CN116179573B - Application of carrot gibberellin oxidase gene DcGA2ox1 in regulation of plant growth and development - Google Patents
Application of carrot gibberellin oxidase gene DcGA2ox1 in regulation of plant growth and development Download PDFInfo
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Abstract
The application discloses application of a carrot gibberellin oxidase gene DcGA2ox1 in regulating and controlling plant growth and development, and belongs to the technical field of plant genetic engineering. The application discloses an application of an isolated carrot gibberellin oxidase gene DcGA2ox1, wherein the nucleotide sequence of the gene is shown as SEQ ID NO. 1; and through cloning DcGA2ox1 expressed in carrot, construction of recombinant expression vector, transformation of arabidopsis and carrot and application of the genes in regulation of transgenic plant height, flowering and seed development, the results show that: the gene can dwarf plants and inhibit the development of flowers and seeds by overexpression, so that the gene can be applied to the regulation and control of plant growth and development, and provides scientific basis for the research of the gene in plant growth.
Description
Technical Field
The application relates to the technical field of plant genetic engineering, in particular to application of carrot gibberellin oxidase gene DcGA2ox1 in regulating and controlling plant growth and development.
Background
Carrot (Daucus carota l.), an herb of the family Umbelliferae (Apiaceae), is one of the important horticultural cash crops. Carrots are also one of the ten vegetables in the world, and the market demand for carrots continues to grow. The carrot fleshy root is rich in nutrient components such as carotenoid, vitamin, natural pigment and the like, and has higher nutritive value.
Gibberellin (GA) is a plant hormone with gibberellin alkyl ring structure, which can promote plant growth, promote cell and stem elongation, leaf expansion, parthenocarpy, fruit growth, break seed dormancy, change male and female flower ratio, influence flowering time, and reduce flower and fruit abscission.
Key enzymes in GA synthesis and metabolism include Ke Bajiao phosphate synthase (CPS), kaurene Synthase (KS), kaurene Oxidase (KO), GA 20-oxidase (GA 20 ox), GA-3 beta-hydroxylase (GA-3 beta-hydroxylase), GA 2-oxidase (GA 2 ox), and the like. GA 2-oxidase encoded by GA2ox gene plays a negative regulation role in GA synthesis, and can decompose bioactive GA and its direct synthesis precursor into inactive GA through 2 beta hydroxylation.
Studies have shown that GA2ox is involved in many processes of plant growth, such as internode elongation, root growth, fruit development, and abiotic stress response. However, there is no report on the GA2ox gene of carrot, which affects the research of regulating carrot growth and development by using the gene.
Disclosure of Invention
The application aims to provide the application of the carrot gibberellin oxidase gene DcGA2ox1 in regulating the growth and development of plants, so as to solve the problems in the prior art, and the transgenic plants containing the DcGA2ox1 obtained by utilizing genetic engineering are dwarfed, stamen and seed development are inhibited, and the gene has wide application prospect in regulating the growth and development of plants.
