CN106749578A

CN106749578A - Dendrobium myb transcription factor and its encoding proteins and application

Info

Publication number: CN106749578A
Application number: CN201710057319.1A
Authority: CN
Inventors: 李崇晖; 尹俊梅; 杨光穗; 丁灵
Original assignee: Tropical Crops Genetic Resources Institute CATAS
Current assignee: Tropical Crops Genetic Resources Institute CATAS
Priority date: 2017-01-26
Filing date: 2017-01-26
Publication date: 2017-05-31

Abstract

The invention discloses dendrobium myb transcription factor and its encoding proteins and application.Dendrobium myb transcription factor provided by the present invention, is following protein a) or b)：A) protein that the amino acid sequence shown in sequence in sequence table 1 is constituted；B) by the amino acid sequence shown in sequence in sequence table 1 by the substitution of one or several amino acid residues and/or missing and/or addition and as a) derived from the protein related to anthocyanidin synthesis.Transcription factor DhMYB2 provided by the present invention is obtained from dendrobium, belongs to the myb transcription factor family of positive regulation and control anthocyanidin synthesis in monocotyledon.By reverse transcription RT PCR, the complete encoder block of the gene is obtained, then build overexpression carrier and homologous instantaneous conversion dendrobium petal, it is found that the transcription factor has the ability for promoting anthocyanin accumulation.

Description

Dendrobium myb transcription factor and its encoding proteins and application

Technical field

The present invention relates to biological technical field, more particularly to dendrobium myb transcription factor and its encoding proteins and application.

Background technology

Dendrobium (Dendrobium) includes that more than 1500 are planted and many artificial hybrids, mainly grows and plants in Asia heat Band and subtropical zone and Oceania east.Stem of noble dendrobium orchid tint is gorgeous, and flower pattern is various, long fresh-keeping period, is a kind of ornamental value pole Flowers high, belong to (Cattleya) and claim with Phalaenopsis (Phalaenopsis), oncidiumLuridum category (Oncidium) and Bowring cattleya For cattleya is viewed and admired greatly by the world four, it is used widely at aspects such as cut-flower, potted flower, corsage, bouquet, pantry flowers.Petal, sepal and The CF of lip is the topmost fancy points of dendrobium.Stem of noble dendrobium orchid tint mainly has kermesinus, aubergine, pink Color, yellow, green and white etc., and have abundant coloring mode, such as pure color, there are decorative pattern, lip color special.Anthocyanidin It is the primary pigments in dendrobium flower, controls the formation of the color such as purplish red, pink, pink.Breeder focuses on changing in recent years Become stem of noble dendrobium orchid tint and flower coloring mode.But breeding resources are comprehensively collected in conventional hybridization breeding requirement very much, and it is difficult Realize expected breeding objective.With developing rapidly for biotechnology, improve pattern using technique for gene engineering and directive breeding is new Pattern stem of noble dendrobium orchid species will be possibly realized.Dendrobium petal/sepal is typically different with the coloring mode of lip, which imply same The regulation and control model of the anthocyanidin synthesis of complexity is there is in dendrobium flower.The molecule mechanism pair of research regulation and control flower colour generation pattern The improvement of stem of noble dendrobium orchid tint has significant contribution.

R2R3-MYB transcription factors are the key factors for determining anthocyanin synthesis spatiotemporal mode in plant flower organ.Such as gold ROSEA1, ROSEA2 and VENOSA of water shield；AN2, DPL, PHZ of petunia etc..In orchid, 3 MYB of iris Transcription factor gene PeMYB2, PeMYB11 and PeMYB12 control the formula of the overall red, punctation and texture of petal respectively Sample, in lip, PeMYB11 determines the formation of lip corpus callosum punctation, and PeMYB12 controls the entirety of sliver in lip Coloring.

Enzyme gene such as CHS, CHI, F3H, F3'5'H, DFR, ANS expression in dendrobium on rarely seen anthocyanidin route of synthesis The correlation research of amount and pattern.However, the transcriptional control research on anthocyanidin biosynthesis in dendrobium is not yet reported.Mesh Before untill, in dendrobium regulate and control anthocyanidin synthesis myb gene sequence and function have not been reported.

The content of the invention

In order to make up the deficiency in above field, the invention provides dendrobium myb transcription factor and its encoding proteins with should With.

