KR20150051692A - BrSTY1 gene in plant dwarfing and their uses - Google Patents

Abstract

The present invention relates to a BrSTY1 gene involved in plant dwarfness and a use thereof. Specifically, chrysanthemum, petunia and tobacco that are difficult to transfect were transfected with a BrSTY1 gene that belongs to short internode (SHI)-family genes from Chinese cabbage. As a result, the transfected plants were identified to have dwarfness without an influence on the flowering time. Therefore, the gene of the present invention can be useful in cultivating dwarf flowering plant for differentiation and dwarf plants.

Description

BrSTY1 gene and its use related to plant dwarf {BrSTY1 gene in plant dwarfing and their uses}

The present invention relates to a BrSTY1 (Brassica rapa STYLISH1) gene involved in plant dwarfism and its use.

Dwarfism refers to a trait whose size is smaller than the standard size of the species, or a breed with such traits. These dwarf traits are important in various aspects of agriculture and horticulture. For example, by reducing plant height or stem length, it is possible to produce ornamental plants with new aesthetic values and to produce crops with new commercial value, such as having a bite-sized size by reducing vegetables or fruits .

In addition to these industrial applications, dwarfing also enables efficient use of laboratory space, as it facilitates handling with the miniaturization of laboratory plants and reduces the space required for growing plants.

In order to make these dwarf properties appear in plants, growth regulators of various chemicals are now being used as topical agents, but these substances are found to be harmful to the environment and human body. In the case of foreign countries, the use of paclobutrazol, daminozide, etc., representative substances of dwarf induction, is prohibited or reevaluated by environmental groups. In addition, although B-9 (Daminozide), Psycocell (Crylcecote), Sumi Seven Liquid (Uniconazole) and Bipiopaabel (Parcobutazole) are used as causative agents, And the like.

These dwarf varieties can not be easily secured by traditional breeding in a short period of time, and development of GM flowers using genes involved in growth regulation has been actively conducted in foreign countries. Among them, Arabidopsis gai -1 gene-assisted petunia was developed or Agrobacterium rhizogenes role Using gene waehwa Callan was developed in co-production was a baby when blooming through the overexpressed genes SHI derived from the pole, leaf shape, etc. to other waehwa Callan nose while holding a flower jean features a small size, derived from Arabidopsis thaliana, poplar SHI Overexpression of the gene induced diminishing dendritic characteristics. In addition, studies are under way on how to identify and control size-related genes that can induce dwarfism in a variety of crops.

Thus, the present inventors have searched for a dendritic gene without processing a causative agent and discovered a BrSTY1 (Brassica rapa STYLISH1) gene, which is one of the SHI (SHORT INTERNODE) genes of Chinese cabbage. As a result, the BrSTY1 gene- Was confirmed to be dwarf without being influenced by the flowering time, thereby completing the present invention.

An object of the present invention is to provide a BrSTY1 (Brassica rapa STYLISH1) gene which induces dwarfism comprising the nucleotide sequence of SEQ ID NO: 1

Another object of the present invention is to provide a protein consisting of the amino acid sequence of SEQ ID NO: 2 deduced from the gene.

It is still another object of the present invention to provide a recombinant vector containing the gene.

Yet another object of the present invention is to provide a microorganism transformed with said recombinant vector.

It is still another object of the present invention to provide a transgenic plant transformed with the above-mentioned transforming microorganism.

It is another object of the present invention to provide a method for producing the transgenic plant.

In order to achieve the above object, the present invention provides BrSTY1 (Brassica rapa STYLISH1) gene which induces dwarfism comprising the nucleotide sequence of SEQ ID NO: 1.

In addition, the present invention provides a protein consisting of the amino acid sequence of SEQ ID NO: 2.

The present invention also provides a recombinant vector comprising the gene.

In addition, the present invention provides a microorganism transformed with said recombinant vector.

The present invention also provides a transgenic plant transformed with the above-mentioned transforming microorganism.

In addition, the present invention provides a method for producing the transgenic plant.

As a result of transformation of a vector containing BrSTY1 (Brassica rapa STYLISH1) gene inducing dwarfism of the present invention into chrysanthemum, petunia and tobacco which are difficult to transform, the overall size of the plant is small, It would be useful to make dwarfed flower seeds and dwarf plants for differentiation.

