CN114561402A

CN114561402A - Malanian indole metabolism related gene BcWRKY1, protein and application thereof

Info

Publication number: CN114561402A
Application number: CN202210307409.2A
Authority: CN
Inventors: 曾美娟; 刁勇
Original assignee: Huaqiao University
Current assignee: Huaqiao University
Priority date: 2022-03-25
Filing date: 2022-03-25
Publication date: 2022-05-31
Anticipated expiration: 2042-03-25
Also published as: CN114561402B

Abstract

The invention provides a cerulenin metabolism related gene BcWRKY1, a protein and an application thereof, wherein the gene BcWRKY1 has a nucleotide sequence shown as SEQ ID No. 1; 1 nucleotide sequence, allele or derivative generated by adding, substituting, inserting or deleting one or more nucleotides in the nucleotide sequence; the protein coded by the kalanchoe metabolic related gene BcWRKY1 has an amino acid sequence shown as SEQ ID No. 2; the gene BcWRKY1 has a nucleotide sequence shown as SEQ ID No.1, and the gene BcWRKY1 and a nucleotide sequence, an allele or a derivative generated by adding, substituting, inserting or deleting one or more nucleotides in the nucleotide sequence are applied to indole acetate for regulating tryptophan metabolism in Arabidopsis.

Description

Malanian indole metabolism related gene BcWRKY1, protein and application thereof

[ technical field ] A method for producing a semiconductor device

The invention relates to the field of genetic engineering, and in particular relates to a cerulenin metabolism related gene BcWRKY1, and a protein and application thereof.

[ background of the invention ]

The Chinese kale (Baphica canthus cuiia (Nee) Bremek) is a plant of acanthaceae, also called south isatis, is a perennial herb, prefers warm and sunny climate environment, has a long clinical medicinal history, underground rhizomes and roots of the Chinese kale are often used as raw materials of medicinal materials and preparations of isatis roots, overground leaves and stems and leaves are often used as main processing raw materials of Chinese medicinal indigo, and are collected in Chinese pharmacopoeia. Indigo naturalis has effects of clearing heat and detoxicating, cooling blood and resolving macula, clearing pathogenic fire and arresting convulsion, relieving pain and resisting inflammation. The south isatis root is dry root and rhizome of Indigowoad root (B.cusia) and is called as the south north isatis root together with isatis root derived from isatis root or rhizome, has good antiviral, antibacterial and anti-inflammatory effects, and plays an important role in preventing and treating epidemic diseases such as SARS, avian influenza, influenza A and the like. The main medicinal components of the kalanchoe comprise indigo blue and indirubin which are related to the synthesis path of indole. In the early stage of a subject group, in the research on the excavation and function of forming key genes of medicinal substances of the kalanchoe odorata, 9 transcription factors including WRKY are found to possibly play an important regulation and control role in the metabolic process related to the kalanchoe odorata.

Transcription factors are an important class of regulatory factors that regulate the progress of life activities. WRKY transcription factors are a superfamily of transcription factors that are predominantly present in plants. The main structural feature of WRKY is that there are one or two WRKY domains in the DNA binding domain of each member. The WRKY transcription factor has close relation with biotic or abiotic stress response, plant growth and development and aging. Studies of Populus chinensis et al in the medicinal plant Catharanthus roseus find that CrWRKY may participate in the synthesis of terpenoid indole alkaloids and stress reaction. Research shows that WRKY can participate in disease-resistant regulation in a model plant Arabidopsis thaliana and terpenoid indole alkaloid metabolic pathway in catharanthus roseus, and WRKY can play related functions by participating in jasmonic acid pathway.

However, in the study on the aspect of the horse blue, no report that the corresponding gene is cloned and the function is studied is found. Considering that there is no mature genetic transformation system of Kalanchoe, the model plant Arabidopsis thaliana was chosen to carry out the relevant research work.

[ summary of the invention ]

One of the technical problems to be solved by the invention is to provide a malkane metabolism related gene BcWRKY1, which can significantly up-regulate indole acetate metabolized by tryptophan in Arabidopsis thaliana and has influence on Arabidopsis thaliana metabolism.

