CN112812164A - Application of rice transcription factor WRKY53 in MAPK cascade signal pathway - Google Patents
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Abstract
An application of a rice transcription factor WRKY53 in a MAPK cascade signal pathway relates to the field of rice genetic breeding, in particular to an application of a rice transcription factor WRKY53 in the MAPK cascade signal pathway. The invention firstly explains that WRKY53 is a substrate of a MAPK cascade signal channel and positively regulates and controls the leaf angle and grain type of rice. Provides an important theoretical basis for rice plant type breeding and has important reference value for screening new high-yield rice plant types. In the regulation and control of the inclination angle of rice leaves, WRKY53 is a direct target gene of MAPK cascade signals; in regulation of granule type, WRKY53 is a micro-effective target gene of MMAPK cascade signal. The invention is applied to the field of genetic breeding of rice.
Description
Technical Field
The invention relates to the field of rice genetic breeding, in particular to application of a rice transcription factor WRKY53 in a MAPK cascade signal pathway.
Background
The inclination angle and grain type of rice leaves are important agronomic characters influencing crop yield, new plant types with high rice yield are screened by taking the inclination angle and grain type of the leaves as indexes, the planting density is reasonably planned, and the improvement of the crop yield is realized. The rice MAPK cascade signals are reported to participate in biological processes such as regulation of pathogen defense reaction and mechanical injury defense response of rice, regulation of rice plant types and the like. In addition, in the aspect of plant type regulation, the plant type regulation is mainly involved in regulating the inclination angle of rice leaves and the growth of grain type. MAPKKK10 mutant smg2-1 shows phenotype of small leaf inclination angle, small grain shape and the like; whereas constitutively activated MAPKKK10 overexpressing transgenic rice exhibited phenotypic characteristics of increased grain type. MAPKK4 mutant smg1-1 shows phenotype of small leaf inclination angle, small grain shape and the like; whereas the constitutively activated MAPKK4 overexpressing transgenic rice showed the phenotypic characteristics of increased leaf inclination and increased grain type. MAPK6 mutant dsg1 shows phenotypes of upright leaf inclination angle, small grain shape and the like; and the constitutively activated MAPK6 overexpression transgenic rice shows the phenotype characteristic of grain type increase, which indicates that MAPKKK10-MAPKK4-MAPK6 can positively regulate the inclination angle and grain type development of rice leaves. The action mechanism of the rice transcription factor OsWRKY53 in MAPKKK10-MAPKK4-MAPK6 cascade signals is not clear.
Disclosure of Invention
The invention aims to firstly clarify the action mechanism of WRKY53 in the MAPKKK10-MAPKK4-MAPK6 signal pathway, provide an important theoretical basis for rice plant type breeding, and have important reference value for screening new high-yield rice plant types.
The invention provides application of a rice transcription factor WRKY53 in a MAPK cascade signal pathway.
Furthermore, the rice transcription factor WRKY53 is a substrate of a MAPK cascade signal channel and can positively regulate and control the inclination angle of rice leaves.
The rice transcription factor WRKY53 is a direct target gene of MAPK cascade signal. The direct target gene refers to a direct acting substrate downstream of the MAPK cascade signal.
Furthermore, the rice transcription factor WRKY53 is a substrate of a MAPK cascade signal pathway and can positively regulate rice grain type.
The rice transcription factor WRKY53 is a weak target gene of MAPK cascade signals.
WRKY53 is defined as weak target gene because it mainly regulates the extension of cells and the amplification of weak regulatory cells in the regulation of glume development
The MAPK cascade signal is MAPKKK10-MAPKK4-MAPK6 cascade signal.
The invention has the beneficial effects that:
the WRKY53 is a substrate of a MAPKKK10-MAPKK4-MAPK6 signal channel and is used for positively regulating and controlling the leaf angle and grain type of rice for the first time.
