在转基因水稻中超量表达wnk激酶基因的的研究

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1、2扬州大学硕士学位论文表达WNK激酶基因的转基因水稻植株对白叶枯病菌混合菌株的抗性均有一定的提高。(5)经常规杂交将含WNK转基因纯合系与含Xa21基因的台北309转基因植株聚合，对聚合两个转基因的阳性植株用强致病菌P6进行接种，调查其抗病性，并与未转化对照比较。结果显示，聚合后部分阳性植株与未转化对照相比，抗病性有了极显著增；戥关键词：水稻；白叶枯病；wNK激酶；遗传转化；过表达钱盈佳在转基因水稻中超量表达WNK激酶基因的研究3Studiesonover-expression0fWNKkinasegenesintransgenicrice●Postgraduate：Ying-jiaQ

2、ianSupervisor：Prof．Qiao—quanLiu(YangzhouUniversity,Yangzhou22500只diangsuProvince)ABSTRACTXa21isthefirstdiseaseresistancegeneclonedfromrice(OryzasativaL．)，whichencodesaspectrumagainstXanthomonasoryzaepv．oryzae(Xoo．)．Basedonyeasttwo-hybridtechnique．ithasbeenidentifiedthatXB3caninteract、)l，itllXA21

3、andtheremaybeinteractionbetweentheWNK(withnolysineⅨ))kinasesandXB3proteins．Inpresentstudies，aseriesofspecificvectorsforover-expressionofWNK二”geneswereconstructed,alltheconstructswereintroducedintoasusceptiblejaponicaricecultivarTaipei309(TP309)．Lotsoftransgenicriceplantshavebeengenerated,andthei

4、rdiseaseresistancesweretheninvestigated．ThepresentresultswillhelpUSunderstandthemolecularmechanismsofXa21signalingpathwaysinrice．Themainresultsareasfollowings：1．Accordingtothepublishedsequences，theC0dil培regionofeachWNKgeneswasidentified，theircodingsequenceswereclonedandcompared，andalsousedforcon

5、structionofover-expressionvectors，respectively．2．ByAgrobacterium—mediatedtransformation,alltheconstructedvectorswereintroducedintoTaipei309，andlotsoftransgenicricelineswereregenemted．TheresultsfrombothPCRanalysisandHygromycinresistancetestshowedthatthetransgenehadintegratedintothegenomeoffleepla

6、nts．Furthermore，severaltransgeniclinescontaininghomozygoustransgenewereselected．3．ThetotalRNAswereextractedforRT-PCRanalysisinsometransgenicriceplants．Theresultsshowedthattheexpressionofendogenoustargetgene(s)was4扬州大学硕士学位论文increasedwithdifferentrangesintheleafoftransgenicplantscontainingthetrans

7、gene．Itindicatedthatthedesignedstructurehasfunctionedintransgenicrice．4．AmixedXoobacteriaWasusedfortreatmentofthetransgenicplantsaSwelIastheirwildtypeTaipei309．TheresultsshowedthattheXooresistanceoftransgenicplantshadbeenimp