The present invention achieves one of the above technical problems:

a cerulean indole metabolism related gene BcWRKY1, wherein the gene BcWRKY1 has a nucleotide sequence shown as SEQ ID No. 1.

Further, the nucleotide sequence, allele or derivative generated by adding, substituting, inserting or deleting one or more nucleotides in the nucleotide sequence of SEQ ID No. 1.

Further, a plasmid, a plant expression vector and a host cell of the cerulean indole metabolism-related gene BcWRKY 1.

The second technical problem to be solved by the invention is to provide a protein encoded by a cerulenin metabolism related gene BcWRKY1, which can significantly up-regulate indole acetate metabolized by tryptophan in Arabidopsis thaliana and has influence on Arabidopsis thaliana metabolism.

The invention realizes the second technical problem in the following way:

a protein coded by a cerulean indole metabolism related gene BcWRKY1 has an amino acid sequence shown as SEQ ID No. 2.

The invention aims to solve the third technical problem and provides application of a cerulenin metabolism related gene BcWRKY 1.

The invention realizes the third technical problem in the following way:

an application of a cerulean indole metabolism related gene BcWRKY1 is provided, wherein the gene BcWRKY1 has a nucleotide sequence shown as SEQ ID No.1, and the gene BcWRKY1 and a nucleotide sequence, allele or derivative generated by adding, substituting, inserting or deleting one or more nucleotides in the nucleotide sequence thereof are applied to indole acetate for regulating tryptophan metabolism in Arabidopsis.

The invention has the following advantages:

the kalimeris indica metabolism related gene or protein can remarkably up-regulate indole acetate metabolized by tryptophan in arabidopsis thaliana, and influences the arabidopsis thaliana metabolism.

[ description of the drawings ]

The invention will be further described with reference to the following examples with reference to the accompanying drawings.

FIG. 1 is an electrophoresis chart showing the result of PCR amplification of the BcWRKY1 gene of example 1 of the present invention.

FIG. 2 is a fluorescent image of transgenic Arabidopsis thaliana overexpressing BcWRKY1 in example 2 of the present invention, wherein: (a) and (c) under the white light channel, (b) and (d) under the fluorescent channel; WT: wild type arabidopsis, BcWRKY 1: transgenic Arabidopsis thaliana overexpressing BcWRKY 1.

FIG. 3 is a KEGG enrichment pathway diagram of tryptophan metabolism of transgenic Arabidopsis thaliana and wild type Arabidopsis thaliana overexpressing BcWRKY1 according to example 3 of the present invention.

FIG. 4 is a comparison histogram of metabolites related to the metabolism of transgenic Arabidopsis overexpressing BcWRKY1 and wild type Arabidopsis indole according to example 3 of the present invention;

wherein: com _889_ pos: N-Acetyl-DL-tryptophan; com _3035_ pos: 5- {4- [ (5-Azidopentyl) carbamoyl ] phenyl } -2,3,3,7,7,8-hexamethyl-12-sulfo-2,3,7, 8-tetrah-dro-1H-pyro [3',2':6,7] chromeno [3,2-f ] indole-9-ium-10-sulfonate; com _2586_ pos: s- (6- {4- [ (1,3-Dioxo-1, 3-dihydro-2H-isoindolol-2-yl) methyl ] phenoxy } hexyl) propanoethioate; com _1515_ pos: (6aS,7aS,12aS) -12-Acetyl-7a- (2-methyl-3-b uten-2-yl) -7,7a,12,12a-tetrahydroindolo [3',2':4,5] pyrrolo [2,1-c ] [1,4] benzodiazepin-6, 14(5H,6aH) -dione; com _252_ pos: indole-3-acetic acid.

[ detailed description ] A

The invention relates to a cerulenin metabolism related gene BcWRKY1, wherein the gene BcWRKY1 has a nucleotide sequence shown as SEQ ID No. 1.

