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,2016,36(7):1337-1342
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犐狊狅犾犪狋犻狅狀犪狀犱犈狓狆狉犲狊狊犻狅狀犃狀犪犾狔狊犻狊狅犳犪狀犃狀狋犺狅犮狔犪狀犻狀犜狉犪狀狊狆狅狉狋
狉犲犾犪狋犲犱犖狋犃犖9犌犲狀犲犳狉狅犿犖犻犮狅狋犻犪狀犪狋犪犫犪犮狌犿
CUIChaojun,LIJiamin,CHENJunyu,ZHANGRunzhao,ZHUQinlong
(ColegeofLifeSciences,SouthChinaAgriculturalUniversity,StateKeyLaboratoryforConservationandUtilizationof
SubtropicalAgrobioresources,KeyLaboratoryofPlantFunctionalGenomicsandBiotechnology,EducationDepartmentof
GuangdongProvince,Guangzhou510642China)
犃犫狊狋狉犪犮狋:Tostudyanthocyanintransport,weisolatedafullengthgene犖狋犃犖9(GenBankAccessionNo.
KX356542)by犻狀狊犻犾犻犮狅狀cloningandRTPCRmethodfrom犖犻犮狅狋犻犪狀犪狋犪犫犪犮狌犿.The犖狋犃犖9containsa
690bpopenreadingframe(ORF)encoding229aminoacidresidues,whichbelongstoaputativephiclass
glutathionestransferase(GST)family.Itsgenomicstructurecontains3exonsand2introns.Multiplese
quencealignmentshowedthatNtAN9hashighestidentities(88%)withpetuniaPhAN9thatisanantho
cyanintransportrelatedGST.Phylogeneticanalysisshowedthat犖狋犃犖9wasclusteredintoananthocya
nintransportrelatedGSTgroup,andwasanorthologof犘犺犃犖11.Theanthocyanincontentanalysisand
realtimequantitationPCR (qRTPCR)showedthat犖狋犃犖9wasexpressedindifferentflowerdevelop
mentalstages,withhighestlevelinstageⅢ,butalmostnoexpressioninroot,stemandleafwhichareal
mostnoanthocyanin.Theseresultssuggestthatthe犖狋犃犖9functionissimilarwith犘犺犃犖9andmightbe
relatedtothetransferandaccumulationofanthocyaninintobaccoplants.
犓犲狔狑狅狉犱狊:犖犻犮狅狋犻犪狀犪狋犪犫犪犮狌犿;anthocyanintransport;glutathionestransferase;geneclone;expression
analysis
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conserveddomainGST_C_Phi(E91~L209)aresingleanddouble
underlines,respectively.TheGSHsitesaremarkedwithgray
background,andthesubstratebindingsitesaremarkedinbox.
Thetwoverticalarrowsindicatethesplicesitesbetweenexons
Fig.1 cDNAsequenceandthededucedaminoacid
sequenceof犖狋犃犖9
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KX356542。:
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withfivepointedstar.Triangleindicatessubstratebindingsites
Fig.3 ThemultiplesequencealignmentofNtAN9withknownfunctionanthocyanintransportrelatedGSTproteins
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Fig.5 Theanthocyanincontentanalysisintobacco
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:
[1] ZHAOJ.Flavonoidtransportmechanisms:howtogo,andwith
whom[J].犜狉犲狀犱狊犘犾犪狀狋犛犮犻犲狀犮犲,2015,20(9):576585.
[2] xbÔ,x y,z{|,F.#${Eãêëæ
åzSj
[J].#$+[%&,2015,51(11):18151820.
ZHANGYZ,ZHANGF,WANGLY,犲狋犪犾.Plantglutathi
one犛transferases:rolesinflavonoidaccumulation[J].犘犾犪狀狋
犘犺狔狊犻狅犾狅犵狔犑狅狌狉狀犪犾,2015,51(11):18151820.
[3] MARRSKA,ALFENITOMR,LLOYDAM,犲狋犪犾.Aglutathi
oneStransferaseinvolvedinvacuolartransferencodedbythemaize
gene犅狉狅狀狕犲2[J].犖犪狋狌狉犲,1995,375(6530):397400.
