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WO2018066898A2 - Protéine mtatpg2 à fonction d'augmentation de productivité, améliorant la résistance au stress et retardant la sénescence d'une plante, gène associé, et utilisation de ladite protéine et dudit gène - Google Patents

Protéine mtatpg2 à fonction d'augmentation de productivité, améliorant la résistance au stress et retardant la sénescence d'une plante, gène associé, et utilisation de ladite protéine et dudit gène Download PDF

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WO2018066898A2
WO2018066898A2 PCT/KR2017/010859 KR2017010859W WO2018066898A2 WO 2018066898 A2 WO2018066898 A2 WO 2018066898A2 KR 2017010859 W KR2017010859 W KR 2017010859W WO 2018066898 A2 WO2018066898 A2 WO 2018066898A2
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plant
gene
mtatpg2
seq
productivity
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WO2018066898A3 (fr
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이동희
김국진
김동수
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제노마인(주)
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    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • 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)
    • 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
    • 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
    • C12N15/8266—Abscission; Dehiscence; Senescence
    • 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
    • C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance

Definitions

  • the present invention relates to a MtATPG2 protein having its productivity enhancing function, stress resistance function and delaying aging function, genes thereof and uses thereof.
  • Plant senescence is considered to be a major step in the plant's lifecycle, and progresses in a highly sophisticated manner at the cellular, tissue, organ and individual levels through genetically regulated reactions. The aging process can occur differently in different organs of the plant. In the case of leaves, the degradation of chloroplasts is followed by the degradation of lipids, proteins and nucleic acids.
  • Cell structures such as cell membranes and intracellular compartments, are maintained until the end and lead to death after rearrangement of the degraded product nitrogen and nutrients (Lohman et al., Physiologia Plantarum, 1994, 92: 322-8 .; Smart, New Phytologist, 1994, 126: 419-48; Pruzinska et al., Plant Physiology, 2005, 139: 52-63).
  • the timing at which plants determine aging is related to nutrient retention and rearrangement, so flowering and seed production are often factors that promote aging. Plants also progress the aging process as a species-wide survival strategy for seasonal or unforeseen external environmental changes.
  • SAGs Senescence Associated Genes
  • Tomatoes have been reported to control the ripening of fruits by inhibiting the ethylene synthesis process (Oeller et al., Science. 1991, 254 (5030): 437-9) and also inhibit the expression of polygalacturonase genes involved in cell wall degradation.
  • Flav-O-Savor which increases the transport and storage properties of tomatoes, may be a typical commercialized example (Giovannoni et al., 1989, Plant Cell 1 (1): 53-63).
  • GmSARK gene and WRKY53 , WRKY6 and AtNAP genes belonging to the NAC family have been reported as genes overexpressed during aging (Miao et al., Plant Mol. Biol. 2004, 55: 853-867; Guo and Gan et al. , Plant J. 2006, 46 (4): 601-12; Li et al., Plant Mol Biol. 2006, 61: 829-844; Besseau et al., J Exp Bot 2012, 63 (7): 2667-79 Genes such as CBF2 and CBF3 are known to inhibit aging (Sobieszczuk-Nowicka et al., Physiol Plant 2007; 130: 590-600).
  • U.S. Pat.No.8420890 discloses a method for delaying aging by inhibiting the expression of AtNAP gene. Recently, studies have been conducted for overexpressing AtNAP to facilitate the harvesting of cotton or the like or to help ripening fruit. (Kou et al., J Exp Bot. 2012, 63 (17): 6139-47).
  • GhCKX a cytokinin regulator gene isolated from cotton
  • TobEA Tobacco Expression Atlas
  • the present invention also discloses a gene having a function of delaying aging and the like, which can improve the productivity of crops and the like.
  • An object of the present invention is to provide a MtATPG2 protein having a productivity enhancing function of a plant, a stress resistance function, and a delaying aging function.
  • Another object of the present invention is to provide a gene encoding the protein.
  • Still another object of the present invention is to provide a method for producing a plant having a yield increasing characteristic.
  • Still another object of the present invention is to provide a method for producing a plant having stress resistance.
  • Still another object of the present invention is to provide a method for producing a plant having aging delay characteristics.
  • the present invention relates to a MtATPG2 protein having a productivity enhancing function of a plant, a stress resistance function, and a delaying aging function.