1 is a schematic diagram showing a pCAMBIA1390 vector containing NPTII , which is a kanamycin resistance gene, as an antibiotic screening gene.
FIG. 2 is a schematic diagram showing a vector for transformation of a plant by fusing a BrSHI gene containing NPTII , which is a kanamycin resistance gene, as an antibiotic screening gene into a CaMV35S promoter.
FIG. 3 is a diagram showing the transformation process of chrysanthemum (A) and petunia (B) for introduction of BrSHI family gene.
4 is an analysis of transgenic petunia into which the BrSTY1 gene has been introduced:
Verification of insertion of BrSTY1 gene by genomic DNA PCR in transgenic plants (A); RT-PCR confirmed BrSTY1 gene expression (B); Morphological features of transgenic petunia (T ₁ ) overexpressing BrSTY1 gene (C); Control (control): non-transformant, # 7-2-2; # 11-1-1; # 12-2-1; # 16-2; # 18-1; # 21-2-2; # 25-1; # 27-1-2; # 30-2; # 40-1; # 45-2-1; # 47-2: Petunia transformant T ₁ system.
FIG. 5 is an analysis of the phenotype of the BrSTY1 transgenic transformed petunia flower: Cont: non-transformant; # 7-2-2; # 11-1-1; # 12-2-1; # 16-2; # 18-1; # 21-2-2; # 25-1; # 27-1-2; # 30-2; # 40-1; # 45-2-1; And # 47-2: Petunia transformants T ₁ system.
Figure 6 is an analysis of transgenic chrysanthemums into which the BrSTY1 gene has been introduced:
Verification of insertion of BrSTY1 gene by genomic DNA PCR in transgenic plants (A); Northern analysis confirmed BrSTY1 gene expression (B); Chrysanthemum transformants overexpressing BrSTY1 gene (C); Control: Non-transgenic, # 13, # 16, # 27: Chrysanthemum transformation system.
Figure 7 is an analysis of tobacco with BrSTY1 gene introduced:
Verification of insertion of BrSTY1 gene by genomic DNA PCR in transgenic plants (A); Tobacco transgenic characteristics overexpressing BrSTY1 gene (B); Control: non-transformant, # 7, # 8, # 12: tobacco transfection system.
8 is a diagram showing the nucleotide sequence of BrSTY1 (Brassica rapa STYLISH1) gene.

Hereinafter, the present invention will be described in detail.

The present invention provides BrSTY1 (Brassica rapa STYLISH1) gene which induces dwarfism consisting of the nucleotide sequence of SEQ ID NO: 1.

The gene is derived from Chinese cabbage (Brassica rapa ssp. Pekinensis).

The gene consists of the nucleotide sequence of SEQ ID NO. 1 and 1120 bp.

Variants of the above base sequences are also included within the scope of the present invention. Specifically, the gene has a nucleotide sequence having a sequence homology of 70% or more, more preferably 80% or more, still more preferably 90% or more, and most preferably 95% or more, with the nucleotide sequence of SEQ ID NO: 1 . "% Of sequence homology to polynucleotides" is ascertained by comparing the comparison region with two optimally aligned sequences, and a portion of the polynucleotide sequence in the comparison region is the reference sequence for the optimal alignment of the two sequences (I. E., A gap) relative to the < / RTI >

In addition, the present invention provides a protein consisting of the amino acid sequence of SEQ ID NO: 2.

The present invention also provides a recombinant vector comprising the gene of SEQ ID NO: 1 according to the present invention.

In order to prepare the recombinant vector, the His-tag region existing in the vector pCAMBIA1390 was treated with Spe I and BstE II to remove the recombinant vector, and the antibiotic kanamycin was selected for the selection of the petunia transformant. , The BrSTY1 gene of the BrSHI sequence was fused to the CaMV 35S promoter to produce a plant transformation vector 35SP :: BrSTY1, which is shown in FIG.

In particular, the vector described in FIG. 2 was used to select an individual having dwarfism characteristics by introducing BrSTY1 gene of BrSHI sequence into Petunia, which is difficult to transform in flower crops.

The term "recombinant" refers to a cell in which a cell replicates a heterologous nucleic acid, expresses the nucleic acid, or expresses a protein encoded by a peptide, heterologous peptide or heterologous nucleic acid. The recombinant cell can express a gene or a gene fragment that is not found in the natural form of the cell in one of the sense or antisense form. In addition, the recombinant cell can express a gene found in a cell in its natural state, but the gene has been modified and reintroduced intracellularly by an artificial means.