According to the invention, the CAMAPK6 wrky53 and MKP1RNAi wrky53 double mutants are obtained through gene knockout and hybridization, and the phenotype of increased CAMAPK6 and MKP1RNAi grain types can be completely restored by the double mutants through phenotype observation, and the inclination angle of the leaves is obviously reduced to a certain extent. Shows that WRKY53 acts on the genetic downstream of MAPK6 and MKP1 in the regulation of leaf inclination angle and grain type.
According to the invention, WRKY53 gene is over-expressed in smg2-1 and dsg1 by means of genetic transformation, so that smg2-1WRKY53OE and dsg 1WRKY53OE double mutants are obtained, and compared with a single mutant, the double mutant can completely recover the phenotype of a single mutant with reduced leaf inclination angle and reduced grain type, and shows the phenotype of increased leaf inclination angle and increased grain type. Shows that WRKY53 acts on the genetic downstream of MAPKKK10 and MAPK6 in the regulation of leaf inclination angle and granule type.
According to the invention, dsg 1WRKY53OE, smg2-1WRKY53OE, CAMAPK6 WRKY53 and MKP1RNAi WRKY53 double mutants and glumes contrasted by the double mutants are observed by a scanning electron microscope technology, and MAPK6 is found to be mainly involved in regulating the number of glume cells and weakly regulating the size of the glume cells; WRKY53 mainly regulates cell size and weakly regulates cell number.
The conclusion obtained by combining cytology and genetics is that WRKY53 is a direct target gene of MAPKKK10-MAPKK4-MAPK6 cascade signals in the regulation and control of the inclination angle of rice leaves; WRKY53 is a micro-effective target gene of MAPKKK10-MAPKK4-MAPK6 cascade signal in regulation of granule type.
The WRKY53 is a direct substrate of the MAPKKK10-MAPKK4-MAPK6 cascade signal, the inclination angle and the grain type of the rice leaf are positively regulated, important clues and theoretical bases are provided for rice plant type breeding, important theoretical bases are provided for modifying the rice plant type and further improving the crop yield, and the WRKY53 has a wide application prospect.
Drawings
FIG. 1 is a general morphology chart of dsg 1WRKY53OE double mutant and its control
FIG. 2 shows the identification result of dsg 1WRKY53OE double mutants
FIG. 3 is a graph of the leaf dip angle morphology of dsg 1WRKY53OE double mutant
FIG. 4 is the statistical result of the leaf inclination angle of dsg 1WRKY53OE double mutant
FIG. 5 is a morphogram of dsg 1WRKY53OE double mutant and its control grain type
FIG. 6 shows the statistical results of dsg 1WRKY53OE double mutants and their control grain length
FIG. 7 shows the statistical results of dsg 1WRKY53OE double mutants and their control particle widths
FIG. 8 is the result of the statistics of the length and number of the cells of the longitudinal cells of dsg 1WRKY53OE double mutant palea
FIG. 9 is a gross morphology of CAMAPK6 wrky53 double mutants and controls thereof
FIG. 10 shows the result of the identification of CAMAPK6 wrky53 double mutants
FIG. 11 is a graph of leaf dip angle morphology of CAMAPK6 wrky53 double mutants
FIG. 12 is the statistics of the leaf inclination of CAMAPK6 wrky53 double mutant
FIG. 13 is a morphogram of CAMAPK6 wrky53 double mutant and its control granule type
FIG. 14 shows the statistics of CAMAPK6 wrky53 double mutant and its control grain length
FIG. 15 shows the statistical results of CAMAPK6 wrky53 double mutants and control particle widths thereof
FIG. 16 is the result of statistics of the cell length and cell number of the longitudinal palea of the CAMAPK6 wrky53 double mutant
