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Cloning and Expression Analysis of Betalain related Transcription Factor Gene AmMYB1 in Amaranthus tricolor L.

苋菜甜菜红素合成相关基因AmMYB1的克隆及表达分析



全 文 :书!"#$%&
,2016,36(6):1080-1090
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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0123

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4565789:;<,=犃犿犕犢犅1
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 (1,) *1,+ ,1,-./1,0122,
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5 61,7891,:;<1,=>?1
(1(de+% ]^#$J$fg:Ih,i350002;2ijOP,%:Ih,i350111)
F G:klRACE-S,M‘+m’nPopqr1s 犕犢犅;tcDNAuvwx,yz{犃犿犕犢犅1(|}~{
KU557504)。犃犿犕犢犅1;t€G‚{723bp,ƒ„…240†‡;ˆ。‰;tŠwx_cDNA‹ŒŽ,
犃犿犕犢犅1;t‘1†’“。J$U”%•–—˜,AmMYB1™‘2†š›œ MYBžŸ, ¡†¢£œ
R2R3MYB;¤c•–Ž,¥;t„…œ‡;ˆwx_¦Pm§¨©BvMYB1œ¡ª«¬0,­r54%。®¯
°±²³Ž
,AmMYB1´µ±²¶¯°·。¸¹º»±¼PCR•–—˜,犃犿犕犢犅1;tF‘+m’nP½¾
m¿À²œ—­¼0¶Á¿À²

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;犃犿犕犢犅1;t;—­•–
JKDL$
:Q785;Q786 !MNOP:A
犆犾狅狀犻狀犵犪狀犱犈狓狆狉犲狊狊犻狅狀犃狀犪犾狔狊犻狊狅犳犅犲狋犪犾犪犻狀狉犲犾犪狋犲犱犜狉犪狀狊犮狉犻狆狋犻狅狀
犉犪犮狋狅狉犌犲狀犲犃犿犕犢犅1犻狀犃犿犪狉犪狀狋犺狌狊狋狉犻犮狅犾狅狉犔.
XIELiyang1,LIUShengcai1,BAIYu1,LIUYan1,LINMi1,CAIShunliang1,ZHENGXueli2,
XIEXiaoqing1,FENGXin1,CHENGChunzhen1,CHENYukun1,LAIZhongxiong1
(1InstituteofHorticulturalBiotechnology,FujianAgricultureandForestryUniversity,Fuzhou350002,China;2Instituteof
VegetableScienceofFuzhou,Fuzhou350111,China)
犃犫狊狋狉犪犮狋:RACEmethodwasusedtoobtainthecompletecDNAsequenceof犃犿犕犢犅1(Genbankaccession
No.KU557504)from‘Dahong’amararanth(犃犿犪狉犪狀狋犺狌狊狋狉犻犮狅犾狅狉L.),whichbelongto犕犢犅gene.The
犃犿犕犢犅1genecontainsanopenreadingframeof723bpthatencodes240aminoacids.Comparisonbe
tweenthecDNAandgenomicDNAsequencesshowedthat犃犿犕犢犅1genecontainsoneintron.Bioinfor
maticalanalysisrevealedthatAmMYB1isatypicalR2R3MYBcontainingtwoconsecutiveMYBdomains.
BlastXanalysisshowedthatAmMYB1isrelativelyclosetobetalainbiosynthesisrelatedBvMYB1,withan
identityof54% .SubcelularlocalizationanalysisrevealedthattheAmMYB1proteinwaslocatedinthenu
cleus.QuantitativerealtimePCRresultsshowedthattheexpressionlevelof犃犿犕犢犅1wassignificantly
higherintheredpartofleavesthanthatinthegreenpart,andwasalsohigherintheleavesandstemswith
highbetalainconcentrationsthanthatintheroots.Theexpressionof犃犿犕犢犅1wasobviouslyincreased
whenexposuretolight.犃犿犕犢犅1wasfoundtobehighlyexpressedintheredleafvarietycomparedwith
thegreenleafvariety.Therefore,the犃犿犕犢犅1mayplaypositiverolesinregulatingbetalainbiosynthesis
in犃犿犪狉犪狀狋犺狌狊.
犓犲狔狑狅狉犱狊:犃犿犪狉犪狀狋犺狌狊狋狉犻犮狅犾狅狉L.;betalain;犃犿犕犢犅1gene;expressionanalysis
  MYBÔ}t“™‘0ÕÖל MYBžŸ,
؆ MYBžŸÙ51~52†‡;ˆÚ;,ÄÛ
‰ MYBžŸ*ÜÝAœÞ¤,MYBßà•{
4†®9,á 4RMYB、3RMYB、R2R3MYB Â
R1MYB[12]。=Mâãoäå框#$MYBÔ
}t“çè
[3],
éêMëQ#$oìí

