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Isolation and Expression Analysis of an Anthocyanin Transport related NtAN9 Gene from Nicotiana tabacum

烟草花青素苷转运相关基因NtAN9的分离与表达分析



全 文 :书!"#$%&
,2016,36(7):1337-1342
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ŒŽ‘{’“
(GST)MN,` ”u犖狋犃犖9(GenBank•–—KX356542)。犖狋犃犖9˜™š‹690bpz›œ
Fž

Œ229‹ŸM ¡M,¢£phi¤GST。犖狋犃犖9MNq¥¦§3‹¨©~2‹ª™~q«。¬­®
¯°ˆ±²³
,NtAN9<´µ¶(犘犲狋狌狀犻犪犺狔犫狉犻犱犪)vwxy{|·¸zGSTMNPhAN9¹º88%zš»¼。
½¾¿Àˆ±©Á
,犖狋犃犖9ˆ±²³
,犖狋犃犖9MNE™ºvwxyzɋvÊËÌ͇κ²Ï,ЇE›vÑzÒⅢÍ(2cm <vÓ<
4cm)²ÏÔÕÏÖ×Ø,ÙEڙvwxyzÛ、܁݇ڲÏ。§Þßà,ˆ‰áÖz犖狋犃犖9â=¹ºãä
犘犺犃犖9zp=,<…†vwxyz{|<åæ·¸。犖狋犃犖9MNzˆ‰<²Ïˆ±,u¿šç:;…†vwx
yz{|èNJMé

=HI

…†

vwxy{|

Ž‘{’“

MN€

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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
SubtropicalAgrobioresources,KeyLaboratoryofPlantFunctionalGenomicsandBiotechnology,EducationDepartmentof
GuangdongProvince,Guangzhou510642China)
犃犫狊狋狉犪犮狋:Tostudyanthocyanintransport,weisolatedafullengthgene犖狋犃犖9(GenBankAccessionNo.
KX356542)by犻狀狊犻犾犻犮狅狀cloningandRTPCRmethodfrom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
glutathionestransferase(GST)family.Itsgenomicstructurecontains3exonsand2introns.Multiplese
quencealignmentshowedthatNtAN9hashighestidentities(88%)withpetuniaPhAN9thatisanantho
cyanintransportrelatedGST.Phylogeneticanalysisshowedthat犖狋犃犖9wasclusteredintoananthocya
nintransportrelatedGSTgroup,andwasanorthologof犘犺犃犖11.Theanthocyanincontentanalysisand
realtimequantitationPCR (qRTPCR)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;glutathionestransferase;geneclone;expression
analysis 
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º
4ã#$,%Ž‘{’“(glutathioneStrans
ferase,GST)、¬&&(¼·¸#$(multidrug
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ì$*+(,
(multidrugandtoxiccompoundex
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“Æ,4
(bilitranslocasehomologue,BTLhomo
logue),â=5GST#$7ÄkJMézšã[1]。
#$GSTĚ㙺¬‹«\zBMN(,,
Û8ÐMNq¥¦#$ŸM Æf¼

ˆuâ9
¼z7ã GST,%φ(phi,F)、τ(tau,U)、θ(theta,
T)、ζ(zeta,Z)、λ(lamda,L)、DHAR(dehydroascor
batereductase) TCHQD(tetrachlorohydroqui
nonedehalogenase),:;Ú9¼<=GST,>?E
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ýþ

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‹€zvwxy·¸犌犛犜ÄLM犅狉狅狀狕犲2MN,
ЌGSTⅢ#$,NOP»vwxyÚ=ELM
Q=‡åæ
[3]。
´µ¶犘犺犃犖9MNŒBronze2
zÆf#$ GSTI,NOP»vwxyÚ=E´µ
¶vR‡åæ

ÐNO²¤=STLM犅狉狅狀狕犲2M
NUV
[4]。


QÿGSTMNNO4WXp=
UV:;

Y€ŠZˆ犌犛犜MN,[\^]犃狋犜犜19[5]、^ _(犘犲狉犻犾犾犪犳狉狌
狋犲狊犮犲狀狊)犘犳犌犛犜1[6]、` a犞狏犌犛犜4[7]、bcd犆犽
犿犌犛犜3[8]、efg犇犮犌犛犜犉2[9]hi犔犮犌犛犜4[10]。
jk4ÑGenBankl8m‡Yº¬I#$GSTM
N

õÚ¬
[1]。
…†ÄšIkJzqrS$

ÆÌJĚIj
£vwxðñst:;zkJuv#$

4х†
vwxyì«òóZˆ¥¦MNZˆ{–stN
~YT€
[1112],
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犌犛犜MN€z&n。G:;ij}~€z‚
ƒ