The nucleotide sequence, allele or derivative generated by adding, substituting, inserting or deleting one or more nucleotides in the nucleotide sequence of SEQ ID No. 1.

The invention also relates to a plasmid, a plant expression vector and a host cell of the cerulenin metabolism related gene BcWRKY 1.

The invention also relates to a protein coded by the cerulenin metabolic related gene BcWRKY1, and the protein has an amino acid sequence shown as SEQ ID No. 2.

The invention also relates to application of the cerulenin metabolism related gene BcWRKY1, wherein the gene BcWRKY1 has a nucleotide sequence shown as SEQ ID No.1, and the gene BcWRKY1 and a nucleotide sequence, allele or derivative generated by adding, substituting, inserting or deleting one or more nucleotides in the nucleotide sequence are applied to indole acetate for regulating tryptophan metabolism in arabidopsis thaliana.

The technical solution of the present invention will be clearly and completely described with reference to the accompanying drawings 1-4 and the detailed description. All other embodiments, which can be obtained by a person skilled in the art without making any creative effort based on the embodiments in the present invention, belong to the protection scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.

Example 1

Construction of kalanchoe metabolic related coding gene BcWRKY1 gene overexpression vector

1. Acquisition of kalanchoe metabolic related coding gene BcWRKY1

The target gene is obtained by PCR amplification with primer1 (shown as SEQ ID No: 3) and primer2 (shown as SEQ ID No: 4) using the reverse transcription cDNA of the kalant as a template:

(primer1)WRKY-for-5941GFP-F：5'-TTACAATTACCATGGGGCGCGCCAT GGAGAGCATGGATGATTA-3'；

(primer2)WRKY-for-5941GFP-R：5'-GCCCTTGCTCACCATGGCGCGCCC AGGTGCAGAAGGTAGATGA-3'；

and (3) PCR system:

PCR amplification System (50. mu.L):

PCR conditions were as follows:

the PCR products were detected by electrophoresis on a 1.0% agarose gel for a total of 38 cycles.

After the PCR product is recovered and purified, the PCR product is connected with a pEASY-Blunt cloning vector (full-scale gold biotechnology), then the connection product is converted into escherichia coli Trans1-T1 competent cells, a single colony grows on a flat plate, colony PCR detection is carried out, positive bacteria are cultured overnight at 37 ℃, plasmids are extracted and identified by plasmid PCR, and the identified positive clone is sent to Fuzhou Shang sub biotechnology limited for sequencing; the electrophoresis chart of the PCR amplification result of the BcWRKY1 gene is shown in FIG. 1.

Sequencing results show that the PCR product obtained by amplification is a nucleic acid sequence with a sequence shown as SEQ ID NO.1, a fragment with the length of 534bp and named as BcWRKY1 gene, and the amino acid sequence of the protein coded by the BcWRKY1 gene is shown as SEQ ID NO. 2.

2. Construction of kalanchoe metabolic related coding gene BcWRKY1 gene overexpression vector

2.1 primer design

When an overexpression vector is constructed, a primer for amplifying the coding region of the ceruleus maryland BcWRKY gene:

WRKY-for-5941GFP-F：5'-TTACAATTACCATGGGGCGCGCCATGGAGAGCATGGATGATTA-3'

WRKY-for-5941GFP-R：5'-GCCCTTGCTCACCATGGCGCGCCCAGGTGCAGAAGGTAGATGA-3'

2.2 PCR amplification System and conditions

PCR amplification System (50. mu.L):

PCR conditions were as follows:

2.3 glue recovery

The operation is carried out by referring to the specification of the agarose gel DNA recovery kit of Beijing all-style gold biotechnology Limited, and the steps are as follows:

(1) cutting a target DNA strip, putting the cut target DNA strip into a clean eppendorf tube, adding 3 times volume of solution GSB (gel SolubilizationBuffer), melting the glue on a thermostatic mixer at 55 ℃ for 10 minutes, turning the glue 5 times up and down at intervals of 2 minutes, and then putting the glue back to the thermostatic mixer to ensure that the glue blocks are completely melted.

