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Cloning of a New Gene TaDRLea3 2 from Wheat and Its Expression under Different Stresses Treatments

小麦TaDRLea3 2基因的克隆及胁迫诱导表达分析



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,2016,36(6):1091-1097
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LIANGYajun1,YUZhengyang1,QIANGZhiquan1,DUYa1,ZHU Weining2,
ZHANGDapeng1,ZHANGLinsheng1
(1ColegeofLifeSciences,NorthwestA&FUniversity,Yangling,Shaanxi712100,China;2ColegeofLifeSciences,North
westUniversity,Xian710069,China)
犃犫狊狋狉犪犮狋:Lateembryogenesisabundantprotein(LEA)areasuiteofimportantfamilyproteinsfoundinor
ganisms,whicharegeneralyinvolvedinosmoregulationandareexpressedabundantlywhensubjectedto
environmentalstresses.Inthisstudy,weisolatedanovel犔犈犃3gene犜犪犇犚犔犲犪32fromwheat‘Zhengyin
No.1’.Thefullengthof犜犪犇犚犔犲犪32is668bp,whoseORFis570bp,encoding189aminoacids,and
consistsof9conserved11merrepeatingmotifs;Proteinpredictionshowsthattheproteinishighlyhydro
philicandlocatedinthecytoplasm,withthreephosphorylationsitesandhasnotransmembranedomainor
signalpeptide;Secondarystructureanalysisshowsthatthemainstructureisalphahelix;Homologycom
parisonandregeneratingphylogenetictreewerecarriedoutbyMEGA5.1.Expressionanalysesrevealed
that犜犪犇犚犔犲犪32genewasinducedbydrought,lowtemperature,highsaltstressandalsomoreover,can
beinducedbyABAtreatment.Theseresultssuggestedthat犜犪犇犚犔犲犪32isanABAdependent犔犈犃3
gene,participatingintheresponseprocessofwheatundertheabioticstressesindifferentmechanismsand
canlayafoundationforelucidating犔犈犃3geneprotectionmechanismunderabioticstressesinwheat.
犓犲狔狑狅狉犱狊:wheat;犜犪犇犚犔犲犪32;homologicalcloning;sequenceanalysis;realtimePCR
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M.DNAMarker;1,2.PCRproductsof犜犪犇犔犲犪32gene;
3.PCRidentificationofpositiveplasmidpMD19T犜犪犇犚犔犲犪32
Fig.1 Cloneandidentifyof犜犪犇犚犔犲犪32
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42 犜犪犇犚犔犲犪321ž¡¢v¤1¥äå
Ninerepeating11meraminoacidmotifsareunderlined
andnumbered
Fig.2 Theaminoacidsequenceof犜犪犇犚犔犲犪32
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狋犺犪犾犻犪狀犪)vHIJr”K(44)。
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Fig.3 SequencealignmentofwheatTaDRLea32proteinandotherLEAproteins
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500replicates;Thescalerepresentsgeneticdistance
Fig.4 PhylogentictreeofwheatTaDRLea32proteinand
otherLEAproteins
45 “”TaDRLea32nov©ª/hª«áÔ
Fig.5 Hydrophilic/hydrophobicityanalysisof
wheatTaDRLea32protein
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Fig.6 Phosphorylationsitesofwheat
TaDRLea32protein
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Fig.7 Secondarystructure(A)andtertiarystructure(B)predictionofwheatTaDRLea32protein
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[1] µ ¶,·´X.3¸]¹8\81ã[M]."Þ:,%º

,1998:618.
[2] SHINOZAKIK,YAMAGUCHISHINOZAKIK.Genenet
worksinvolvedindroughtstressresponseandtolerance[J].
犑狅狌狉狀犪犾狅犳犈狓狆犲狉犻犿犲狀狋犪犾犅狅狋犪狀狔,2007,58(2):221227.
[3] HUNDERMARKM,HINCHADK.LEA(lateembryogene
6901 ! " # $ % &                   36ö
sisabundant)proteinsandtheirencodinggenesin犃狉犪犫犻犱狅狆狊犻狊
狋犺犪犾犻犪狀犪[J].犅犕犆犌犲狀狅犿犻犮s,2008,9:118.
[4] ¼hU,TU8,½¾U,(.“”>ª¿1ž犠犣犢2vŽ
Â!äåO§
[J].!"]U,^_%%&(æç,%1),
2009,37(2):9399.
HUANGFP,ZHANGLS,WANGLP,犲狋犪犾.Cloningand
sequenceanalysisofanewdehydringene(犠犣犢2)fromwheat
[J].犑狅狌狉狀犪犾狅犳犖狅狉狋犺狑犲狊狋犃牔犉犝狀犻狏犲狉狊犻狋狔(NaturalScience
Edition),2009,37(2):9399.