:´µ¶犘犺犃犖9z ­®u{|,}~…†
ESTl8m,€‚MN­®ö,jRTPCR‚
ƒƒ„ˆ‰…á…†vwxy{|·¸z犌犛犜 M
N犖狋犃犖9,†°ÐEÚÆq‡ÚÆvÊË÷øz
²ÏCˆ¿GŠ:;

:Íun‰ŠMNp=¿Ù
‹±vwxy{|ŒèÇMé

1 Ž‚ƒ
1.1 QRST
„ Q … †
(犖犻犮狅狋犻犪狀犪狋犪犫犪犮狌犿) ‘ ’ I
K326,§mnog“,E”•z–p—˜™+‚。
š›”•ÊËz…†vӁvœ

:;Û

܁Ýg
“£-80℃žŸ ,j£RNA¡¢。
1.2 UV
1.2.1 45,>W犇犖犃XY、6犚犖犃@AZ犮犇
犖犃[\]^_ jSDS‚ƒƒ„¡¢…†£ÝM
Nq DNA;jInvitrogenz Trizol¤¥¡¢…†
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Ü

݁vz4‹ÌÍz¦RNA,§¨’j1μg
¦RNAuu©,šjPromegaz MMuLVª{–
“OligodTPrimer¿G«{–,ì«cDNAҚ
¬

«{–—˜u
:42℃ 40min,50 ℃ 30min,
99℃5min,5℃5min。
1.2.2 456789?XYB`a …†vÊË
z4‹ÌÍ5­PattanaikFz®¯&n[11]2ˆ:
ⅠÍ(vÓ<1cm)、ⅡÍ(1cm<vÓ<2cm)、ⅢÍ
(2cm<vÓ<4cm)ⅣÍ(°vÍ)。…†vw
xyz¡¢šjZhuF&nz‚ƒ[13],j1% ± 
²³4℃´µ¶¡ÿ·,pöjˆµµÕ¸ˆ¹à
Ȩ’E530nm650nmzµº»¼A,½8v
[(A5300.25×A650)/¾¿k]1Àvwxyz·
°™È

1.2.3 45犖狋犃犖9,>bcde?fg šj}
~€z‚ƒ

j´µ¶犘犺犃犖9MN(Y07721)
cDNA­®Su{|,E NCBIÁÂzexpressed
sequencetags(EST)l8mgenomicsurveyse
quence(GSS)l8m¿Gz ­®}~<¯°,á
8331 ! " # $ % &                   36Ã
Ö<犘犺犃犖9ØÕ·äz…†ESTGSS­®,Ä
jÅÆ­®EÇÈl8m‡¿šç¿G}~

×É
…ázʺ­®jDNAman˘°Ê…ázEST
GSS­®¿G¬k¯°Contig€。Û8¬
k¯°€z¥Ì

@15′3′ÍÎCDÏ$
FNtAN9(5′ATGGTAGTGAAGGTGTATGGT
TCAG3′)RNtAN9(5′TTAAAGTTTTACT
TCTTTAAATAGGGGTGCAG3′)j£ÐÑ 犖狋
犃犖9‚cDNAŒÒ­®°ÓzMNq­®。
RTPCR:…†vcDNA Қ¬1μLuu
©

šj TOYOBO ØgÔz KOD FX “¿G
50μLÕÖPCR4½ÐÑ,5l[™:95℃ ×O¼
2min,35‹ÐÑØÙ(95℃ O¼1min,58℃ ÚÛ
1min,72℃ ÜÝ2min),72℃ ÄÜÝ5min。犖狋
犃犖9MNqDNAÐÑ,Þj…†£ÝDNAuu
ߨ

Ð>Ðї˜ÆRTPCR—˜。àÇȅá
zPCRá$âã»ö,qÿ TA €,ä€~
PCRåàö,Xæ¼€~à­。
1.2.4 45犖狋犃犖9,>?hRijk@E 犖狋
犃犖9MNzˆ~Cˆˆ±IJšj VectorNTI
Suite10.0˘˜¿GOpenReadingFrame(ORF)
çèéê

#$ûMG¼û
。NCBIÁÂEëO
¹BLASTˆ±#$g쥦í}~。Ój Ex
PASy、Softberry、CBSPsortFÁÇîì˘˜,
¿GŒ#$zMG¼û

¥¦ClFz×àˆ±

HjCLUSTALX(1.83)˘°ÚÆGST#$­
®¿G¬k¯°:ö

ÄHj Mega6.0˘‡zï
3·ðƒ
(NJƒ),šjBootstrap1000A,ñ«½
¾¿Àòz¦?