In order to achieve the above object, the present application provides the following solutions:
the application provides application of a carrot gibberellin oxidase gene DcGA2ox1 in regulating plant growth and development, wherein the nucleotide sequence of the DcGA2ox1 is shown as SEQ ID NO. 1:
ATGGTGCTCGAGCAACCACAGTTGACTACTCCCACATTGCTTCATGGCGTTCCTGTCATTGACCTCTCAAACCCTGACTCTATTCCATGCCTTGTCAAGGCCTGTGAAGACTATGGTTTCTTCAAAGTTGTCAACCATGGCATCCCTGCTCATTTCATCTCCACTCTTGAGTCACAAGCCATCGATTTCTTTTCTTTACCTCTACATGAAAAAGAAAAGGCAGGACCTCCTCACCCTTTTGGCTATGGAAACAAGACCATTGGACGTAACGGCGACTTCGGCTGGCTTGAATATCTCCTCTTAACAACCGACCATGCCTTTGATTACCACAATCTTGCTTCTGTCTCTGCTCAGACCCCGGAAACTTTTAGGCGGGCTGTGAACGATTATGTATTTGCTGTGAAGAATATGGCATGTCAGATTCTGGAGCTTTTGGCGGATGGTTTGAGGATTCAAGAGAAGAATGTGTTTAGTAAACTTCTGATGGATGAACAGAGTGACTCTGTTTTTAGGCTGAATCACTATCCTCCCTCTCCAGATAAAAATTTGATTGGGTTCGGAGAACACACCGACCCACAAATCATATCTGTTCTGAGGTCTAATAACACCTCAGGCCTTGAAATCTGTTTGAAAGATGACAGCTCTTGGTTTTCAGTTCCTCCTGACCAAGACTCCTTCTTCATCAACGTTGGAGACTCTTTACAGGTGATGACTAATGTGAGGTTTAAGAGTGTGAAGCATAGGGTTTTGGCAAATAGTGAGAAATCAAGGGTGTCAATGATTTATTTCGGAGGACCACCACTAAGTGAAAAGATAGCTCCATTGCCTTCTTTGCTCATGGAAGGAGAGAACAGCAGTTTGTACAAGGAGTTTACTTGGTTCGAGTACAAAAAGTCTGCCTACAAGTCAAGGCTTGCTGACAATAGGCTCTGCCTGTTTGAGAAAATCGCAGCCTCATAA。
the application also provides application of the protein expressed by the carrot gibberellin oxidase gene DcGA2ox1 in regulating plant growth and development, wherein the nucleotide sequence of the DcGA2ox1 is shown as SEQ ID NO. 1, and the amino acid sequence of the protein is shown as SEQ ID NO. 2.
SEQ ID NO. 2 is shown below:
MVLEQPQLTTPTLLHGVPVIDLSNPDSIPCLVKACEDYGFFKVVNHGIPAHFISTLESQAIDFFSLPLHEKEKAGPPHPFGYGNKTIGRNGDFGWLEYLLLTTDHAFDYHNLASVSAQTPETFRRAVNDYVFAVKNMACQILELLADGLRIQEKNVFSKLLMDEQSDSVFRLNHYPPSPDKNLIGFGEHTDPQIISVLRSNNTSGLEICLKDDSSWFSVPPDQDSFFINVGDSLQVMTNVRFKSVKHRVLANSEKSRVSMIYFGGPPLSEKIAPLPSLLMEGENSSLYKEFTWFEYKKSAYKSRLADNRLCLFEKIAAS; ". Times." is terminator.
The application also provides application of the recombinant vector in regulating plant growth and development, wherein the recombinant vector comprises the DcGA2ox1.
The application also provides application of the recombinant genetically engineered cell in regulating plant growth and development, wherein the recombinant genetically engineered cell comprises the recombinant vector.
Preferably, the regulating plant growth includes regulating plant height and regulating flower and seed development.
Preferably, the plant comprises arabidopsis thaliana and carrot.
The application also provides a method for regulating the growth and development of plants, which comprises the steps of transferring a carrot gibberellin oxidase gene DcGA2ox1 into a receptor plant, and regulating the growth and development of the receptor plant; the nucleotide sequence of DcGA2ox1 is shown as SEQ ID NO. 1.
Preferably, overexpression of DcGA2ox1 reduces plant height of the recipient plant and inhibits flower and seed development.
Preferably, the recipient plant comprises arabidopsis thaliana and carrot.
The application discloses the following technical effects:
the gene DcGA2ox1 responding to the plant growth and development process is obtained from carrot variety 'five inches in black field', and is a novel gibberellin oxidase gene of the plant, and experiments prove that the gene has functions in plant growth regulation and control, and the result shows that: the transgenic plant containing the gene obtained by genetic engineering has short plant height, delayed plant stamen development and reduced seed quantity, and has wide application prospect in the aspect of regulating and controlling plant growth.