[5] DUREL,GREENWAYSC,GALAUGA.Developmental
biochemistryofcottonseedembryogenesisandgermination:
changingmessengerribonucleicacidpopulationsasshownby
犻狀狏犻狋狉狅and犻狀狏犻狏狅proteinsynthesis[J].犅犻狅犮犺犲犿犻狊狋狉狔,1981,
20(14):41624168.
[6] RIEDJL,WALKERSIMMONSMK.Group3lateembryo
genesisabundantproteinsindesiccationtolerantseedlingsof
wheat(犜狉犻狋犻犮狌犿犪犲狊狋犻狏狌犿L.)[J].犘犾犪狀狋犘犺狔狊犻狅犾狅犵狔,1993,
102(1):125131.
[7] HSINGYC,CHENZY,犲狋犪犾.Unusualsequencesofgroup3
LEA mRNAinducibleby maturationordryinginsoybean
seeds[J].犘犾犪狀狋犕狅犾犲犮狌犾犪狉犅犻狅犾狅犵狔,1995,29(4):863868.
[8] ZEGZOUTIH,JONESB,犲狋犪犾.ER5,atomatocDNAenco
dinganethyleneresponsiveLEAlikepeotein:characterization
andexpressioninresponsetodrought,ABAandwounding
[J].犘犾犪狀狋犕狅犾犲犮狌犾犪狉犅犻狅犾狅犵狔,1997,35(6):847854.
[9] CAMPBELLSA,CRONEDE,犲狋犪犾.Aca.40kDamaize(犣犲犪
犿犪狔狊犔.)embryodehydrinisencodedbythedhn2locusonchro
mosome9[J].犘犾犪狀狋犕狅犾犲犮狌犾犪狉犅犻狅犾狅犵狔,1998,38(3):417423.
[10] MINAMIA,NAGAOM,犲狋犪犾.Coldacclimationinbryophytes:
lowtemperatureinducedfreezingtolerancein犘犺狔狊犮狅犿犻狋狉犲犾犾犪狆犪狋
犲狀狊isassociatedwithincreasesinexpressionlevelsofstressrelat
edgenesbutnotwithincreaseinlevelofendogenousabscisicacid
[J].犘犾犪狀狋犪,2005,220(3):414423.
[11] TUNNACLIFFEA,LAPINSKIJ,MCGEEB.AputativeLEA
protein,butnotrehalose,ispresentinanhydrobioticbdeloidro
tifers[J].犎狔犱狉狅犫犻狅犾狅犵犻犪,2005,546(1):315321.
[12] ÀÁU,Âß.LEAnovOP8$%0JõÄ[J].8
$^Q&
,2003,(6):613.
SUNLP,LIDQ.ProgressinmolecularbiologyofLEA
protein[J].犅犻狅狋犲犮犺狀狅犾狅犵狔犅狌犾犾犲狋犻狀,2003,(6):613.
[13] ÅÆÇ,c)È,(.Z3€LEAnoÂ!1ž0JõÄ
[J].!eÉ\%a%&(æç,%1),2006,9(4):2730.
ZHAOYM,YANGJX,犲狋犪犾.Progressinresearchofgroup3
LEAproteinandgene[J].犑狅狌狉狀犪犾狅犳犡犻犪狀犝狀犻狏犲狉狊犻狋狔狅犳犃狉狋狊
牔犛犮犻犲狀犮犲(NaturalScienceEdition),2006,9(4):2730.
[14] LIUY,ZHENGYZ.PM2,agroup3LEAproteinfrom
soybean,andits22merrepeatingregionconfersalttolerance
inEscherichiacoli[J].犅犻狅犮犺犲犿犻犮犪犾犪狀犱 犅犻狅狆犺狔狊犻犮犪犾犚犲
狊犲犪狉犮犺犆狅犿犿狌狀犻犮犪狋犻狅狀狊,2005,331(1):325332.
[15] · Ê, Ë,Â2,,(.#$LEAnoÂ!üÆ[J].
r9]%Q&
,2009,25(24):143146.
CHENL,LIL,LIJH,犲狋犪犾.PlantLEAProteinsandtheir
functions[J].犆犺犻狀犲狊犲犃犵狉犻犮狌犾狋狌狉犪犾犛犮犻犲狀犮犲犅狌犾犾犲狋犻狀,2009,
25(24):143146.