F#
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 #$o¬+œÔ}t“òóô¡

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™
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DEFG
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n,
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n,
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[1213],R2R3
MYB"_%,-§œÍÎ。¬6:Iäå[25],
MYBé"_%¦Pm§œÍÎ。Hatlestad1[2526]
\] MYBƒÌé"_%¦Pm§œÍÎ^_`
a

ìí%框ÒÓ¦Pm§œ MYBÔ}t“
„…;t犅狏犕犢犅1。:Iäå:¥;t„…¡†
R2R39Ô}t“,F¦PÂbcno/¼—­
dÌef¦Pm§œÍÎ
;BvMYB1 _,-§
R2R3MYBƒÌ™‘¨¤œfýgc,S‰Vl>
üh¶,-§ R2R3MYB。ij,Hatlestad1ä
å
,犅狏犕犢犅1;tFklmo/¼—­,ÞÌe
f,-§œÍÎ

A0

©¶¦Pm§nB/gœ
:Ioo¶nBpÐo¡P©qr
[2728],
s@F¦
Poäå1s"_¦Pm§nBœ犕犢犅;t[25],
tF‰Y$Qouvwr_¦Pm§nB¨©
犕犢犅;tœ:I&x。
nP
(犃犿犪狉犪狀狋犺狌狊狋狉犻犮狅犾狅狉L.) n,n2¡
yJz{#$

ý|¦Pm§
[29],^
™‘}
~€Â[30]。
nPJv

J$¼+¿
§¼0

 ‚ƒÂ:I¦Pm§œ*Lbc

MYBV{#$o¡9òó„+、…Ì܆œÔ}


F¦Pm§nBÒӇú>?ˆ‰Lf¡Š
‹Œ

{:IŒ¡†¦Pm§犕犢犅;tf%:
I

çŽ{è‘‹Œ¦Pm§nBœÒӇÊ
’±;“

1 ”•Â>–
1.1 Q R
nPQ+bc—(de+%]^#$J$f
g:IhÖñ

ç

+m

nP
、‘RRC241’、‘RRC
110’、‘LalSag’{”•,ƒ˜l™š›ñ^Öñ
¶-80℃œl¶·ˆÂ¦Pm§‚ƒ。
1.2 S T
1.2.1 45U犚犖犃、,=V犇犖犃WXA犮犇犖犃Y
Z[9: žlëŸë RNA‚ƒ`¡¢("£
¤¥pJ$-S‘¦§¨

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#$;tŠDNA`¡¢("£¤¥pJ$-S‘
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nP;tŠDNAœ‚ƒ。l
ª«¼•»»Õ¬
(ThermoElectronCorp.,USA)
­®DNARRNA˜Ë¯Õ,^ l1% °±Ÿ²³
´µ­®‰R¶«

žl GeneRacerTM `¡¢
(TaKaRa)êþ‚ƒœ© RNA·Ô}{cDNA
l¶;tpq

lSYBRExScriptTM¸Ô}`¡¢
(TaKaRa)êþ‚ƒœ© RNA·Ô}{cDNA
l¶±¼PCR•–。
1.2.2 犃犿犕犢犅1,=?@ çNBCIo|}œ¦
P犅狏犕犢犅1;t(|}~JF432080)·¹ˆwxV
{"ºwx

_{¸a»ž(œnPÔ}ŠÝÛ¼

|}~ SRR924089、SRR924090、SRR924091 Â
SRR924092)f‹Œ•–,½¾¿¡s0Õ¡ªœ
cDNA¾À(+Á{264bp)。>ÄÛ¥¾À•Â
18016Ž          BCD,1:nP¦Pm§ÍΨ©;t犃犿犕犢犅1œpqR—­•–
ì2s3’RACEÄhÅ$Â5’RACEÄhÅ
$

