一个新的水稻低温应答类糖基转移酶基因OsCrGtl的遗传转化及其功能的初步研究.pdf

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1、分类号学校代码密级学号10542200814020095一个新的水稻低温应答类糖基转移酶基因OsCrGtl的遗传转化及其功能的初步研究TransformationandFunctionalAnalysisofaNovelColdResponsiveGlycosyltransferase--likeGene(OsCrGtl)inRice研究生姓名张俊指导教师姓名、职称．盆耋壹堑窒基．湖南师范大学学位评定委员会办公室二零一一年四月个糖基转移酶结构域，故该基因为一类糖基转移酶基因。为了确认该基因是否具有耐冷作用或其它作用，构建了由2xCaMV35S组

2、成型强启动子驱动的pCAMBIAl300．OsCrGtl双元表达载体，并通过农杆菌介导法转化了耐冷性弱的籼稻品种93．1l，获得9个独立的转OsCrGtl基因cDNA的93．11To代植株。对To代与Tl代转基因植株的耐冷性进行了鉴定，结果表明：在较低温度(15℃)下发芽，转OsCrGtl基因93．11发芽势显著大于非转基因的93．1l；经低温处理后，转OsCrGtl基因93．11To代再生苗与Tl代实生苗的相对电导率较非转基因的93．11低，游离脯氨酸积累较非转基因93一11多，Tl代实生苗可溶性糖积累较非转基因93．11多，过氧化物酶活性也

3、较非转基因93．11高，说明转OsCrGtl基因93．11耐冷性强于非转基因的93．11。此外，在非逆境下，对转基因93．1lT0代植株与野生型93．11的株高、剑叶长、单株有效穗数、每穗总粒数、结实率与千粒重等主要农艺性状也进行了比较观测，结果表明，二者主要农艺性状无显著性差异。关键词：糖基转移酶；转基因；低温；耐冷性；水稻ABSTRACTOsCrGtl(OryzasativaL．coldresponsiveglycosyltmnsferase—likegene)wasanovelstressresponsivegeneidentifiedi

4、ntheindicacultivarPei’ai64S．Itishighlyinducedbyabioticstresses，especiallybycold，inbothleafandpanicleatallthedevelopmentstagesanalyzedinrice．Itencodedaproteincontainingaglycosyltransferasedomain，SOitisaglycosyltransferase-likegene．Inordertoanalyzeitsfunctionsincoldtoleranceor

5、others，abinaryvectorpCAMBIAl300·OsCrGtl，drivenbystrongconstitutivepromoter2xCaMV35S，wasconstructed，andOsCrGt／Wasthentransformedintoriceindicavarieties93．11whichissensitivetocoldthroughagrobacterium-mediatedmethod．Nineindependenttransgenicriceplantswereobtained．Thecoldtoleran

6、ceoftmnsgenicriceplantswereassessedfortheirToandTIgenerationsseedlings．Theresultssuggestthatthetransgenicriceseeds(T1)posseshighergerminatingpotentialitythanwildtypeonesunderlowtemperature(15"C)althoughtheyshowsnosignificantdifferenceintheirgerminatingrates．Theelectrolytelea

7、kage(％)ofthetransgenicriceplant(To)increasessloweranditsfreeprolinecontentaccumulatesfasterthanthoseofthewild·-type93··11aftercoldtreatmentinchamber．TheresultsalsoshowedthattheelectrolyteleakageoftheT1generationtransgenicseedlingsgrowsslower,itssolublesugarandfreeprolineIIco

8、ntentaccumulatefaster,andtheactivityofitsPOD．oneofROS‘scavengingenzymes，ish