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In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings that are needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present application, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a diagram showing the screening result of transgenic DcGA2ox1 Arabidopsis plants;
FIG. 2 is a graph showing the growth state of a WT Arabidopsis plant of the present application and a DcGA2ox 1-transgenic Arabidopsis T3 generation plant, wherein WT represents a wild type control group and DcGA2ox1 represents a DcGA2ox 1-transgenic Arabidopsis plant;
FIG. 3 is a graph showing the rosette leaf growth status and plant height comparison of a WT Arabidopsis plant and a DcGA2ox1 transgenic Arabidopsis T3 generation plant according to the present application, wherein WT represents a wild control group, and OE-11, OE-14 and OE-34 represent different strains of DcGA2ox1 transgenic Arabidopsis;
FIG. 4 is a graph showing a comparison of flower development of a WT Arabidopsis plant of the present application and a DcGA2ox 1-transgenic Arabidopsis T3 generation plant, wherein WT represents a wild control group, and OE-11, OE-14 and OE-34 represent different DcGA2ox 1-transgenic Arabidopsis lines;
FIG. 5 is a comparative plot of seed development of a WT Arabidopsis plant of the present application versus a DcGA2ox1 transgenic Arabidopsis T3 generation plant, wherein WT represents a wild control group and OE-11, OE-14 and OE-34 represent different DcGA2ox1 transgenic Arabidopsis lines;
FIG. 6 is a graph showing the results of fluorescent quantitative expression verification of a WT Arabidopsis plant and a DcGA2ox 1-transgenic Arabidopsis T3 generation plant of the present application, wherein WT represents a wild control group, and OE-11, OE-14 and OE-34 represent different DcGA2ox 1-transgenic Arabidopsis lines;
FIG. 7 is a diagram showing the screening result of transgenic carrot plants transformed with DcGA2ox1 gene according to the present application;
FIG. 8 is a graph showing the growth status of a control carrot plant according to the application and a DcGA2ox1 transgenic carrot plant, wherein CK represents the control group and DcGA2ox1 represents the DcGA2ox1 transgenic carrot plant.
Detailed Description
Various exemplary embodiments of the application will now be described in detail, which should not be considered as limiting the application, but rather as more detailed descriptions of certain aspects, features and embodiments of the application.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. In addition, for numerical ranges in this disclosure, it is understood that each intermediate value between the upper and lower limits of the ranges is also specifically disclosed. Every smaller range between any stated value or stated range, and any other stated value or intermediate value within the stated range, is also encompassed within the application. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application. All documents mentioned in this specification are incorporated by reference for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the application described herein without departing from the scope or spirit of the application. Other embodiments will be apparent to those skilled in the art from consideration of the specification of the present application. The specification and examples of the present application are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are intended to be inclusive and mean an inclusion, but not limited to.
Example 1 carrot gibberellin oxidase gene DcGA2ox1 and application thereof
1. Test method
1.1 extraction of total RNA from carrot and Synthesis of cDNA
Total RNA was extracted from five inch ' carrot ' Heiyan ' samples using RNA Simple Total RNA Kit (Beijing Tiangen Biochemical technology Co., ltd.). The extracted total RNA was reverse transcribed into cDNA using Prime Script RT reagent Kit (Nanjinouzan Biotechnology Co., ltd.).
1.2 cloning of carrot gibberellin oxidase Gene DcGA2ox1
Based on carrot genome and transcriptome sequencing information, taking an Arabidopsis GA2ox family as an information probe, and carrying out search analysis to obtain the gene sequence of the carrot DcGA2ox1. Designing a pair of primers according to the sequence:
forward primer (SEQ ID NO: 3): 5'-ATGGTGCTCGAGCAACCACAG-3';
reverse primer (SEQ ID NO: 4): 5'-TTATGAGGCTGCGATTTTCTC-3'.
The cDNA of five inches in the black field is used as a template for amplification, and a PCR reaction system is as follows: template DNA 2. Mu.L, upstream primer 1.5. Mu.L, downstream primer 1.5. Mu.L, primerStar Max Premix (2X) 15. Mu.L, ddH 2 O10. Mu.L; the PCR reaction conditions were: pre-denaturation at 98℃for 3min; denaturation at 98℃for 10s, annealing at 55℃for 30s, extension at 72℃for 30s for 35 cycles; extending at 72℃for 5min. The PCR product is recovered after agarose gel electrophoresis with the mass and volume fraction of 1.2 percentThe target band was ligated to pMD19-T vector (TaKaRa bioengineering Co., ltd.) and transformed into E.coli DH 5. Alpha. And the extracted plasmid was subjected to PCR identification and then sequenced by Nanjing Jinsri Biotechnology Co., ltd.