ÍGeneRacerTM3’Â3’ÆüÅ$R5’Â5’
ÆüÅ$

ç

+m

nPcDNA {ûÇfÆü
PCR·k,•ÂÈÉ3’Â5’Êõwx。DNAMAN
Œìí¾ÀfuvËÌ
,^
ìËÌa[Å$
犃犿犕犢犅1ORFFÂ犃犿犕犢犅1ORFR(—1),f
PCR·kça[Ë̝³œÑÍ«。¤¹ç‘+
m

nP;tŠDNA{ûÇ,ç犃犿犕犢犅1ORFF
Â犃犿犕犢犅1ORFR(—1){Å$,PCRÈÉ¥;
tDNAwx。çÎÅ$d—"£ÏÍÐ+;t,
-Ñґ¦§¨ÍÎ

PCR·kÈÉÓÔ"ÅLATaqr(TaKaRa)
î˜ÕÖú
。PCR·ksÆ{:94℃×Ø«2min,
94℃ Ø«30s,55~59℃ ÙÚ30s,72℃ÛÜ
1min,30†ÝV,72℃ÛÜ10min。ìíœA
œ¾ÀÞ³ßà
,TApqáƒâ«pqã9,>
f9™PCRÈÉ,‘Aœsäœ9™åæç
uJ$-S

Îè

‘¦§¨®w

1.2.3 \]^_`ab žlDNAMAN6.0é
ƌêþìíœCDSwxf¦ú«’Þr•–,
ì•ÂärÞ²5犅犵犾ⅡÂ犛狆犲ⅠœÎÇêÅ$
犃犿犕犢犅11302FÂ犃犿犕犢犅11302R(—1),T/
PCR>–ìíärÞ²5œAëwx。PCRì
$š‘1302+c^ԑ+íî9DH5α,Fïð
ñ§
(50mg/L)œLBòÓó~;ν¾â«pq
“
,PCR$±Â+crÞa[*Š+cœÑÍ«,
¤¹åçu§¨f®w

žl›ô–*Š+c
\‘EHA105dî9,õldî9ö\–÷øDù
—úf®¯°±²ûü

1.2.4 cdefgDE õl ExPASyProt
param×®„…´µœÊý«+;õlPORT×®
´µ®¯°±²

õlSignalP3.0Server×®´
µ+U~ý

õlNetPhos2.0×®´µ+þˆý
²5

õlDNAMAN6.0éÆfwxËÌR‹
Œ

ÿl Primer5.0éÆfÅ$ì。õl
MEGA5.1éÆoœ NeighborJoining(NJ–)ž
( MYB´µœÔ!fý"。ÿl GeneStucture
DisplayServerF#éÆ(http://gsds.pku.edu.
cn/)•–;tœž。
1.2.5 狇犚犜犘犆犚DE ç犈犉犔1{’ë;t(±
¼Å$ 犈犉犔1F  犈犉犔1R w— 1),žl$%
LightCycler480&,T/ qRTPCR ­® 犃犿
犕犢犅1;tF‘+m’nP½¾œm¿À²ÂÁ¿À
²

Þ¤Š(À²

Þ¤»ÅÉÊÂÞ¤ËQœ—­
B1 hdij
Table1 Listofprimersused
Å$z)
Primername
Å$wx
Primersequence(5’→3’)
ÙÚ*Õ
TM/℃

Description
犃犿犕犢犅13’SP1 GATGAATTGCTTCGTAAGTG
犃犿犕犢犅13P GCTGTCAACGATACGCTACGTAAC
犃犿犕犢犅13’SP2 CGCTACGTAACGGCATGACAGTG
AmMYB13NP CGCTACGTAACGGCATGACAGTG
犃犿犕犢犅15’SP1 TAACCATCGCCATCTACAAC
犃犿犕犢犅15P CGACTGGAGCACGAGGACACTGA
犃犿犕犢犅15’SP2 GGAAGTCTATCTGCTATCAAGG
犃犿犕犢犅15NP GGACACTGACATGGACTGAAGGAGTA
犃犿犕犢犅1ORFF GGCTGGTCGGAAAAATGATACAAC
犃犿犕犢犅1ORFR CGAAGAGATGAGAAAATTAGTATCTCC
RT犃犿犕犢犅1F GGGTCGTTTCACGAAGAGATGAG
RT犃犿犕犢犅1R GCCTTCATTGTTTGGGCC
犈犉犔1F GGGATGCTGGTATGGGAA
犈犉犔1R ACGGGTCATTTCTTCTTCTGGAG
犃犿犕犢犅11302F GAAGATCTATGATACAACCGTTCGTGTCGACA
犃犿犕犢犅11302R GGACTAGTCGAAGAGATGAGAAAATTAGT
55
56
55
57
59
52
59
59
3’RACE
5’RACE
ORFa[ÂgDNAÈÉ
ORFverificationandgDNAamplification
qRTPCR
®¯°±²
Subcelularlocalization
+