1.2.5 45犖狋犃犖9,>?CDlm@E ij
Primer5.0˘,@1犖狋犃犖9MNzmÌóµÇ
È PCR Ï $ FqRTNtAN9(5′CATCTATGAA
CAAAGGTTGTCC3′)RqRTNtAN9(5′CAAA
CACTTTCTCTCCGTTACC3′),: 犖狋犌犃犘犇犎 M
NSuª5

Ï$uFGAPDH(5′CTGCTCACT
TGAAGGGTGGT3′)RGAPDH (5′GGGAG
CAAGGCAATTTGTGG3′)。:…†Û、Ü、݁v
z4‹ÊËÌÍcDNA uuß,šjô+SYBR
GreenMasterMix¤¥õz«Ó4½«ÓP­,
EBioRadIQ5realtimePCR¸Çñ«óµÇÈ
PCR。«ÓP­u:95℃ ×O¼15s,58℃ ÚÛ
15s,72℃ ÜÝ15s,40‹ØÙ。¾¨’kö3A,
šj2-ΔΔCtƒ1ÀMNz·°²Ï。
2 ¥Ìˆ±
2.1 犖狋犃犖9,>?fgBnodepq
Qÿ 犘犺犃犖9MN­® BLASTnˆ±…†
ESTl8mGGSl8mö,÷…á…†犃犖9M
Nz2—EST­®6—GSS­®。šjDAN
man˘¿G EST GSS­®¬k¯°€
ö

…áš—‚2317bpzContig­®。Qÿ¿š
ç}~Contig­®ŒÒORF,Êøš‹º3‹¨
©~2‹ª™~¦«z ORF。BLASTnˆ±²
³Š­®ŒÒ<´µ¶犘犺犃犖9MNz ¹º
89%zš»¼,â:@çùÇÐu…†犃犖9MN

ú1)。
:FNtAN9RNtAN9ÐÑÏ$,RTPCR
Ðх†vcDNAö,…á1—û600bpzCD—
d

âã»
、TA€à­。୥̲³,…†
犃犖9MNcDNAORF‚u690bp,Œ229‹
ŸM 

ú1),` ”u犖狋犃犖9,GenBank•–—u
üýþ~ATGjÿ4²Á,É!~TAAjÿ4"—²Á;
NÎg쥦íGST_N_Phi(M10~K77)jä™2ë²Á;
CÎg쥦íGST_C_Phi(E91~L209)j#™2ë²Á;
$%ugìzGSH&¼3l,‚žugìzB$¥ì3l。
ú‡(l)ð²z l)

*+3,²Á¨©~-z./0
ú1 犖狋犃犖9MNzz ­®;ßǟM ­®
TheATGinitiationcodonisbold,andtheTAAterminationcodon
isinboldandindicatedbyfivepointedstar.TheNterminal
conserveddomainGST_N_Phi(M10~K77)andtheCterminal
conserveddomainGST_C_Phi(E91~L209)aresingleanddouble
underlines,respectively.TheGSHsitesaremarkedwithgray
background,andthesubstratebindingsitesaremarkedinbox.
Thetwoverticalarrowsindicatethesplicesitesbetweenexons
Fig.1 cDNAsequenceandthededucedaminoacid
sequenceof犖狋犃犖9
93317Í          ABC,F:…†vwxy{|·¸MN犖狋犃犖9zˆ‰<²Ïˆ±
KX356542。:…†£ÝgDNA uuß,ÐÑ 犖狋
犃犖9MNqDNA。ÐgDNAKüýþ~ÖÉ!
þ~‚u1538bp,™º3‹¨©~2‹ª™
~

ª™~¦‚u848bp,Їª™~Ⅰu225
bp,ª™~Ⅱu623bp(ú2),­®1ìÕÖzGT
AGª™~2€ƒ3。ÐMNq¥¦<*¬lvw
xy{|·¸zGSTMNz¥¦š»。
2.2 犖狋犃犖9,>#Prs?pqtuvw@E
NtAN9#$‚Õu229‹ŸM (aa),ˆ~È
(Mw)u26.34kDa,F}l(pI)u5.76,Ї4¼
ŸM ¯520.96%,6¼ŸM ¯5u13.97%,
 ¼ŸM ¯5u12.22%。™È×Ô7zu8Ÿ
 Leu,9¦ŸM z13.57%,ÐAÄValLys,
ˆ¹99.17%  8.73%。NtAN9 #$Cˆ<
PhAN9#$ØÕ·ä。Blastpzgìí}~²³,
NtAN9¹ºGST_N_Phi(M10~K77)GST_C_
Phi(E91~L209):‹g쥦í(ú1),;³犖狋
犃犖9MN¢£phiã¤犌犛犜 BMN(,#$«