Dendrobium myb transcription factor provided by the present invention, entitled DhMYB2, from stem of noble dendrobium orchid species Dendrobium Blue sapphine No.3。

It is following protein a) or b) it is an object of the present invention to provide a kind of albumen：

A) protein that the amino acid sequence shown in sequence in sequence table 1 is constituted；

B) by the amino acid sequence shown in sequence in sequence table 1 by the substitution of one or several amino acid residues and/or Missing and/or addition and as a) derived from the protein related to anthocyanidin synthesis.

The encoding gene of the albumen falls within protection scope of the present invention.

The encoding gene of the albumen is following 1) or 2) or 3) or 4) shown：

1) its nucleotide sequence is DNA molecular shown in sequence 2 in sequence table；

2) its nucleotide sequence is DNA molecular shown in sequence 3 in sequence table；

1) or 2) 3) DNA molecular for hybridizing with the DNA molecular for limiting under strict conditions；

1) or 2) or 3) 4) there is the DNA molecular of more than 90% homology with the DNA molecular for limiting.

The encoding gene is named as DhMYB2, the albumen for being encoded is named as DhMYB2.The DhMYB2 genes of acquisition The cDNA of total length is 881bp, as shown in sequence 2 in sequence table；Wherein, in sequence table sequence 2 from 5 ' ends the 3rd to the 872nd Position is ORFs, and ORFs part is 870bp, as shown in sequence 3 in sequence table；Sequence 1 in its polynucleotide Shown albumen, sequence 1 is made up of 290 amino acid sequences, and this amino acid sequence has conservative R2R3 repetitive sequences, with list Cotyledon plant anthocyanidin synthesis correlation MYB has conservative higher, and C- ends possess the base of conservative " KAx [K/R] C [S/T] " Sequence.

Expression cassette, recombinant expression carrier, transgenic cell line or recombinant bacterium containing the encoding gene fall within this hair Bright protection domain.

The primer pair for expanding the encoding gene total length or its any fragment falls within protection scope of the present invention；It is described to draw Thing centering, as shown in sequence 4 in sequence table, another primer sequence is as shown in sequence 5 in sequence table for a primer sequence.

Application of the albumen in as transcription factor falls within protection scope of the present invention.

The application of the albumen, the encoding gene in regulation and control anthocyanidin synthesis falls within protection scope of the present invention.

Transcription factor DhMYB2 provided by the present invention is obtained from dendrobium, is belonged to and just regulate and control in monocotyledon The myb transcription factor family of anthocyanidin synthesis.By reverse transcription RT-PCR, the complete encoder block of the gene, subsequent structure are obtained Overexpression carrier and homologous instantaneous conversion dendrobium petal are built, it is found that the transcription factor has the energy for promoting anthocyanin accumulation Power.

Brief description of the drawings

Fig. 1 is the MYB amino acid alignment figures of DhMYB2 and the monocotyledon regulation and control anthocyanidin synthesis reported. PeMYB2, iris Phalaenopsis equestris；PsUMYB6, iris Phalaenopsis schilleriana； OhMYB1, oncidiumLuridum Oncidium hybrid, ZmC1, corn Zea mays.

Fig. 2 is expression of the DhMYB2 in different colours dendrobium petal.BS No.3,‘Blue sapphine No.3’； RB,‘Red bull’；PS,‘Pink stripe’；BWD,‘Burana white dove’.

Fig. 3 is particle gun instantaneous conversion result.A, conversion recombinant plasmid pCXSN-DhMYB；B, converts empty plasmid pCXSN；C, conversion empty plasmid pCAMBIA (1303)；Scale is 200 μm.

Fig. 4 is detected for DhMYB2 transcriptional activation activities.

Specific embodiment

Following embodiment facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiments Method, unless otherwise specified, is conventional method.Test material used in following embodiments, unless otherwise specified, is certainly What routine biochemistry reagent shop was commercially available.Quantitative test in following examples, is respectively provided with three repetitions and tests, and as a result makes even Average.