  • the inventor (s) is based on the nucleotide sequence of the AT-hook motif nuclear localized protein (GeneBank accession number NC_016414.1) of Medicago truncatula , as identified in the following examples
  • the productivity increase characteristics such as the increase in the biomass and / or the seed productivity of the individual were clearly shown, and the resistance to oxidative stress was also clearly shown, and the aging of the plant was delayed. It was confirmed that the phenomenon appeared clearly.
  • MtATPG2 M edicago t runcatula AT -hook p rotein of G enomine 2
  • MtATPG2 M edicago t runcatula AT -hook p rotein of G enomine 2
  • the MtATPG2 protein of the invention is one of the polypeptides of (a), (b) and (c) below.
  • protein are used interchangeably with each other in the same sense as a polypeptide
  • gene is used interchangeably with each other in the same sense as a polynucleotide.
  • a "polypeptide comprising a substantial portion of the amino acid sequence set forth in SEQ ID NO: 2" still retains productivity enhancement and stress tolerance and plant aging delayed functions as compared to the polypeptide consisting of the amino acid sequence set forth in SEQ ID NO: It is defined as a polypeptide comprising a portion of an amino acid sequence of SEQ ID NO: 2 sufficient to the following.
  • the length of the polypeptide and the degree of activity of such a polypeptide is not a problem since it still needs to be sufficient to retain productivity and stress resistance and to delay plant aging.
  • polypeptide As such a polypeptide, the polypeptide which deleted the N-terminal part or C-terminal part in the polypeptide containing the amino acid sequence of SEQ ID NO: 2 is mentioned. This is because it is generally known in the art that even if the N- or C-terminal portion is deleted, such a polypeptide has the function of the original polypeptide. Of course, in some cases, the N-terminal or C-terminal moiety is necessary for maintaining the function of the protein, so that a polypeptide deleted with the N-terminal or C-terminal moiety does not exhibit this function. It is within the ordinary skill of one of ordinary skill in the art to distinguish and detect inactive polypeptides from active polypeptides.
  • the deletion of the N-terminal or C-terminal moiety as well as other moieties may still have the function of the original polypeptide.
  • one of ordinary skill in the art will be able to ascertain whether such deleted polypeptide still has the function of the original polypeptide within the scope of its usual ability.
  • the present specification discloses the nucleotide sequence of SEQ ID NO: 1 and the amino acid sequence of SEQ ID NO: 2, furthermore, a polypeptide encoded by the nucleotide sequence of SEQ ID NO: 1 and consisting of the amino acid sequence of SEQ ID NO: 2 increases productivity and stress resistance of plants.
  • the present invention discloses an embodiment confirming whether the plant has a delaying function of aging, and thus, a polypeptide having a partial deletion in the amino acid sequence of SEQ ID NO: 2 may still retain the function of the polypeptide comprising the amino acid sequence of SEQ ID NO: 2 It is very clear that those skilled in the art can fully confirm within the ordinary capability range. Therefore, in the present invention, the "polypeptide comprising a substantial part of the amino acid sequence set forth in SEQ ID NO: 2" as described above, based on the disclosure of the present specification, the productivity of a plant that can be manufactured by those skilled in the art within the range of its usual capacity can be increased. It is to be understood as meaning including all polypeptides in a deleted form having a function, a stress resistance function, and a plant aging delay function.
  • polypeptide substantially similar to the polypeptide of (a) and (b) includes a function comprising one or more substituted amino acids, but comprising the amino acid sequence of SEQ ID NO: 2, ie, the productivity of a plant. It refers to a polypeptide possessing augmentation and stress resistance functions and a delay in plant aging.
  • the degree of activity or amino acid substitution of the polypeptide is not a problem as long as the polypeptide containing at least one substituted amino acid retains the productivity of the plant, the stress resistance function, and the delayed plant aging function.
  • a polypeptide comprising one or more substituted amino acids contains a large number of substituted amino acids, even if its activity is lower than that of a polypeptide comprising the amino acid sequence of SEQ ID NO: 2, the polypeptide is a plant productivity. It is included in the present invention as long as it possesses augmentation and stress tolerance functions, and a plant aging delay function. Even if one or more amino acids are substituted, if the amino acid before substitution is chemically equivalent to the substituted amino acid, the polypeptide comprising such substituted amino acid will still retain the function of the original polypeptide.
