CN113684211B - Specific expression promoter of chestnut male flowers and application thereof - Google Patents
Specific expression promoter of chestnut male flowers and application thereof Download PDFInfo
- Publication number
- CN113684211B CN113684211B CN202110928218.3A CN202110928218A CN113684211B CN 113684211 B CN113684211 B CN 113684211B CN 202110928218 A CN202110928218 A CN 202110928218A CN 113684211 B CN113684211 B CN 113684211B
- Authority
- CN
- China
- Prior art keywords
- chestnut
- promoter
- male
- flowers
- specific expression
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8201—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation
- C12N15/8202—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation by biological means, e.g. cell mediated or natural vector
- C12N15/8205—Agrobacterium mediated transformation
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
- C12N15/8222—Developmentally regulated expression systems, tissue, organ specific, temporal or spatial regulation
- C12N15/823—Reproductive tissue-specific promoters
- C12N15/8231—Male-specific, e.g. anther, tapetum, pollen
Abstract
The invention discloses a chestnut male flower specific expression promoter and application thereof. The chestnut male flower specific expression promoter is named as follows: cmP28, the nucleotide sequence of which is shown in SEQ ID NO. 1. According to quantitative PCR analysis of different tissues of the Chinese chestnut, the invention discovers that the CmP promoter regulates the downstream CmLTP gene to have strong expression activity in male flowers of the Chinese chestnut, and to be expressed in tissues such as stems, leaves, female flowers and the like; the CmP promoter is fused with a marker gene GUS, transient transformation of the leaf of the bentazone is mediated by agrobacterium GV3101, and the arabidopsis thaliana is stably transformed, and the result proves that the CmP promoter has similar expression activity to a positive control (strong promoter CaMV35 s). The CmP promoter disclosed by the invention can be used for constructing transgenic plants for specific expression of chestnut male flowers.
Description
Technical Field
The invention belongs to the technical field of plant genetic engineering, and particularly relates to a chestnut male flower specific expression promoter and application thereof.
Background
Chestnut (Castanea mollissima Blume) is a tree of the genus chestnut of the family Fagaceae, and is native to China. The chestnut kernels are rich in starch, crude protein and minerals, are important woody grains, and according to 2019 statistics of international grain and agriculture organizations, the yield of Chinese chestnut reaches 184.9 ten thousand tons, and accounts for 76.8% of the global edible chestnut yield. Chestnut is a hermaphrodite heteroHua plant, and flower buds can be divided into mixed flower buds and incompletely mixed flower buds. Wherein, the mixed flower buds firstly differentiate to form male inflorescences, then differentiate female inflorescences at the basal parts of the flower branches, and the female flowers pollinate and fertilize and develop into chestnut fruits. The incompletely mixed flower buds can only differentiate to form pure male inflorescences, usually 5-7 flowers are clustered, and the male flower clusters of hundreds of flowers form the cotton bud inflorescences. The normal fruiting branch of chestnut tree has more than 10 male flowers, and the mixed inflorescences have only 1-3 flowers, and the number of female flowers is far smaller than that of male flowers. In production, the Chinese chestnut has excessive male flowers and insufficient female flowers, and is an important limiting factor for high yield and high quality of Chinese chestnut.
The research shows that the chestnut flower bud primordium is amphoteric and has stronger differentiation plasticity. Theoretically, the sex differentiation of chestnut buds can be regulated by regulating and controlling the genes related to the differentiation and development of female flowers, so that the purposes of increasing female and suppressing male, increasing yield and improving efficiency are achieved. On the other hand, the chestnut male flowers are rich in flavonoid substances, have higher antioxidant activity, and the crude extracts of the chestnut male flowers, volatile essential oil and the like have better utilization value. At present, the research on the sex differentiation and development genetic engineering of Chinese chestnut is less, and the applicant develops SSR molecular markers suitable for Chinese chestnut genetic analysis by utilizing male and female flower differential expression genes screened by the male and female flower transcriptome in the earlier stage, but has not been reported on male and female flower specific expression genes or promoters.