FIG. 17 is a gross morphology chart of MKP1RNAi wrky53 double mutants and their control
FIG. 18 shows the result of identifying MKP1RNAi wrky53 double mutants
FIG. 19 is a graph showing leaf inclination angle morphology of double mutants of MKP1RNAi wrky53
FIG. 20 shows the statistics of leaf inclination of MKP1RNAi wrky53 double mutants
FIG. 21 is a morphogram of MKP1RNAi wrky53 double mutant and its control grain type
FIG. 22 shows the statistics of MKP1RNAi wrky53 double mutants and their control grain length
FIG. 23 shows statistical results of MKP1RNAi wrky53 double mutants and control grain widths thereof
FIG. 24 is a total morphology of smg2-1WRKY53OE double mutants and controls thereof
FIG. 25 shows the identification result of smg2-1WRKY53OE double mutants
FIG. 26 is an inclination angle morphogram of smg2-1WRKY53OE double mutant leaf
FIG. 27 is the statistic result of leaf inclination angle of smg2-1WRKY53OE double mutant
FIG. 28 is a morphogram of smg2-1WRKY53OE double mutant and its control granule type
FIG. 29 shows the statistical results of smg2-1WRKY53OE double mutants and control grain length thereof
FIG. 30 shows the statistical results of smg2-1WRKY53OE double mutants and control particle widths thereof
FIG. 31 is the result of the statistics of the length and number of the cells of the longitudinal palea of the smg2-1WRKY53OE double mutant
Detailed Description
The following examples are given to illustrate the present invention, and the following examples are carried out on the premise of the technical solution of the present invention, and give detailed embodiments and specific procedures, but the scope of the present invention is not limited to the following examples.
Acquisition of one, dsg 1WRKY53OE double mutants
1. Vector construction: using Nipponbare cDNA as a template, see TaKaRa
The HS DNA Polymerase operation instruction uses a forward primer F1 and a reverse primer R1 as amplification primers to amplify a coding region of a WRKY53 gene, and clones an amplified fragment into a plant over-expression vector PC1390U to form a WRKY53 gene over-expression vector driven by a Ubiquitin promoter.
Forward primer F1: 5'-GTTACTTCTGCACTAGGTACCATGGCGTCCTCGACGGGG-3'
Reverse primer R1: 5'-TCTTAGAATTCCCGGGGATCCCTAGCAGAGGAGCGACTCGACG-3'
2. MAPK6 mutant dsg1 is taken as an experimental material, and an agrobacterium-mediated genetic transformation method is adopted to obtain a dsg 1WRKY53OE double mutant, wherein the transformation method is as follows:
(1) PC1390U-WRKY53 transformed Agrobacterium tumefaciens competent EHA 105: taking out EHA105 from a refrigerator at-80 deg.C, and thawing on ice; 500 ng-1 mu g of the target plasmid is added into 100ul EHA105 competence and placed on ice for 30 min; rapidly placing in liquid nitrogen for 5 min; taking out from liquid nitrogen, and rapidly placing in water pre-pot at 37 deg.C for 5 min; ice for 2 min; adding 800 mul of liquid LB culture medium, placing in a full-temperature oscillator (purchased from MKN company), and incubating at 28 ℃ and 120rpm for 4-5 h; after centrifugation, most of the supernatant was discarded, and the remaining bacterial solution was applied to LB solid medium containing kanamycin (50. mu.g/ml) (obtained from Amresco) and rifampicin (50. mu.g/ml) (obtained from Amresco), and cultured at 28 ℃ for about 3 days.
(2) Colony PCR identification: identifying positive clones by means of PCR; the positive clones were picked up in liquid LB medium with the corresponding antibiotics and rifampicin and cultured at 28 ℃ for about 16h at 180 rpm.