1.3 construction of recombinant expression vector of DcGA2ox1 Gene
(1) Firstly, obtaining a linearization vector of pCAMBIA1301 by using a double enzyme digestion BamHI and SacI (Thermo Scientific) method, and then purifying by agarose gel electrophoresis and a gel recovery kit (Hangzhou virtute Biochemical technology Co., ltd.) to obtain the linearization vector of pCAMBIA1301 with high purity;
(2) Adding target fragment DNA and a linearization vector pCAMBIA1301 into a 1.5mL centrifuge tube in a molar ratio of 3:1 for recombination reaction, uniformly mixing, connecting at room temperature for about 30min, adding 10 mu L of reaction solution into 50 mu L of DH5a competent cells, lightly mixing by a pipette, incubating for 30min on ice, rapidly placing on ice for cooling after heat shock for 45s in a 42 ℃ water bath;
(3) 300. Mu.L of LB liquid medium was added and incubated at 37℃for 45-60min. Centrifuging at 5000rpm for 2min, collecting thallus, discarding part of supernatant, suspending the thallus with the rest culture medium, lightly coating with sterile coating rod on LB solid culture medium containing 50mg/L Kan resistance, and culturing in 37 deg.C incubator for 16-24 hr;
(4) Several clones on the recombinant reaction conversion plate are selected for colony PCR identification, positive colonies are identified, corresponding single colonies are selected for culture in a liquid LB culture medium containing 50mg/L Kan antibiotics at 37 ℃ and 200rpm incubator overnight, plasmids are extracted or bacterial liquid is directly sequenced to identify the accuracy of the vectors.
(5) After successful identification, the plasmid pCAMBIA1301-DcGA2ox1 was stored.
1.4 transfer of the recombinant vector into Agrobacterium GV3101
(1) 2 mug of recombinant vector pCAMBIA1301-DcGA2ox1 is added into each 100 mug of GV3101 agrobacterium competent cells, the mixture is stirred by hands at the bottom of a tube and mixed uniformly, and then the mixture is placed on ice for 5min, liquid nitrogen for 5min, water bath at 37 ℃ for 5min and ice bath for 5min. Adding 700 mu L of LB liquid medium without antibiotics, and culturing for 2h at 28 ℃ in a shaking way;
(2) The cells were collected by centrifugation at 6000rpm for 1min, 100. Mu.L of the supernatant was left, resuspended cells were gently blown, and the cells were uniformly spread on a YEB solid medium containing 50mg/L Kan and 100mg/L Rif, and were placed upside down in a 28℃incubator for 2 days, and positive clones were picked up for verification.
1.5 Arabidopsis transformation
1.5.1 cultivation of Arabidopsis thaliana
An appropriate amount of wild type Arabidopsis seeds were placed in a 1.5mL centrifuge tube, washed 3 times with 75% alcohol and 1 time with 5% NaClO. After rinsing 4-5 times with ultra-pure water on an ultra-clean bench, seeds are evenly spread on MS solid culture medium by a sterilized inoculating loop. The spread seeds were vernalized at 4℃for 3d and incubated in an illuminated incubator for about 10d. Then, selecting seedlings with consistent growth vigor, planting the seedlings in sterilized nutrient soil and continuously culturing the seedlings in a light culture box. Arabidopsis seedlings of about one month were selected as materials for transformation experiments.