à#À•{rÞ²5
Note:Theunderlinedsectionshowrestrictionenzymedigestionsites
2801 ! " # $ % &                   36,
-.

ÄÛº»Å$ìœ/0

Fìí犃犿犕犢犅1
;tuvœ;“ÎìÄhÅ$ RT犃犿犕犢犅1F
ÂRT犃犿犕犢犅1R(—1)。PCR·ksÆ{:95℃
×Ø«30s,95℃Ø«10s,58℃ÙÚ30s,72℃
ÛÜ15s,45†ÝV。õl‘LalSag’‘RRC241’
cDNA˜{1Í$f323ÕÔx45,•ÂŒ
犃犿犕犢犅1;t’";tfÈÉ,ìíë67
#

çͱÅ$Äh«ÂÈÉ89

çÞ¤ÉÊ
cDNA10245™{ûÇfÈÉ。ÄÛ2-ΔΔCt¬
:;t¨Œ—­¼

1.2.6 6578>WX、ka )ƒ0.05g™š
:;œnPŠ(<Ê

=‘1mL75% >),?@A
B1C
,4 ℃ € D 30 min,10000r/min E F
10min,GÎH,lIJ3*KLM,4℃30min,
12000r/minEF10min,ƒÎH3¨,lKjƒw
•»»Õ¬
(UV900,ÎèN–&‘¦§¨)®±
‰F538nmÇO»€。ÄÛ¦Pm§œPQ7»
ÔÝ5.66×104 ¬:¦Pm§œ¼[40]。
2 ³_•–
2.1 45犃犿犕犢犅1,=?@lcdefgDE
ç

+m

nPcDNA{ûÇ,犃犿犕犢犅13’SP1
Â犃犿犕犢犅13’SP2{ÄhÅ$,þ3’RACEÆü
PCRÈÉ,´ µ­®ìí1sR600bpsä(S1,
A);ç犃犿犕犢犅15’SP1Â犃犿犕犢犅15’SP2{Ä
hÅ$

þ5’RACEÈÉ,´ µ­®Žìí1s
R300bpsä(S1,B),®w³—˜,T2†¾
À•Â{600bpÂ317bp。ìíœ5’RACEÊ
õ
、3’RACEÊõÂÔ}ŠoêUœ264bpoV
¾ÀfËÌ

Wír985bpcDNAuv。ÄÛË
Ìwxì犃犿犕犢犅1ORFFR犃犿犕犢犅1ORFR
Å$

fRTPCRÈÉça[Ë̝³ÑÍ«,
³Ž

ÈÉír723bpX¡sä(S1,C),®w
³—˜_Ë̝³¡ª

¥wxyz{犃犿犕犢犅1,
|}~{KU557504。ç‘+m’nPgDNA{ûÇ,
犃犿犕犢犅1ORFFÂ犃犿犕犢犅1ORFR{Å$ÈÉì
í828bpAësä(S1,D)。ìíœ犕犢犅;t
cDNAwx_gDNAwxfF#‹Œ,³Ž¥
;t‘1†’“Â2†jŽ“(S2)。
éƕ–—˜犃犿犕犢犅1;t ORFYvÕ{
723bp,„…240†‡;ˆ(S3)。klProtparam
•–‰Êý«+Ž