(3) Infecting rice calluses: the inoculum from step 2 was incubated to a color that visually looked like orange juice (OD ═ 1.0 or so) and was available for transformation. Taking about 500 mu l of bacterial liquid into a 1.5ml centrifuge tube, centrifuging for 3min at the temperature of 28 ℃ and 5000rpm, and discarding supernatant to see white bacterial colonies at the bottom of the tube; gently pipetting the subterminal pellet with 300. mu.l of liquid coculture medium containing acetosyringone (purchased from Aldrich) at a final concentration of 20ug/ml to uniformly suspend the subterminal pellet in the liquid medium; selecting the callus with good growth state to a 50ml centrifuge tube, wherein the volume of the callus is about 5ml of the scale of the centrifuge tube; adding 20ml of liquid co-culture medium containing 20 mu g/ml acetosyringone, and then adding the suspended 300ul of bacterial liquid into a 50ml centrifuge tube; and (4) continuously and softly mixing for 2-3 min for infection. Pouring the liquid co-culture medium, transferring the infected callus into a culture dish paved with filter paper, and adsorbing the redundant culture medium, wherein the process is about 1 min; spreading a layer of filter paper on the solid co-culture medium to soak the filter paper, and transferring the infected callus to the solid culture medium; dark culture is carried out for 2-3 days at 28 ℃.
(4) And (3) recovery culture: after the infected callus is cultured in dark for 2-3 days, transferring the callus particles into a 50ml centrifuge tube; washing the callus with sterile water containing carbenicillin (purchased from Amresco) with a final concentration of 400 μ g/ml for 4-5 times, each time lasting for about 1min, and sterilizing; then, cleaning the callus with sterile water for 2-3 times, transferring the callus to a culture dish paved with filter paper, and sucking off excessive water; and transferring the callus onto a recovery culture medium containing 400 mu g/ml carbenicillin, and performing recovery culture in a climatic incubator (24h light culture) at 28 ℃ for 4-5 days.
(5) Screening and culturing: after 4-5 days of recovery culture, transferring the callus on the recovery culture medium to a screening culture medium containing 400 μ g/ml carbenicillin and 50 μ g/ml hygromycin (purchased from Roche); the cells were transferred to a 28 ℃ climatic incubator (24h light culture) and cultured for about 30 days.
(6) Differentiation culture: transferring the resistant callus on the screening culture medium to a differentiation culture medium, and transferring each bottle to a cluster of callus; culturing in a 28 deg.C artificial climate incubator (24h light culture) for about 30 days to obtain transgenic seedling.
3. Identification of dsg 1WRKY53OE double mutant
And F2 and R2 are used as primers, and the dsg 1WRKY53OE double mutant is identified by a qRT-PCR mode.
Forward primer F2: 5'-GAGCGACATCGACATCCT-3'
Reverse primer R2: 5'-TTGTGCTTGCCCTCGTAG-3'
As shown in FIGS. 1 and 2, the overall morphology of dsg 1WRKY53OE and its control and the identification result thereof are shown. As can be seen from the figure, the dsg1 has upright leaf inclination angle and smaller grain shape; and the inclination angle of the double-mutant leaf of dsg 1WRKY53OE is larger, and the grain type is larger.
Statistical analysis of leaf inclination and grain type of double-mutant leaf dsg 1WRKY53OE
Selecting the sword leaves of the rice with consistent growth states and in the heading stage as experimental materials, measuring the inclination angle of the leaves by using ImageJ, counting 50 leaves for each material, and finally comparing every two leaves by using one-factor variance analysis. In the measurement of grain type, mature and plump rice seeds are selected and photographed, then the length and the width of the rice seeds are measured by ImageJ, 50 grains are counted for each material, and finally, pairwise comparison is carried out by utilizing single-factor variance analysis.
And thirdly, analyzing the dsg 1WRKY53OE double mutant and a control glume development mechanism thereof by cytology.
1. Mature and plump dsg 1WRKY53OE double mutants and control seeds thereof are selected, fixed on an experiment table of a scanning electron microscope by conductive adhesive, and sprayed with gold for 1min by adopting gold spraying equipment.
2. And (4) placing the processed sample under a scanning electron microscope for scanning observation.
3. Selecting the center part of the rice husk, counting the number of longitudinal cells by ImageJ, and dividing the total length under the corresponding number of cells by the number of cells to obtain the average longitudinal cell length.