(2) Cooling the melted gel solution to room temperature, sucking into the centrifugal column with a pipette, standing for 1min, centrifuging at 12000rpm for 1min, pouring the effluent back into the centrifugal column, centrifuging at 12000rpm for 1min, and pouring the effluent.

(3) Pipet 650. mu.L of WB (Wash buffer) solution into the spin column, centrifuge at 12000rpm for 1min, and pour off the effluent.

(4) Centrifuging at 12000rpm for 2min to remove the residual WB in the column.

(5) Placing the centrifugal column into a clean eppendorf tube, opening the eppendorf tube, standing for 1min to volatilize the residual ethanol, adding 40 μ L of deionized water preheated by a constant temperature mixer at 65 ℃ into the center of the column, and standing for 1min at room temperature.

(6) The DNA was eluted by centrifugation at 12000rpm for 1 min.

The plasmid is connected with a pEASY-Blunt cloning vector (all-purpose gold biotechnology), then the connection product is transformed into escherichia coli Trans1-T1 competent cells, single colonies are selected and grown on a plate, colony PCR detection is carried out, positive bacteria are cultured overnight at 37 ℃, plasmids are extracted and identified by plasmid PCR, and positive clones are sequenced by Fuzhou Shanghai sub biotechnology limited.

2.4 Single enzyme digestion 5941-35S-GFP vector

The enzyme is cut for 2h at 37 ℃, and then the enzyme cutting product is recovered by glue.

2.5 one-step cloning

Referring to the operation instruction of the rapid and efficient cloning kit of Nanjing NuoZan Biotechnology GmbH, the single enzyme digestion vector and the PCR product are mixed and added into the reaction system, which is as follows:

(1) preparing a recombination reaction system:

(2) and (4) lightly sucking and uniformly mixing by using a pipette gun, and collecting the reaction solution to the bottom of the tube by short-time centrifugation.

(3) At 50 ℃ for 5min, immediately cooled on ice.

(4) mu.L of the recombinant product was taken and transformed into 30. mu.L of E.coli Trans-T1.

2.6 transformation of E.coli Trans-T1

With reference to the specification of Trans1-T1 Phage resist chemical company Unit Cell of King's Biotechnology Limited, the specific method is as follows:

(1) the competent cells were thawed on ice, and 5. mu.L of the recombinant product was pipetted into 30. mu.L of the competent cells and gently whipped into a uniform mixture and ice-washed for 30 min.

(2) The heat shock was applied to the water bath at 42 ℃ for 40s and immediately placed on ice for 3min without shaking the centrifuge tube.

(3) 600 μ LLB broth was added and shaken at 37 ℃ and 200rpm for 1 h.

(4) Centrifugation was carried out at 7000rpm for 5min, and the cells were collected. 500. mu.L of the supernatant was aspirated in an ultraclean bench, and the remaining cells and supernatant were resuspended, and plated on LB solid medium containing Kan.

(5) The cells were cultured overnight at 37 ℃ by inversion.

2.7 identification of recombinants

And after the single bacterium grows on the LB plate, carrying out PCR identification on the recombinant. The method comprises the following steps:

(1) single colonies were randomly picked on the plates and picked with a pipette tip into 600. mu.L of LB liquid medium containing Kan resistance, shaken at 37 ℃ and 200rpm for 4 h.

(2) PCR amplification is carried out by using a primer WRKY-for-5941GFP-F/WRKY-for-5941GFP-R and 2 xTaq MasterMix and taking a bacterial solution as a template. The PCR amplification reaction (10. mu.L) was as follows:

PCR conditions were as follows:

(3) 10. mu.L of the positive bacterial suspension was aspirated, and the mixture was added to 5mL of LB liquid medium containing Kan, and cultured overnight at 37 ℃ and 200 rpm.

2.8 plasmid extraction

Plasmid extraction is carried out by adopting a plasmid miniextraction kit of TianGen with slight modification, and the method comprises the following steps:

(1) 5mL of overnight-cultured bacterial solution was aspirated into an eppendorf tube by a pipette, centrifuged at 12000rpm for 1min to collect the cells, and the supernatant was gently aspirated.