¥´µ•“ü{ C1213H1916
N342O369S9,¨ Œ•“+¼{27483.1Da,1´5{
6.33,2¶ˆ«´µ。SignalP3.0ServerU~ý×
®³—˜AmMYB1ZU~ý。InterProÝÛ¼
$±³Ž
,犃犿犕犢犅1„…œ‡;ˆwx™‘2
†¢£œ MYBDNAžŸ,T—˜犃犿犕犢犅1„
…1†R2R3MYBÔ}t“。
2.2 犃犿犕犢犅1m,nijopAqrsDE
FNCBIÝÛ¼of¤c«•–Ž犃犿
犕犢犅1„…œ‡;ˆwx_¦P犅狏犕犢犅1„…œ
‡;ˆwx¡ª«¬0

­r54%。犃犿犕犢犅1
„…œ‡;ˆwx_¦P犅狏犕犢犅1R,-§¨©
犚2犚3犕犢犅 ;t„…œ‡;ˆwxfë*‹Œ,
³ŽAmMYB1_¦PBvMYB1FR3žŸ
’ñFbHLH;w。¥;w_,-§R2R3MYB
obHLH;w^ÞRu¡ª,ñF¡P[hœ‡;
ˆÚ;

,-§R2R3MYB;t„…œ‡;ˆwx
S2 犃犿犕犢犅1œ;tž
Fig.2 Genomicstructureof犃犿犕犢犅1gene
A.3’RACE;B.5’RACE;C.cDNA;D.gDNA;M.DL2000;\]h^{Aësä
S1 犃犿犕犢犅1;tœPCRÈÉ
A.3’RACE;B.5’RACE;C.cDNA;D.gDNA;M.DL2000;Arrowsshowthetargetbands
Fig.1 PCRamplificationof犃犿犕犢犅1
38016Ž          BCD,1:nP¦Pm§ÍΨ©;t犃犿犕犢犅1œpqR—­•–
_#À•—R2žŸ;` #À•—R3žŸ;ATG{abc…“;TGA{dec…“
S3 犃犿犕犢犅1;tœ·¹ˆwxÂf®œ‡;ˆwx
ThesectionunderlinedbystraightlinerepresentstheR2domain;thesectionunderlinedbybreaklinerepresentstheR3domain;
ATGpresentsinitiationcodon;TGApresentsstopcodon
Fig.3 cDNAsequenceanddeducedaminoacidsequenceof犃犿犕犢犅1
BvMYB1(M1ETK3.1){¦P MYB‡;ˆwx;PAP1(NP_176057.1)、AtMYB90(NP_176813.1)、AtMYB113
(NP_176811.1)、AtMYB114(NP_176812.1){klm MYB‡;ˆwx;>‚’{bHLH;w
S4 #$R2R3MYB‡;ˆwx‹Œ
BvMYB1representsaminoacidsequenceofMYBfrom犅犲狋犪狏狌犾犵犪狉犻狊;PAP1(NP_176057.1),AtMYB90(NP_176813.1),
A tMYB113(NP_176811.1),AtMYB114(NP_176812.1)representaminoacidsequencesofMYBfrom犃狉犪犫犻犱狅狆狊犻狊狋犺犪犾犻犪狀犪;
TheboxsectionshowsbHLHmotif
Fig.4 AlignmentofaminoacidsequenceofMYBinplants
4801 ! " # $ % &                   36,
S5 #$R2R3MYBÔ}t“fý"•–
Fig.5 MultiplealignmentofdeducedaminoacidsequencesofAmMYB1andotherR2R3MYBtranscriptionfactors
g5{¦PBvMYB1R{:IoœAmMYB1,‰h‹Œwxd{klmR2R3MYB;ig{klmR2R3MYB•9oœæÏ®ó
S6 AmMYB1_klmR2R3MYBÔ}t“œÔ!fý©Ô
ThedotpointsindicateBvMYB1of犅犲狋犪狏狌犾犵犪狉犻狊andAmMYB1,othersareR2R3MYBfrom犃.狋犺犪犾犻犪狀犪;Elipseshows犃狉犪犫犻犱狅狆狊
MYBfamilysubgroup6
Fig.6 PhylogenetictreecomparingtheaminoacidsequenceofAmMYB1and犃.狋犺犪犾犻犪狀犪R2R3MYBfamilytranscriptionfactors
58016Ž          BCD,1:nP¦Pm§ÍΨ©;t犃犿犕犢犅1œpqR—­•–
FCõ‘(A/S)NDV(I);w,tAmMYB1¥É{
INDV(I),_BvMYB1¡ª(S4)。
nP AmMYB1 _è=Þ¤#$œ R2R3
MYBœfý•–³—˜ AmMYB1_BvMYB1
œfý©Ô¬j
;AmMYB1_,-§ R2R3MYB
k{¡9

