIP2promoters,andPGIP1genepromoterscontainedmethyljasmonate-responsiveelements,whilePGIP2genepromotersowedsalicylicacidresponsiveelements.It’sspeculatedthattherearedifferentdiseaseresp
6、onsepathwaysofPGIP1andPGIP2.PGIPpromoteractivitywereevaluatedbypromoter︰︰GUSfusionexpressionintransgenictobaccoleavesandtheresultsshowedthatthePGIP1promotersactivitywasnotsignificantbetweencultivarswhilePGIP2promoterof‘Qinguan’washigherthanthatof‘PacificRose’,andtheformer’sactivit
7、ywas2.37foldsofthelatter,whichwasprobablyrelatedtotheresistancedifference.Inthesameapplecultivar,PGIP1genepromoters’收稿日期:2014–06–27;修回日期:2014–09–12基金項(xiàng)目:國(guó)家現(xiàn)代農(nóng)業(yè)產(chǎn)業(yè)技術(shù)體系建設(shè)專(zhuān)項(xiàng)資金項(xiàng)目(CARS-28)*通信作者Authorforcorrespondence(E-mail:zhangjk@nwsuaf.edu.cn)2170園藝學(xué)報(bào)41卷activitiesweresignificantlyhigh
8、erthanthatofPGIP2.Keywords:apple;