As shown in FIGS. 3 and 4, the photographs and the statistical results show that the WRKY53 can completely recover the phenotype of dsg1 with reduced leaf inclination angle, and WRKY53 is located at the genetic downstream of MAPK6 in the regulation of leaf inclination angle. FIGS. 5, 6 and 7 are photographs of the grain type and the statistics of grain length and grain width for dsg 1WRKY53OE and its control. As can be seen from the figure, compared with the single dsg1 mutant, the dsg 1WRKY53OE double mutant has significantly longer grain length and grain width, which indicates that WRKY53 can completely recover the phenotype of dsg1 with reduced grain size; fig. 8 is a scanning electron microscope result of dsg 1WRKY53OE and its control lemma, and statistical results show that the number of dsg1 mutant cells is significantly reduced, while after over-expressing WRKY53 gene in dsg1, the cell length is significantly increased, and the contribution rate of the cell number is relatively lower than that of the cell length. The WRKY53 mainly regulates the cell extension and slightly regulates the cell amplification in the regulation of the grain type; according to previous reports that MAPK6 is mainly involved in regulating cell expansion, we conclude that WRKY53 is a fine-tuning target gene of MAPK6 in regulation of granuloma.
Fourth, the acquisition of CAMAPK6 wrky53 double mutant
1. The CDS sequence of the WRKY53 gene is input into CRISPR Primer Designer software, 2 pairs of target site primers (F3 and R3; F4 and R4) are designed, and a knockout vector CRISPR/Cas9-WRKY53 is constructed.
Forward primer F3: 5'-GGCATTCCAGTCGTACCTCTGAGC-3'
Reverse primer R3: 5'-AAACGCTCAGAGGTACGACTGGAA-3'
Forward primer F4: 5'-GCCGAGCTGGAGGACGGGTACAAC-3'
Reverse primer R4: 5'-AAACGTTGTACCCGTCCTCCAGCT-3'
2. The CAMAPK6 transgenic rice is used as an experimental material, and an agrobacterium-mediated genetic transformation method is adopted to obtain a CAMAPK6 wrky53 double mutant. (Agrobacterium-mediated genetic transformation methods are as above)
3. MAPK6 gene identification primers (F5 and R5) are designed, and qRT-PCR identification is carried out on the double mutants; designing a pair of sequencing primers (F6 and R6) covering a target sequence, amplifying the WRKY53 gene target sequence, and sequencing a PCR product; until a homozygous double mutant CAMAPK6 wrky53 was obtained.
Forward primer F5: 5'-CTTTATCAGATTCTCCGTGGCTT-3'
Reverse primer R5: 5'-TCATAAAATCGGTTTCTGAGGTG-3'
Forward primer F6: 5'-CGGGGTGCCCAAGTTCAAGTC-3'
Reverse primer R6: 5'-ATGGAGCAGCCGTTGTAGGTG-3'
As shown in fig. 9 and 10, the overall morphology of campk 6 wrky53 and its control and the results of their identification are shown. As can be seen from the figure, the CAMAPK6 wrky53 double mutant is similar to the wrky53 single mutant, and the inclination angle of the leaves is smaller and the grain type is smaller. As shown in fig. 11 and 12, which are photographs and statistics of the leaf inclination angle of campk 6 WRKY53 and its control, the results show that the knock-out of WRKY53 gene in campk 6 results in a significant decrease of the leaf inclination angle of campk 6, and again, it is demonstrated that WRKY53 is located genetically downstream of MAPK6 in the regulation of leaf inclination angle. Fig. 13, 14, 15 are photographs of the grain types and statistics of grain length and grain width for campk 6 wrky53 and its control. The results show that compared with a single CAMAPK6 mutant, the CAMAPK6 wrky53 double mutant has obviously reduced grain length and grain width, which indicates that wrky53 can completely recover the enlarged phenotype of the CAMAPK6 grain type; fig. 16 is a scanning electron microscope result of campk 6 wrky53 and its control lemma, showing that the cell length of campk 6 is significantly longer; the cell length of the WRKY53 mutant is obviously shortened, and the number of the WRKY53 mutant is reduced to a lower degree, so that the WRKY53 is mainly involved in regulating cell extension and weakly regulating cell division; and the cytological statistical results of combining dsg1 and CAMAPK6 show that MAPK6 mainly regulates cell division and weakly regulates cell extension. Again, WRKY53 is a micro-effective target gene for MAPK6 in regulation of granulotype.