t()R/,-§R2R3MYBXlk
{¡9

S5)。{%f¡Šf®犃犿犕犢犅1œmF
…Ì

ž(%AmMYB1R¦PBvMYB1_klm
A、D{GFP´µœÁ¿º»öäSn;B、E{˜oS;C、F{p=S
S7 1302犃犿犕犢犅1GFP´µFDùoœ®¯°±²
AandDimagepresentthegreenfluorescenceofGFP;BandEimagepresentoutlookofonioncelunderbrightfield;
CandFimagepresentthemergedimages
Fig.7 1302犃犿犕犢犅1GFPfusionproteinlocalizedinthenucleioftheonionepidermalcels
A.‘+m’nP½¾;B.»ÅÉÊ(q)ÂÈ»ÉÊ(r)œ7st‘+m’nPuv;C1~C3.‘RRC241’、‘RRC110’‘LalSag’
S8 {`ahlnP”•
A.Leavesof‘Dahong’Amranth;B.7dayold‘Dahong’Amaranthseedlingswithnormallight(left)anddark(right)treatments;
C1C3.RRC241’,‘RRC110’and‘LalSag’
Fig.8 Amaranthmaterialsusedinthisstudy
6801 ! " # $ % &                   36,
R2R3MYBœÔ!fý"(S6),³—˜ Am
MYB1ÂBvMYB1d_klmoæÏ®ók{¡
9

¥óœÎ[ AtMYB113、AtMYB114、PAP1、
AtMYB90d"_ÒÓ,-§ÍÎ,2¶ R2R3
MYB。çΝ³î˜ AmMYB1Þ@_BvMYB1
‘W¤œfýgc

é_ÒÓ,-§ÍΨ©œ
R2R3MYB™‘W¤œfýgc。
2.3 45犃犿犕犢犅1>^_`ab
PORT®¯°×®³—˜AmMYB1ƒÌ±
²¶¯°·

wÍÔ}t“œ®¯°±²Äx

ÿ
lDù—úœy¹—­Œ犃犿犕犢犅1;tf®¯
°±²

³—˜¥;t±²F¯°·

_×®œ
³¡ª

S7)。
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犃犿犕犢犅1BCDE
  ŒnP½¾Þ¤À²o¦Pm§¼œ•–Ž


m½À²œ¦Pm§¼z0

_Á½À²¦P
m§¼[h­r{Ž|3}

S9)。T/º»±
¼•–犃犿犕犢犅1;tFnP½¾m¿À²ÂÁ¿
À²œ±¼—­-.

³—˜
,犃犿犕犢犅1FT2
†À²od‘—­

S9),SFm¿À•œ—­¼
 Á¿À•œ20ë2,~WôV[hŽ|,s­r
{Ž|3}
。犃犿犕犢犅1;tFnP½¾Þ¤À²
—­¼œØý€_nP¦Pm§œ¼Øý€
¡ª

Tî˜犃犿犕犢犅1;tF½¾m¿À²0—
­ƒÌ‘ÿ¶¦Pm§œÍÎ

 —[h{Ž|(犘<0.01)
S9 m½À²ÂÁ½À²¦Pm§¼Â犃犿犕犢犅1
—­¼[h
indicatesignificantdifferenceamongsamplesat0.01level
Fig.9 Differencesofbetalaincontentsand犃犿犕犢犅1
expressionlevelsbetweentheredpartleafand
thegreenpartleaf
2.5 |}~J6578z{w犃犿犕犢犅1B
CDE
  ŒnPÞ¤Š(À²¦Pm§¼œ•–³
Ž