  • the polypeptide having such substituted amino acid (s) may be It will still retain the function of the original polypeptide.
  • a negatively charged amino acid such as glutamic acid
  • another negatively charged amino acid such as aspartic acid
  • polypeptide having such substituted amino acid (s) will still retain the function of the original polypeptide even if its activity is low. will be.
  • a polypeptide comprising amino acid (s) substituted at the N-terminal or C-terminal portion of the polypeptide will still retain the function of the original polypeptide.
  • One skilled in the art would prepare a polypeptide that contains one or more substituted amino acids as described above, but still retains the productivity and stress resistance functions of plants comprising the amino acid sequence of SEQ ID NO. can do.
  • polypeptides comprising one or more substituted amino acids still has this function.
  • the present disclosure discloses the nucleotide sequence of SEQ ID NO: 1 and the amino acid sequence of SEQ ID NO: 2, and also the polypeptide comprising the amino acid sequence of SEQ ID NO: 2 has a function of increasing productivity and stress resistance of plants, and delaying plant aging. Since the confirmed examples are disclosed, it is evident that the "polypeptides substantially similar to the polypeptides of (a) and (b)" of the present invention can be easily implemented by those skilled in the art.
  • polypeptide substantially similar to the polypeptide of (a) or (b) above is meant to include all polypeptides that contain one or more substituted amino acids but still have plant productivity and stress resistance and delayed aging of plants. Should be understood as. As such, the term “polypeptide substantially similar to the polypeptide of (a) or (b)” includes all polypeptides that contain one or more substituted amino acids but still have plant productivity and stress resistance functions, and plant aging function. In view of the degree of activity, however, the polypeptide is preferably higher in sequence homology with the amino acid sequence of SEQ ID NO.
  • the polypeptide has at least 60% sequence homology at the lower end of sequence homology, while at the upper limit of sequence homology, it is preferred that the polypeptide has 100% sequence homology. More specifically, the above sequence homology is 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90% , 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% in order of higher.
  • polypeptides substantially similar to the polypeptides of (a) and (b) above are not only “polypeptides substantially similar to polypeptides comprising the entire amino acid sequence of SEQ ID NO: 2", All descriptions above are intended to include “substantially similar to polypeptides comprising the entire amino acid sequence of SEQ ID NO: 2" as well as “amino acids of SEQ ID NO: 2", since the polypeptide comprising substantially part thereof is included. The same applies to polypeptides that are substantially similar to polypeptides comprising substantial portions of the sequence.
  • polypeptide as described above refers to a polypeptide comprising the entire amino acid sequence of SEQ ID NO: 2, a substantial portion of the amino acid sequence of SEQ ID NO. Including polypeptides, and polypeptides substantially similar to the above polypeptides, as well as all polypeptides of the preferred embodiments as described above.
  • the polynucleotide of the present invention is an isolated polynucleotide encoding a polypeptide comprising all or a substantial part of the amino acid sequence of SEQ ID NO: 2 and having a function of increasing productivity and stress resistance of plants, and delaying aging of plants
  • Polypeptides encoding polypeptides that are substantially similar to the polypeptides include isolated polynucleotides and, in a preferred embodiment, the sequences in the order of sequence homology as described above, with increasing plant productivity and stress resistance, and delayed plant aging. It includes an isolated polynucleotide encoding all polypeptides with homology. When amino acid sequences are found, those skilled in the art can readily prepare polynucleotides encoding such amino acid sequences based on those amino acid sequences.
  • isolated polynucleotide includes both chemically synthesized polynucleotides, polynucleotides isolated from organisms, especially Medicago truncatula , and polynucleotides containing modified nucleotides, It is defined as including all polymers of stranded or double stranded RNA or DNA.
  • the present invention relates to a method for producing a plant having productivity enhancing properties.
  • Method for producing a plant having a productivity enhancing feature of the present invention comprises the steps of (a) expressing a gene having a nucleotide sequence of SEQ ID NO: 1 or a gene having a sequence similar to the nucleotide sequence of SEQ ID NO: 1 in the plant and (b) increased productivity And selecting the plant having the characteristic.
  • productivity enhancing properties means that the biomass (size and / or mass) of the whole, stem, root and / or leaves of the plant is increased compared to wild-type plants, and / or the productivity of the plant's seeds (plants) Number and / or mass of seeds per individual) is increased compared to wild type plants.