The term "vector" is used to refer to a DNA fragment (s), nucleic acid molecule, which is transferred into a cell. The vector replicates the DNA and can be independently regenerated in the host cell. The term "expression vector" is often used interchangeably with a "recombinant vector ". The term "recombinant vector" means a recombinant DNA molecule comprising a desired coding sequence and a suitable nucleic acid sequence necessary for expressing a coding sequence operably linked in a particular host organism. Promoters, enhancers, termination signals and polyadenylation signals available in eukaryotic cells are known.

The vector of the present invention can typically be constructed as a vector for cloning or expression. In addition, the vector of the present invention can be constructed by using prokaryotic cells or eukaryotic cells as hosts. For example, when the recombinant vector of the present invention is an expression vector and a prokaryotic cell is used as a host, a strong promoter (for example, pL? Promoter, trp promoter, lac promoter, T7 promoter, tac promoter, etc.) , Ribosome binding sites for initiation of detoxification, and transcription / translation termination sequences.

The expression vector will preferably comprise one or more selectable markers. The marker is typically a nucleic acid sequence having a property that can be selected by a chemical method, and includes all genes capable of distinguishing a transformed cell from a non-transformed cell. Examples include herbicide resistance genes such as glyphosate, glufosinate ammonium or phosphinothricin, kanamycin, G418, Bleomycin, hygromycin, ), Chloramphenicol (chloramphenicol), but are not limited thereto.

In the recombinant vector of the present invention, the promoter may be CaMV 35S, actin, ubiquitin, pEMU, MAS, or histone promoter, but is not limited thereto. The term "promoter " refers to the region of DNA upstream from the structural gene and refers to a DNA molecule to which an RNA polymerase binds to initiate transcription. A "plant promoter" is a promoter capable of initiating transcription in plant cells. A "constitutive promoter" is a promoter that is active under most environmental conditions and developmental conditions or cell differentiation. Constructive promoters may be preferred in the present invention because the choice of transformants can be made by various tissues at various stages. Thus, constitutive promoters do not limit selectivity.

In addition, the present invention provides a microorganism transformed with said recombinant vector.

The host cell is preferably a microorganism, and Agrobacterium tumefaciens GV3101 was used in the examples of the present invention.

The present invention also provides a transgenic plant transformed with the above-mentioned transforming microorganism.

The plant is preferably a flower crop, and it is most preferably petunia.

The plant is dwarfed by BrSTY1 gene expression and is not affected by flowering time.

In addition,

(1) preparing a recombinant vector comprising the BrSTY1 gene of claim 1;

(2) infecting the plant with the vector using Agrobacterium; And

(3) selecting a transformant in the above-mentioned plant.

The plant of step (2) is preferably a flower crop, and it is most preferably petunia.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are intended to illustrate the contents of the present invention, but the scope of the present invention is not limited to the following examples. The embodiments of the present invention are provided to more fully explain the present invention to those skilled in the art.

< Example  1> BrSTY1  Vectors for introducing genes

The pCAMBIA1390 vector was used to construct a vector that fused the BrSTY1 gene.

Specifically, the His-tag region present in the pCAMBIA1390 vector was removed by treatment with Spe I and BstE II, and the antibiotic kanamycin was selected for selection of the petunia transformant.

In order to construct a transformed vector containing the BrSHI gene, the BrSTY1 gene of the BrSHI family was fused to the CaMV 35S promoter to produce a plant transformation vector 35SP :: BrSTY1, which is shown in FIG.

< Example  2> BrSTY1  Transformation to introduce genes Petunia  making

In Example 1, the plant transformation vector 35SP :: BrSTY1 prepared by fusing the BrSTY1 gene to the CaMV 35S promoter was transformed into Dream Red (PanAmerican Seed) petunia leaves using Agrobacterium GV3101.