½o¦Pm§¼¬0

Ãoô

Äo¬‚

_Ão¦Pm§¼™‘Ž|[h

t_½o¦P
m§¼œ[h­r{Ž|3}

žl¸¹º»±
¼•–犃犿犕犢犅1FnPÞ¤ƒ(Ä、Ã、½)œ—
­ûü

³—˜犃犿犕犢犅1FnPœÄ、ý
od‘—­
。犃犿犕犢犅1—­¼F½o¬0;FÃ
oô

FÄo—­¼¬‚

_Ão—­¼™‘Ž|
[h

_½o—­¼œ[h­r{Ž|3}
。犃犿
犕犢犅1FnPÞ¤ƒ(Ä、Ã、½)—­¼œØý
€_nPm§œ¼Øý ¡ªœ

S10)。
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犃犿犕犢犅1BCDE
  nPuvFÞ¤»ÅÉÊǦPm§¼•–
—˜

»ÅÉÊefnPuv¦Pm§œÍÎ

È»
ÉÊùúnPuv¦Pm§œÍÎ

S8,B),t„,
»ÅÉÊnPuv¦Pm§œ¼0¶È»ÉÊn
Puv¦Pm§œ¼

S11),[h­r{Ž|3
}

¸¹º»±¼•–犃犿犕犢犅1;tF»ÅÉÊ
…†Éʜ—­ûü

äå犃犿犕犢犅1œ—­¼
F»ÅÉÊoÎÒ

È»ÉÊoÇÒ

»ÅÉÊ
犃犿犕犢犅1—­¼ È»Éʗ­¼œ5.72,~W
ôV[h­r{Ž|3}

FȻ»ÅÉÊo

nP犃犿犕犢犅1—­¼Øý€_‰¦Pm§ÍÎ
Â•Â—F0.05Â0.013}[hŽ|;Ǥ
S10 nPÞ¤ƒo¦Pm§¼Â犃犿犕犢犅1
—­¼[h
andindicatesignificantdifferenceamongsamplesat0.05
and0.01level,respectively;Thesameasbelow
Fig.10 DifferencesofbetalaincontentsandAmMYB1
expressionlevelsindifferentorgans
78016Ž          BCD,1:nP¦Pm§ÍΨ©;t犃犿犕犢犅1œpqR—­•–
S11 »ÅÂÈ»Éʌ¦Pm§¼Â犃犿犕犢犅1
—­œTU
Fig.11 Effectsoflightanddarktreatmentsonbetalain
contentsand犃犿犕犢犅1expression
S12 Þ¤nPËQo¦Pm§¼Â犃犿犕犢犅1
—­¼[h
Fig.12 Differencesofbetalaincontentsand犃犿犕犢犅1
expressionlevelsindifferentAmaranthcultivars
Øý€¨X

S11)。f®»ƒÌT/‡\犃犿
犕犢犅1œ—­ÒÓ¦Pm§œÍÎ。
2.7 |}45‡ˆJ6578z{w犃犿犕犢犅1B
CDE
  ‘RRC241’‘RRC110’2†nPËQF¶†J
vä?/go½¾u{Á¿

S8,C1ÂC2),¦P
m§¼{‚

S12);t‘LalSag’F¶†Jvä?
/go½¾um

S8,C3),|¦Pm§(S12)。
žl¸¹º»±¼•–犃犿犕犢犅1;tFnP½¾
uÁËQ
(‘RRC241’‘RRC110’)ÂnP½¾u
mËQœ—­ûü

³äå犃犿犕犢犅1F½¾u
mœËQ
(‘LalSag’)œ—­¼0¶F½¾uÁœ
ËQ
(‘RRC241’‘RRC110’),[h{Ž|。犃犿
犕犢犅1FT3†Þ¤ËQœ—­¼Øý€_ˆË
Qoœ¦Pm§¼Øý€‰Ñ¨©

S12),î
˜犃犿犕犢犅1ƒÌ_nP¦Pm§ÍÎcÞ¨©。
3 Œ Š
3.1 犃犿犕犢犅1l‰Š8犚2犚3犕犢犅>0‹SŒ
Ž|}

1Ž‘’;}>qr“”
  Hatlestad1[26]äåBvMYB1ÒÓ¦Pm§
œÍÎ

SBvMYB1_,-§ MYBŒ¿§œVl
> ü ^ Þ ¨ ¤

, - § R2R3MYB T / _
bHLH、WD40‹V>üÒÓ,-§ÍÎ,FR3
ž’‘¡†_bHLH´µ‹Vœ(D/E)Lx2(R/
K)x3Lx6Lx3R ;w[3132]。>t,BvMYB1 —¶
bHLH;w²5œŒØ,^ ÞÌ_bHLH‹V[26]。
{`aT/‡;ˆwx‹Œäå
,AmMYB1FR3
žŸ’ñFbHLH;w,S‰;w’œ¡PÍ
²5äJŒØ