The clone of embodiment 1, dendrobium DhMYB2 genes

Dongfang Shi Canaan Orchis planting peasant (is purchased from stem of noble dendrobium orchid species Dendrobium Blue sapphine No.3 Specialty Co-operative Organization, catalog number is D20) flower be material, using Tiangeng polysaccharide polyphenol plant total RNA extraction reagent box (purchase From TIANGEN Biotech (Beijing) Co., Ltd., catalog number is DP441) method extraction total serum IgE, use Thermo Scientific reverse transcription reagent box (purchased from the silent winged generation that science and technology of match, catalog number is K1622) synthesis cDNA, as mould Plate.Design full-length cDNA amplification primer：F:5'-CAATGGGAAAAAATCCGTGCT-3'；R:5'- TTGTCCCTAGAATTCACTTCCTAT-3'。

Expanded with above-mentioned template, the response procedures be 94 DEG C of 3min, 94 DEG C of 30S, 56 DEG C of 30S, 72 DEG C of 1min 30S, 30 circulations, 72 DEG C.Archaeal dna polymerase used is Takara ExTaq enzymes.Amplification system：Extaq enzymes 0.4 μ L, 10 × Extaq The μ L of 5 μ L, dNTP mix of buffer 4, forward primer and each 1 μ L of reverse primer, template cDNA1 μ L, totality is supplied with sterilized water It is 50 μ L.Amplified production carries out 1% agarose electrophoresis, reclaims purpose band and is cloned into pMD 18-T carriers (purchased from precious biological Engineering (Dalian) Co., Ltd, catalog number be 6011) on be sequenced.

Result is shown in Fig. 1.The cDNA of the DhMYB2 full length genes of acquisition is 881bp, as shown in sequence 2 in sequence table；Wherein, Sequence 2 is ORFs from 5 ' ends the 3rd to the 872nd in sequence table, and ORFs part is 870bp, such as sequence In table shown in sequence 3；Albumen in its polynucleotide shown in sequence 1, sequence 1 is made up of 290 amino acid sequences, this amino Acid sequence has conservative R2R3 repetitive sequences, has conservative higher, C- to the related MYB of monocotyledon anthocyanidin synthesis End possesses the motif of conservative " KAx [K/R] C [S/T] ".

Through sequence alignment, DhMYB2 is respectively with the iris PeMYB2 and the amino acid identity of corn ZmC1 that have delivered 62.0%th, 43.9%.It is DhMYB2 by the unnamed gene, the albumen for being encoded is named as DhMYB2.

The expression of embodiment 2, DhMYB2 genes in the dendrobium petal of different colours compares

Choose 3 stages during dendrobium flower growth：First stage is small flower bud phase (~0.5 × 1.0-1.5cm: It is wide × high), second stage be big flower bud phase (1.5-2.0cm), phase III just to open the phase (flower is just opened).It is many using Tiangeng Sugared polyphenol plant total RNA extraction reagent box method extract respectively stem of noble dendrobium orchid species ' Blue sapphine No.3 ' (bluish violet), ' Red bull ' (aubergine) (being purchased from Dongfang Shi Canaan Orchis planting farmer specialized cooperative society, catalog number is D4), ' Pink Stripe ' (pink) (being purchased from Dongfang Shi Canaan Orchis planting farmer specialized cooperative society, catalog number is D10), ' Burana 3 hairs of white dove ' (white) (being purchased from Dongfang Shi Canaan Orchis planting farmer specialized cooperative society, catalog number is D16) Total serum IgE in the petal in stage is educated, is that cDNA (uses Thermo Scientific reverse transcription reagent box, closes by total serum IgE reverse transcription Into cDNA).It is ((big purchased from precious bioengineering using Takara SYBR Premix Ex TaqTM II (Tli RNaseH Plus) Even) Co., Ltd, catalog number is RR420A) fluorescence quantitative kit analyze DhMYB2 genes in different colours stem of noble dendrobium orchid Expression in valve.Primer used by real-time fluorescence quantitative PCR detection DhMYB2 genes is as follows：

DhMYB2-qF：5′-TGATTGCTGGAAGGCTACCC-3′；

DhMYB2-qR：5′-TCTTTGTATGATGCTTGGGCG-3′.

The amplified reaction program of real-time fluorescence quantitative PCR：95℃3min；95 DEG C of 10s, 56 DEG C of 30s, 72 DEG C of 30s, 40 are followed Ring.

The relative expression levels of DhMYB2 genes are normalized by internal reference of 18S genes, with 2^–△CTMethod represents gene Relative quantification.

Result is shown in Fig. 2.In bluish violet, aubergine and pink petal, DhMYB2 genes expression quantity before flowering Increase, when being opened at the beginning of phase III flower, expression quantity drastically declines.And DhMYB2 genes are in bluish violet and aubergine petal Expression quantity is more much higher than its expression quantity in pink and white petal.

