WO2011108794A2 - Gène régulant la cytocinèse, plantes transformées par le gène et procédé pour la régulation de la croissance de plantes l'utilisant - Google Patents

Gène régulant la cytocinèse, plantes transformées par le gène et procédé pour la régulation de la croissance de plantes l'utilisant Download PDF

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WO2011108794A2
WO2011108794A2 PCT/KR2010/007322 KR2010007322W WO2011108794A2 WO 2011108794 A2 WO2011108794 A2 WO 2011108794A2 KR 2010007322 W KR2010007322 W KR 2010007322W WO 2011108794 A2 WO2011108794 A2 WO 2011108794A2
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gene
atsh3p
plant
represented
recombinant vector
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WO2011108794A3 (fr
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황인환
김혜란
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포항공과대학교 산학협력단
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8262—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield involving plant development
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8201—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation
    • C12N15/8202—Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation by biological means, e.g. cell mediated or natural vector
    • C12N15/8205—Agrobacterium mediated transformation
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
    • C12N15/8222—Developmentally regulated expression systems, tissue, organ specific, temporal or spatial regulation
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield

Definitions

  • the present invention relates to a gene for controlling cytoplasmic division, a plant transformed with the gene, and a method for controlling growth of a plant using the same.
  • the AtSH3P-P2 gene is able to regulate the growth of a plant when the gene is involved in cellular division and regulates its expression. Revealed to complete the present invention.
  • one example of the present invention provides an AISH3P-P2 protein represented by the amino acid sequence of SEQ ID NO: 3.
  • Another example provides an AtSH3P-P2 gene encoding the AtSH3P-P2 protein.
  • Another example provides a recombinant vector comprising the AtSH3P-P2 gene.
  • Another example provides a plant transformed with the recombinant vector. Another example provides seeds of the transformed plant.
  • Another example provides a plant growth promoter containing at least one AtSH3P-P2 protein, an AtSH3P-P2 gene encoding the same, and a recombinant vector comprising the gene as an active ingredient.
  • AtSH3P-P2 protein Another example provides the use of the AtSH3P-P2 protein, the AtSH3P-P2 gene encoding the same, and at least one plant growth promoter selected from the group consisting of a recombinant vector comprising the gene.
  • AtSH3P-P2 protein of the present invention includes a protein having the amino acid sequence of SEQ ID NO: 3 and a functional equivalent of the protein.
  • the functional equivalent is at least 70%, preferably at least 80%, more preferably at least 90%, even more preferably at least 70% of the amino acid sequence represented by SEQ ID NO: 3.
  • substantially homogeneous physiological activity refers to the growth of plants when overexpressed in the plant. When promoted or when expression is suppressed in a plant, it means an activity in which plant growth is inhibited.
  • AtSH3P-P2 gene of the present invention is not limited thereto as long as it can encode the AtSH3P-P2 protein, and includes both genomic DNA and cDNA.
  • the AISH3P-P2 gene represented by the nucleotide sequence of SEQ ID NO: 2 and variants thereof.
  • the AtSH3P-P2 gene variant has a nucleotide sequence having at least 70%, more preferably at least 80% , even more preferably at least 90% , most preferably at least 95% homology with the nucleotide sequence of SEQ ID NO: 2. It may include.
  • the '% sequence homology' of the nucleotide sequence is identified by comparing two optimally arranged sequences with a comparison region, wherein a portion of the polynucleotide sequence in the comparison region is the reference sequence (addition or deletion) for the optimal alignment of the two sequences. It may include the addition or deletion (ie, gap) compared to).
  • the AtSH3P-P2 gene of the present invention is not limited thereto. It is preferred to have the nucleotide sequence of the number 2 (see ⁇ Example 1>).
  • the recombinant vector of the present invention comprises the AtSH3P-P2 gene.
  • said recombinant vector is a recombinant plant expression vector.