Transcriptional regulation is one of the main modes of plant gene expression regulation. Wherein the transcription factor is coordinated with the promoter, and the transcription of the promoter gene is the key of transcription control under the action of factors such as RNA polymerase and the like. Promoters commonly used in plants are constitutive expression promoters, tissue specific expression promoters and conditional inducible expression promoters. At present, constitutive promoters such as cauliflower mosaic virus (CaMV) 35S promoters are mainly used in genetic engineering and genetic improvement researches. Because the constitutive promoter continuously over-expresses the target gene in the whole plant, not only is substances and energy wasted, but also normal growth and development of the plant can be influenced. Therefore, the development of tissue-specific or conditionally inducible expression promoters is particularly important in practical applications. For the Chinese chestnut, the promoter for the specific expression of the male flowers can be used for adjusting the sex differentiation of the buds of the Chinese chestnut on one hand, so that the purposes of 'reducing male and increasing yield' are achieved; on the other hand, the functional components of the male flowers can be increased by specifically expressing the functional genes, which is beneficial to the fine and deep processing and comprehensive utilization of the male flowers.
Disclosure of Invention
The first object of the invention is to provide a chestnut male flower specific expression promoter, wherein the nucleotide sequence of the chestnut male flower specific expression promoter is shown as SEQ ID NO. 1.
The length of the chestnut male flower specific expression promoter sequence is 1117bp, and the chestnut male flower specific expression promoter sequence is named CmP.
The second object of the invention is to provide an expression vector which contains the chestnut male flower specific expression promoter.
A third object of the present invention is to provide a host bacterium containing the above-mentioned expression vector.
Preferably, the host bacteria are E.coli DH5 alpha or Agrobacterium tumefaciens Agrobacterium tumefaciens GV3101 monoclonal cell lines.
The fourth object of the invention is to provide the application of the promoter in constructing transgenic plants for specific expression of chestnut male flowers.
According to the invention, a male flower specific expression gene CmLTP (temporary name) is screened and identified through the analysis of a Chinese chestnut male and female flower differential transcriptome, the expression characteristics of the CmLTP in different Chinese chestnut tissues are analyzed through real-time fluorescence quantitative PCR, and the fact that CmP promoter regulated downstream CmLTP only has strong expression activity in Chinese chestnut male flowers and extremely weak expression in tissues such as stems, leaves, female flowers and the like is found (figure 1). The CmP promoter is fused with a reporter gene GUS, and the agrobacterium tumefaciens GV3101 mediates transient transformation of the benthamiana to stably transform the arabidopsis thaliana, so that the CmP promoter has stronger promoter activity. The results show that the chestnut male flower specific expression promoter can be used for constructing transgenic plants for male flower specific expression, and plays an important role in genetic engineering improvement and utilization of chestnut male flowers and the like.
Drawings
FIG. 1 shows the relative expression level of the downstream CmLTP gene in different tissues of chestnut regulated by the CmP promoter. Male flower 1: about 2cm for male inflorescences after 4 months and 13 days; male flower 2: day 4, 29; male flower 3:5 months 13 days; male flower 4:5 months and 25 days, and the full bloom stage of the male flowers. Female flowers 1 and 2 correspond to sampling periods of male flowers 3 and 4, respectively.
FIG. 2A shows the results of double digestion with HindIII and BamH I after the CmP promoter was ligated to the TA cloning vector, and FIG. 2B shows the results of double digestion with HindIII and BamH I for pCAMBIA 2300-CmP-GUS-eGFP.
FIG. 3 shows the results of GFP-induced fluorescence (A) and GUS staining (B) of transient transformation of leaf blades of Nicotiana benthamiana with Agrobacterium GV3101 containing pCAMBIA2300-CmP28-GUS-eGFP plasmid and pCAMBIA2300-CaMV35s-GUS-eGFP plasmid (control) by injection. The left leaf of each lamina of benthamiana shows the results of the CaMV35s plasmid and the right leaf shows the results of the CmP plasmid.