Embodiment 3, the structure of dendrobium DhMYB2 expression vectors and functional analysis

It is template with the pMD 18-T-DhMYB2 plasmids being sequenced, (has purchased from precious bioengineering (Dalian) with Extaq enzymes Limit company, catalog number is RR001A) amplification (amplification primers F used:5'-CAATGGGAAAAAATCCGTGCT-3'；R： 5'-TTGTCCCTAGAATTCACTTCCTAT-3', primer is with embodiment 1) go out comprising complete ORF and terminator codon DhMYB2 fragments, are cloned by TA and are connected to pCXSN (T- carriers) (Chen S, Songkumarn P, Liu J, Wang G.A versatile zero background T-Vector system for gene cloning and functional genomics.Plant Physiology,2009,150:1111-1121) on, and be sequenced confirmation connection it is errorless, extraction builds Overexpression plasmid pCXSN-DhMYB2 it is standby.

Using GJ-1000 high pneumatic gene guns by pCXSN-DhMYB2 and control plasmid pCXSN it is unloaded and pCAMBIA1303(Hajdukiewicz P,Svab Z,Maliga P.The small,versatile pPZP family of Agrobacterium binary vectors for plant transformation.Plant Mol.Biol.,1994,25 (6):989-994) ' Burana charming ' (are purchased from Dongfang Shi Canaan Orchis planting agriculture to unloaded bombardment importing stem of noble dendrobium orchid species People Specialty Co-operative Organization, catalog number is D5) white petal in.Biolistic bombardment parameter is：Pressure 7MPa, target distance 55cm, vacuum is -0.085MPa.Petal is transferred on MS culture mediums after bombardment, is sealed.At 25 DEG C, 16h light/8h is dark Photoperiod under cultivate 2 days, observed with Stereo microscope.

Result is shown in Fig. 3.Transient expression DhMYB2 genes cause white Petal to generate the pigment spot of purple.Conversion PCXSN unloaded petal does not produce any pigment spot.Petal after conversion pCAMBIA1303, dyes through GUS and produces Blue spot, shows transient expression success.

The transcriptional activation activity analysis of embodiment 4, DhMYB2 albumen

DhMYB2 genes are subcloned in pGBKT7 carriers (clontech, product ID 630489), step is as follows：

DhMYB2-pMD 18-T plasmids are as template with embodiment 1, with primer：F：5'- CCCATATGGGAAAAAATCCGTGCT-3'；R：5'-CATGCCATGGCTAGAATTCACTTCCTATATC-3' amplifications are included The DhMYB2 fragments of NdeI and NcoI restriction enzyme sites, complete ORF and terminator codon.

DNA fragmentation and pGBKT7 carriers NdeI (purchased from the silent winged generation that science and technology of match, product ID FD0583) and NcoI After (purchased from the silent winged generation that science and technology of match, product ID FD0573) restriction enzymes double zyme cutting, (it is purchased from T4DNA ligases Precious bioengineering (Dalian) Co., Ltd, catalog number is 2011A) connection structure fusion vector, it is sequenced and confirms that connection is errorless, The plasmid pGBK-DhMYB2 that extraction builds is standby.

Using the complete conversion reagent box of a small amount of yeast cells of outstanding U.S.'s gene, pGBK-DhMYB2 is imported according to specification Expression, is negative control to convert pGBKT7 zero loads in yeast Y2HGold bacterial strains (clontech).

The yeast Y2HGold inoculations after genes of interest and empty carrier will be converted and lack ferment to SD-Trp (tryptophan) On female defect culture medium, screening positive clone.Positive colony is transferred to SD-Trp-Ade (adenine)-His (histidine) three Lack yeast defect culture medium further to screen, finally lacking culture medium with containing X- α-gal three verifies that α-galactosidase live The method of property, detects its transcriptional activation activity.

SD culture medium prescriptions：0.67%YNB (Yeast Nitrogen Base without Amino Acids), 2% Portugal Grape sugar, 0.06%Ade/-His/-Leu/-Trp DO supplement, 1%100 × AA (amino acid) solution, 2% agar.

Wherein 100 × AA is formulated：Adenine 0.5g is weighed respectively, and histidine 2.0g, leucine 10.0g is dissolved in 1000ml's In water.