1.5.2 genetic transformation of Arabidopsis thaliana
10 mu L of Rif and 20 mu L of Kan are respectively added into 20mL of LB liquid medium, shaking is carried out, bacteria are connected, and shaking and activation are carried out for 8-10h at 220rpm at 28 ℃ to obtain an activated bacterial liquid of agrobacterium. 100 mu L of Rif and 200 mu L of Kan are respectively added into 200mL of YEB liquid culture medium, 5-10mL of activated bacterial liquid is added, shake culture is carried out for 14-16h at 220rpm at 28 ℃ until the OD value is 1.6-2.0, centrifugation is carried out for 10min at 4500rpm, supernatant is discarded from the precipitate bacterial body, and the precipitate bacterial body is naturally dried. 100mL of 5% sucrose and 20. Mu.L of SILWETL-77 surfactant solution were added to the precipitated cells to resuspend the cells, and a pipette was used to blow the cells uniformly and resuspend the cells. Adding the bacterial liquid in the centrifugal bottle into a plate, folding the arabidopsis inflorescence, immersing the plate, gently shaking for 15s, after the transformation, uniformly stirring the bacterial liquid, sleeving the plant by using a black bag, and keeping moisture for 24h in a dark place. The transformation was repeated once more after one week.
1.5.3 Positive plant selection
Planting the seeds harvested in the T0 generation of arabidopsis thaliana, sterilizing the T0 generation of seeds, inoculating hygromycin-containing MS screening culture medium, culturing for 10 days at 22 ℃ by illumination, and screening to obtain T1 generation positive plants. And (3) carrying out single plant seed collection on the T1 generation positive plants to obtain T1 generation seeds, continuing to carry out hygromycin screening to obtain T2 generation positive plants, taking out the T2 generation positive plants by forceps when the plants grow to 10d, and staining the T2 generation positive plants and the WT control by using a GUS staining kit. The obtained positive plants are transplanted and grown, leaf genome DNA is extracted as a template, SEQ ID NO. 3 and SEQ ID NO. 4 are used as upstream and downstream primers, PCR molecular identification is carried out by referring to the description of Green Taq Mix kit (Nanjinouzan biotechnology Co., ltd.) to determine T2 generation positive plants, and the PCR identification result is shown in figure 1.
Phenotype observation of 1.5.4 Arabidopsis positive plants
Wild WT arabidopsis and T2 generation seeds were simultaneously sown in the sterilized vegetative soil seeds, vernalized at 4 ℃ for 3d, transferred into a plant growth room until 45d of growth, and observed (the results are shown in fig. 2). By the time of growth for 70d, the length of the main stem was measured as the plant height of Arabidopsis thaliana (the result is shown in FIG. 3).
1.5.5 determination of flower development of Arabidopsis thaliana positive plants
The T3 generation plants obtained after the sowing of the wild type WT arabidopsis and the T2 generation seeds are respectively observed for the flower development states of the wild type and the transgenic arabidopsis plants, and the difference between the top ends of the filaments and the top ends of the stigma is measured, and the result is shown in figure 4.
1.5.6 observation of seed development of Arabidopsis thaliana positive plants
The wild type WT arabidopsis and the T3 generation plants obtained after the T2 generation seed sowing are respectively observed and counted for the seed development condition and the seed number in the silique in the later growth and development period of the wild type and the transgenic arabidopsis plants, and the result is shown in figure 5.
Quantitative expression verification of 1.5.7 Arabidopsis positive plants
(1) Taking leaf samples of Wild (WT) and transgenic Arabidopsis lines OE-11, OE-14 and OE-34, immediately quick-freezing with liquid nitrogen, and storing in a refrigerator at-80 ℃, and obtaining RNA and cDNA according to the method in the method of extracting total RNA of 1.1 carrot and synthesizing cDNA;
(2) Expression detection primers were designed for the reference sequences selected from carrot DcGA2ox1, 19 genes of Arabidopsis thaliana, and the transcriptional expression level of Actin in Arabidopsis thaliana, as shown in Table 1.
TABLE 1 primer sequences
(3) Real-time quantitative PCR was performed using a ChamQ SYBR qPCR Master Mix kit provided by Nanjinopran Biotechnology Co., ltd.
(4) The calculation formula of the relative transcription expression level of the target gene is 2 -ΔΔCt ，ΔΔCt＝(Ct Target gene -Ct Actin ) Treatment group- (Ct) Target gene -Ct Actin ) Control group.