š<â9¼GSTÄ:#$=Â4z>vGH
p=

¾‹û26kDbM?˜™@Az GSH CD
¥ì3l
(G 3l)¥ìBCB$z3l(H 3
l
),
Q•G3lgì,H3l¥¦âO¼í*。Nt
AN9‡zG3lEÆã¤GST#$‡ÄØÕgì
š»z

ÙH3lgì¼·°í(ú1,ú3)。
¬­®¯°ˆ±Êø
,NtAN9zGST#$­®¹ºíØzš»¼,Ї<´µ¶
zPhAN9¹º×Øzš»¼,Ï88%;<\^]
AtTT19、^ _ PfGST1、` a VvGST4  h i
LcGST4FJ¹ºíØz­®š»¼,ÐDEE
53%~70% ô-。NtAN9<ÅÆ GST#$E“
&p=·¸zgìŸM p=¥¦íDzø+Ø
՚»¼

ú3),;³>?â=¹º·äzp=。
2.3 犖狋犃犖9,>?xyz{|@E
E¬­®¯°zMéÇ

j MEGA6.0˘,
ú2 …†犖狋犃犖9MNq¥¦
Fig.2 Thegenomicstructureofcommontobacco犖狋犃犖9gene
FGHI²Áñš»zŸM ­®

JK"²ÁgìzGSH&¼3l,LK>²ÁB$¥ì3l
ú3 …†NtAN9<Ð>#$YMvwxy{|·¸zGST#$z¬­®¯°
Shadesinblackrepresentidenticalaminoacidsamongthehomologousproteins.HighlyconservedGSHactivesitesaremarked
withfivepointedstar.Triangleindicatessubstratebindingsites
Fig.3 ThemultiplesequencealignmentofNtAN9withknownfunctionanthocyanintransportrelatedGSTproteins
0431 ! " # $ % &                   36Ã
Nlð²G:;zNtAN9E½¾Ê+ò‡z3,
ú4 …†NtAN9#$<Ð>#$ÚÆã¤GST#$z½¾¿Àò
ThedotindicatesNtAN9positioninthephylogenetictree
Fig.4 NeighborjoiningphylogenetictreeofthecompletesequencesofNtAN9andotherplantGST
Ⅰ.vÓ<1cm;Ⅱ.1cm<vÓ<2cm;Ⅲ.2cm<vÓ<4cm;
Ⅳ.°vÍ。™Æ
ú5 …†ÚÆq‡vwxy™Èˆ±
Ⅰ.flowerbud<1cm;Ⅱ.1cm<flowerbud<2cm;Ⅲ.2cm<
flowerbud<4cm;Ⅳ.fulbloomstage.Thesameasbelow
Fig.5 Theanthocyanincontentanalysisintobacco
differenttissues
Xj NJƒ,bootstrap1000A,¦?Š NtAN9M
Nã¤
zGSTMNz½¾¿Àò(ú4)。°¦?zNJò
ˆ±Êø

O‹½¾òˆu3‹*Ã,Ї NtAN9
KP<ÆuQHz´µ¶PhAN9ÂEšü,Ä<
^_PfGST1、` azVvGST4、hiLcGST4^]AtTT19ÂãEšü>«@Azš‹*Ã,Ù
Ð>ã¤z GSTˆ¹>«ÚÆzÃ。ÅRST
NtAN9¢£vwxy{|·¸zGSTMN(,,
¹º·äzvwxy{|p=

îìBlasts、#$
­®¬­®¯°ˆ±½¾¿Àˆ±z¥Ì


犖狋犃犖9MNUÄ´µ¶犘犺犃犖9MNzŽÆf
MN

VÁTйº<ô·Æz{|p=

ú6 犖狋犃犖9MNE…†ÚÆq‡vÚÆ
ÊËÌÍz²Ïˆ±
Fig.6 犖狋犃犖9expressionpatternanalysisindifferent
t obaccotissuesanddifferentflowerdevelopmentalstages
2.4 犖狋犃犖9,>CDlm@E
°…†Û