Wherein YNB gives birth to work, product serial number A610507 purchased from Shanghai；Glucose is purchased from Sigma, product serial number G8270； Ade/-His/-Leu/-Trp DO supplement are purchased from Clontech, product serial number 630428；L-Leu, purchased from Shanghai Raw work, product serial number A100811-0025；Adenine, work, product serial number A600013-0025 are given birth to purchased from Shanghai；L-Histidine, Work, product serial number 5934-29-2 are given birth to purchased from Shanghai.

Result is shown in Fig. 4.Lack in Trp- mono- and converted on culture medium pGBKT7 zero loads and recombinant plasmid pGBKT7-DhMYB2 With normal growth, show to convert successfully.Lack on culture medium three, conversion pGBKT7 unloaded yeast strain can not grow, and turn The yeast strain for changing recombinant plasmid pGBKT7-DhMYB2 can be with normal growth, and through X-gal Activity determinations, can be anti-with X-gal Should develop the color.The above results prove that DhMYB2 has transcriptional activation activity.

Sequence table

<110>Tropical Crop Variety Resource Institute of Chinese Academy of Tropical Agricultural Sciences

<120>Dendrobium myb transcription factor and its encoding proteins and application

<130> P160686/RKY

<160> 5

<170> PatentIn version 3.5

<210> 1

<211> 290

<212> PRT

<213>Stem of noble dendrobium orchid species Dendrobium Blue sapphine No. 3

<400> 1

Met Gly Lys Asn Pro Cys Cys Phe Lys Glu Gly Leu Asn Lys Gly Ala

1 5 10 15

Trp Thr Thr Ser Glu Asp Met Leu Leu Lys Ala Phe Val Asn Ile His

20 25 30

Gly Glu Gly Lys Trp Thr Thr Val Pro His Lys Ala Gly Leu Arg Arg

35 40 45

Ser Gly Lys Ser Cys Arg Leu Arg Trp Leu Asn Tyr Leu Arg Pro Asn

50 55 60

Val Lys His Gly Asn Phe Ser Glu Glu Glu Asp Asp Leu Ile Ile Arg

65 70 75 80

Leu His Lys Leu Leu Gly Asn Arg Trp Ser Leu Ile Ala Gly Arg Leu

85 90 95

Pro Gly Arg Thr Asp Asn Glu Ile Lys Asn Tyr Trp Asn Ile Thr Leu

100 105 110

Cys Lys Lys Ala Ser Phe Gln His Gln Met His Gln Pro Ser Arg Pro

115 120 125

Ser Ile Ile Gln Arg Pro Pro Thr Ser Asn Leu Gly Phe Pro Thr Ser

130 135 140

Ser Ser Thr Thr Gln Ser Gln Ala Ala Thr Asn Asp Thr Thr Leu Ile

145 150 155 160

Leu Thr Thr Ala Ile Arg Cys Asn Lys Leu Ala Ile Pro Met Leu Leu

165 170 175

Pro Ser Ser Ser Thr Ser Lys Gln Glu Ile Pro Asn Met Gln Leu Ala

180 185 190

Glu Glu Ser Ile Ala Lys Val Ala Ser Met Ile Pro Glu Ser Ser Lys

195 200 205

Val Gly Pro Leu Val Glu Glu Asp Leu Phe Lys Glu Leu Phe Gln Val

210 215 220

Glu Glu Asn Met Val Leu Asn Asn Asp Lys Phe Asn Asp Asp Asn Ile

225 230 235 240

Val Ser Phe Pro Asp Gln Ala Ala Val Met Glu Phe Glu Arg Leu Gln

245 250 255

Asp Phe Glu Lys Trp Met Leu Asn Asp Glu Asp Val Asp Cys Leu Pro

260 265 270

Pro Asn Asp Gln Met Arg Met Leu Thr Ser Leu Phe Asp Ile Gly Ser

275 280 285

Glu Phe

290

<210> 2

<211> 881

<212> DNA

<213>Stem of noble dendrobium orchid species Dendrobium Blue sapphine No. 3

<400> 2

caatgggaaa aaatccgtgc tgcttcaagg aagggcttaa caagggagca tggactactt 60

cggaagacat gctcttgaag gctttcgtca atattcatgg agaaggcaaa tggacgacgg 120

tgccacacaa agcagggctg agaaggtcgg ggaagagctg tcgactccga tggctaaatt 180

acctgaggcc caacgttaaa catggaaact tttccgagga agaggacgac ctcatcatca 240