[16] BATTAGLIAM,OLVERACARRILLOY,GARCIARRU
BIO A,犲狋犪犾.TheenigmaticLEAproteinsandotherhy
drophilins[J].犘犾犪狀狋犘犺狔狊犻狅犾狅犵狔,2008,148(1):624.
[17] Ì Í,ÂÎÏ,(.#$Z6no(LEA3)22¤1¥ƒÎ®
¯ºGÐÑÉÊrv9Ç
[J].#$0J,2009,29(1):7479.
LIUY,LIR H,犲狋犪犾.Functionalidentificationofplant
stressresistantprotein(LEA3)andits22mermotifsinyeast
[J].犅狌犾犾犲狋犻狀狅犳犅狅狋犪狀犻犮犪犾犚犲狊犲犪狉犮犺,2009,29(1):7479.
[18] HSINGYI,CHENZY,CHOWTY.Nucleotidesequences
ofasoybeancomplementaryDNAencodinga50kilodalton
lateembryogenesisabundantprotein[J].犘犾犪狀狋犘犺狔狊犻狅犾狅犵狔,
1992,99(1):354355.
[19] WANGL,LIX,CHENS,犲狋犪犾.Enhanceddroughttoler
anceintransgenic犔犲狔犿狌狊犮犺犻狀犲狀狊犻狊plantswithconstitutively
expressedwheatTaLEA3[J].犅犻狅狋犲犮犺狀狅犾狅犵狔犔犲狋狋犲狉狊,2009,
31(2):313319.
[20] |}~,TU8,(.“”ƒ61ž犜犪犔犈犃3vŽÂGÐ
ÑrvüÆO§
[J].8$,Þ%&,2004,20(4):832838.
YUJN,ZHANGLS,犲狋犪犾.Cloningofanovelstresstoler
antgene犜犪犔犈犃3fromwheatandthefunctionalanalysisin
yeast[J].犑狅狌狉狀犪犾狅犳犅犻狅狋犲犮犺狀狅犾狅犵狔,2004,20(4):832838.
[21] XUD,DUANX,WANGB,犲狋犪犾.Expressionofalateem
bryogenesisabundantproteingene,HVA1,frombarleycon
ferstolerancetowaterdeficitandsaltstressintransgenicrice
[J].犘犾犪狀狋犘犺狔狊犻狅犾狅犵狔,1996,110(1):249257.
[22] DUREL.Arepeating11meraminoacidmotifandplantdes
iccation[J].犘犾犪狀狋,1993,3(3):363369.
[23] WOODSWS,BOETTCHERJM,ZHOUDH,犲狋犪犾.Con
formationspecificbindingofalphasynucleintonovelprotein
partnersdetectedbyphagedisplayandNMRspectroscopy
[J].犅犻狅犾狅犵犻犮犪犾犆犺犲犿犻狊狋狉狔,2007,282(47):3455534567.
[24] ÒÓ~,Å Ô,(.“”ƒ„~1ž 犜犪犔犈犃犔3vŽ
ÂOPW«O§
[J].5$%&,2012,38(10):18471855.
MINDH,ZHAOY,CHENY,犲狋犪犾.IsolationandMolecu
larcharacterizationofstressrelated犜犪犔犈犃犔3geneinwheat
[J].犃犮狋犪犃犵狉狅狀狅犿犻犮犪犛犻狀犻犮犪,2012,38(10):18471855.
[25] DUREL,CROUCHM,犲狋犪犾.Commonaminoacidsequence
demainsamongtheLEAproteinsofhigherplants[J].犘犾犪狀狋
犕狅犾犲犮狌犾犪狉犅犻狅犾狅犵狔,1989,12(5):475486.
[26] INGRAMJ,BARTELSD.Themolecularbasisofdehydra
tiontoleranceinplants[J].犃狀狀狌犪犾犚犲狏犻犲狑狅犳犘犾犪狀狋犘犺狔狊犻
狅犾狅犵狔犪狀犱犘犾犪狀狋犕狅犾犲犮狌犾犪狉犅犻狅犾狅犵狔,1996,47:377403.
[27] WOLKERSWF,MCCREADYS,BRANDTWF,犲狋犪犾.I
solationandcharacterizationofaD7LEAproteinfrompol
lenthatstabilizesglasses犻狀狏犻狋狉狅[J].犅犻狅犮犺犻犿犻犮犪犲狋犅犻狅狆犺狔狊
犻犮犪犃犮狋犪,2001,1544(12):196206.
[28] SUNX,RIKKERINKEHA,JONESWT,犲狋犪犾.Multifa
riousrolesofintrinsicdisorderinproteinsilustrateitsbroad
impactonplantbiology[J].犘犾犪狀狋犆犲犾犾,2013,25(1):3855.

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