FIG. 4 shows GUS staining results of stable transformation of Agrobacterium GV3101 containing CmP plasmid constructed according to the invention and CaMV35s plasmid (control) into Arabidopsis thaliana by dip-flower method. The left leaf of each lamina of benthamiana shows the results of staining with the CaMV35s plasmid GUS, and the right leaf shows the results of staining with the CmP plasmid GUS.
Detailed Description
The following examples are further illustrative of the invention and are not intended to be limiting thereof.
Example 1 construction of pCAMBIA 2300-CmP-GUS-eGFP
The applicant utilizes a chestnut female-male flower difference transcriptome database to screen a gene CmLTP specifically expressed by male flowers, and the function is unknown. Further adopts real-time fluorescenceAnd (3) analyzing the expression patterns of different tissues by quantitative PCR, and collecting chestnut stems, leaves and male flowers (male flowers 1:4 months and 13 days, male flowers 2cm or so, male flowers 2:4 months and 29 days, male flowers 3:5 months and 13 days, male flowers 4:5 months and 25 days, and male flowers full bloom) in different development periods (female flowers 1 and 2 correspond to the sampling periods of male flowers 3 and 4 respectively). The sample cDNA is used as a template, and CmLTP specific primers qF are adopted: 5'-CACAACCTTGACCACTGCAC-3' and qR:5'-GTGAGGTCAATGTGCTTGGC-3' quantitative PCR amplification was performed. The Chinese chestnut housekeeping gene beta-actin is taken as an internal reference, and the primer is as follows: aF:5'-ATTCACGAGACCACCTACA-3' and aR:5'-TGCCACAACCTTAATCTTCAT-3'. The reaction was carried out on an ABI 7500 real-time quantitative PCR apparatus (Applied Biosystems, USA), 10. Mu.L of the reaction system, SYBER Green PCR Mix. Mu.L, 0.5. Mu.L (10. Mu. Mol. L) of each primer -1 ) Approximately 200ng of cDNA template was used, 3 replicates per reaction. The reaction procedure is 2min at 50℃and 10min at 95 ℃; 15s at 95℃and 1min at 60℃for 40 cycles. The relative expression level of the gene was 2 -ΔΔC(t) The formula calculation is that the relative expression quantity is calculated by taking the expression level in the stem as 1 after the correction of the internal reference beta-actin. The PCR reaction system results prove that the CmP promoter regulates the downstream CmLTP to have lower expression in the early stage of chestnut male flower development (male flowers 1 and 2), extremely obviously up-regulates the expression in the middle and later stages of male flower development (male flowers 3 and 4), especially has the highest relative expression level in male flowers 3, and has relatively lower expression level in chestnut stems, leaves and female flowers (figure 1).
Based on chestnut genome data, a primer is designed by taking a sequence about 1500bp upstream of a CmLTP gene as a reference sequence, chestnut genome DNA is taken as a template, and a specific primer F with an enzyme cutting site is utilized: 5' -CCAAGCTTTGAATGGCAGTTAATCCCTAAT-3' and R:5' -CGGGATCCATCTGAATCTGATATCTCTCAC-3' (underlined indicates the cleavage sites HindIII and BamHI, respectively) were amplified by PCR. The PCR amplification uses 50 μl reaction system containing 200ng template DNA;2 XEs Taq Master mix (premix Es Taq polymerase, 3mM MgCl) 2 400 μm dNTPs) (kang is century, beijing) 25. Mu.l; 2. Mu.l each of the forward primer (10. Mu.M) and the reverse primer (10. Mu.M); the volume was set to 50. Mu.l with double distilled water. The PCR procedure was: pre-denaturation at 94℃for 3min; denaturation at 94℃for 30s, annealing at 56.5℃for 30s, extension at 72℃for 1min,35 cycles; finally, the temperature is 72 ℃ for 5min. The PCR product was subjected to electrophoresis on a 1.0% agarose gel, and a target band of about 1.2kb was excised, followed by using QiA
Gel Extraction kit (QIAGEN, hilden, germany) the target band DNA was recovered, ligated with the pCloneEZ-NRS-TA cloning vector (Zhongmeitai and Biotechnology (Beijing) Co., ltd.) and heat-shocked to E.coli DH 5. Alpha.