(2) Mu.l of LBuffer P1 (containing RNaseA) was pipetted into an eppendorf tube containing the pellet and the pellet was suspended with a shaker.

(3) The 250. mu.L of LBuffer P2 was pipetted into an eppendorf tube using a pipette and gently turned up and down 8 times, so that the suspended cells could be lysed sufficiently.

(4) The eppendorf tube was pipetted with a pipette at 350. mu.L BufferP3 and gently turned up and down 8 times until white flocculent precipitate appeared in the eppendorf tube, and centrifuged at 12000rpm for 10 min.

(5) The supernatant in the eppendorf tube after centrifugation was pipetted into an adsorption column CP3 which was placed in a clean eppendorf tube, without aspirating the pellet, and centrifuged at 12000rpm for 2 min. Pouring the effluent back to adsorption column CP3, centrifuging at 12000rpm for 2min, discarding the effluent, and returning adsorption column CP3 to the collection tube.

(6) Pipetting 600 μ LBuffer PW into adsorption column CP3 with pipette, centrifuging at 12000rpm for 1min, discarding the effluent, and returning adsorption column CP3 to the collection tube. This step was repeated once.

(7) Centrifuging at 12000rpm for 2min to remove residual rinse solution. The adsorption column CP3 was placed in a homomixer at a constant temperature of 65 deg.C and dried for 2min to completely remove ethanol.

(8) The adsorption column CP3 was placed in a clean eppendorf tube, and 40. mu. LddH preheated to 65 ℃ was aspirated by a pipette gun₂O to the center of the adsorption column. Standing for 2min, centrifuging at 12000rpm for 2min, collecting plasmid solution to eppendorf tube, and placing in a refrigerator at-20 deg.C for use. 3 mu L of the extracted plasmid is taken and electrophoresed in 1% agarose gel, if a target band is obtained, sequencing is carried out, and after the sequencing is correct, the 5941-35S-WRKY-GFP vector is obtained.

2.9 Agrobacterium tumefaciens EHA105

The agrobacterium-infected competent cells stored in a refrigerator at the temperature of-80 ℃ are placed on ice to be melted, and 4 mu L of the recombinant plasmid 5941-35S-BcWRKY-GFP is added into 50 mu L of the agrobacterium EHA105 competent cells and ice-cooled for 30 min. Freezing with liquid nitrogen for 1min, heating in 37 deg.C water bath for 5 min. Subsequently, 600. mu.L of LB liquid medium without antibiotic was added to the centrifuge tube, and the mixture was mixed by pipetting, incubated at 28 ℃ and 200rpm for 4 hours at constant temperature. Centrifuging at 5000rpm for 6min, discarding supernatant, and adding 200 μ LLB liquid culture medium to resuspend thallus. The resuspended cells were spread on LB solid medium containing 10mg/LRif and 50mg/L Kan, and inverted cultured at 28 ℃ for 18-36 h. After the bacterial colonies grow out, selecting a single bacterial colony, performing overnight oscillation culture on an LB liquid culture medium containing 10mg/LRif and 50mg/Lkan, performing PCR verification on bacterial liquid, adding glycerol into the correctly verified bacterial liquid, and storing in a refrigerator at the temperature of 80 ℃ below zero.

Example 2

Identification of breeding over-expression BcWRKY1 transgenic arabidopsis thaliana with increased expression level of kalanchoe metabolism related coding gene BcWRKY1 gene and transgenic plant

1. Culturing transgenic arabidopsis plant with overexpressed BcWRKY1 and improved expression level of kalanchoe metabolism related coding gene BcWRKY1 gene

The 5941-35S-BcWRKY-GFP recombinant vector is transformed into Rdr6 wild type Arabidopsis thaliana by an agrobacterium-mediated inflorescence dip method. The collected T1 generation Arabidopsis seeds were dibbled on LB plate containing hygromycin, and the solid LB plate was cultured in an artificial light incubator (22 ℃ C. for 18h in the daytime and 20 ℃ C. for 6h in the dark). The arabidopsis thaliana which can normally grow on the LB plate containing the hygromycin can be a positive plant of the transgenic arabidopsis thaliana, then is transplanted into the soil with equal volume and uniform mixing of organic soil and vermiculite, and is placed in an artificial illumination incubator to be cultured for a period of time.