[4] ALFENITO MR,SOUERE,GOODMANCD,犲狋犪犾.Func
tionalcomplementationofanthocyaninsequestrationinthe
vacuolebywidelydivergentglutathioneStransferases[J].
犘犾犪狀狋犆犲犾犾,1998,10(7):11351149.
[5] KITAMURAS,SHIKAZONO N,TANAKA A.犜犚犃犖犛
犘犃犚犈犖犜犜犈犛犜犃19isinvolvedintheaccumulationofboth
anthocyaninsandproanthocyanidinsin犃狉犪犫犻犱狅狆狊犻狊[J].犘犾犪狀狋
犑狅狌狉狀犪犾.,2004,37(1):104114.
[6] YAMAZAKIM,SHIBATA M,NISHIYAMAY,犲狋犪犾.Differenti
algeneexpressionprofilesofredandgreenformsof犘犲狉犻犾犾犪犳狉狌
狋犲狊犮犲狀犮犲leadingtocomprehensiveidentificationofanthocyaninbio
syntheticgenes[J].犉犈犅犛犑狅狌狉狀犪犾,2008,275(13):34943502.
[7] CONNS,CURTINC,B?ZIERA,犲狋犪犾.Purification,molecu
larcloning,andcharacterizationofglutathioneStransferases
(GSTs)frompigmented犞犻狋犻狊狏犻狀犻犳犲狉犪L.celsuspensioncul
turesasputativeanthocyanintransportproteins[J].犑狅狌狉狀犪犾
犈狓狆.犅狅狋.,2008,59(13):36213634.
[8] KITAMURAS,AKITAY,ISHIZAKA H,犲狋犪犾.Molecular
characterizationofananthocyaninrelatedglutathioneStrans
ferasegeneincyclamen[J].犑狅狌狉狀犪犾狅犳犘犾犪狀狋犘犺狔狊犻狅犾狅犵狔,
2012,169(6):636642.
[9] SASAKIN,NISHIZAKIY,犲狋犪犾.Identificationoftheglutathione
Stransferasegeneresponsibleforflowercolorintensityincarna
tions[J].犘犾犪狀狋犅犻狅狋犲犮犺狀狅犾狅犵狔,2012,29(3):223227.
[10] HUB,ZHAOJ,LAIB,犲狋犪犾.犔犮犌犛犜4isananthocyaninre
latedglutathioneStransferasegenein犔犻狋犮犺犻犮犺犻狀犲狀狊犻狊Sonn
[J].犘犾犪狀狋犆犲犾犾犚犲狆.,2016,35(4):831843.
[11] PATTANAIKS,KONG Q,犲狋犪犾.Isolationandfunctional
characterizationofafloraltissuespecificR2R3MYBregula
torfromtobacco[J].犘犾犪狀狋犪,2010,231(5):10611076.
[12] BAIY,PATTANAIKS,PATRAB,犲狋犪犾.Flavonoidrelated
basichelixloophelixregulators,NtAn1aandNtAn1b,ofto
baccohaveoriginatedfromtwoancestorsandarefunctionaly
active[J].犘犾犪狀狋犪,2011,234(2):363375.
[13] ZHUQ,SUIS,LEIX,犲狋犪犾.EctopicexpressionoftheColeus
R2R3MYBTypeproanthocyanidinregulatorgene犛狊犕犢犅3
alterstheflowercolorintransgenictobacco[J].犘犔狅犛犗狀犲,
2015,10(10):e0139392.
[14] YAMAZAKIM,NAKAJIMAJ,YAMANASHIM,犲狋犪犾.
Metabolomicsanddifferentialgeneexpressioninanthocyanin
chemovarietalformsof犘犲狉犻犾犾犪犳狉狌狋犲狊犮犲狀狊[J].犘犺狔狋狅犮犺犲犿犻狊
狋狉狔,2003,62(6):987995.
[15] }~v, ,F.rÝs{MN犌犛犜 z
;²Ï±
[J].%&,2013,40(6):11291138.
JINXH,HONGY,犲狋犪犾.Isolationandexpressionanalysisof
犌犛犜geneencodingglutathioneStransferasefrom犛犲狀犲犮犻狅犮狉狌犲狀
狋狌狊[J].犃犮狋犪犎狅狉狋犻犮狌犾狋狌狉犪犲犛犻狀犻犮犪,2013,40(6):11291138.
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