The monoclonal bacteria were picked for propagation and plasmids were extracted and positive by double digestion with Hind III and Bam HI (FIG. 2A). The positive plasmid was sequenced by Wuhan Tianhua, a gene technology Co., ltd. Comparing the sequence (SEQ ID NO. 1) with the chestnut genome reference sequence, selecting the plasmid with correct sequence, and storing at-20deg.C for use.
The plasmid and pCAMBIA2300-CaMV35s-GUS-eGFP target vector (stored in this laboratory) with correct sequence were subjected to double digestion with HindIII and BamHI, respectively (FIG. 2B), and the products were subjected to 1.0% agarose gel electrophoresis to obtain target bands of about 1.2kb and 12kb, respectively, and QiA
Gel Extraction kit (QIAGEN, hilden, germany) the DNA of the band of interest is recovered. The insert (about 1.2 kb) and the vector (about 12 kb) were ligated using T4 DNA library (Thermo Fisher Scientific, US) according to the instructions. The reaction system is as follows: 1 mu L T DNA library, 2 mu L T4 DNA Ligase Buffer (10X), vector DNA about 100ng, insert DNA about 45ng, ddH 2 O was added to a final volume of 20. Mu.L; after reaction at 16℃for 12-16h (overnight), the reaction was terminated by treatment at 65℃for 10min to give a ligation reaction product.
mu.L of the ligation reaction product was used and heat-shock transformed into E.coli DH 5. Alpha. Picking up monoclonal bacteria for propagation, and utilizing a specific primer F:5' -CCAAGCTTTGAATGGCAGTTAATCCCTAAT-3' and R:5' -CGGGATCCATCTGAATCTGATATCTCTCAC-3' to perform PCR positive verification. The positive bacterial liquid is used for extracting plasmid, hind III and Bam HI double enzyme digestion is used for further confirming whether the plasmid is constructed successfully, and the result of 1.0% agarose gel electrophoresis is shown in figure 2As shown. As can be seen from FIG. 2, the pCAMBIA 2300-CmP-GUS-eGFP plasmid vector was constructed successfully, and the electrophoresis lane contained the target band of about 1.2kb fragment. Finally, 30% glycerol is added into the positive clone bacterial liquid, and the positive clone bacterial liquid is preserved at the temperature of minus 80 ℃, and pCAMBIA 2300-CmP-GUS-eGFP plasmid is preserved at the temperature of minus 20 ℃.
Example 2 Agrobacterium-mediated transient transformation of leaf discs of Benzonum and genetic transformation of Arabidopsis thaliana by the dip method
About 1. Mu.g of pCAMBIA 2300-CmP-GUS-eGFP vector plasmid is taken and added into 200. Mu.L of Agrobacterium GV3101 competent cells, the mixture is placed on ice for 30min and then treated by liquid nitrogen for 5min, and is placed on ice for 2min after heat shock at 37 ℃ for 5min, 500. Mu.L of LB liquid medium is added, and shaking culture is carried out at 28 ℃ and 180rpm for 4-5h. The bacterial liquid was collected by centrifugation at 3000rpm for 5min at 4℃and resuspended in 200. Mu.L of LB liquid medium, and 100. Mu.L of the suspension was spread on LB solid medium (containing 100mg/L kanamycin and 25mg/L rifampicin). After the culture dish is sealed by a Parafilim sealing film, culturing for 2-3d at 28 ℃, picking up monoclonal bacteria for propagation, and utilizing a specific primer F:5' -CCAAGCTTTGAATGGCAGTTAATCCCTAAT-3' and R:5' -CGGGATCCATCTGAATCTGATATCTCTCAC-3' PCR was performed to screen positive clones. The verified agrobacterium GV3101 is stored at-80 ℃ with 30% glycerol for later use.