¥-.¤ BvMYB1¨X;,-§
R2R3MYB;t„…œ‡;ˆwxFCõ‘(A/
S)NDV(I);w,t AmMYB1_BvMYB1F¥É
{INDV(I)。f®,AmMYB1_,-§ R2R3
MYBœVl>üƒÌÞ¤。AmMYB1ƒÌ—¶
bHLH;woÍ²5äJŒØ,ÞÌ_bHLH
‹V

¦Pm§BvMYB1_,-§R2R3MYB™‘
¨¤œfýgc
[26]。R2R3MYBÒÓ,-§Í
Î
[33],
Fklmoêþ•E¿ë†ÒÓ(9„…
R2R3MYB Ô } t “ œ ; t[34]。At犕犢犅90/
犘犃犘2、At犕犢犅75/犘犃犘1、At犕犢犅113、At犕犢犅114
ÒÓ,-§œÍÎ
[32],
hZ2¶klmœæÏ9®
ó
;At犕犢犅11/犘犉犌2、At犕犢犅12/犘犉犌1、At犕犢犅111/
犘犉犌3ÒÓ()ÍÎ[35],2¶klm据ó;
At犕犢犅123/犜犜2"_/,-§œÍÎ[36],2¶k
lm掮ó

{`aT/•–AmMYB1_,-
§

()R/,-§ R2R3MYBœfý©Ôä
å
:AmMYB1_,-§ R2R3MYBfý©Ô¬
j

T_ Hatlestad1œ³¨X。¤¹•–%
AmMYB1_klmR2R3MYBœfý©Ô,äå
AmMYB1_klmœæÏ9®ófý©Ô¬j,
î˜AmMYB1ƒÌ_,-§ R2R3MYB™‘¨
¤œfýgc

ij

ÿlDù—úœy¹—­Œ AmMYB1
f®¯°±²

³—˜ AmMYB1±²F¯°
·

¥³wÍ MYBÔ}t“œÄx。
3.2 犃犿犕犢犅1•6578>9:JŽ‘’–G
>—˜™š0‹
  犅狏犕犢犅1F|¦Pm§œËQo™‘€
—­
[26]。
,-§犚2犚3犕犢犅 陑9Xœ—­Ä
8801 ! " # $ % &                   36,
«,
hZF,-§¼|œŠ(RËQo—­¼
+

‘ ¤ Í o œ 犔犺犕犢犅6[37]、Á ’ 犜犪犕犢犅3
4犇[38]、“”犔犮犕犢犅1[39]1;犜犪犕犢犅34犇KLF
,-§œQú•–——­
;犔犮犕犢犅1F,-§
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|¦Pm§œ¤¡½¾œm½À²

ƒ

ÃÂ
½
)、
ËQo—­¼z0

tF¦Pm§¼‚œÁ
½À•

Ä

½¾{uÁËQœ—­¼z‚

¥³
_犅狏犕犢犅1R,-§犚2犚3犕犢犅 —­Ä«¨X。
» TU¦Pm§œ*Lt“
[4041]。Wohlpart
1
[42]
äå¦Pm§œÍÎEÞ»

F»ÅÉÊÇ
¦Pm§œ¼ …†Éʜ52。{:Iäå,
»ÅÉÊǦPm§¼0¶È»ÉʦPm§
¼

_ Wohlpart1œ³¡ª,s犃犿犕犢犅1Œ»
UkØý_¦Pm§¼Øý€¡ª

»ƒÌÒ
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犃犿犕犢犅1F¦Pm§œÍÎoƒÌ™‘*LœÑ
˜ÒÓVl

¦Pm§nBpÐ܆



¦Pm§nBp
МÒÓt“R¨©‡úˆ‘™f¡Š:I‹Œ

犃犿犕犢犅1™ÓJ$%…̈‰f¡Ša[,{%
‘š¥;tF¦Pm§ÍÎoœVl

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