(5) The data were analyzed and plotted to verify the relative expression level as shown in fig. 6.
1.6 carrot transformation
1.6.1 cultivation of carrot
Selecting carrot seeds with embryo, soaking in 5ml centrifuge tube for 12 hr, washing with 75% alcohol for 1 time, washing with 40% NaClO for 1 time, and soaking in 40% NaClO for 45min. Use of sterilized ddH in ultra clean bench 2 O is washed 3-5 times and placed on filter paper to absorb water. Seed is spread on B5 solid culture medium to accelerate germination, and dark culture is carried out for about 10d. The hypocotyl of the seedling was cut into 3-5 mm long sections and used as explants.
1.6.2 genetic transformation of carrot
Agrobacterium was shake cultivated in YEB medium containing 100mg/L Rif and 50mg/L Kan at 28℃for 18h. The culture was centrifuged at 4000rpm for 15min, resuspended at a density of od600=0.4 in B5 medium containing 3% sucrose and 200 μm acetosyringone (ph 5.2) and shake-cultured for 1h. The hypocotyl sections were immersed in the agrobacterium suspension for 15min, the explants transferred to B5 solid medium and then co-cultured for 2d in the dark at 25 ℃.
Transferring the infected explant into a conical flask, and sterilizing with ddH 2 O-washing for 4 timesAdding sterile ddH containing carbenicillin sodium (Cb) 2 O, place on 100rpm shaking table, wash for 10min. The explants were then transferred to B5 screening solid medium for dark culture for 30d.
1.6.3 culture of callus and differentiation of transgenic regenerated plants
Explants with callus differentiation are transferred into new screening media for subculture until the callus is sufficiently large. It was then transferred to hormone-free B5 solid medium to induce bud differentiation. The differentiated embryogenic buds are placed for light culture, and after true leaves grow out, observation is carried out.
1.6.4 identification of transgenic carrot
And (3) extracting DNA from the leaves of the screened positive plants, identifying that the leaves contain DcGA2ox1 genes by using a PCR method, carrying out molecular verification of target genes of transgenic plants, and finally confirming that the genes are transferred into the positive plants. Positive plants and control plants were stained using GUS staining kit (Shanghai Pu Di Biotechnology Co., ltd.) and the results are shown in FIG. 7.
Phenotype observation of 1.6.5 carrot positive plants
Control plants and transgenic carrot plants were sampled simultaneously after 3 months of growth and the results are shown in fig. 8.
2. Test results
(1) As shown in figures 2 and 3, the result shows that the positive plant obtained by transferring the carrot DcGA2ox1 gene obtained by cloning of the application into an Arabidopsis plant has slow rosette leaf growth, fewer and smaller leaves and obviously shorter plant height than a control plant.
(2) As shown in fig. 4, stamen development of transgenic plants was significantly inhibited, indicating that expression of DcGA2ox1 gene altered flower development of recipient plants.
(3) As shown in FIG. 5, the number of seeds of the transgenic plants was significantly less than the control, indicating that expression of the DcGA2ox1 gene altered seed development of the recipient plants.
(4) As shown in FIG. 6, the results of fluorescent quantitative PCR showed that the expression level of 19 genes in the transgenic plants was affected by the overexpression of DcGA2ox1 gene.
The analysis result of the fluorescence quantitative expression and the physiological index can prove that: the carrot DcGA2ox1 gene can obviously dwarf receptor plants and inhibit flower development and seed development.
(5) As shown in FIG. 7, both PCR and GUS staining showed that the carrot DcGA2ox1 gene had been transferred into the recipient plant carrot.
(6) As shown in FIG. 8, the carrot DcGA2ox1 gene obtained by cloning the application is transferred into a positive plant obtained in a carrot plant, the overground part grows slowly, and the plant height is obviously lower than that of a control.
The above embodiments are only illustrative of the preferred embodiments of the present application and are not intended to limit the scope of the present application, and various modifications and improvements made by those skilled in the art to the technical solutions of the present application should fall within the protection scope defined by the claims of the present application without departing from the design spirit of the present application.