  • the term "recombinant” refers to a cell replicating, expressing a heterologous nucleic acid or expressing a protein encoded by the heterologous nucleic acid.
  • the 'vector' is used when referring to DNA fragment (s), nucleic acid molecules that are delivered into cells.
  • the vector replicates DNA and can be reproduced independently in a host cell.
  • the expression vector or recombinant vector refers to a recombinant DNA molecule comprising a coding sequence of interest and an appropriate nucleic acid sequence necessary to express a coding sequence operably linked in a particular host organism.
  • the 'plant expression vector (recombinant vector expressible in plants)' is not limited thereto, but when present in a suitable host such as Agrobacterium tumerfaciens, a portion of itself, a so-called T-region that can transfer T-regions to plant cells
  • a plasmid vector and a preferred form may be the so-called binary vector described in EP0120516B1 and US Pat. No. 4,940,838.
  • Other suitable plant expression vectors can also be selected from viral vectors, such as those that may be derived from double stranded plant viruses (eg CaMV) and single stranded viruses, gemini viruses, etc., for example incomplete plant virus vectors. .
  • the use of such vectors can be advantageous especially when it is difficult to properly transform a plant host.
  • the plant expression vector is used frequently in the art pBI101 (Cat #: 6018-1, Clontech, USA), pBIN19 (Genbank Accession No. U09365), pBI121 and pCAMBIA vector. Most preferably it is a pCAMBIA vector (see ⁇ Example 2>).
  • the recombinant vector of the invention preferably comprises one or more selectable markers.
  • the marker is typically a nucleic acid sequence having properties that can be selected by chemical methods, and all genes that can distinguish transformed cells from non-transformed cells. Examples include herbicide resistance genes such as glyphosate or phosphinothricin (phosphinothricin), kanamycin, G418, bleomycin, hygromycin, chloramphenicol There are antibiotic resistance genes such as, but not limited to.
  • the promoter that may be included in the recombinant vector of the present invention is a promoter capable of initiating transcription in plant cells, and preferably may be capable of overexpressing a gene inserted into the plant. More preferably, it may be a 'constitutive promoter' which is active under most environmental conditions and developmental conditions or cell differentiation. That is, the constitutive promoter is preferable because the selection of the transformant can be made by various tissues at various stages. Even more preferably the promoters that may be included in the expression vector of the invention are CaMV 35S, actin, ubiquitin, pEMU, MAS or histone promoters. May be, but is not limited thereto.
  • 'promoter' refers to a region of DNA upstream from a structural gene and refers to a DNA molecule to which an RNA polymerase binds to initiate transcription.
  • the terminator that can be included in the recombinant vector of the present invention may use a conventional terminator, such as nopalin synthase (NOS), rice ⁇ -amylase RAmyl A terminator, phaseoline (phaseoline) terminator, agrobacterium Terminator of the octopine gene of agrobacterium tumefaciens, but is not limited thereto.
  • NOS nopalin synthase
  • rice ⁇ -amylase RAmyl A terminator phaseoline (phaseoline) terminator
  • phaseoline phaseoline terminator
  • agrobacterium Terminator of the octopine gene of agrobacterium tumefaciens but is not limited thereto.
  • such regions are generally known to increase the certainty and efficiency of transcription in plant cells.
  • the recombinant vector in one embodiment of the present invention is prepared by inserting the AtSH3P-P2 gene having the nucleotide sequence of SEQ ID NO: 2 into a pCAMBIA vector using CaMV 35S as a promoter and nopaline synthase (NOS) as a terminator. . More specifically, the recombinant vector was prepared by removing a stop codon (TGA) of the AtSH3P-P2 gene and inserting a construct linking the initiation codon (AUG) of the green fluorescent protein (GFP) into a pCAMBIA vector. It can be displayed on a map, it can be confirmed whether the expression of the AtSH3P-P2 gene through GFP (see ⁇ Example 2>).
  • the transformed plant of the present invention is transformed with the recombinant vector will be.
  • Any transformation method can be used for transferring the nucleic acid to the plant for transforming the plant.
  • the transformation method is not limited thereto but is preferably a calcium / polyethylene glycol method for protoplasts (Krens, FA et al., 1982, Nature 296, 72-74; Negrutiu I. et al., June 1987, Plant Mol. Biol. 8, 363-373), electroporation of protoplasts (Shillito RD et al., 1985 Bio / Technol. 3, 1099-1102), microscopic injection into plant elements (Crossway A. et al., 1986, Mol Gen. Genet.
  • Agrobacterium tumerfaciens mediated gene transfer by transformation can be used as appropriately selected from (incomplete) viral infection (EP 0 301 316) and the like.
  • Preferred methods according to the invention may be Agrobacterium mediated transformation methods (see ⁇ Example 3>).
  • Plants that can be transformed by introducing the AtSH3P-P2 gene in the present invention is not limited thereto, but may be food crops, vegetable crops, special crops, fruit trees, flowers and feed crops, preferably rice, wheat, barley, Corn, beans, potatoes, eight, oats, sorghum, baby pole, cabbage, radish, pepper, strawberry, tomato, watermelon, cucumber, cabbage, melon, pumpkin, green onion, onion, carrot, ginseng, tobacco, cotton, sesame, candy Sorghum, Beet, Perilla, Peanut, Rape, Apple tree, Pear, Jujube, Peach, Yard, Grape, Citrus, Persimmon, Plum, Apricot, Banana, Rose, Gladiolus, Gerbera, Carnation, It may be selected from the group consisting of chrysanthemum, lily, lily, lygras, red clover, orchardgrass, alfalfa, pescue and perennial lice.
  • AtSH3P-P2 gene in the present invention can be effectively expressed in the plant, including the baby pole, to promote plant growth, the transformed plant of the present invention may be much better than the non-transformed plant.
  • the seed of the present invention is a seed produced by culturing the transformed plant.
  • the seed of the transformed plant may be introduced by the AtSH3P-P2 gene, thereby promoting the growth of the seed.
  • the plant growth promoter of the present invention contains at least one selected from the group consisting of the AtSH3P-P2 protein of the present invention, the AtSH3P-P2 gene encoding the same, and a recombinant vector comprising the gene as an active ingredient.
  • the AtSH3P-P2 protein, the AtSH3P-P2 gene encoding the same, and a recombinant vector comprising the gene can effectively promote plant growth by inducing cytoplasmic division of the plant.
  • AtSH3P-P2 gene is not limited thereto but preferably It may be represented by the nucleotide sequence of SEQ ID NO: 2.
  • the recombinant vector is not limited thereto, but may be preferably represented by the cleavage map of FIG. 1.
  • the AtSH3P-P2 gene is expressed at the cell plate position, that is, the AtSH3P-P2 gene is present in the cell plate appearing at the cytoplasmic division after the cell division stage of nuclear division, the AtSH3P-P2 gene
  • the AtSH3P-P2 gene is involved in cytoplasmic division, and furthermore, the AtSH3P-P2 gene maintains the cytoplasmic division since the position where the AtSH3P ⁇ P2 gene is expressed is maintained from the initial cell plate formation until the cell plate growth progresses and the cellular division is completed. There is inducing activity (see ⁇ Example 4>).
  • the growth of the leaves and roots in the transgenic plant is significantly superior to the wild type, the root length is longer than 30% compared to the wild type, the transgenic plant to the wild type Larger leaf sizes and longer root lengths indicate that the AtSH3P-P2 gene effectively induces cellular division and promotes plant growth (see Example 5).
  • the AtSH3P-P2 protein, the AtSH3P-P2 gene encoding the same, and a recombinant vector including the gene can effectively promote plant growth by inducing cytoplasmic division of the plant.
  • the method of controlling the growth of the plant of the present invention is the AtSH3P-P2 It is a way to control the growth of plants by controlling the expression of genes.
  • the plant growth regulation includes not only promoting plant growth by inducing expression of the AtSH3P-P2 gene, but also inhibiting plant growth by inhibiting the expression of the gene.
  • the plant growth may be promoted by introducing the AtSH3P-P2 gene into a recombinant vector and expressing or overexpressing the AtSH3P-P2 gene by transforming the plant with the recombinant vector.
  • the 'overexpression of the gene' means that the AISH3P-P2 gene is expressed above the level expressed in wild-type plants.
  • Recombinant vectors and transformation methods capable of expressing or overexpressing the gene are as described above, and the recombinant vector is not limited thereto but may be represented by the cleavage map of FIG. 1.
  • the method of controlling the growth of the plant of the present invention includes the step of inducing the expression of the AtSH3P-P2 protein coding gene represented by the amino acid sequence of SEQ ID NO: 3, it may be characterized by promoting the growth of the plant have.
  • Transforming the plant with the recombinant vector may be performed by the signed plant transformation method.
  • Plants that may be transformed by introducing the AtSH3P-P2 gene may be food crops, vegetable crops, special crops, fruit trees, flowers and feed crops, preferably rice, wheat, barley, corn, Beans, potatoes, arms, oats, sorghum, baby poles, cabbage, radish, peppers, strawberries, tomatoes, watermelons, cucumbers, cabbage, melons, pumpkins, green onions, onions, carrots, ginseng, tobacco, cotton, sesame seeds, sugar cane, Beet, Perilla, Peanut, Rapeseed, Apple tree, Pear, Jujube, Peach, Yard, Grape, Chisel, Persimmon, Plum, Apricot, Banana, Rose, Gladiolus, Gerbera, Carnation, Chrysanthemum, Lily, Yurilip, Ryegrass It may be selected from the group consisting of, red clover, orchard grass, alpha wave, vesque cue and perennial grass.
  • the AtSH3P-P2 gene in the present invention can be effectively expressed in the plant, including the baby pole, to promote plant growth, the transformed plant was able to grow much better than the untransformed plant.
  • the AtSH3P-P2 gene is expressed at the cell plate position, that is, the AtSH3P-P2 gene is after nuclear division during the cell division stage. Since the AtSH3P-P2 gene is involved in cytoplasmic division, the position where the AtSH3P-P2 gene is expressed is at the initial cell plate formation and cell plate growth proceeds to complete the cell division.
  • the AtSH3P-P2 gene has an activity of inducing cytoplasmic division since it is maintained until it is maintained (see ⁇ Example 4>).
  • the growth of leaves and roots in the transgenic plant is much superior to the wild type, the root length is longer than 30% compared to the wild type, the transgenic plant to the wild type Larger leaf sizes and longer root lengths indicate that the AtSH3P-P2 gene effectively induces cellular division and promotes plant growth (see Example 5).
  • plant growth can be suppressed by suppressing the expression of the AtSH3P-P2 gene.
  • dsRNA can be used to inhibit expression of the AtSH3P-P2 gene.
  • the dsRNA is a high specificity target of the AtSH3P-P2 gene.
  • RNA (primary or processed) can be specifically targeted and the dsRNA can be used to effectively inhibit the expression of these AtSH3P-P2 genes.
  • a vector capable of effectively expressing the dsRNA in plants and Transformation methods using the same are known in the art, and preferably
  • the pTA7002 expression vector represented by the cleavage map of 6 can be used.
  • dsRNA was prepared by expressing 300 bases of the N-terminus in the base sequence of the AtSH3P-P2 gene, and the expression of the AtSH3P-P2 gene was suppressed using the AtSH3P-P2 gene.
  • Expression of the AtSH3P-P2 protein is not able to function as a system, and cytoplasmic division does not occur normally after nuclear division. Furthermore, less differentiated cell walls appear due to the failure of cytoplasmic division, which prevents normal cell layer formation, thereby inhibiting plant growth (see Example 6).