Specifically, in order to introduce the prepared vector 35SP :: BrSTY1 into the petunia, Agrobacterium tumefaciens GV3101 was repeated two times with freeze and thaw, and then the antibiotic kanamycin (kanamycin) ), 50 mg / l of rifampicin, 10 mg / l of rifampicin, and 40 mg / l of gentamycin. The introduction of the carrier in the GV3101 strain was confirmed by colony PCR, and the colonies in which the gene introduction was confirmed were used for the transformation of the petunia. The seeds were germinated in germination medium (MS basal medium + 2% sucrose + 0.9% Phytagar) and then cut into 0.5 cm when they were grown. MS basal medium + 3% sucrose + 1.0 mg / l BA, 2.0 mg / l IAA, 0.9% Phytagar) and co-cultured with agrobacterium containing 35SP :: BrSTY1 for 3 days at 25 ° C and dark. After co-cultivation, the petunia leaves were gently washed 5 times or more with sterilized water, and then cultured in selective medium (MS basal medium + 3% sucrose + 1.0 mg / l BA, 2.0 mg / l IAA, 100 mg / L carbenicillin, cefotaxim 250 mg / 50 mg / l kanamycin, 0.9% Phytagar). After about 2 to 3 weeks of cultivation, the callus was separated from the end of the section and then cultured subcultured in a selection medium and cultured until a shoot was obtained. When shoots were induced, roots were transferred to roots organic medium (MS basal medium + 2% sucrose 100 mg / l carbenicillin, cefotaxim 250 mg / l, 50 mg / l kanamycin, 0.9% Phytagar). The transformed organisms with roots were cultivated in a greenhouse through a purification step, and succeeded seeds were obtained through crossing (FIG. 3B).

< Example  3> BrSTY1  Gene-introduced Petunia  Generation and Morphological Characterization

In the case of the petunia transformant prepared by introducing the BrSTY1 gene in Example 2, 35 independent T ₀ seeds were sown in MS base medium containing 50 mg / l of kanamycin for generation progress, and antibiotic resistance The emerging individuals were selected and three transgenic plants were transferred from each individual to the greenhouse. After transferring to the greenhouse, transgenic petunia showing morphological characteristics were selected and developed into T ₁ . Transformants were identified by genomic PCR and RT-PCR using gene-specific primers. T ₁ selected from kanamycin antibiotics Transgenic petunia showing morphological characteristics were selected and analyzed in the transformants.

As a result, the number of transgenic plants developed with the BrSTY1 transgenic petunia transformant (T ₁ ) was finally 60, which is shown in Table 1 below.

Status of BrSHI gene transfected petunia transgenic (T ₁ ) kind gene Transgenic plants
Number of Objects Seed harvest completion
Number of individuals (T ₀ ) T ₁ Selection and Analysis
Population Dream Red 35S :: BrSTY1 37 35 T ₁ -60

< Example  4> BrSTY1  Gene introduction Petunia  Morphological characterization of transformant

Transgenic petunia showing morphological characteristics were selected and analyzed in T ₁ transformants selected from kanamycin antibiotics.

Specifically, BrSTY1 transgenic petunia Genomic DNA PCR and RT-PCR confirmed the presence of gene insertion and gene expression.

To confirm the introduction of the gene, Chinese cabbage BrSTY1 gene was introduced and genomic DNA was isolated from a petunia plant selected as an antibiotic (DNeasy Plant Kit, Qiagene). BrSTY1-Forward: 5 'ATG GCG GGT TTT TTC TCG TTA GGC 3' (SEQ ID NO: 3), BrSTY1-Reverse: 5, which can identify the transgene using 500 ng of isolated genomic DNA (1 min at 95 ° C, 1 min at 95 ° C, 1 min at 55 ° C, 1 min at 72 ° C, 35 cycles, 1 min at 95 ° C) using 'TCA TGA TCT TGG GTT GGG AAA GAA 3' 72 ° C for 10 minutes and 1 cycle).

As a result, no PCR amplification product was found in the non-transformant (Wt), and the PCR product was well amplified in the petunia transformants surviving the antibiotic selection, indicating that the Chinese cabbage BrSTY1 gene was introduced (FIG.

