一个新水稻根尖特异表达启动子的分离与鉴定精品.doc

一个新水稻根尖特异表达启动子的分离与鉴定 赵红玉１,２　徐磊２　魏溪涓１　邓敏娟２　王芳２,　易可可２ (１浙江师范大学化学与生命科学学院,浙江金华３２１００４;２浙江省农业科学院病毒学与生物技术研究所,杭州３１００２１;通讯联 系人,EＧmail:wffsy＠mail．zaas．ac．cn) IsolationandCharacterizationofaNovelRootTipＧspecificPromoterinRice ZHAOHongＧyu１,２,XULei２,WEIXiＧjuan１,DENGMinＧjuan２,WANGFang２,,YIKeＧke２ (１CollegeofChemistryandLifeSciences,ZhejiangNormalUniversity,Jinhua３２１００４,China;２InstituteofVirologyand Biotechnology,ZhejiangAcademyofAgriculturalSciences,Hangzhou３１００２１,China;Correspondingauthor,EＧmail:wffsy＠ mail．zaas．ac．cn) ZHAOHongyu,XULei,WEIXijuan,etal．IsolationandcharacterizationofanovelroottipＧspecificpromoterinrice． ChinJRiceSci,２０１４,２８(４):３５１Ｇ３５７． Abstract:ToobtaintherootＧspecificpromotersinrice,arootＧspecificgeneOs０４g２４４６９wasselectedbyanalyzingthe publicmicroarraydataandfurtherconfirmedbyRTＧPCR．Todeterminetheexpressionpatternofthenovelpromoter, theputativepromoterwasisolatedandfusedwiththeGUSreportergenetotransformintothericevarietyNipponbare． SubsequentGUSstainingofthetransgenicplantsshowedthattheOs０４g２４４６９promoterwasonlyactivatedinriceroot tip．UsingthisroottipＧspecificpromoter,wecharacterizedthefunctionofaubiquitouslyexpressedgeneOsSRGinroot development．TheabsenceofOsSRGledtoashortrootphenotypeinthesrgmutant,theroottipＧspecificexpressionof GUSＧOsSRGinsrgplantssignificantlyreversedtherootgrowthdefect,indicatingthattheexpressionofOsSRGinroot tip,notinshoot,isimportantforrootdevelopmentinrice．ThepresentstudyshowedthatOs０４g２４４６９promoteris highlyactiveintheroottipandcanbeusefulfortheroottipＧspecificexpressionoftargetgenesandgenefunction analysisinriceroot． Keywords:rice;promoter;roottipＧspecificpromoter;GUSreportergene;SRGgene 赵红玉,徐磊,魏溪涓,等．一个新水稻根尖特异表达启动子的分离与鉴定．中国水稻科学,２０１４,２８(４):３５１Ｇ３５７． 摘　要:为了挖掘水稻中的根系特异表达启动子,通过生物信息学分析和RTＧPCR验证获得１个在根尖特异表达的水稻基 因Os０４g２４４６９.用分离出的该基因上游启动子,与GUS报告基因构建表达载体并转化到受体水稻日本晴中.通过对转基 因后代中不同组织器官中的GUS活性检测,证明该启动子在水稻根尖特异性强烈表达.结合该根尖特异表达启动子,我们 分析了一个在水稻地上部及地下部都组成表达的功能基因OsSRG对水稻根系发育的贡献.该基因的缺失导致水稻突变体 srg根系变短,而利用分离的根尖特异性Os０４g２４４６９启动子在根尖驱动GUSＧOsSRG的表达,能回复突变体根系生长,表明 OsSRG在根尖的