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,2016,36(4):0648-0654
犃犮狋犪犅狅狋.犅狅狉犲犪犾.犗犮犮犻犱犲狀狋.犛犻狀.
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犛狋狉犲狊狊犜狅犾犲狉犪狀犮犲狅犳犜狉犪狀狊犵犲狀犻犮犃狉犪犫犻犱狅狆狊犻狊犈狓狆狉犲狊狊犻狀犵犆犽犔犈犃4犌犲狀犲
犱狌狉犻狀犵犛犲犲犱犌犲狉犿犻狀犪狋犻狅狀犛狋犪犵犲
YUXiumin,ZHANGYanna,YUEWenran,YANGQi,WANYongqing,LIGuojing,WANGRuigang
(ColegeofLifeSciences,InnerMongoliaAgriculturalUniversity,Hohhot010018,China)
犃犫狊狋狉犪犮狋:Basedonourpreviousstudy,犆犽犔犈犃4,a犆犪狉犪犵犪狀犪犽狅狉狊犺犻狀狊犽犻犻LEAgeneinducedbydehydration,
saltandABAtreatments,wastransferredinto犃狉犪犫犻犱狅狆狊犻狊.ThetransgeneexpressionofeightT3trans
genicplantswasdetectedbyquantitativerealtimePCRandthreeindependentoverexpressionlineswith
differenttransgeneexpressionlevelwereselectedforfurtheranalysis.Forthegerminationassays,犆犽犔犈犃4
overexpressionlinesgerminatedmorequicklythanthatofwildtypeonmediumcontainingdifferentcon
centrationsofNaCl,mannitolorABA.Thegerminationrateofwildtypeand犆犽犔犈犃4overexpressionlines
werealdecreasedwiththeincreasedconcentrationsofNaCl,mannitolorABA,whilethegerminationrate
ofwildtypedecreasedmoredramaticalythanthatoftheoverexpressionlines.Thecotyledongreening
ratesof犆犽犔犈犃4overexpressionlineswerealsohigherthanthoseofwildtypeunder200mmol/LNaCl,
400mmol/LmannitolorlowconcentrationsofABA.Altogether,ourresultssuggestedthat犆犽犔犈犃4im
proved犃狉犪犫犻犱狅狆狊犻狊tolerancetosalt,ABAandosmoticstressduringseedgerminationstage.
犓犲狔狑狅狉犱狊:犆犪狉犪犵犪狀犪犽狅狉狊犺犻狀狊犽犻犻;lateembryogenesisabundant(LEA)proteins;abioticstress;transgenic
犃狉犪犫犻犱狅狆狊犻狊
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1,2.犆犽犔犈犃4cDNA;3.Vectorcontrol;
4.Digestedwith犛犪犾Ⅰand犛犪犮Ⅰ
Fig.1 TheconstructionandidentificationofpCanGCkLEA4
recombinantvector
ö2 犆犽犔犈犃4rÜl
gPCRtu
Fig.2 Theexpressionlevelof犆犽犔犈犃4intransgenic
犃狉犪犫犻犱狅狆狊犻sbyrealtimequantitativePCR
mannitoll1/2MS^T,,íR¡¢£。
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cJ¡¢£þÿ
;
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300c400mmol/L,¡¢£¶ÇÄ
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B.200mmol/LNaCl ¼RÂÃO£í;ý犘 <0.01
ö3 Jc犆犽犔犈犃4DNaCl l¡¢£tu
A.Seedgerminationrateofthewildtypeand犆犽犔犈犃4overexpressionlinesunderdifferent
concentrationsofNaCltreatment;B.Cotyledongreeningrateofwildtypeand犆犽犔犈犃4
overexpressionlinesunder200mmol/LNaCltreatment;indicatesignificantdifferenceamongsamplesat0.01level
Fig.3 Germinationassaysofwildtypeand犆犽犔犈犃4overexpressionlinesunderNaClstress
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A.°±mannitol Jcl¡¢£í;B.400mmol/Lmannitol RÂÃO£í;
cQáý犘<0.05c犘<0.01
ö4 Jc犆犽犔犈犃4Dmannitol l¡¢£tu
A.Seedgerminationrateofthewildtypeand犆犽犔犈犃4overexpressionlineswithdifferentconcentrationsofmannitol
treatment;B.Cotyledongreeningrateofwildtypeand犆犽犔犈犃4overexpressionlineswith400mmol/Lmannitoltreatment;
andindicatesignificantdifferenceamongsamplesat0.05and0.01level,respectively.
Fig.4 Germinationassaysofwildtypeand犆犽犔犈犃4overexpressionlineswithmannitoltreatment
|D#$§¨ØJ$tuå¹OLlU
。
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1、犃狋犔犈犃42c犃狋犔犈犃45¬ûºK#$Ì5h
tul§¨&
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â
[19]
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[21]。
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[14]。
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,
D°±l ABA ,
犆犽犔犈犃4¡¢£¶·@J,D0.2
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A.°±ABA Jcl¡¢£í;B.0.2c0.3μmol/LABA RÂÃO£í
ö5 Jc犆犽犔犈犃4DABA l¡¢£tu
A.Seedgerminationrateofwildtypeand犆犽犔犈犃4overexpressionlineswithdifferentconcentrationsofABAtreatment;
B.Cotyledongreeningrateofwildtypeand犆犽犔犈犃4overexpressionlineswith0.2or0.3μmol/LABAtreatment
Fig.5 Germinationassaysofwildtypeand犆犽犔犈犃4overexpressionlineswithABAtreatment
J
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Ð
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MÜÈ·É¡¢£
。
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,
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3564k @AB,â:}犆犽犔犈犃4,|R¡¢kl¨©ÐQ
tu!K
:
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犻狀狏犻狋狉狅andinvivoproteinsynthesis[J].犅犻狅犮犺犲犿犻狊狋狉狔,1981,
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犆狅犿犿狌狀犻犮犪狋犻狅狀狊,2006,348(1):5661.
[4] BROWNEJ,TUNNACLIFFEA,BURNELLA.Anhydrobio
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[5] BATTISTA JR,PARK MJ,MCLEMOREAE.Inactivationof
twohomologuesofproteinspresumedtobeinvolvedinthedesicca
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desiccation[J].犆狉狔狅犫犻狅犾狅犵狔,2001,43(2):133139.
[6] STACYRA,AALENRB.Identificationofsequencehomolo
gybetweentheinternalhydrophilicrepeatedmotifsofgroup1
lateembryogenesisabundantproteinsinplantsandhydrophilic
repeatsofthegeneralstressproteinGsiBof犅犪犮犻犾犾狌狊狊狌犫狋犻犾犻狊
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