2. Identification of transgenic Arabidopsis with over-expression BcWRKY1

Observing an arabidopsis plant with hygromycin resistance by using a fluorescence microscope, judging the expression level of BcWRKY1 according to the intensity of fluorescence of the plant, and selecting the transgenic arabidopsis with strong fluorescence intensity (high expression level). The results are shown in FIG. 2.

Example 3

Metabolites of wild type Arabidopsis and over-expressed BcWRKY1 Arabidopsis were analyzed by LC-MS.

Firstly, the seeds of arabidopsis thaliana are disinfected, the seeds of arabidopsis thaliana are put into a 1.5mL centrifugal tube, 1mL 75% ethanol is added to shake for 5min, the seeds are precipitated at the bottom of the centrifugal tube through slow centrifugation, the ethanol is absorbed, 2% NaClO is added to treat for 5min, and the seeds are precipitated at the bottom of the centrifugal tube through slow centrifugation, and the NaClO is absorbed. Washing with sterile water for 5 times, and after each washing, centrifuging at low speed to settle the seeds at the bottom of the centrifuge tube, and absorbing the sterile water. The Arabidopsis seeds are dibbled on MS solid plates, and after 3-4 days of culture at 4 ℃, the solid plates are placed in an artificial light incubator for culture (22 ℃ for 18h in the daytime and 20 ℃ for 6h in the dark) for about 1 week. And (3) transferring the arabidopsis seedlings from the MS solid flat plate into the soil in which the organic soil and the vermiculite are mixed in equal volume. 4 seeds are planted in each pot, 10 seeds are repeatedly planted, the illumination is carried out for 18 hours, and the seeds are cultured for four weeks at 22 ℃. Two groups of Arabidopsis thaliana were analyzed by LC-MS.

The experimental results are as follows:

since the main pharmacodynamic substance indirubin in the medicinal plant, namely the bluegrass, is related to the metabolic pathway derived from tryptophan-indoleacetic acid, the research analyzes the influence of over-expression of BcWRKY1 on metabolites in the tryptophan-indoleacetic acid metabolic pathway in Arabidopsis thaliana. By drawing a metabolic pathway network diagram, the synthesis and decomposition of each metabolic compound can be more clearly found. From the KEGG enrichment pathway map (shown in fig. 3) of tryptophan metabolism of transgenic arabidopsis thaliana overexpressing BcWRKY1 and wild-type arabidopsis thaliana, it can be known that indole acetate (indolacetate) overexpressing tryptophan metabolism in the transgenic arabidopsis thaliana overexpressing BcWRKY1 is a significant up-regulated metabolite compared with the wild-type arabidopsis thaliana. As can be seen from FIG. 4, the indole-related metabolites of transgenic Arabidopsis overexpressing Bcw RKY1 (Com _889_ pos; Com _3035_ pos; Com _2586_ pos; Com _1515_ pos; Com _252_ pos) were significantly higher than those of wild type Arabidopsis.

Although specific embodiments of the invention have been described above, it will be understood by those skilled in the art that the specific embodiments described are illustrative only and are not limiting upon the scope of the invention, and that equivalent modifications and variations can be made by those skilled in the art without departing from the spirit of the invention, which is to be limited only by the appended claims.

