Application of a gibberellin oxidase and its encoding gene in regulating plant growth

Abstract

The present invention provides an application of a gibberellin oxidase and a coding gene thereof in regulating plant height. The gibberellin oxidase protein has an amino acid sequence shown in SEQ ID NO: 2. The protein is overexpressed in plants to increase plant height.

Description

Technical Field

[0001] The invention relates to the field of biotechnology, and in particular to application of a gibberellin oxidase and a coding gene thereof in regulating plant strains. Background Art

[0002] Gibberellins are growth regulators in plants and are tetracyclic diterpene carboxylic acid compounds widely found in higher plants. They play a very important role in seed germination, stem elongation, leaf expansion, flower and fruit development, and wood formation. Gibberellin oxidase plays a key role in regulating the balance of gibberellins in plants. It is the main regulatory site for catalyzing the late stages of GA biosynthesis and decomposition. It helps plants adapt to complex and changing environments by regulating the content of active and inactive gibberellins in plants to maintain a dynamic balance. Research on the function and expression regulation of gibberellin oxidase can effectively regulate the growth and development of plants to adapt to different environmental conditions and agricultural production needs.

[0003] Studies have shown that gibberellin oxidase is involved in many processes of plant growth and development, such as internode elongation, root growth, fruit development, and abiotic stress response. Currently, gibberellin oxidase genes have been cloned from a variety of plants, such as Arabidopsis, rice, and poplar.

[0004] However, there are no reports on the cloning of the gibberellin oxidase gene in Lonicera japonica and its application in increasing plant height. Summary of the invention

[0005] The present invention aims to solve one of the technical problems in the related art at least to a certain extent. The present invention provides a gibberellin oxidase protein isolated from Lonicera japonica and its encoding gene, which is obtained from Lonicera japonica for the first time, filling the gap of isolating and cloning gibberellin oxidase genes in Lonicera japonica. It is also found that the expression level of the gene in the transgenic strain of Lonicera japonica overexpressing gibberellin oxidase is significantly higher than that of the wild-type plant, and the plant height of the transgenic strain of gibberellin oxidase overexpressing is significantly greater than that of the wild-type, confirming the application of the gibberellin oxidase gene in increasing plant height.

[0006] Specifically, the present invention provides the following technical solutions:

[0007] The first aspect of the present invention provides a Lonicera japonica gibberellin oxidase protein having an amino acid sequence shown in SEQ ID NO:2.

[0008] According to an embodiment of the present invention, the protein is encoded by the sequence shown in SEQ ID NO:1.

[0009] A second aspect of the present invention provides an isolated nucleic acid molecule having a sequence as shown in SEQ ID NO: 1 or a sequence complementary to the sequence as shown in SEQ ID NO: 1.

[0010] According to an embodiment of the present invention, the nucleic acid molecule is cDNA. The cDNA molecule has a sequence shown in SEQ ID NO: 1, and can be used to encode the Lonicera japonica gibberellin oxidase protein.

[0011] The third aspect of the present invention provides a Lonicera japonica gibberellin oxidase gene encoding gibberellin oxidase, wherein the gene has a sequence shown in SEQ ID NO:1.

[0012] The fourth aspect of the present invention provides a construct comprising the nucleic acid molecule described in the second aspect or the gene described in the third aspect.

[0013] According to an embodiment of the present invention, the construct is at least one of a plasmid, a virus, and a bacteriophage.

[0014] The fifth aspect of the present invention provides a host cell, comprising the nucleic acid molecule described in the second aspect, the gene described in the third aspect, or the construct described in the fourth aspect.

[0015] The sixth aspect of the present invention provides a transgenic plant, comprising the nucleic acid molecule described in the second aspect, or the gene described in the third aspect, or the construct described in the fourth aspect, or the host cell described in the fifth aspect.

[0016] According to an embodiment of the present invention, the transgenic plant includes at least one of poplar, locust, willow, paulownia, ash, Arabidopsis, tobacco, rice, spinach, celery, lettuce, marigold, soybean, corn, sugarcane, peanut, potato, tomato, and sorghum. According to a specific embodiment, the transgenic plant is poplar. For example, it can be 84k poplar. 84k poplar is a deciduous tree of the genus Populus of the Salicaceae family, and has a variety of excellent characteristics, including rapid growth, strong disease resistance, wide adaptability, no flying catkins pollution, etc., and therefore plays an important role in ecological improvement and urban greening.

[0017] A seventh aspect of the present invention provides a method for regulating plant height, comprising:

[0018] Introducing the nucleic acid molecule described in the second aspect, or the gene described in the fourth aspect, or the construct described in the fourth aspect, or the host cell described in the fifth aspect into the plant;

[0019] Plants overexpressing the gibberellin oxidase protein of Lonicera japonica were screened and obtained.

[0020] According to an embodiment of the present invention, the nucleic acid molecule, or the gene, or the construct, or the recombinant cell is introduced into the plant by Agrobacterium transformation.

[0021] The eighth aspect of the present invention provides an application of the above-mentioned protein or nucleic acid molecule or gene in regulating the height of a plant.

[0022] The beneficial effects achieved by the present invention are at least:

[0023] The invention provides a honeysuckle gibberellin oxidase protein and a coding gene. The gene and the protein are obtained from honeysuckle for the first time, filling the gap of isolating and cloning gibberellin oxidase protein and gibberellin oxidase gene from honeysuckle. The relative gene expression amount and plant height of the transgenic strain of honeysuckle gibberellin oxidase protein overexpressed in the honeysuckle are significantly higher than those of the wild type, and the gibberellin oxidase gene can be used to increase plant height. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 It is a map of the recombinant vector provided according to an embodiment of the present invention.

[0025] Figure 2 It is the relative expression result of the gibberellin oxidase gene of the wild type and transgenic plants provided according to the embodiments of the present invention.

[0026] Figure 3 It is a schematic diagram of plant height phenotypes of wild-type and transgenic poplars grown in a greenhouse for one month according to an embodiment of the present invention.

[0027] Figure 4 It is a statistical graph of plant heights of wild-type and transgenic poplars grown in a greenhouse for one month according to an embodiment of the present invention. DETAILED DESCRIPTION

[0028] Embodiments of the present invention are described in detail below. Examples of the embodiments are shown in the accompanying drawings. The embodiments described below with reference to the accompanying drawings are exemplary and are intended to be used to explain the present invention, but should not be construed as limiting the present invention.

[0029] The invention provides a honeysuckle gibberellin oxidase protein, which has an amino acid sequence shown in SEQ ID NO:2.

[0030] SEQ ID NO:2

[0031] MVIDCMVTKMSQPPKDEHEKEKQLIFDASVLQHESNIPTQFIWPDHEKPCATTRPLPVPLIDLAGFLSGNPTAATEASKLVGEACQEHGFFLVVNHGVDSNLINHAHKYMDLFFELPLAEKKRAQRQMGEHCGYASSFTGRFSSKLPWKETLSFQYSAEKDSSNIVEDYFRTKMGDDFSKLGKVYQDYSNAMSTLSLGIMELLGMSLGVSQAHFKEFFEENNSIMRLNYYPPCQKPDLTLGTGPHCDPTSLTILHQDTVGGLEVFVDDEWRSISPNYSAFVVNIGDTFMALSNGRYKSCLHRAVVNNKTPRKSLAFFLCPKKDKVVSPPSELVDTKNPRIYPDFTWPMLLEFTQKHYRADMNTLQAFSNWLQQHKTSQLVL (SEQ ID NO:2)

[0032] The protein is encoded by the sequence shown in SEQ ID NO:1.

[0033] SEQ ID NO:1

[0034]

[0035] The present invention also provides an isolated nucleic acid molecule, wherein the nucleic acid molecule has a sequence shown in SEQ ID NO: 1 or a sequence complementary to the sequence shown in SEQ ID NO: 1.

[0036] According to an embodiment of the present invention, the nucleic acid molecule is cDNA. The cDNA molecule has a sequence shown in SEQ ID NO: 1, and can be used to encode the Lonicera japonica gibberellin oxidase protein.

[0037] The invention provides a honeysuckle gibberellin oxidase gene, which encodes the honeysuckle gibberellin oxidase. The gene has a sequence shown in SEQ ID NO:1.

[0038] The present invention provides a kind of construct again, including the nucleic acid molecule described above. The construct mentioned can be a cloning vector such as a T vector, a lambda phage vector, a P1 phage vector, a cosmid vector, a bacterial artificial chromosome, a yeast artificial chromosome, pGEM-T, pUC18, or an expression vector such as an adenovirus vector, a retrovirus vector or a plasmid vector. The plasmid vector can be a plant expression vector such as an expression vector selected from plant expression vectors pBIN19, pBI121, pBI221, pCAMBIA1300, pGreen, etc., and the type of the vector can be selected according to actual needs. According to an embodiment of the present invention, the construct is at least one of a plasmid, a virus, and a phage.

[0039] The present invention further provides a host cell, comprising the above-mentioned nucleic acid molecule or the above-mentioned construct.

[0040] The host cell includes but is not limited to plant, bacteria, yeast or insect cells. According to a specific embodiment, the recombinant cell is a plant cell.

[0041] The present invention further provides a transgenic plant, comprising the nucleic acid molecule, or the construct, or the host cell.

[0042] The present invention further provides a method for increasing plant height, comprising:

[0043] Introducing the above-mentioned nucleic acid, or the construct, or the host cell into the plant;

[0044] Plants overexpressing the gibberellin oxidase protein of Lonicera japonica were screened and obtained.

[0045] The overexpression mentioned refers to the phenomenon that the amount of protein synthesized, accumulated or released in the cell exceeds the normal level. Specifically in this article, overexpression means that the expression amount of the gibberellin oxidase protein of Lonicera japonica shows a significant difference compared with plants or wild plants without the introduction of exogenous genes or exogenous proteins. According to a specific embodiment, the significant difference mentioned can be p<0.5.

[0046] For example, the isolated nucleic acid molecule or gene provided by the present invention as described above can be transformed into a recipient plant cell by Agrobacterium-mediated method or gene gun method, and differentiated into a complete plant to obtain a plant capable of highly expressing the gibberellin oxidase protein of Lonicera japonica.

[0047] Of course, gene editing techniques such as zinc finger nuclease technology (ZincFingerNuclease, ZFN), TALEN technology (Transcription activator-like effectornucleases), CRISPR / Cas (Clustered Regularly Interspaced Short palindromicRepeat Sequences) technology, etc. can also be used to modify the genomic sequence of the recipient plant genome so that it contains the isolated nucleic acid molecule or gene as described above, using the isolated nucleic acid molecule or the gene as described above as a template.

[0048] The nucleic acid molecule or gene or protein described above can be used to increase the content of the gibberellin oxidase protein of Lonicera japonica in transgenic plants. The nucleic acid molecule or gene provided by the present invention can increase the content of the gibberellin oxidase protein of Lonicera japonica in transgenic plants, thereby increasing the plant height. Taking poplar as an example, compared with wild-type poplars that have not been transferred with the gene, the plant height of the transgenic poplar plants is at least increased by 1.4 to 1.8 times, for example, by 1.5 to 1.6 times.

[0049] The protection scheme of the present invention is described below by specific examples. It should be noted that these examples are only used to facilitate the understanding of those skilled in the art and should not be regarded as limiting the scope of protection of the present invention. Unless otherwise specified, the reagents mentioned in the examples can be obtained commercially.

[0050] Example 1: Cloning of the coding sequence of the Lonicera japonica gibberellin oxidase gene

[0051] The gibberellin oxidase (EVM0024577) gene sequence was obtained from the Lonicera japonica database, and its amplification primers were analyzed and designed according to the base sequence of its coding region to amplify the gene. The gene sequence is shown in SEQ ID NO: 1, and the amino acid sequence of its encoded protein is shown in SEQ ID NO: 2.

[0052] Specifically, the PCR method was used to reverse transcribe the extracted RNA from the leaves of Lonicera japonica, and then the obtained cDNA was used as a template to amplify the coding region sequence of the EVM0024577 gene, including:

[0053] (1) Total RNA extraction from leaves of Populus chinensis

[0054] The leaves of the tissue culture seedlings of Lonicera japonica were cut and ground into powder under liquid nitrogen. Total RNA was extracted according to the instructions of the Plant RNA Rapid Extraction Kit (RN38-EASYspin Plus) of Beijing Adlai Biotechnology Co., Ltd.

[0055] (2) cDNA synthesis

[0056] 1) Prepare the reverse transcription mixture according to the following system and place it in a 200µL RNase free PCR tube. The system is shown in Table 1:

[0057] Table 1: Reverse transcription system

[0058] Components Dosage Total RNA 50 ng-5 µg Anchored oligo (dT) 18 primer 1 µL 2 × TS Reaction Mix 10 µL TransScript RT / RI Enzyme Mix 1 µL gDNA Remover 1 µL RNase-free Water to 20 µL

[0059] 2) Mix thoroughly and incubate at 42°C for 30 min;

[0060] 3) Heat at 85℃ for 5 seconds to inactivate TransScript RT / RI and gDNA Remover;

[0061] 4) The obtained cDNA was stored at -20°C. All reagents used in the reaction were purchased from Beijing Quanshijin Biotechnology Co., Ltd.

[0062] (3) Amplification of target gene

[0063] 1) Based on the CDS sequence of the EVM0024577 gene of Lonicera japonica, the homologous amplification primers for its coding region were analyzed and designed:

[0064] EVM0024577-F1:ATGGTCATTGATTGTATGGTCACG (SEQ ID NO:3)

[0065] EVM0024577-R1: CAAGACAAGCTGAGACGTTTTATGTTG (SEQ ID NO: 4)

[0066] 2) Using the cDNA sequence of Lonicera japonica as a template, the cDNA sequence of Lonicera japonica EVM0024577 was amplified according to the following PCR reaction system.

[0067] The following reagents and dosages are referred to the PrimeSTAR MaxDNA Polymerase Reagent Instructions of Bio-Rad Biotechnology (Beijing) Co., Ltd., as shown in Table 2 below:

[0068] Table 2: PCR amplification system

[0069] Components Dosage cDNA 1 μL EVM0024577-F1 2 μL EVM0024577-R1 2 μL PrimeSTAR Max Premix 50μL Enzyme-free sterile water 45μL Total volume 100μL

[0070] The corresponding PCR reaction program is as follows:

[0071] Pre-denaturation: 98°C for 2 min;

[0072] (denaturation: 98°C 10 s, annealing: 55°C 15 s, extension: 72°C 1 min 30 s) × 35 cycles;

[0073] Extension: 72℃ for 5 min; keep warm at 16℃.

[0074] The amplified EVM0024577 gene sequence is shown in SEQ ID NO:1.

[0075] (4) Agarose gel electrophoresis detection

[0076] Weigh 0.5 g agarose, add 50 mL 1×TAE, heat in a microwave oven, add 5 μL GoldView (purchased from Zhongke Ruitai Biotechnology Co., Ltd.), shake well, and pour into the gel plate. After the agarose gel solidifies, add the mixture of PCR product and loading buffer (purchased from Beijing Quanshijin Biotechnology Co., Ltd.) into the gel well for electrophoresis detection, and recover the cDNA fragment with a length of 1143 bp.

[0077] Example 2

[0078] Example 2 The EVM0024577 gene overexpression vector was constructed by the following method, including:

[0079] (1) Use Primer Premier 5 to analyze the EVM0024577 gene and design primers with Kpn I and BamH I restriction sites (the restriction sites are in bold italics):

[0080] EVM0024577-F2: ACGAATTCGAGCTCGGTACCATGGTCATTGATTGTATGGTCACG (SEQ IDNO: 5)

[0081] EVM0024577-R2:CAGGTCGACTCTAGAGGATCCCAAGACAAGCTGAGACGTTTTATGTTG (SEQ IDNO: 6)

[0082] (2) Amplify the EVM0024577 gene fragment with restriction endonuclease sites according to the method described in Example 1, and recover the PCR product.

[0083] (3) Construction of pCAMBIA2300-35S::EVM0024577:

[0084] The expression vector pCAMBIA2300 with restriction sites was double-digested with endonucleases Kpn I and BamH I. The endonucleases Kpn I and BamH I were purchased from Bio-Rad Biotechnology Beijing Co., Ltd. The pCAMBIA2300 vector structure has a 35S promoter, which can achieve overexpression of the transformed gene.

[0085] Recover the digested products separately, and connect the target gene fragment and the expression vector with homologous recombinase (purchased from Beijing Jinsha Biotechnology Co., Ltd.). The recombinant vector map is shown in Figure 1 shown.

[0086] (4) The ligation product was transformed into competent E. coli DH5α (purchased from Shanghai Weidi Biotechnology Co., Ltd.), 700 μL of LB liquid culture medium was added, and then placed in a constant temperature shaker for 30 min at 37°C and 150 rpm. The revived E. coli was centrifuged at 5,000 rpm for 1 min, and the supernatant was poured out and discarded. The remaining 100 μL was mixed by pipetting with a pipette tip, and evenly spread on LB solid culture medium containing 50 mg / mL Kan with a sterile coating rod. The culture medium was inverted in a constant temperature incubator and grown at 37°C for 12-16 h. After the colonies grew, single clones were picked and grown in 1 mL of LB liquid culture medium containing 50 mg / mL Kan at 37°C for 4 h. The picked single clones were identified by PCR. After running the gel, the bacteria with the correct bands were selected for testing to screen positive clones. The plasmid was extracted using a kit (purchased from Beijing Quanshijin Biotechnology Co., Ltd.), and the vector was named pCAMBIA2300-35S::EVM0024577.

[0087] The LB liquid culture medium was prepared as follows: 5 g of tryptone, 5 g of NaCl, and 2.5 g of yeast extract were weighed, ddH2O was added to make the total volume 500 mL, sterilized by high-pressure steam at 121°C for 20 min, and cooled to room temperature.

[0088] The preparation method of LB solid culture medium refers to LB liquid culture medium. Before making up the volume, add 5g agar powder, sterilize at 121℃ high pressure steam for 20 min, and cool to room temperature.

[0089] Example 3: Poplar genetic transformation

[0090] According to the reagent instructions, the overexpression vector 35S::EVM0024577 was introduced into poplar leaves through the genetic transformation method mediated by Agrobacterium competent GV3101 (purchased from Shanghai Weidi Biotechnology Co., Ltd.), and then transplanted into the greenhouse after a series of processes including pre-culture, infection, dark culture, induction of adventitious buds, induction of bud rooting, propagation, and seedling hardening. Specifically including:

[0091] (1) Tissue culture of 84K poplar: Cut 1-2 cm long stem segments of 84K poplar tissue culture seedlings of 5-7 weeks old and place them in subculture medium for subculture. The culture temperature is about 25℃ and the light intensity is 50 µmol·m -2 s -1 , the photoperiod is 16 h / 8 h.

[0092] The formulas of the pre-culture and subculture culture media were: MS powder 4.43 g / L, sucrose 30 g / L, and agar 6 g / L.

[0093] (2) Leaf pre-culture: Cut the 3rd to 6th leaf sequence of the sterile tissue culture seedlings, use a sterilized scalpel to make 4 to 5 horizontal cuts on the main vein of the leaf, and then place the leaf upside down on the differentiation medium without antibiotics and culture for 1 to 2 days.

[0094] The formula of the differentiation medium without antibiotics is: MS powder 4.43 g / L, sucrose 30 g / L, agar 6 g / L, NAA 0.05 mg / L, 6-BA 0.5 mg / L.

[0095] (3) Preparation of Agrobacterium infection solution: Pipette 300 µL of Agrobacterium containing pCAMBIA2300-35S::EVM0024577 vector into 100 mL LB liquid medium containing antibiotics (50 mg / L kanamycin, 50 mg / L rifampicin), mix well, and culture overnight at 28°C and 180 rpm / min. 600 When it is 0.6-0.8, infection and transformation are carried out.

[0096] (4) Infection and dark culture: In a clean bench, soak the pretreated leaves in the Agrobacterium bacterial solution for 10-15 minutes, and shake the bacterial solution every 3-5 minutes to ensure that the injured part of the leaf is fully in contact with the bacterial solution. After 10-15 minutes of infection, remove the leaves, absorb the excess bacterial solution on the leaves with sterile filter paper, and then spread the leaves with the leaf surface facing up on the differentiation medium without antibiotics, and culture them in the dark for 3-4 days.

[0097] (5) Inducing resistant adventitious buds: In a clean bench, transfer the dark-treated leaves to a differentiation medium containing 50 mg / L kanamycin and 0.2 g / L timentin for selective culture. The culture conditions are a light cycle (16 h light / 8 h dark) and a temperature of 25°C. Adventitious buds will grow from the wounds of the leaves in about 2 weeks. When the adventitious buds grow to 0.5-1 cm, they will be transferred to a new differentiation medium to promote their continued growth.

[0098] The formula of the differentiation medium with added antibiotics is: MS powder 4.43 g / L, sucrose 30 g / L, agar 6 g / L, NAA 0.05 mg / L, 6-BA 0.5 mg / L, Kan 50 mg / L, and timentin 0.2 g / L.

[0099] (6) Rooting culture: Place a single adventitious bud longer than 1 cm in a rooting medium containing 25 mg / L kanamycin and 200 mg / L timentin to induce rooting and develop into a complete plant.

[0100] The formula of the rooting medium is: MS powder 2.215 g / L, sucrose 30 g / L, agar 6 g / L, NAA 0.02 mg / L, IBA 0.05 mg / L, Kan 30 mg / L, and Timentin 0.2 g / L.

[0101] Greenhouse transplanting: When the seedlings grow to about 8 cm and have a well-developed root system, transplant them after 6-8 days of hardening. Wash the culture medium at the roots of the poplar seedlings with clean water, transplant them into sterilized nutrient soil, and place them in the greenhouse for cultivation.

[0102] Example 4

[0103] Example 4 The relative gene expression and plant height of the transgenic poplar prepared in Example 3 were tested, specifically including:

[0104] (1) In order to further screen the overexpressed transgenic strains and detect the relative expression level of the EVM0024577 gene in each transgenic strain, quantitative primers of the EVM0024577 gene were designed using the real-time fluorescence quantitative PCR (Quantitative Real-time PCR) technology, and quantitative analysis of the specific cDNA sequences in the test samples was performed using internal references.

[0105] Design primers as follows:

[0106] EVM0024577-F3: TTCACTAGGGATCATGGAG (SEQ ID NO:7),

[0107] EVM0024577-R3: GGCGGGTAATAGTTGAGT (SEQ ID NO: 8).

[0108] Gene expression analysis Specific primers for real-time fluorescence quantitative PCR are used to amplify the target gene. Actin is used as an internal reference to analyze the expression pattern of the target gene.

[0109] qRT-PCR reaction system:

[0110] SYBR Master Mix 5μL Upstream primer (EVM0024577-F3) 0.2μL Downstream primer (EVM0024577-R3) 0.2μL cDNA 1μL <![CDATA[ddH2O]]> 3.6μL Total volume 10µL

[0111] Reaction program: 95°C for 5 min; 95°C for 10 s, 60°C for 10 s, 72°C for 20 s, for 45 cycles.

[0112] The experiment was repeated three times, and the data obtained were analyzed using 2 -△△CT Methods The relative expression levels were calculated.

[0113] Figure 2 is the relative expression level of the EVM0024577 gene in the transgenic lines. The results showed that the relative expression level of the EVM0024577 gene in L1, L4, and L7 transgenic poplar plants was greatly increased, and L1, L4, and L7 respectively represent different transgenic lines of EVM0024577 overexpressing transgenic poplars.

[0114] (2) Statistics of transgenic poplar plant height:

[0115] The first two internodes and about 1-2 cm including the stem tip of the identified wild-type and transgenic poplar tissue culture seedlings were cut and placed in a rooting medium without antibiotics for 5-8 weeks. After hardening, the tissue culture seedlings were transplanted to a greenhouse for growth. After one month of growth, the heights of the three transgenic lines L1, L4, and L7 with relatively high expression levels of EVM0024577 were observed.

[0116] Figure 3This is a schematic diagram of the plant height phenotype of wild-type (WT is wild-type) and 35S::EVM0024577 transgenic poplars grown in the greenhouse for one month. It can be clearly seen that the 35S::EVM0024577 transgenic poplars are taller than the wild-type poplars. The specific data statistics are as follows: Figure 4 shown.

[0117] Figure 4 The plant height statistics of wild-type and 35S::EVM0024577 transgenic poplars grown in the greenhouse for one month. From left to right, 35S::EVM0024577-1 / 4 / 7 correspond to three different strains of transgenic poplars. The average plant height of the transgenic poplars can reach 23.43 cm, while the average plant height of the wild-type is only 15.3 cm, and there is a significant difference between the two.

[0118] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific implementation", or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Although the embodiments of the present invention have been shown and described above, it can be understood that the above embodiments are exemplary and cannot be construed as limitations of the present invention. Those of ordinary skill in the art can change, modify, replace and modify the above embodiments within the scope of the present invention.

Claims

1. A honeysuckle gibberellin oxidase protein, characterized in that: The sequence of the protein is shown in SEQ ID NO:

2.

2. The protein according to claim 1, characterized in that The protein is encoded by the sequence shown in SEQ ID NO:

1.

3. An isolated nucleic acid molecule, characterized in that The sequence of the nucleic acid molecule is as shown in SEQ ID NO: 1 or is a sequence complementary to the sequence shown in SEQ ID NO:

1.

4. A construct, characterized in that Comprising the nucleic acid molecule of claim 3.

5. A host cell, characterized in that Comprising the nucleic acid molecule of claim 3 or the construct of claim 4.

6. A method for regulating plant height, characterized in that: include: Introducing the nucleic acid molecule of claim 3, or the construct of claim 4, or the host cell of claim 5 into the plant; Screening and obtaining plants that overexpress the gibberellin oxidase protein of Lonicera japonica; The plant is poplar.

7. The method according to claim 6, characterized in that The nucleic acid molecule, the construct, or the host cell is introduced into the plant by Agrobacterium transformation.

8. Use of the protein according to claim 1 or 2, or the nucleic acid molecule according to claim 3 in regulating the height of a plant, wherein the plant is poplar.

Images