The pCAMBIA 2300-CmP-GUS-eGFP expression vector is used, pCAMBIA2300-CaMV35s-GUS-eGFP is used as a control, and the injection method is adopted to transiently transform the leaf of the Nicotiana benthamiana (Nicotiana benthamiana), and the specific steps are as follows:
(1) Preparing agrobacterium infection liquid: the positive Agrobacterium GV3101 was streaked on LB medium (containing 100mg/L kanamycin and 25mg/L rifampicin), single clones were picked up and cultured on 5mL YEB broth (containing 100mg/L kanamycin and 25mg/L rifampicin) at 180rpm with shaking at 28℃for 1-2d, and then 1mL of the bacterial liquid was picked up and cultured in 50mL YEB broth (containing 100mg/L kanamycin and 25mg/L rifampicin) overnight to OD 600 The value is about 1.0. The cells were collected by centrifugation at 6000rpm for 8min and then treated with 25mL of induction buffer (10 mM MgCl) 2 Resuspension of 10mM MES, pH 5.8), re-centrifugation to collect the cells, use infection buffer (10 mM MgCl) 2 10mM MES, 200. Mu.M acetosyringone, pH 5.8) resuspended cells to OD 600 The value is about 1.0. Can be used after standing for 3-5h at room temperatureTransient transformation.
(2) Instantaneous conversion of raw tobacco: tobacco plants which are sown for about 45 days are used, the leaves are transformed by a penetration method, water is controlled 1d before the experiment, the core is removed, and the necrotic old leaves at the lower layer are removed. The contaminated solution was aspirated with a 1mL syringe with the needle removed, and the bacterial solution was injected into the leaf by infiltration into the back of the leaf. After 2-3d of illumination culture at room temperature, taking the transformed leaves for standby.
(3) Arabidopsis thaliana floral dip genetic transformation: when the main inflorescence of the arabidopsis is pod-bearing, a small amount of flowers are bloomed in the secondary inflorescence, and most of the flowers are white and not bloomed, flower dipping is carried out, and the upper inflorescence tissue is immersed into an invasion dye solution (1/2 MS,5.0% sucrose, 0.05% Silwet L-77, pH=5.7) for about 10min; keeping humidity and avoiding light, horizontally placing for 24 hours, transferring into an incubator for normal growth, collecting T0 generation seeds, screening a resistant culture medium (1/2 MS, containing 100mg/L kanamycin) and obtaining transgenic T1 generation plants.
The transformed leaf of Benzonum and Arabidopsis were histochemically stained by GUS. GSU staining solution (formula: 100mM Tis HCl buffer pH7.0, 50mM sodium chloride, 0.5mM potassium ferrites, 0.5mM potassium ferricyanide, 0.01% Triton-X,2mM X-Gluc) was prepared. Immersing the material in the dyeing liquid, pumping for about 5min, and placing in a 37 ℃ incubator for more than 1 h. And (3) taking out the material after dyeing, and rinsing with 75% ethanol for 3-5 times, wherein each time lasts about 30min until the material is completely decolorized. The decolorized material can be soaked in 70% ethanol for long-term storage. The staining results of the transformed material were observed under a microscope and photographed, and the results are shown in fig. 3 and 4. From fig. 3 and 4, it can be seen that the CmP promoter has an expression activity with a stronger promoter in both the lamina of benthamiana and arabidopsis thaliana.