Ü

݁4‹ÊËÌÍzv(Ⅰ、Ⅱ、
ⅢⅣ)zvwxy™Èˆ±²³(ú5),…†vw
xyIJ“E£v‡

EⅠÍ(vÓ<1cm)ⅡÍ
(1cm<vÓ<2cm)v‡™Èí,WⅢÍ(2cm
<vÓ<4cm)̙ȩXÑY,ÖⅣÍ(°vÍ)Ì
ÏÖ×ؙÈ

ÙEہÐ>ZGq‡‡™È"

qRTPCRˆ±犖狋犃犖9MNE…†§q‡z
²Ï[DÊø

E™ºvwxyz4‹ÊËÌÍz
v‡¹ºíØÔÕ²Ï

ÙEڙvwxy\™È
4]zÛ

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ËzⅠ́Ⅱ͇jkvwxy™Èí,õ犖狋
犃犖9MNz²ÏÔՃ„ÑY,ÖÒⅢÍÌÏÖ×
*²Ï¼

ÙEvwxy™È×ØzⅣ°vÍ,²Ï
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Ś¥Ì;³
,犖狋犃犖9MNz²Ïuv<
vwxyåæuvþ_·¸

J<åævwxyz
14317Í          ABC,F:…†vwxy{|·¸MN犖狋犃犖9zˆ‰<²Ïˆ±
q‡zÊË·¸

3 ` a
vwxyz+$ì«IJÄEùúûªñ«

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cd

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G

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#$GSTB(,ziÆ«\Å
€5<Švwxyz{|

G:;ij}~€j
kRTPCR‚ƒ,K…†v‡ˆ‰Š{|·¸zGST#$MN犖狋犃犖9。
¬­®¯°ˆ±²³
,NtAN9xy{|·¸zGST¹º"Øzš»¼,:;·Æ
zg쥦í
Ї<ÆuQHz´µ
¶PhAN9[4]­®š»¼×ØÏ88%。½¾¿Àò
ˆ±©Á

vwxy{|·¸GSTE$Iô-¹º
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[1] ZHAOJ.Flavonoidtransportmechanisms:howtogo,andwith
whom[J].犜狉犲狀犱狊犘犾犪狀狋犛犮犻犲狀犮犲,2015,20(9):576585.
[2] xbÔ,x y,z{|,F.#$Ž‘{’“Eãêëæ
å‡zSj
[J].#$+[%&,2015,51(11):18151820.
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):18151820.
[3] MARRSKA,ALFENITOMR,LLOYDAM,犲狋犪犾.Aglutathi
oneStransferaseinvolvedinvacuolartransferencodedbythemaize
gene犅狉狅狀狕犲2[J].犖犪狋狌狉犲,1995,375(6530):397400.
[4] ALFENITO MR,SOUERE,GOODMANCD,犲狋犪犾.Func
tionalcomplementationofanthocyaninsequestrationinthe
vacuolebywidelydivergentglutathioneStransferases[J].
犘犾犪狀狋犆犲犾犾,1998,10(7):11351149.
[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):104114.
[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):34943502.
[7] CONNS,CURTINC,B?ZIERA,犲狋犪犾.Purification,molecu
larcloning,andcharacterizationofglutathioneStransferases
(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):36213634.
[8] KITAMURAS,AKITAY,ISHIZAKA H,犲狋犪犾.Molecular
characterizationofananthocyaninrelatedglutathioneStrans
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):636642.
[9] SASAKIN,NISHIZAKIY,犲狋犪犾.Identificationoftheglutathione
Stransferasegeneresponsibleforflowercolorintensityincarna
tions[J].犘犾犪狀狋犅犻狅狋犲犮犺狀狅犾狅犵狔,2012,29(3):223227.
[10] HUB,ZHAOJ,LAIB,犲狋犪犾.犔犮犌犛犜4isananthocyaninre
latedglutathioneStransferasegenein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):831843.
[11] PATTANAIKS,KONG Q,犲狋犪犾.Isolationandfunctional
characterizationofafloraltissuespecificR2R3MYBregula
torfromtobacco[J].犘犾犪狀狋犪,2010,231(5):10611076.
[12] BAIY,PATTANAIKS,PATRAB,犲狋犪犾.Flavonoidrelated
basichelixloophelixregulators,NtAn1aandNtAn1b,ofto
baccohaveoriginatedfromtwoancestorsandarefunctionaly
active[J].犘犾犪狀狋犪,2011,234(2):363375.
[13] ZHUQ,SUIS,LEIX,犲狋犪犾.EctopicexpressionoftheColeus
R2R3MYBTypeproanthocyanidinregulatorgene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
chemovarietalformsof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):987995.
[15] }~v, €,F.rÝsŽ‘{’“MN犌犛犜 zˆ
‰;²Ïˆ±
[J].‚%&,2013,40(6):11291138.
JINXH,HONGY,犲狋犪犾.Isolationandexpressionanalysisof
犌犛犜geneencodingglutathioneStransferasefrom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):11291138.

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