  • plant is meant to include mature plants, immature plants (plants), plant seeds, plant cells, plant tissues and the like.
  • plant cells or plant tissues are described in European Patent EP0116718, European Patent EP0270822, International Patent WO 84/02913, Gould et al. 1991, Plant Physiol 95,426-434, etc., can be used to develop and grow into mature plants.
  • plant includes all plants for which productivity gains can give useful results to humans. Therefore, the meaning of the plant includes crops (specifically, crops such as edible crops, feed crops, craft crops, and horticultural crops), forest trees, and ornamental plants. Specifically, rice, wheat, barley, corn, soybeans, potatoes, red beans, oats, sorghum, legumes (specifically knotweed, hoe belonging to Medicago truncatula ) from which the MtATPG2 gene of the present invention is isolated.
  • crops specifically, crops such as edible crops, feed crops, craft crops, and horticultural crops
  • forest trees and ornamental plants. Specifically, rice, wheat, barley, corn, soybeans, potatoes, red beans, oats, sorghum, legumes (specifically knotweed, hoe belonging to Medicago truncatula ) from which the MtATPG2 gene of the present invention is isolated.
  • gene consisting of a sequence similar to the nucleotide sequence of SEQ ID NO: 1 is a gene encoding the first amino acid of SEQ ID NO: 2 while having a base sequence different from that of the gene of SEQ ID NO: 1 due to codon degeneracy
  • homologue of genes consisting of the nucleotide sequence of SEQ ID NO: 1 having the characteristics of increasing the productivity of the plant, etc. due to the evolutionary pathways different according to the type of plant consisting of the nucleotide sequence of SEQ ID NO: 1 and other nucleotide sequences It is meant to include all genes.
  • the gene consisting of a sequence similar to the nucleotide sequence of SEQ ID NO: 1 is preferably higher in sequence homology with the nucleotide sequence of SEQ ID NO: 1, and most preferably, having 100% sequence homology.
  • the gene has a sequence homology of 60% or more with the nucleotide sequence of SEQ ID NO: 1.
  • sequence homology is 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73 %, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, Higher in order of 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% and 99% is preferred.
  • step (a) may be performed by a genetic engineering method.
  • Genetic engineering method includes the steps of (i) inserting the gene of SEQ ID NO: 1 or a gene having a sequence similar to the nucleotide sequence of SEQ ID NO: 1 into an expression vector to be operably linked to a regulatory sequence capable of expressing it, and ( ii) transforming the expression vector into a plant.
  • operably means that the transcription and / or translation of a gene is linked to be affected. For example, if a promoter influences the transcription of a gene linked to it, the promoter and the gene are operably linked.
  • regulatory sequence is meant to include all sequences whose presence may affect the transcription and / or translation of a gene linked thereto, and such regulatory sequences include a promoter sequence and a polyadenylation signal. ), The replication start point .
  • promoter follows the conventional meaning known in the art, specifically located at the top (5 'side) based on the transcription initiation point of a gene, binding to DNA-dependent RNA polymerase By nucleic acid sequences having the function of controlling transcription of one or more genes, including sites, transcriptional initiation sites, transcription factor binding sites, and the like.
  • This promoter may be a TATA box (usually located at the transcription start point (+1) -20 to -30 position) above the transcription initiation point if it is of eukaryotes, and a CAAT box (typically about -75 position compared to the transcription start site) Present), a 5 'enhancer, a transcription repression factor, and the like.
  • Usable promoters include constitutive promoters (promoters which induce expression in all plant tissues at all times), inducible promoters (expression of target genes in response to specific external stimuli) as long as they are capable of expressing the gene of SEQ ID NO. 1 linked thereto. Promoters that induce expression or promoters that specifically induce expression in specific developmental periods or specific tissues). Representative examples of constitutive promoters that can be used include the promoter of the 35S RNA gene of cauliflower mosaic virus (CaMV), and the ubiquitin family of promoters (Christensen et al., 1992, Plant Mol). Biol. 18, 675-689; EP0342926; Cornejo et al., 1993, Plant Mol. Biol.