The foregoing is merely a preferred embodiment of the present invention, and it should be noted that the above-mentioned preferred embodiment should not be construed as limiting the invention, and the scope of the invention should be defined by the appended claims. It will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the spirit and scope of the invention, and such modifications and adaptations are intended to be comprehended within the scope of the invention.
Sequence listing
<110> institute of fruit tree tea at academy of agricultural sciences in Hubei province
<120> Chinese chestnut male flower specific expression promoter and application thereof
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1117
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 1
tgaatggcag ttaatcccta atatatccac acacaaaaaa gaaaaagaaa aaaaaacaat 60
tcttaattaa ctaagaatag gggcaataca agtgattaca accccatttg gatagcgttt 120
tgctcctcac ttttggcgtt tttgcctttt ttgtttttgt tttttttttt gaagagtgcc 180
ttttacattg ttcatgagac atgaacagtg cattaagaca aatgtatagt aattcagcag 240
tgagcagtac taactcataa attttttttt attttatttt cagtaataaa ttttcagttt 300
tcagtaaaat aagtggtatc caaacacaca ctacatatga tgatttacac ccccaatcac 360
cacaaaccat gtgcatcgac aagtaaaact tagaatctct gatactttat ggtttgtttg 420
gattgaaaga aagggaggag gagtagcata ggaaaggaga gtagagtttg tccaaaatta 480
gcctattttc agccaatttt acttctctct ctccactttg cctcccttcc tcttcaatcc 540
aaacatgccc ttaatttgcc acttcaattt tacttaatag aatcagtgac taaaacttct 600
tttgataaca tcatttgatt atatatatat atatatatta taaatattat acgtgtgtgt 660
gaaactttca catgtaattt tccttccgcg attggctaat ttttgtaggt tatattcata 720
tttacacaat cttctttctc caacgcaaag acagtactaa agacaaacgt cacatgaaat 780
gaggagcagc aacctcgtga ttctgtcatg tagattgtct tcattagcat tttattataa 840
ggcaaattgt ctgatcatat tggttagtat ttcaataatc tctaccttcc gaaaatcacc 900
ccaacccacc ttacatccca tttagcacta tacgcttcaa tccaatccca attctgtcat 960
ttcaatcaag caaataaaca ttacatgaca aataacttgt gaacccaaag caccttactc 1020
actctcttct atatatatat aggccaacat taattacagg ttaattactg cgtaaacgag 1080
caaataaggt ttaaagtgag agatatcaga ttcagat 1117
Claims (4)
1. A chestnut male flower specific expression promoter is characterized in that the nucleotide sequence is shown as SEQ ID NO. 1.
2. An expression vector, which is characterized by comprising the chestnut male flower specific expression promoter of claim 1.
3. A host bacterium comprising the expression vector according to claim 2.
4. The use of the promoter of claim 1 in constructing transgenic plants specifically expressed by chestnut flowers.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110928218.3A CN113684211B (en) | 2021-08-12 | 2021-08-12 | Specific expression promoter of chestnut male flowers and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110928218.3A CN113684211B (en) | 2021-08-12 | 2021-08-12 | Specific expression promoter of chestnut male flowers and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113684211A CN113684211A (en) | 2021-11-23 |
| CN113684211B true CN113684211B (en) | 2023-04-28 |
Family
ID=78579708
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110928218.3A Active CN113684211B (en) | 2021-08-12 | 2021-08-12 | Specific expression promoter of chestnut male flowers and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113684211B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104611362A (en) * | 2015-01-05 | 2015-05-13 | 北京农学院 | In-vivo expression method of woody plant |
| CN105145340A (en) * | 2015-09-08 | 2015-12-16 | 北京市农林科学院 | Molecular breeding method of short catkin chestnut varieties |
| CN110791586A (en) * | 2019-12-06 | 2020-02-14 | 湖北省农业科学院果树茶叶研究所 | SSR (simple sequence repeat) marker primer group for identifying Chinese chestnut varieties and application thereof |
-
2021