Five, acquisition of MKP1RNAi wrky53 double mutant
1. MKP1RNAi and wrky53 are used as experimental materials to carry out hybridization to obtain hybrid particles.
2. And accelerating germination, planting and identifying the hybrid grains to obtain F1 generation seeds.
3. The obtained F1 generation seeds are subjected to pregermination and planted, and WT, MKP1RNAi, wrky53 and MKP1RNAi wrky53 are identified through separation. wrky53 identified the primers as above, and MKP1 identified the primers as (F7 and R7). And phenotypically observing the separated offspring.
Forward primer F7: 5'-ACCGTAGGGTGGAATCTTATG-3'
Reverse primer R7: 5'-AACTTGCGCCTTAGTGAACTC-3'
As shown in FIGS. 17 and 18, the overall morphology of MKP1RNAi wrky53 and its control and the result of its identification are shown. As can be seen from the figure, the MKP1RNAi wrky53 double mutant is similar to the wrky53 single mutant in that the leaf inclination angle is smaller and the grain type is smaller. As shown in fig. 19 and 20, which are photographs of leaf inclination angles of MKP1RNAi WRKY53 and its control and statistical results, the statistical results show that after knocking-out WRKY53 gene in MKP1RNAi, the leaf inclination angles of MKP1RNAi become significantly smaller, and since MKP1 is a phosphatase inhibiting MAPK6, it is shown again that WRKY53 is located genetically downstream of MAPK6 in the regulation of leaf inclination angles. FIGS. 21, 22 and 23 are photographs of the grain type and the statistics of grain length and grain width of MKP1RNAi wrky53 and its control. As can be seen from the figure, compared with the single MKP1RNAi mutant, the MKP1RNAi wrky53 double mutant has obviously reduced grain length and grain width, which indicates that wrky53 can completely restore the phenotype of the increased grain type of the MKP1 RNAi; WRKY53 was further confirmed to be a micro-effective target gene for MAPK cascade signaling.
Acquisition of six, smg2-1WRKY53OE double mutants
1. Smg2-1(MAPKKK10 mutant) is used as an experimental material, and an agrobacterium-mediated genetic transformation method is adopted to obtain the smg2-1WRKY53OE double mutant. (Agrobacterium-mediated genetic transformation methods and vector construction are as above)
2. The double mutants were identified by means of qRT-PCR.
As shown in FIGS. 24 and 25, the overall morphology of smg2-1WRKY53OE and its control and the identification result thereof are shown. As can be seen from the figure, smg2-1 leaves have upright inclination angle and smaller grain shape; and the inclination angle of the smg2-1WRKY53OE double mutant leaf is larger, and the grain type is larger. As shown in FIGS. 26 and 27, the photographs and the statistical results show that the leaf inclination photographs of dsg 1WRKY53OE and the control thereof show that WRKY53 can completely restore the phenotype of smg2-1 leaf inclination reduction, which indicates that WRKY53 is located downstream of the genetics of MAPK cascade signals in the regulation of leaf inclination. FIGS. 28, 29 and 30 are photographs of the grain type smg2-1WRKY53OE and its control, and statistics of grain length and grain width. As can be seen from the figure, compared with the smg2-1 single mutant, the smg2-1WRKY53OE double mutant has significantly longer grain length and grain width, which indicates that WRKY53 can completely recover the phenotype of smg2-1 with smaller grain size. FIG. 31 is a scanning electron microscope of smg2-1WRKY53OE and its control lemma, and statistical results show that the smg2-1 mutant significantly decreased in cell number, whereas after over-expression of WRKY53 in smg2-1, the cell length was significantly increased and the contribution rate was relatively low compared to the cell number and cell length. Again, WRKY53 is a micro-effective target gene downstream of the MAPK cascade signaling in regulation of granulotype.