  • constitutive promoters include the promoter of the 35S RNA gene of cauliflower mosaic virus (CaMV), and the ubiquitin family of promoters (Christensen et al., 1992, Plant Mol). Biol. 18, 675-689; EP0342926; Cornejo et
  • rice actin promoter Zhang et al. 1991, The Plant Cell 3, 1155-1165
  • rice actin promoter Zhang et al. 1991, The Plant Cell 3, 1155-1165
  • inducible promoters include the yeast metallothionein promoter (Mett et al., Proc. Natl. Acad. Sci., USA, 90: 4567, 1993), which is activated by copper ions, substituted by substituted benzenesulfonamides.
  • In2-1 and In2-2 promoters (Hershey et al., Plant Mol. Biol., 17: 679, 1991), GRE regulatory sequences regulated by glucocorticoids (Schena et al., Proc. Natl. Acad.
  • Transcription termination sequence is poly (A ) addition signal ( polyadenylation As a sequence acting as a signal ), it is intended to enhance the integrity and efficiency of transcription.
  • An example of a transcription termination sequence that can be used is nopalin Synthase (NOS) a transcription termination sequence, rice ⁇ - amylase RAmy1 A transcription termination sequence, Agrobacterium Tome Pacific Enschede of the gene of the gene Transcription termination sequence of octopine gene, transcription termination sequence of wheat heat shock protein 17, transcription termination sequence of wheat ubiquitin gene, transcription termination sequence of rice gluterin gene, transcription termination sequence of rice lactate dehydrogenase gene Etc. can be mentioned.
  • NOS nopalin Synthase
  • the expression vector may include a selection marker gene.
  • screening Marker gene means a gene encoding a trait that enables the selection of a plant comprising such a marker gene selectable marker gene may be a number of days antibiotic resistance genes and herbicide tolerance genes. Examples of antibiotic resistance genes Examples of antibiotic resistance genes include puromycin resistance genes (eg Streptomyces).
  • the herbicide resistance gene may include a vasta herbicide resistance bar gene.
  • the term "transformation” refers to a modification of the genotype of a host plant by the introduction of a hereditary gene, and regardless of the method used for the transformation, the herb gene is a host plant, more precisely a cell of the host plant. Introduced into and integrated into the genome of a cell.
  • the hereditary genes include homologous and heterologous genes, wherein “homologous genes” refer to endogenous genes of a host organism or the same species, and “heterologous genes” are genes that do not exist in the organism to which they are transformed.
  • the MtATPG2 gene of the present invention is homologous to Medicago truncatula from which it is isolated, but is heterologous to Arabidopsis and tomato plants.
  • a method of transforming a plant with an exogenous gene may use a method known in the art, such as a direct gene transfer method using a gene gun, an in planta transformation method using a floral dip, pollen mediation, and the like. Transformation methods, protoplast transformation methods, viral mediated transformation methods, liposome mediated transformation methods, and the like can be used.
  • a transformation method suitable for a specific plant for example, a method for transforming corn is described in US Pat. No.
  • Generally used in transforming plants is a method of infecting seedlings, plant seeds and the like with the transformed Agrobacterium.
  • Such Agrobacterium mediated transformation methods are well known in the art (Chilton et al., 1977, Cell 11: 263: 271; European Patent EP 0116718; US Patent US 4,940,838), and methods suitable for particular plants are also known in the art.
  • the Agrobacterium mediated transformation method uses Ti-plasmid, which will contain left and right border sequences that allow the integration of T-DNA into the genome of plant cells.
  • the selection step (b) may be selected through the characteristics of the inserted gene by growing and growing the transformed plant, or, if the selection marker gene is transformed together during transformation, the selection marker gene may be selected. Can be.
  • the characteristics of the inserted gene include the biomass of the plant and / or the productivity of the seed.
  • the present invention relates to a method for producing a plant having stress resistance properties of the present invention.
  • the method for producing a stress resistant plant of the present invention comprises the steps of: (a) expressing a gene having a nucleotide sequence of SEQ ID NO: 1 or a gene having a sequence similar to the nucleotide sequence of SEQ ID NO: 1 in the plant, and (b) having a stress resistant phenotype Comprising plant screening.
  • stress means oxidative stress
  • Step (a) may be performed genetically, as for the genetic engineering method, as described with reference to the method of manufacturing a plant having the productivity increasing characteristic of the present invention.
  • the step (b) is selected by comparing the stress resistance of the plant that is characteristic of the inserted gene (for example, the degree of progress of leaf yellowing or necrosis, the biomass of the leaves and / or stems, chlorophyll content, photosynthetic efficiency, etc.).