  • the AtSH3P-P2 gene has an activity of inducing cytoplasmic division, and thus the transformed plant can be promoted for growth, and thus can be effectively used for improving the biomass of the plant.
  • FIG. 1 shows a cleavage map of a pCAMBIA expression vector to which a gene of AtSH3P-P2 and a GFP coupled to confirm the expression of the gene.
  • Figure 2 is the result of performing Western blot to confirm whether the gene of AtSH3P-P2 is expressed in plants transformed with AtSH3P-P2.
  • FIG. 3 is a result confirming that the AtSH3P-P2 protein is expressed in the cell plate position in plants transformed with AtSH3P-P2.
  • Figure 4 is a result of comparing the development of plants and wild type leaves and roots transformed with AtSH3P-P2 and wild type.
  • 5 is a result of comparing the root length change of plants transformed with AtSH3P-P2 to wild type after 6 days and 10 days after sowing.
  • 2P # 1 Transgenic Plant Corresponding to Line 1 in Western Blots
  • 2P # 2 Transgenic Plant Corresponding to Line 2 in Western Blots
  • FIG. 6 shows pTA7002 expression inhibiting AtSH3P-P2 gene expression. The cleavage map of the vector is shown.
  • Figure 7 is the result of observing the intracellular nucleus (a) and cell wall formation (b) in plants in which AtSH3P-P2 gene expression is suppressed.
  • the cDNA of AtSH3P-P2 is composed of 18 nucleotide sequences in 5 'ATG and 18 nucleotide sequences in front of 3' TGA as sense and antisense primers, respectively.
  • cDNA of AtSH3P-P2 was extracted by PCR amplification (30 cycles consisting of 30 seconds at 94 ° C, 30 seconds at 55 ° C, and 30 seconds at 72 ° C), and the sequence was determined. It was confirmed by sequencing known in the art. ⁇ 1-2> Experimental Results
  • the gene sequence of AtSH3P-P2 has the nucleotide sequence represented by SEQ ID NO: 1, AtSH3P-P2 It was found that cDNA had a nucleotide sequence represented by SEQ ID NO: 2. In addition, according to SEQ ID NO: 2 it can be seen that the amino acid sequence of AtSH3P-P2 is represented by SEQ ID NO: 3.
  • Example 1> Based on the cDNA of the AtSH3P-P2 gene analyzed in ⁇ Example 1> was prepared an expression vector that can be used for the production of transgenic plants of the gene.
  • the termination codon (TGA) of the AtSH3P-P2 gene was removed, and the initiation codon (AUG) of the green fluorescent protein (GFP) was linked, and inserted between the 35s promoter of pCAMBIA 1300 vector (Cambia Labs) and the nos (nopaline synthease).
  • TGA termination codon
  • AVG initiation codon
  • GFP green fluorescent protein
  • the cDNA of AtSH3P-P2 was cloned into the 326-sGFP vector.
  • a 5 'AtSH3P-2 sGFP Xbal primer of SEQ ID NO: 4 was prepared by adding an Xbal restriction enzyme sequence in front of 5' ATG, and a BamHl restriction enzyme sequence was added to remove the 3 'termination codon (TGA).
  • 5 '3' AtSH3P-2 sGFPBamHl primers were prepared.
  • AtSH3P-P2 cDNA library using the two primers (30 seconds at 94 ° C, 30 seconds at 55 ° C, 30 cycles consisting of 30 seconds at 72 ° C) of AtSH3P-P2 cDNA was extracted and cloned by cutting the 326-sGFP vector with two Xbal, BamHl restriction enzymes (Bioneer, Korea or NEB).
  • the prepared AtSH3P-P2: sGFP sequence was again confirmed through sequencing.