Claims (10)
1. The application of the carrot gibberellin oxidase gene DcGA2ox1 in regulating the growth and development of plants is characterized in that the nucleotide sequence of the DcGA2ox1 is shown as SEQ ID NO. 1, the regulation of the growth and development of plants is realized by reducing plant height, inhibiting stamen development and reducing seed number, and the plants are arabidopsis.
2. The application of the carrot gibberellin oxidase gene DcGA2ox1 in regulating the growth and development of plants is characterized in that the nucleotide sequence of the DcGA2ox1 is shown as SEQ ID NO. 1, the regulation of the growth and development of plants is to reduce the plant height, and the plants are carrots.
3. The application of the protein expressed by the carrot gibberellin oxidase gene DcGA2ox1 in regulating the growth and development of plants is characterized in that the nucleotide sequence of the DcGA2ox1 is shown in SEQ ID NO. 1, the amino acid sequence of the protein is shown in SEQ ID NO. 2, the regulation and the control of the growth and the development of the plants are realized by reducing the plant height, inhibiting the stamen development and reducing the seed number, and the plants are arabidopsis thaliana.
4. The application of the protein expressed by the carrot gibberellin oxidase gene DcGA2ox1 in regulating the growth and development of plants is characterized in that the nucleotide sequence of the DcGA2ox1 is shown in SEQ ID NO. 1, the amino acid sequence of the protein is shown in SEQ ID NO. 2, the plant growth and development is regulated to reduce the plant height, and the plants are carrots.
5. Use of a recombinant vector comprising DcGA2ox1 of claim 1 for regulating plant growth to reduce plant height, inhibit stamen development and reduce seed number, wherein the plant is arabidopsis.
6. Use of a recombinant vector comprising DcGA2ox1 of claim 1 for regulating plant growth to reduce plant height, said plant being carrot.
7. Use of a recombinant genetically engineered cell comprising the recombinant vector of claim 5 for regulating plant growth to reduce plant height, inhibit stamen development and reduce seed number, wherein the plant is arabidopsis.
8. Use of a recombinant genetically engineered cell comprising the recombinant vector of claim 5 for regulating plant growth to reduce plant height, wherein the plant is carrot.
9. A method for regulating the growth and development of plants, which is characterized by comprising the steps of transferring a carrot gibberellin oxidase gene DcGA2ox1 into a receptor plant, and regulating the growth and development of the receptor plant; the nucleotide sequence of DcGA2ox1 is shown as SEQ ID NO. 1; the regulation and control of plant growth and development is to reduce plant height, inhibit stamen development and reduce seed number, and the plant is Arabidopsis thaliana.
10. A method for regulating the growth and development of plants, which is characterized by comprising the steps of transferring a carrot gibberellin oxidase gene DcGA2ox1 into a receptor plant, and regulating the growth and development of the receptor plant; the nucleotide sequence of DcGA2ox1 is shown as SEQ ID NO. 1; the regulation of plant growth and development is to regulate and reduce plant height, and the plant is carrot.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102757487A (en) * | 2011-04-27 | 2012-10-31 | 中国农业大学 | Plant dwarfing related protein GA2ox, and encoding gene and application thereof |
| CN111718940A (en) * | 2019-03-22 | 2020-09-29 | 南京农业大学 | Sequence of carrot exogenous hormone-responsive related DcWRKY69 gene and application thereof |
| CN113215180A (en) * | 2021-06-08 | 2021-08-06 | 吉林大学 | Corn 9-cis-epoxy carotenoid dioxygenase protein gene ZmVP14 and application |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102757487A (en) * | 2011-04-27 | 2012-10-31 | 中国农业大学 | Plant dwarfing related protein GA2ox, and encoding gene and application thereof |
| CN111718940A (en) * | 2019-03-22 | 2020-09-29 | 南京农业大学 | Sequence of carrot exogenous hormone-responsive related DcWRKY69 gene and application thereof |
| CN113215180A (en) * | 2021-06-08 | 2021-08-06 | 吉林大学 | Corn 9-cis-epoxy carotenoid dioxygenase protein gene ZmVP14 and application |
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