gacttcataa actccttggc aatagatggt ctctgattgc tggaaggcta cccggtcgga 300

cagataatga aataaaaaac tattggaaca taaccttatg taagaaagca agctttcaac 360

atcaaatgca tcagccaagc cgcccaagca tcatacaaag acctccaact tctaatcttg 420

gatttccaac atcatcatct acaacacaat cacaagctgc cacaaatgat acaactttga 480

tcctaacaac ggccataagg tgcaataaac tggctattcc aatgctactt ccatcttctt 540

caacaagtaa gcaggagatc ccaaacatgc aattagcaga agagtcaata gctaaagtgg 600

ctagtatgat tcctgaaagc agcaaagttg gtcccttggt agaagaagat ctttttaagg 660

aactatttca ggtggaggag aatatggttt tgaataacga caaatttaat gatgacaata 720

ttgtttcatt tcctgatcaa gcggctgtaa tggagtttga aagattacaa gattttgaga 780

aatggatgct gaatgatgaa gatgttgatt gccttcctcc taatgatcaa atgcgtatgt 840

tgacctcttt atttgatata ggaagtgaat tctagggaca a 881

<210> 3

<211> 870

<212> DNA

<213>Stem of noble dendrobium orchid species Dendrobium Blue sapphine No. 3

<400> 3

atgggaaaaa atccgtgctg cttcaaggaa gggcttaaca agggagcatg gactacttcg 60

gaagacatgc tcttgaaggc tttcgtcaat attcatggag aaggcaaatg gacgacggtg 120

ccacacaaag cagggctgag aaggtcgggg aagagctgtc gactccgatg gctaaattac 180

ctgaggccca acgttaaaca tggaaacttt tccgaggaag aggacgacct catcatcaga 240

cttcataaac tccttggcaa tagatggtct ctgattgctg gaaggctacc cggtcggaca 300

gataatgaaa taaaaaacta ttggaacata accttatgta agaaagcaag ctttcaacat 360

caaatgcatc agccaagccg cccaagcatc atacaaagac ctccaacttc taatcttgga 420

tttccaacat catcatctac aacacaatca caagctgcca caaatgatac aactttgatc 480

ctaacaacgg ccataaggtg caataaactg gctattccaa tgctacttcc atcttcttca 540

acaagtaagc aggagatccc aaacatgcaa ttagcagaag agtcaatagc taaagtggct 600

agtatgattc ctgaaagcag caaagttggt cccttggtag aagaagatct ttttaaggaa 660

ctatttcagg tggaggagaa tatggttttg aataacgaca aatttaatga tgacaatatt 720

gtttcatttc ctgatcaagc agctgtaatg gagtttgaaa gattacaaga ttttgagaaa 780

tggatgctga atgatgaaga tgttgattgc cttcctccta atgatcaaat gcgtatgttg 840

acctctttat ttgatatagg aagtgaattc 870

<210> 4

<211> 21

<212> DNA

<213> Artificial Sequence

<220>

<223>CDNA amplimers F

<400> 4

caatgggaaa aaatccgtgc t 21

<210> 5

<211> 22

<212> DNA

<213> Artificial Sequence

<220>

<223>CDNA amplimers R

<400> 5

caaaacttca tccctagcca ca 22

Claims

1. a kind of albumen, is following protein a) or b)：

B) substitution by the amino acid sequence shown in sequence in sequence table 1 by one or several amino acid residues and/or missing And/or addition and as a) derived from the protein related to anthocyanidin synthesis.

2. the encoding gene of albumen described in claim 1.

3. encoding gene according to claim 2, it is characterised in that：The encoding gene for it is following 1) or 2) or 3) or 4) It is shown：

4. expression cassette, recombinant expression carrier, transgenic cell line or recombinant bacterium containing encoding gene described in Claims 2 or 3.

5. the primer pair of encoding gene total length or its any fragment described in Claims 2 or 3, in the primer pair, one are expanded As shown in sequence 4 in sequence table, another primer sequence is as shown in sequence 5 in sequence table for primer sequence.

6. application of the albumen described in claim 1 in as transcription factor.

7. application of the albumen, encoding gene described in Claims 2 or 3 described in claim 1 in regulation and control anthocyanidin synthesis.