In order to analyze gene expression through RT-PCR, total RNA was isolated from transgenic petunia leaves selected from antibiotics (Trizol, Invitrogen). 3 μg of the separated total RNA was prepared by using reverse transcriptase (MMLV transcriptase RNase free, Toyobo). Each cDNA sample was diluted 3-fold and 2 μl was used for PCR amplification. (BrSTY1-forward: 5 'ATG GCG GGT TTT TTC TCG TTA GGC 3' (SEQ ID NO: 3), BrSTY1-reverse: 5 'TCA TGA TCT TGG GTT GGG AAA GAA 3' (1 cycle at 95 ° C for 1 minute, 95 ° C for 1 minute, 55 ° C for 1 minute, 72 ° C for 1 minute, 35 cycles, 72 ° C for 10 minutes, 1 cycle) Expression of the BrSTY1 gene introduced from the petunia was confirmed. Actin (actin-forward: 5 'TGG CAT CAC ACT TTT CTA CAA 3' (SEQ ID NO: 5), Actin-reverse: 5 ': CAA CGG AAT CTC TCA GCT CC 3' cDNA samples were used as quantitative control measures to ensure that cDNA was generated from the same amount of RNA. PCR amplification products were confirmed by electrophoresis and finally confirmed by DNA sequencing analysis.

As a result, it was confirmed that BrSTY1 was overexpressed in the BrSTY1 transgenic petunia transformant (FIGS. 4A and 4B). In addition, the BrSTY1 transgenic petunia transgenic plants exhibited small dendritic features of whole plant size, with dwarf and up-ward curled phenomena. The transgenic plants exhibiting dwarfed characteristics than the control (control) were consistently dwarfed and strong until the flowers of the control group, and the height of the transgenic petunia overexpressing BrSTY1 was lower than that of the non-transformants (Fig. 4C). The size of the BrSTY1 transformant flower was smaller or similar in size as compared with the control group, and the red flower color of the flower was darker than that of the control flower (Fig. 5).

< Example  5> BrSTY1  Production of transgenic chrysanthemums to introduce genes

The leaves of the chrysanthemum grown in the cabin were cut into squares of about 5 mm by using a leaf disk method modified by the method of Horsh et al. (A Simple and general method for transferring genes into plants, 1985) Lt; / RTI &gt; for 2 days. Agrobacterium containing the vector is pre-cultured in a liquid medium (2-3 days) and then about 1.5 mL is inoculated again in 100 mL of medium and harvested to an OD 600 of about 1.0. After centrifugation, the pellet was diluted in the co-culture medium and then immersed in the pre-cultured plant for about 10 minutes, and the water was removed and the cancer was cultured for 2 days in callus organic medium. Two days later, the piece of leaf co-cultured was transferred to the antibiotic medium, removed Agrobacterium for 7 days, and transferred to a shoot selection medium to select a transforming shoot. The hormones used in the culture were IAA 1 mg / L, BA 1 mg / L, and the first antibiotic was kanamycin 15 mg / L and cefotaxime 400 mg / L. In chrysanthemum transformation, secondary selection was carried out in roots medium after shoot selection, with the concentration of antibiotic used being 20 mg / L (FIG. 3A).

&Lt; Example 6 > Characterization of transgenic maize transformed with BrSTY1 gene

Transgenic chrysanthemums showing dwarfism characteristics were selected and analyzed in the transformants selected from kanamycin antibiotics.

Specifically, the BrSTY1 gene introduced chrysanthemum Genomic DNA PCR and Northern analysis confirmed gene insertion and gene expression.

In order to confirm the gene introduction, the brassiere BrSTY1 gene was introduced and genomic DNA was isolated from the chrysanthemum plant selected as an antibiotic (DNeasy Plant Kit, Qiagene). BrSTY1-forward: 5 'ATG GCG GGT TTT TTC TCG TTA GGC 3' (SEQ ID NO: 3), BrSTY1-reverse: 5 'TCA TGA (1 min at 95 ° C, 1 min at 95 ° C, 1 min at 55 ° C, 1 min at 72 ° C, 35 cycles, 72 ° C) using TCT TGG GTT GGG AAA GAA 3 ' 10 min. 1 cycle condition) to confirm the introduction of the gene in the plant.

