24334.兩個野大麥cbls基因家族成員的研究

1、分類號Q75學(xué)校代碼10129UDC577學(xué)號08210032兩個野大麥CBLs基因家族成員的研究StudiesofTwoMembersofCBLsGeneFamilyfromHdeumbrevisubulatum申請人：吳廣宇學(xué)科門類：理學(xué)學(xué)科專業(yè)：生物化學(xué)與分子生物學(xué)研究方向：植物生物化學(xué)與分子生物學(xué)指導(dǎo)教師：李國婧教授李瑞芬研究員論文提交日期：二〇一一年六月StudiesofTwoMembersofCBLsGeneFamilyfr

2、omHdeumbrevisubulatumAbstractCBLs(calcineurinBlikeproteins)aclassofplantspecificCa2senswhichwereiginallyidentifiedinA.thalianawerenamedastheyaresimilartoCNBNCSinanimals.CBLsplayimptantrolesinplant’sspecificgrowthdevelopm

3、entprocessesstressresponseswithitsinteractionproteinCIPK(CBLinteractingproteinkinase).ResearchontheCBLsiscurrentlyfocusedinArabidopsisricebuttherewasverylittlestudyontheotherplants.Shtsubulatebarley(Hdeumbrevisubulatum(T

4、rin.)Link)isoneofthemostsalttoleranthalophytesinTriticeaewhichhasevolvedthemechanismofadaptationtosalinityduringlongtermofevolution.Itisvaluablefresearchapplication.CloningCBLsinShtsubulatebarleynotonlyshowstheeticalmean

5、ingsbutalsobenefitstopracticalapplicationprovidingpotentialcidategenesfimprovingstressestlerancebygeictransfmationinplant.HereweisolatedtwogenesinShtsubulatebarleynamethemHbCBL1HbCBL2.Weanalyzedtheresponseofthegenestodro

6、ughtstressessubcellularlocationtheirfunctionthemainprogressesareasfollows:1.ClonesequenceanalysisofHbCBLs:RTPCRwasusedtoclonefulllengthofHbCBL1HbCBL2.UsingsoftwareanalysisweknowthatbothHbCBL1HbCBL2hasCBLspeicificCa2bindi

7、ngsiteEFhsHbCBL1hasaNmyristoylationsiteatitsNterminaldomain.2.Fussionexpressvectswereconstructedtransfmedintoonionepidermalcellsbyparticlebombardmentrespectively.ThesevectswerealsotransfmedintoArabidopsisbyflaldip.Confoc

8、alobservationsshowedthatHbCBL1accumulatedinplasmamembraneaswellasinnucleusHbCBL2wasaccumulatedintonoplast.3.UsingNouthernblottoanalysistheexpressionofHbCBLswhileunderdroughtstess.WithoutanystressHbCBL1HbCBL2haveasmallamo

9、utofbackgroundespressionexpressioninrootsismestrongthanintheleaf.BythetreatmentofPEG6000mannitolespressionsofHbCBL1HbCBL2startedtodiminishastheprocessingtimewentby.4.IndertoexplethebiologicalfunctionofHbCBLsseveraloverex