  • the pCAMBIA expression vector to which the gene of AtSH3P-P2 and GFP which can confirm the expression of the gene is bound is represented by the cleavage map described in FIG. 1.
  • the Arabidopsis thaliana plants transformed by the Agrobacterium mage transformation method were prepared using the pCAMBIA vector prepared in ⁇ Example 2>.
  • the transformation method is the Floral dip method (The Plant Journal (1998) 16 (6), 735-743).
  • the hygromycin resistance test was carried out to select first.
  • the Western blot was used for 10% SDS-PAGE and the expression was examined using a GFP specific antibody that recognizes GFP at the end of AtSH3-P2: sGFP.
  • FIG. 2 The results of performing the western blot are shown in FIG. 2, and as shown in FIG. 2, it can be seen that the gene of AtSH3-P2 is effectively expressed in the form of a fusion protein of GFP. That is, it was confirmed that a protein of the correct size of 70 kDa was synthesized. Through this, it was possible to secure a stable transgenic plant line.
  • Each line in FIG. 2 means each plant transformed.
  • AtSH3P-P2 protein is expressed at the cell plate position.
  • the AtSH3P-P2 protein is present in the cell plate appearing at the time of cytoplasmic division after nuclear division and nuclear division. Therefore, the AISH3P-P2 protein is involved in cytoplasmic division, Since the position to be expressed is maintained from the initial cell plate formation until cell plate growth proceeds to complete cell division, AtSH3P-P2 protein can be seen that it can play an essential function in cytoplasmic division.
  • the root length of the transgenic plants was measured on the 6th and 10th day after seeding, respectively, and randomly screened 30 times with wild type. Deviations were obtained and plotted.
  • the transgenic baby pole plant Root length is longer than 30% compared to wild type.
  • the size of the leaves and the length of the roots of the transgenic baby pole plants are larger than those of the wild type in the same time period, and the AtSH3-P2 protein effectively induces cellular division and promotes plant growth.
  • AtSH3P-P2 residues were suppressed using a gene expression suppression system that induced double stranded RNA.
  • pTA7002 expression vector having a cleavage map as shown in FIG. was prepared.
  • the expression vector can inhibit the expression of the AtSH3-P2 gene as needed by dexamethasone (dexamethasone).
  • the baby pole plants were transformed in the same manner as in ⁇ Example 3-1>.
  • the transformed plants were isolated and screened by the hygromycin resistance test, and the isolated and screened plants were germinated in 1/2 MS medium, and 30M dexamethasone was able to act on the gene expression suppression system on the 4th day after germination. Transfer to this containing 1/2 MS medium and incubated for 3 more days.

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Abstract

La présente invention concerne un gène régulant la cytocinèse, des plantes transformées par le gène et un procédé de régulation de la croissance des plantes l'utilisant. Comme cela a été examiné ci-dessus, un gène AtSH3P-P2 a une activité d'induction de cytocinèse, favorisant ainsi la croissance des plantes transformées par celui-ci, le gène pouvant ainsi être utilisé de manière efficace pour améliorer la biomasse des plantes.
PCT/KR2010/007322 2010-03-03 2010-10-25 Gène régulant la cytocinèse, plantes transformées par le gène et procédé pour la régulation de la croissance de plantes l'utilisant WO2011108794A2 (fr)

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KR1020100019042A KR20110099993A (ko) 2010-03-03 2010-03-03 세포질 분열을 조절하는 유전자, 상기 유전자로 형질 전환된 식물 및 이를 이용한 식물의 생장을 조절하는 방법
KR10-2010-0019042 2010-03-03

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KR101334408B1 (ko) * 2012-06-08 2013-11-29 포항공과대학교 산학협력단 바이오매스 생산 증가 유전자 및 이를 이용한 형질 전환 식물체

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CN102851280A (zh) * 2012-09-18 2013-01-02 中国科学院遗传与发育生物学研究所 一种rna及产生该rna的基因在调控水稻根系发育中的应用

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