Sequence listing

<110> university of Chinese

<120> cerulenin metabolism related gene BcWRKY1, and protein and application thereof

<130> 100

<160> 4

<170> SIPOSequenceListing 1.0

<210> 1

<211> 534

<212> DNA

<213> （Baphicacanthus cusia (Nee) Bremek）

<400> 1

atggagagca tggatgatta ttattgttat ctggatgcat atgtgtcctt gctgtcggac 60

tcccgctgca gctcacctct tcctcacacc acgtctcagc tgatgggctt tgcagttgca 120

gatcatgatc atgagcatca ccaccttcat caacctcaac cttctcctct caataatcac 180

tggattcaca attactcttc aatctctact acctccaccc cttccccgac cccgaccccg 240

accccgcccg atgctggagc aggaggaccc ataataattg ataacaggga gaagaaacta 300

ggggcaaagg ttgctttcaa gaccaagtct caaattgatg tactggatga tggattcaag 360

tggagaaagt atgggaaaaa gatggtcaag aacagcccta atcccaggaa ctactatagg 420

tgctcaattc aaggctgcgc cgtgaagaag agggtggaga gggacaatga cgactcgcat 480

tacgttctca caacctatga gggcatccac aatcatctac cttctgcacc ttaa 534

<210> 2

<211> 177

<212> PRT

<213> （Baphicacanthus cusia (Nee) Bremek）

<400> 2

Met Glu Ser Met Asp Asp Tyr Tyr Cys Tyr Leu Asp Ala Tyr Val Ser

1 5 10 15

Leu Leu Ser Asp Ser Arg Cys Ser Ser Pro Leu Pro His Thr Thr Ser

20 25 30

Gln Leu Met Gly Phe Ala Val Ala Asp His Asp His Glu His His His

35 40 45

Leu His Gln Pro Gln Pro Ser Pro Leu Asn Asn His Trp Ile His Asn

50 55 60

Tyr Ser Ser Ile Ser Thr Thr Ser Thr Pro Ser Pro Thr Pro Thr Pro

65 70 75 80

Thr Pro Pro Asp Ala Gly Ala Gly Gly Pro Ile Ile Ile Asp Asn Arg

85 90 95

Glu Lys Lys Leu Gly Ala Lys Val Ala Phe Lys Thr Lys Ser Gln Ile

100 105 110

Asp Val Leu Asp Asp Gly Phe Lys Trp Arg Lys Tyr Gly Lys Lys Met

115 120 125

Val Lys Asn Ser Pro Asn Pro Arg Asn Tyr Tyr Arg Cys Ser Ile Gln

130 135 140

Gly Cys Ala Val Lys Lys Arg Val Glu Arg Asp Asn Asp Asp Ser His

145 150 155 160

Tyr Val Leu Thr Thr Tyr Glu Gly Ile His Asn His Leu Pro Ser Ala

165 170 175

Pro

<210> 3

<211> 43

<212> DNA

<213> (Artificial sequence)

<400> 3

ttacaattac catggggcgc gccatggaga gcatggatga tta 43

<210> 4

<211> 43

<212> DNA

<213> (Artificial sequence)

<400> 4

gcccttgctc accatggcgc gcccaggtgc agaaggtaga tga 43

Claims

1. A cerulenin metabolism related gene BcWRKY1 is characterized in that: the gene BcWRKY1 has a nucleotide sequence shown as SEQ ID No. 1.

2. The cerulenin metabolic related gene BcWRKY1 as claimed in claim 1, wherein: the nucleotide sequence, allele or derivative generated by adding, substituting, inserting or deleting one or more nucleotides in the nucleotide sequence of SEQ ID No. 1.

3. A plasmid, a plant expression vector and a host cell containing the cerulenin metabolism-related gene BcWRKY1 as claimed in claim 1 or 2.

4. A protein coded by a cerulenine metabolism-related gene BcWRKY1, which is characterized in that: the protein has an amino acid sequence shown as SEQ ID No. 2.

5. The application of the cerulenin metabolic related gene BcWRKY1 is characterized in that: the gene BcWRKY1 has a nucleotide sequence shown as SEQ ID No.1, and the gene BcWRKY1 and a nucleotide sequence, an allele or a derivative generated by adding, substituting, inserting or deleting one or more nucleotides in the nucleotide sequence are applied to indole acetate for regulating tryptophan metabolism in Arabidopsis.