- 2021-08-12 CN CN202110928218.3A patent/CN113684211B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104611362A (en) * | 2015-01-05 | 2015-05-13 | 北京农学院 | In-vivo expression method of woody plant |
| CN105145340A (en) * | 2015-09-08 | 2015-12-16 | 北京市农林科学院 | Molecular breeding method of short catkin chestnut varieties |
| CN110791586A (en) * | 2019-12-06 | 2020-02-14 | 湖北省农业科学院果树茶叶研究所 | SSR (simple sequence repeat) marker primer group for identifying Chinese chestnut varieties and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| Chen Guosong等 .Roles of the GA-mediated SPL Gene Family and miR156 in the Floral Development of Chinese Chestnut (Castanea mollissima).Int. J. Mol. Sci..2019,第20卷第1-17页. * |
| Staton M等.CMCMBd0158E07f CMCMBd Chinese Chestnut BAC Library (EcoRI) Castanea mollissima genomic, genomic survey sequence.GeneBank .2013,第1-2页. * |
| 李琳玲 ; 陈小玲 ; 袁红慧 ; 程水源 ; 程华 ; .板栗IFRs启动子分离及表达载体构建.北方园艺.2017,(11),第107-114页. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113684211A (en) | 2021-11-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107435047B (en) | Low-phosphorus-resistant key gene GmPHR25 in plant phosphorus signal network and application thereof | |
| CN112625103B (en) | Alfalfa WRKY transcription factor and application thereof in aluminum toxicity and salt stress resistance | |
| CN114014917A (en) | FvbHLH36 protein and coding gene and application thereof | |
| CN111944818B (en) | Promoter Arah1-P of peanut allergen gene Arah1 as well as cloning and application thereof | |
| CN113684211B (en) | Specific expression promoter of chestnut male flowers and application thereof | |
| CN113151306B (en) | Gene PmWRKY57 for improving cold resistance of plum blossom petals and application thereof | |
| CN115772212A (en) | Alfalfa chloroplast MsSAP22 gene and application thereof in improving drought resistance of plants | |
| CN115724930A (en) | Tobacco NtMYB transcription factor and application thereof | |
| CN114672494A (en) | Application of tobacco NtEXB1 gene in plant branch development regulation | |
| CN114181941A (en) | Peanut promoter P28 and application thereof | |
| CN110117594B (en) | Efficient expression promoter for citrus fruit epidermis and application thereof | |
| KR101494191B1 (en) | Promoter for directing the guard-cell specific expression | |
| CN111718938B (en) | High-temperature inducible promoter specifically expressed by plant green tissue and application thereof | |
| CN104560906B (en) | Specifically expressed protein C YP734A1 like 1 and its application in fibrocyte | |
| CN114561387B (en) | Peanut promoter and application thereof | |
| KR102527047B1 (en) | A novel promoter based on the characteristics of drought stress specific cis-regulatory element and its manufacturing method | |
| CN112626081B (en) | Longan flowering regulation gene DlWRKY25, and regulation protein and application thereof | |
| CN110904110B (en) | Application of OsHAP3C gene expression reduction in rice variety with shortened heading period and prolonged growth period | |
| CN113416732B (en) | Dendrobium officinale salt inducible promoter proDoMYB75 and application thereof | |
| CN116478998B (en) | Rice phloem specific expression promoter POs04g0452500 and application thereof | |
| CN112322621B (en) | Eucommia DIR1 gene MeJA response promoter and application thereof | |
| CN112725353B (en) | Recombinant vector, transformant, primer for amplifying AtNAC58 gene and preparation method and application thereof | |
| CN110819640B (en) | Tobacco NtNRP1 gene and application thereof | |
| CN113789327B (en) | Dendrobium officinale constitutive strong promoter ProDoWOX1 and application thereof | |
| KR102390866B1 (en) | Constitutive promoter derived from chrysanthemum and use thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |