CN113150092A

CN113150092A - CsHD1 protein related to apical development and dwarfing, gene and application thereof

Info

Publication number: CN113150092A
Application number: CN202110190577.3A
Authority: CN
Inventors: 张金智; 胡春根; 曾仁芳; 付黎明
Original assignee: Huazhong Agricultural University
Current assignee: Huazhong Agricultural University
Priority date: 2021-02-18
Filing date: 2021-02-18
Publication date: 2021-07-23

Abstract

The invention relates to the technical field of plant genetic engineering, in particular to CsHD1 protein related to apical development, late flowering and dwarfing, a gene and application thereof. The CsHD1 gene is separated from the sweet orange for the first time and is subjected to functional identification, and the gene is found to be capable of promoting the apical development of the orange, causing the dwarfing of plants and prolonging the vegetative growth period of the apical of transgenic plants. In addition, it can make the transgenic plant show dwarfing, blade wrinkling phenotype and other phenotype, and the gene can prolong the vegetative growth stage of plant tip and delay flowering time. The invention analyzes the regulation and control mechanism of citrus apical development and dwarfing, is beneficial to realizing the regulation and control and improvement of citrus plant types in actual production, and is beneficial to utilizing a genetic engineering means to finally achieve the purposes of increasing both production and income.

Description

CsHD1 protein related to apical development and dwarfing, gene and application thereof

Technical Field

The invention relates to the technical field of plant genetic engineering, in particular to a CsHD1 protein related to apical development and dwarfing, a gene and application thereof.

Background

The citrus is the first fruit tree in the world, belongs to perennial woody plants, three types of new shoots are usually generated in the growth period of the citrus in one year, grow in spring, summer and autumn respectively, and are called as spring shoots, summer shoots and autumn shoots respectively, the top ends of the new shoots can drop after developing to a certain degree, and the phenomenon generally exists in all citrus varieties and is called as the top bud self-shearing process. The self-pruning of citrus is a physiological process of active shedding in the growing area at the top of the branch tip, and although the self-pruning process has important significance in the growing and developing processes of various plants, so far, no detailed report has made a reasonable explanation on the rationality of the phenomenon and the aspects of evolution, molecular regulation mechanism and the like.

Disclosure of Invention

One purpose of the invention is to provide application of the CsHD1 protein in regulation of plant apical development and plant dwarfing, wherein the amino acid sequence of the CsHD1 protein is shown as SEQ ID NO: 1.

The other purpose of the invention is to provide the application of the CsHD1 gene of the encoded CsHD1 protein in regulating and controlling the plant apical development and plant dwarfing, and the amino acid sequence of the CsHD1 protein is shown as SEQ ID NO: 1.

Wherein the CsHD1 gene is the DNA molecule described in any one of the following (1) to (2):

(1) the nucleotide sequence of the DNA molecule is shown as SEQ ID NO. 2;

(2) a DNA molecule which is hybridized with the DNA sequence defined in (1) under strict conditions and encodes CsHD1 protein.

The third purpose of the invention is to provide an expression vector containing CsHD1 gene or application of agrobacterium to regulation of plant apical development and plant dwarfing, wherein the nucleotide sequence of the CsHD1 gene is shown as SEQ ID NO. 2.

Wherein, the expression vector is a PBI121 expression vector.

Wherein the plant is Arabidopsis, tobacco and citrus.

In the research of early sweet orange new self-pruning, 1378 genes differentially expressed before and after self-pruning are identified, the genes are related to Programmed Cell Death (PCD) and cell wall biosynthesis or metabolism, and genes closely related to the development of plant apical meristems, a plurality of Homeobox genes containing a Homeobox structural domain are separated through an expression spectrum, one KNOX (KNOTTED 1-like) family member gene CsHD1 has an important regulation function in the apical development of plants, and the gene is presumed to have an important regulation function in the apical development of citrus and influence the self-pruning process.

The CsHD1 gene is separated from sweet orange for the first time and is subjected to functional identification, the nucleotide sequence of the separated gene is shown as SEQ ID NO. 2, the amino acid sequence of the coded protein is shown as SEQ ID NO. 1, the gene is found to be capable of promoting apical development, prolonging the apical development time, prolonging the vegetative growth period of branch tips, enabling transgenic plants to show phenotypes such as dwarfing and leaf wrinkling, and the gene is capable of greatly prolonging the vegetative growth stage of the plant tips and delaying the flowering time of the plant tips. The invention analyzes the regulation and control mechanism of citrus apical development and dwarfing, is beneficial to realizing the regulation and control and improvement of citrus plant types in actual production, and is beneficial to utilizing a genetic engineering means to finally achieve the purposes of increasing both production and income.

Drawings

FIG. 1 is a flow chart of citrus genetic transformation;

FIG. 2 shows the phenotypic results of wild type Arabidopsis thaliana and Arabidopsis thaliana into which the CsHD1 gene has been transferred;

FIG. 3 shows the results of the phenotype of normal citrus and citrus transformed with CsHD1 gene;

FIG. 4 shows the results of the phenotype of normal lemon and lemon with the CsHD1 gene transferred.

Detailed Description

The principles and features of this invention are described below in conjunction with specific embodiments, the examples given are intended to illustrate the invention and are not intended to limit the scope of the invention. Unless defined otherwise, 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 invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

Example 1 cloning of CsHD1 Gene

The embodiment provides a method for cloning a CsHD1 gene, which comprises the following steps:

(1) extraction of plant RNA

The collection of the plant tissue sample (sweet orange top tissue) needs an ice box or liquid nitrogen, and the collected plant tissue sample is immediately stored in a refrigerator at minus 80 ℃ for standby. The whole process of RNA extraction needs to be carried out in an ultra-clean workbench, and the reagent adopts Trizol kit of Takara company, and the main steps are as follows:

a. putting 1mLRNAiso Plus reagent into a centrifuge tube of 2mL RNAase free, and putting on ice;

b. liquid nitrogen sample grinding: placing a proper amount of sample in a mortar, pouring liquid nitrogen, and quickly grinding the sample into powder;

c. taking a proper amount of ground sample, placing the ground sample in an RNAioso Plus solution, standing at room temperature, adding 1/5 volumes of chloroform, shaking, uniformly mixing, standing at room temperature for 5min, and centrifuging at 12000r/min at 4 ℃ for 10 min;

d. optional steps are as follows: transferring the supernatant into a new 2mL RNAase free centrifuge tube, adding chloroform with the same volume, and repeating the step c once;

e. transferring the supernatant (about 600 μ L) into a new 1.5mL RNAase free centrifuge tube, adding isopropanol with the same volume, shaking, mixing, standing at room temperature for 10min, and centrifuging at 12000r/min at 4 deg.C for 10 min;

f. discarding the supernatant, adding 500 μ L of pre-cooled 75% ethanol (prepared with DEPC water), suspending and precipitating, and washing to remove salt;

g. removing alcohol, adding appropriate amount of RNase Free ddH after the precipitate is dried₂Dissolution of OPrecipitating;

h. 2.5 mul RNA solution is taken for electrophoresis detection of the quality, and the rest RNA can be directly used for reverse transcription or stored at minus 80 ℃ for standby.

(2) Reverse transcription reaction

The reagent for reverse transcription reaction is PrimeScript of reverse transcription kit of Takara company^TMRT reagent Kit with gDNA Eraser (cat No. RR047A), all procedures were performed in an ultraclean bench. The reaction steps are as follows:

a. reaction process for removing genome DNA

The reaction mixture was prepared as follows, and carefully performed on ice using a RNase-free PCR tube.

TABLE 1 genome removal reaction System

Reacting at 42 deg.C for 2min or standing at room temperature for 5-10min, and performing the next step.

b. Reverse transcription reaction

TABLE 2 reverse transcription reaction System

Reacting the prepared system at 37 ℃ for 15-20min, then reacting at 85 ℃ for 5s, and storing at 4 ℃ for later use, wherein the prepared system is stored in a refrigerator at-80 ℃ for a long time.

(3) Cloning of genes

The experiment mainly uses the product obtained by the reverse transcription of the RNA of the sweet orange tissue as a PCR amplification template and adopts high-fidelity DNA polymerase of Nanjing Nuojingzan company

Max Super-Fidelity DNA Polymerase kit for PCR amplification reaction. The system is as follows:

wherein: the forward and reverse primers are: 5'-GATAGACAGACTGCCGTTTC-3', respectively;

5’-AGAACACCTGCTGAATTTGC-3’。

TABLE 3 Gene amplification System

TABLE 4 Gene amplification PCR program

And after the reaction is finished, taking out the PCR product, and carrying out electrophoresis detection or storing the PCR product in a refrigerator at 4 ℃ for a short time.

(4) Vector construction

All reagents used for vector construction adopt a one-step cloning method of Nanjing NuoZan Biotechnology limited

The kit II is constructed by fusing a target gene and a carrier through recombination reaction, and the specific experimental operation steps are as follows:

a. firstly, carrying out plasmid extraction on escherichia coli carrying a PBI121 vector, carrying out double enzyme digestion on the obtained vector plasmid by using restriction enzymes BamHI and SacI under the condition of enzyme digestion at 37 ℃ for 30 minutes, designing a recombinant primer to carry out PCR reaction, adding recombinant recognition sites 5'-ACGGGGGACTCTAGAGGATCC-3' and 5'-CGATCGGGGAAATTCGAGCTC-3' to two ends of a target fragment, measuring the concentration of the vector and the target fragment after the vector and the target fragment are prepared, and adjusting the proportion.

b. The ligation reaction of the vector and the target fragment is carried out at 25 ℃ or 37 ℃ according to requirements, and the specific ligation reaction system is as follows:

TABLE 5 ligation reaction System

(5) Transformation of Agrobacterium

After the vector is successfully constructed, the plasmid is introduced into an agrobacterium-infected competent cell for agrobacterium transformation, and the specific operation steps are as follows:

a. taking out Agrobacterium GV3101 competence from-80 deg.C refrigerator, freeze thawing on ice, adding 3-4 μ L plasmid, quick freezing in liquid nitrogen for 1min, warm bathing in 37 deg.C thermostat for 5min, and standing on ice for 5 min;

b. adding 600 plus 800 mu L of liquid non-resistant LB culture medium, shaking the bacteria for 2-3h under the condition of 28 ℃ and 220r/min, and recovering competent cells;

c. centrifuging at high speed for 1min, removing supernatant, spreading the bacterial solution on solid LB + Rif solid culture medium with corresponding resistance, and culturing at 28 deg.C for 2-3 days;

d. after the long spots, monoclonal colonies are picked for PCR detection, and positive colonies can be used for plant transformation.

Example 2 transformation of Arabidopsis thaliana

The transformation procedure of Arabidopsis thaliana was as follows: when the arabidopsis in a culture room grows to blossom, the genetic transformation of the arabidopsis is carried out by adopting a inflorescence infection method, firstly, the required agrobacterium is subjected to shake or streak activation, and the activated bacterium liquid is subjected to the transformation experiment of the arabidopsis, and the specific operation steps are as follows:

a. preparing an osmotic culture solution: 1/2MS solution or 5% sucrose solution, adding 0.5g MES per liter, and adjusting pH to about 5.7;

b. the agrobacterium is prepared for overnight culture one day before transformation until bacterial liquid OD₆₀₀1.2-1.6;

c. centrifuging at room temperature and 5000rpm for 15min to collect thallus;

d. discarding the supernatant, suspending the cells with the prepared permeate to make OD₆₀₀Adding an arabidopsis transformation auxiliary reagent Silwet L-77 into 50mL of suspension liquid at a ratio of 10 mu L at about 0.8;

e. directly spraying the agrobacterium tumefaciens suspension onto the arabidopsis flowers or placing the flowers in the suspension for about 20s, pouring a proper amount of water into a basin, covering a transparent plastic cover or a preservative film to keep enough humidity, opening the cover after culturing for one day under a light-avoiding condition, and culturing under normal illumination;

f. the transformation can be carried out again after a period of about one week, in the same manner as above;

g. after the seeds are mature, collecting the seeds, placing the seeds in a drying oven at 37 ℃ for drying for several days, and then carrying out positive screening, wherein positive seedlings are firstly eluted in 75% alcohol for about 8min and then eluted by 100% alcohol for about 5min, and are absorbed by a sterile sheared gun head to be dried on sterile filter paper, and are spread on 1/2MS culture medium with corresponding resistance (the step is carried out in an ultra-clean workbench of a tissue culture room), and the seedlings are cultured under normal illumination after vernalization at 4 ℃ for three days until the seedlings grow out, and the normal growth seedlings are positive seedlings;

h. after the positive seedlings are transplanted into soil and grow for several days, DNA or RNA can be extracted to carry out further positive detection on the molecular level and observe the phenotype, and the result is shown in figure 2: the CsHD1 transgenic Arabidopsis mainly shows the characters of blade wrinkling, dwarfism, slow growth, late flowering, long vegetative growth cycle and the like.

Example 3 genetic transformation of tobacco

One day ahead of time, Agrobacterium was prepared and cultured Overnight (OD) in a shaker at 28 ℃₆₀₀About 1.2), the next day, the cultured cell suspension was centrifuged to collect cells, and the obtained cells were suspended in a non-anti-MS liquid medium, OD₆₀₀The value is adjusted to be 0.8-1.0 for standby, the whole transformation process is carried out in a clean bench, and the detailed transformation operation steps are as follows:

a. cutting healthy wild tobacco leaves into small pieces with the length of about 1cm (a small amount of wound is properly made for agrobacterium infection), placing the tobacco leaves with the back face facing upwards in an infection solution, and suspending for about 10min while continuously shaking;

b. taking out the leaves with sterile forceps, placing on sterile filter paper, blotting residual liquid, spreading in a dish containing empty solid MS culture medium, sealing the dish, and culturing in dark for three days;

c. shaking and cleaning with cephalosporin water (concentration 400mg/L) for 3 times, each time for about 5min, and then shaking and cleaning with clear water for two times, each time for about 10 min;

d. sucking residual liquid with filter paper, placing the liquid on a differentiation culture medium (MS +6-BA + NAA + Cef) with corresponding antibiotics with the back side facing upwards, culturing in an aseptic illumination culture room for about 30 days to perform bud cutting and rooting culture (the culture medium for rooting culture is MS + Cef with corresponding antibiotics), and replacing the culture medium every 20 days to prevent pollution if no bud grows;

e. after the seedlings grow out, DNA or RNA is extracted for further positive detection on the molecular level and the phenotype is observed, and the result is shown in figure 3, and in the obtained transgenic tobacco, compared with a wild control, the transgenic tobacco shows obvious dwarfing and leaf shrinkage phenotype.

Example 4 genetic transformation of lemon

The genetic transformation procedure for lemon was as follows:

a. preparation of transformation staining solution: 2 days before transformation, streaking activation is carried out on LB solid medium by using an inoculating loop, the cells are cultured in an incubator at 28 ℃ until the cells grow out (the monoclonals can be picked up to ensure the activity and streaking activation is carried out once again), the cells are scraped by using a sterile pipette tip or other tools and are put into 50mL of MT suspension medium containing 20mg/LAS (50 mu LAS mother liquor is added per 50 mL), and shaking culture is carried out for 2 hours at 28 ℃. Measure its OD₆₀₀Adjusting the value to be in the range of 0.6-0.8, and then carrying out the next conversion process. The mother solution concentration of AS is 20mg/mL, and the preparation method comprises the following steps: weighing 2g of powder, dissolving, fixing the volume to 100mL of DMSO, subpackaging the dissolved liquid into a 1.5 or 2.0mL centrifuge tube, and storing at-20 ℃;

b. preparation of the conversion material: taking out healthy fresh lemon seeds from fruits, soaking in 1mol/L NaOH solution for 15min to remove pectin, sterilizing with 2% NaClO in a super clean bench for about 20min, and finally washing with sterile water for 3-5 times, wherein the sterilized seeds can be stored in a refrigerator at 4 deg.C for later use (can be stored in the refrigerator for half a year). When seeds are stripped, the inner and outer seed coats of the seeds are removed by using sterile forceps in a superclean workbench, the seeds are sowed in an MT solid culture medium, and the seeds are cultured in a tissue culture room for 20 to 40 days in a shading mode and are cultured in the light for about one week, so that the seeds can be used for genetic transformation;

c. and (3) transformation process: taking the epicotyl of a well-grown and strong lemon seedling, cutting into stem sections with the length of about 1cm in a super clean workbench by using a sterile scalpel, inclining the cut as far as possible to increase the cut area, temporarily storing the cut stem sections in a sterile triangular flask, paying attention to the fact that a small amount of MT suspension culture medium is put in the flask for moisture preservation, pouring all agrobacterium infection liquid into the triangular flask after all stem sections are cut, infecting for 15-20min, and continuously shaking during the period;

d. culturing and regenerating: after the infection process is finished, absorbing bacteria liquid remained on the surface of the stem section by using sterile filter paper, transferring the bacteria liquid to a co-culture medium (MT + AS) with a layer of sterile filter paper paved on the surface, culturing for 3 days under a dark condition, washing for 3-5 times by using sterile filter paper, absorbing water by using the sterile filter paper, placing the mixture into a screening culture medium, culturing in a culture chamber for 30-50 days in a shading mode, and then transferring to a normal illumination condition for culturing. When the regenerated bud grows to about 0.5cm, the regenerated bud is cut off and transferred to a germination culture medium for continuous growth, and when the regenerated bud grows to about 3cm, the regenerated bud can be transferred to a rooting culture medium for growth. The culture medium needs to be replaced by subculture every 30-40 days in the culture process;

e. DNA of a transgenic material is extracted as a template, PCR is carried out to identify the positive condition (see technical process) and the phenotype is observed, because the transgenic lemon grows very slowly, the test adopts a grafting method to propagate the transgenic lemon, the result is shown in figure 4, the CsHD1 transgenic lemon shows the phenotypes such as internode shortening, leaf wrinkling and the like, wherein the most obvious characteristic is that the top end of the transgenic lemon keeps a continuous growth and development state, and the phenotype of the whole plant shows a dwarfing character which is consistent with the transgenic phenotype of tobacco.

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.

Sequence listing

<120> CsHD1 protein related to apical development and dwarfing, gene and application thereof

<160> 2

<170> SIPOSequenceListing 1.0

<210> 1

<211> 379

<212> PRT

<213> orange (Citrus)

<400> 1

Met Glu Gly Tyr Asn Ser Leu Asn Glu Asn Thr Thr Pro Arg Gly Asn

1 5 10 15

Phe Leu Tyr Ser Ser Ser Gln Val Val Gly Ser Ser Ala Tyr Gly Arg

20 25 30

Ala Ser Asp Gly Gln Met Pro Ile Arg Ser Asn Ser Ser Asn Ser Phe

35 40 45

His Leu Gln Ser Gly Gly Ser Thr Cys Phe Gln Ser Glu Arg Ala Pro

50 55 60

Pro Pro Glu Ala His Thr His His Pro Thr Ile Val Lys Thr Glu Ala

65 70 75 80

Ser Thr Ser Gln Leu Ile Tyr Ser Leu Asn Met Met Arg Glu Gly His

85 90 95

His His His His Gln Ser Gln Gln Glu Gly Asn Glu Asn Ser Asp Thr

100 105 110

Glu Ala Ile Lys Ala Lys Ile Ile Ser His Pro Gln Tyr Ser Ser Leu

115 120 125

Leu Glu Ala Tyr Val Asp Cys Gln Lys Val Gly Ala Pro Pro Glu Val

130 135 140

Val Ala Arg Leu Ala Ala Ala Arg Gln Glu Phe Glu Ser Arg Gln Arg

145 150 155 160

Ser Ser Leu Asn Ser Arg Asp Ser Thr Asn Lys Asp Pro Glu Leu Asp

165 170 175

Gln Phe Met Glu Ala Tyr Tyr Asp Met Leu Val Lys Tyr Arg Glu Glu

180 185 190

Leu Thr Arg Pro Ile Gln Glu Ala Met Asp Phe Ile Arg Arg Ile Glu

195 200 205

Thr Gln Leu Asn Met Leu Ser Asn Gly Pro Val Arg Ile Phe Asn Ser

210 215 220

Thr Asp Asp Gln Lys Cys Glu Gly Val Gly Ser Ser Glu Glu Asp Gln

225 230 235 240

Glu Asn Ser Gly Gly Glu Thr Glu Leu Pro Glu Ile Asp Pro Arg Ala

245 250 255

Glu Asp Arg Glu Leu Lys Asn His Leu Leu Arg Lys Tyr Ser Gly Tyr

260 265 270

Leu Ser Ser Leu Lys Gln Glu Leu Ser Lys Lys Lys Lys Lys Gly Lys

275 280 285

Leu Pro Lys Glu Ala Arg Gln Lys Leu Leu Asn Trp Trp Glu Leu His

290 295 300

Tyr Lys Trp Pro Tyr Pro Ser Glu Thr Glu Lys Val Ala Leu Ala Glu

305 310 315 320

Ser Thr Gly Leu Asp Gln Lys Gln Ile Asn Asn Trp Phe Ile Asn Gln

325 330 335

Arg Lys Arg His Trp Lys Pro Ser Glu Asp Met Gln Phe Met Val Met

340 345 350

Asp Gly Leu His Pro Gln Asn Ala Ala Ser Leu Tyr Met Asp Gly His

355 360 365

Tyr Met Ala Asp Gly Pro Tyr Arg Leu Gly Pro

370 375

<210> 2

<211> 1140

<212> DNA

<213> orange (Citrus)

<400> 2

atggagggtt acaacagtct aaacgagaac acgactccta gaggaaattt cttgtattcg 60

tcttcacaag ttgttggatc atcagcatat ggtcgagcaa gtgatggtca gatgcctata 120

agaagcaaca gcagcaacag ctttcaccta caatcaggtg gcagtacttg tttccaatct 180

gagcgggcac cgccgcctga ggcgcacaca catcatccga cgattgtgaa aactgaagcc 240

agcacttcgc agcttatcta ctctcttaat atgatgagag aagggcatca tcatcatcat 300

caatcacagc aagaaggaaa tgagaactct gatacagaag ctattaaagc caagatcatc 360

tcccatcctc agtattccag ccttttggaa gcctacgtgg attgccaaaa ggtcggagct 420

ccgcctgaag ttgtggctcg gcttgcagct gctagacagg agttcgagtc aaggcagcga 480

tcgtcgttga attcgagaga ctctactaat aaagacccag aactcgatca atttatggag 540

gcttattacg acatgctagt gaaatatcgg gaagagctaa caaggccaat acaagaagcc 600

atggatttca tccggcggat cgaaactcag cttaatatgc ttagcaatgg ccccgttcgg 660

atcttcaact ccactgatga tcagaaatgt gaaggcgttg gctcctccga agaagatcaa 720

gaaaatagtg gtggagagac agagttacct gaaattgatc cccgggctga agatcgagaa 780

ctgaagaacc acctattgag gaaatacagt ggctacttaa gtagcctcaa acaagaactg 840

tccaagaaaa agaagaaagg gaaactaccc aaagaagcca ggcaaaagct acttaactgg 900

tgggaattac actataaatg gccttatcct tcggagacag agaaggtggc attggctgaa 960

tcaactggct tggatcagaa acagataaac aactggttta tcaaccaaag gaagcggcat 1020

tggaaacctt ctgaagatat gcaattcatg gtgatggatg gccttcatcc tcagaatgct 1080

gcttctcttt atatggatgg acactacatg gctgatggtc cataccgttt gggaccatga 1140

Claims

The application of the CsHD1 protein in regulation and control of plant apical development and plant dwarfing is characterized in that the amino acid sequence of the CsHD1 protein is shown as SEQ ID NO. 1.
2. The application of the CsHD1 gene for encoding the CsHD1 protein in regulating and controlling the plant apical development and plant dwarfing is characterized in that the amino acid sequence of the CsHD1 protein is shown as SEQ ID NO. 1.
3. The use of the CsHD1 gene encoding a CsHD1 protein of claim 2 for regulating plant apical development and plant stunting, wherein the CsHD1 gene is a DNA molecule according to any one of the following (1) to (2):

(1) the nucleotide sequence of the DNA molecule is shown as SEQ ID NO. 2;

(2) a DNA molecule which is hybridized with the DNA sequence defined in (1) under strict conditions and encodes CsHD1 protein.
4. The application of the expression vector containing the CsHD1 gene or the agrobacterium tumefaciens in regulating and controlling the plant apical development is characterized in that the nucleotide sequence of the CsHD1 gene is shown as SEQ ID NO. 2.
5. The application of the expression vector or the agrobacterium containing the CsHD1 gene in regulation of plant apical development and plant dwarfing as claimed in claim 4, wherein the expression vector is a PBI121 expression vector.
6. The use of the expression vector or the agrobacterium containing the CsHD1 gene according to claim 4 for regulating plant apical development and plant stunting, wherein the plant is arabidopsis, tobacco and citrus.