Sequence listing
<110> institute of geography and agroecology of northeast China academy of sciences
Application of rice transcription factor WRKY53 in MAPK cascade signal pathway
<160> 14
<210> 1
<211> 39
<212> DNA
<213> Artificial sequence
<220>
<223> Forward primer F1
<400> 1
gttacttctgcactaggtaccatggcgtcctcgacgggg 39
<210> 2
<211> 43
<212> DNA
<213> Artificial sequence
<220>
<223> reverse primer R1
<400> 2
tcttagaattcccggggatccctagcagaggagcgactcgacg 43
<210> 3
<211> 18
<212> DNA
<213> Artificial sequence
<220>
<223> Forward primer F2
<400> 3
gagcgacatcgacatcct 18
<210> 4
<211> 18
<212> DNA
<213> Artificial sequence
<220>
<223> reverse primer R2
<400> 4
ttgtgcttgccctcgtag 18
<210> 5
<211> 24
<212> DNA
<213> Artificial sequence
<220>
<223> Forward primer F3
<400> 5
ggcattccagtcgtacctctgagc 24
<210> 6
<211> 24
<212> DNA
<213> Artificial sequence
<220>
<223> reverse primer R3
<400> 6
aaacgctcagaggtacgactggaa 24
<210> 7
<211> 24
<212> DNA
<213> Artificial sequence
<220>
<223> Forward primer F4
<400> 7
gccgagctggaggacgggtacaac 24
<210> 8
<211> 24
<212> DNA
<213> Artificial sequence
<220>
<223> reverse primer R4
<400> 8
aaacgttgtacccgtcctccagct 24
<210> 9
<211> 23
<212> DNA
<213> Artificial sequence
<220>
<223> Forward primer F5
<400> 9
ctttatcagattctccgtggctt 23
<210> 10
<211> 23
<212> DNA
<213> Artificial sequence
<220>
<223> reverse primer R5
<400> 10
tcataaaatcggtttctgaggtg 23
<210> 11
<211> 21
<212> DNA
<213> Artificial sequence
<220>
<223> Forward primer F6
<400> 11
cggggtgcccaagttcaagtc 21
<210> 12
<211> 21
<212> DNA
<213> Artificial sequence
<220>
<223> reverse primer R6
<400> 12
atggagcagccgttgtaggtg 21
<210> 13
<211> 21
<212> DNA
<213> Artificial sequence
<220>
<223> Forward primer F7
<400> 13
accgtagggtggaatcttatg 21
<210> 14
<211> 21
<212> DNA
<213> Artificial sequence
<220>
<223> reverse primer R7
<400> 14
aacttgcgccttagtgaactc 21
Claims (5)
1. The application of the rice transcription factor WRKY53 in MAPK cascade signal pathway.
2. The use as claimed in claim 1, wherein the rice transcription factor WRKY53 is a substrate of MAPK cascade signaling pathway and can positively regulate rice leaf inclination.
3. The use as claimed in claim 2, wherein the rice transcription factor WRKY53 is a direct target gene for MAPK cascade signaling.
4. The use as claimed in claim 1, wherein the rice transcription factor WRKY53 is a substrate of MAPK cascade signaling pathway and can positively regulate rice grain type.
5. The use as claimed in claim 4, wherein the rice transcription factor WRKY53 is a weak target gene for MAPK cascade signaling.
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