In order to confirm gene expression by Northern analysis, 3 ~ 4 leaves of Chrysanthemum chrysanthemum chrysanthemum, selected as a transformant by genomic PCR, were finely ground with liquid nitrogen and put into a 2-mL screw cap tube so as not to dissolve. Then RNAiso Add about 1 mL and let go. Centrifuge and transfer the supernatant to a new tube so that the layers do not mix, then add 200 μL of chloroform, mix well and centrifuge again. The supernatant was separated and equilibrated with isopropanol. The pellet was centrifuged at -20 ° C for 24 hours and centrifuged. The pellet was dissolved in DEPC-treated DW, and 8M LiCl was added thereto for 4 to 24 hours. Separate the obtained pellet well and measure it. Northern analysis was performed with formaldehyde method (Genecloning protocol 7) using 10 쨉 g of the separated total RNA. The probe was labeled with P32 isotope using the Rediprime ™ II random Prime Labeling System (Amersham) and then used for the hybridization reaction.

As a result, it was confirmed that BrSTY1 was overexpressed in BrSTY1 transfected chrysanthemum transformants (FIGS. 6A and 6B). In addition, the BrSTY1 transgenic chrysanthemum transformants exhibited overall densities of the whole plants smaller than that of the control, and the leaf shape also showed a drying characteristic (Fig. 6C).

<Example 7> Characterization of BrSTY1 transgenic tobacco transformant

Transgenic tobacco showing dwarf characteristics was selected and analyzed in the transformants selected from kanamycin antibiotics.

Specifically, the BrSTY1 gene introduction was carried out in the same manner as in the above-mentioned petunia and chrysanthemum Genomic DNA PCR confirmed the presence of gene insertion

As a result, it was confirmed that BrSTY1 was overexpressed in the BrSTY1 gene-introduced chrysanthemum transformant (Fig. 7A). In addition, BrSTY1 transgenic tobacco transformants showed dendritic characteristics in which the overall plant size was generally smaller than the control (Fig. 7B).

<110> RURAL DEVELOPMENT ADMINISTRATION <120> BrSTY1 gene in plant dwarfing and their uses <130> P131023 <160> 6 <170> Kopatentin 2.0 <210> 1 <211> 1101 <212> DNA <213> Brassica rapa <400> 1 atggcgggtt ttttctcgtt aggcggcgga ggaggcagag gcggaggagg aaacggtcaa 60 gaagaacaca ggaccaacac gaatcctcct ccgcctgtat ctgactcttg gctttggtac 120 agaaacccta acgtcaacgc aaacgcaaac gcaaacgcga acgctccttc ctcttcaaac 180 gctgctgctt taggcaccct tgagctatgg caaaaccaca accagcaaga aatcatgttc 240 cagcatcaac agcatcagca aaggttggat ctatactcct ccgccgctgg tttaggagta 300 ggaccaagca gccatagcca attcaatatc tccggcgaaa cttcaaacgc cggcgccgga 360 ggaacggctg cggcggcact catgatgatg cgaagcggcg gaggaggagg aaccggtgtg 420 agctgtcaag actgtgggaa tcaggccaag aaagactgcg ctcacgttag gtgtcgaact 480 tgctgcaaga gccgtggctt ccagtgtcct actcacgtga ggagcacgtg ggttcctgcc 540 gccaaacgcc gtgagagaca acagcagtta ggcacggttc agcctcaaac gcacctgcca 600 cgcggtgata gcggcgtacc gaaacgccta cgggaaaatt taccggcaac ttcttcgtct 660 cttgtctgca ctcgcgtacc tactcatcac aacgcttcag gtttggaggt tggagatttc 720 ccagcggagg tgagctcacc ggcagtgttt agacgtgtgc gagtgagctc agtggaagac 780 ggagaagaag agtttgctta ccaaacagct gtaagcatcg gtggtcacgt ttttaaaggc 840 attctctacg acaacggtcc tggaagtatt ggtggcggtg gctacaacgt tggtgagagc 900 tcttctggtg gcggaggagc tcagcagatg aatctcataa cagctggatc tgtgacggtt 960 gcaaccgctt cttcctccac tccaaacgct ggtggtattg gcggctcctc cgcggcttac 1020 actgatccag ccgctcttta tccaactccg atcaacactt tcatggccgg tacacaattc 1080 tttcccaacc caagatcatg a 1101 <210> 2 <211> 366 <212> PRT <213> Brassica rapa <400> 2 Met Ala Gly Phe Phe Ser Leu Gly Gly Gly Gly Gly Gly Arg Gly Gly Gly   1 5 10 15 Gly Asn Gly Gln Glu Glu His Arg Thr Asn Thr Asn Pro Pro Pro              20 25 30 Val Ser Asp Ser Trp Leu Trp Tyr Arg Asn Pro Asn Val Asn Ala Asn          35 40 45 Ala Asn Ala Asn Ala Asp Ala Asp      50 55 60 Gly Thr Leu Glu Leu Trp Gln Asn His Asn Gln Gln Glu Ile Met Phe  65 70 75 80 Gln His Gln Gln His Gln Gln Arg Leu Asp Leu Tyr Ser Ser Ala Ala                  85 90 95 Gly Leu Gly Val Gly Pro Ser Ser His Ser Gln Phe Asn Ile Ser Gly             100 105 110 Glu Thr Ser Asn Ala Gly Aly Gly Gly Thr Ala Ala Ala Ala Leu Met         115 120 125 Met Met Arg Ser Gly Gly Gly Gly Gly Thr Gly Val Ser Cys Gln Asp     130 135 140 Cys Gly Asn Gln Ala Lys Lys Asp Cys Ala His Val Arg Cys Arg Thr 145 150 155 160 Cys Cys Lys Ser Arg Gly Phe Gln Cys Pro Thr His Val Arg Ser Thr                 165 170 175 Trp Val Pro Ala Ala Lys Arg Arg Glu Arg Gln Gln Gln Leu Gly Thr             180 185 190 Val Gln Pro Gln Thr His Leu Pro Arg Gly Asp Ser Gly Val Pro Lys         195 200 205 Arg Leu Arg Glu Asn Leu Pro Ala Thr Ser Ser Leu Val Cys Thr     210 215 220 Arg Val Pro Thr His His Asn Ala Ser Gly Leu Glu Val Gly Asp Phe 225 230 235 240 Pro Ala Glu Val Ser Ser Pro Ala Val Phe Arg Arg Val Val Arg Ser                 245 250 255 Ser Val Glu Asp Gly Glu Glu Glu Phe Ala Tyr Gln Thr Ala Val Ser             260 265 270 Ile Gly Gly His Val Phe Lys Gly Ile Leu Tyr Asp Asn Gly Pro Gly         275 280 285 Ser Ile Gly Gly Gly Gly Tyr Asn Val Gly Glu Ser Ser Ser Gly Gly     290 295 300 Gly Gly Ala Gln Gln Met Asn Leu Ile Thr Ala Gly Ser Val Thr Val 305 310 315 320 Ala Thr Ala Ser Ser Ser Thr Pro Asn Ala Gly Gly Ile Gly Gly Ser                 325 330 335 Ser Ala Tyr Thr Asp Pro Ala Ala Leu Tyr Pro Thr Pro Ile Asn             340 345 350 Thr Phe Met Ala Gly Thr Gln Phe Phe Pro Asn Pro Arg Ser         355 360 365 <210> 3 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> BrSTY1-Forward primer <400> 3 atggcgggtt ttttctcgtt aggc 24 <210> 4 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> BrSTY1-Reverse primer <400> 4 tcatgatctt gggttgggaa agaa 24 <210> 5 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Actin-forward <400> 5 tggcatcaca cttttctaca a 21 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Actin-reverse <400> 6 caacggaatc tctcagctcc 20

Claims (11)

BrSTY1 (Brassica rapa STYLISH1) gene inducing dwarfism consisting of the nucleotide sequence of SEQ ID NO: 1.
The gene according to claim 1, wherein the gene is derived from Chinese cabbage (Brassica rapa ssp. Pekinensis).
A protein consisting of the amino acid sequence of SEQ ID NO: 2 inferred from the gene of claim 1.
A recombinant vector comprising the gene of claim 1.
A microorganism transformed with the recombinant vector of claim 4.
6. The transformed microorganism according to claim 5, wherein the microorganism is Agrobacterium tumefaciens.
A transgenic plant transformed with the microorganism of claim 5.
8. The transgenic plant according to claim 7, wherein the plant is selected from the group consisting of tobacco, petunia, and chrysanthemum.
8. The transgenic plant according to claim 7, wherein the plant is dwarfed by BrSTY1 gene expression.
(1) preparing a recombinant vector comprising the BrSTY1 gene of claim 1;
(2) infecting the plant with the vector using Agrobacterium; And
(3) selecting a transformant in the plant.
[Claim 10] The method according to claim 10, wherein the plant of step (2) is selected from the group consisting of tobacco, petunia, and chrysanthemum.

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