  • a selection marker gene When a selection marker gene is transformed together during transformation, it may be selected using a selection marker gene, or may be selected by mixing these methods.
  • the present invention relates to a method for producing a plant having aging delay properties.
  • the method for producing a plant having a aging retardation property of the present invention comprises the steps of: (a) expressing a gene having a nucleotide sequence of SEQ ID NO: 1 or a gene having a sequence similar to the nucleotide sequence of SEQ ID NO: 1 in the plant, and (b) Selecting the plant with the delayed phenotype.
  • aging delay refers to a property of prolonged plant life as compared to wild-type plants, and specifically, the yellowing and / or necrosis of leaves and / or stems is delayed compared to wild-type plants or the chlorophyll content of plants is wild-type. It is more characteristic than plants or photosynthetic efficiency of plants is higher than wild type plants.
  • Step (a) may be performed genetically, as for the genetic engineering method, as described with reference to a method for preparing a plant having productivity enhancing characteristics of the present invention.
  • the selected plant is selected using delayed aging characteristics, which are characteristics of the gene inserted by developing and growing a transformed plant (quantitative evaluation of leaf yellowing or necrosis, chlorophyll content, photosynthetic efficiency, etc.). Method, a method of mixing the above methods, etc.), and when the selection marker gene is transformed together during transformation, the selection may be performed using the selection marker gene.
  • the present invention relates to a method for increasing productivity of a plant.
  • the method for increasing the productivity of a plant of the present invention includes (a) an expression vector such that a gene having a nucleotide sequence of SEQ ID NO: 1 or a gene having a sequence similar to that of SEQ ID NO: 1 is operably linked to a regulatory sequence capable of expressing it And (b) transforming the expression vector into a plant.
  • the present invention relates to a method for increasing stress resistance of a plant.
  • the method of increasing the stress resistance of a plant of the present invention is (a) operably linking a gene having a nucleotide sequence of SEQ ID NO: 1 or a gene having a sequence similar to that of SEQ ID NO: 1 to a regulatory sequence capable of expressing it Inserting into the expression vector preferably and (b) transforming the expression vector into a plant.
  • the present invention relates to a method for delaying aging of a plant.
  • the method for delaying aging of a plant of the present invention is to (a) operably link a gene having a nucleotide sequence of SEQ ID NO. 1 or a gene having a sequence similar to that of SEQ ID NO. 1 to a regulatory sequence capable of expressing it. Inserting into the expression vector and (b) transforming the expression vector into a plant.
  • Steps (a) and (b) in the above methods are as described with reference to the method for producing a plant having the productivity enhancing properties of the present invention.
  • the present invention is a gene having a nucleotide sequence of SEQ ID NO: 1, or a gene having a sequence similar to the nucleotide sequence of SEQ ID NO: 1 obtained by the method for producing a plant having the productivity enhancing characteristics of the present invention
  • the present invention relates to a transgenic plant having improved productivity.
  • the plant is a transgenic plant having productivity enhancing properties by introducing a gene encoding the MtATPG2 protein consisting of the amino acid sequence of SEQ ID NO: 2, in particular the gene MtATPG2 having the nucleotide sequence of SEQ ID NO: 1, into the plant .
  • the present invention is a stress resistance that is expressed by a gene having a nucleotide sequence of SEQ ID NO: 1 or a gene having a sequence similar to the nucleotide sequence of SEQ ID NO: 1 obtained by the method for producing a stress resistant plant of the present invention
  • the present invention relates to a transgenic plant having characteristics.
  • the plant is a transgenic plant having a stress resistance by introducing a gene encoding the MtATPG2 protein consisting of the amino acid sequence of SEQ ID NO: 2, in particular the gene MtATPG2 having the nucleotide sequence of SEQ ID NO: 1, into the plant.
  • the present invention is a gene having a nucleotide sequence of SEQ ID NO: 1 or a gene having a sequence similar to the nucleotide sequence of SEQ ID NO: 1 obtained by the method for producing a plant having the aging delay characteristics of the present invention
  • the present invention relates to a transgenic plant having delayed aging characteristics.
  • the plant is a transgenic plant having delayed aging characteristics by introducing a gene encoding the MtATPG2 protein consisting of the amino acid sequence of SEQ ID NO: 2, in particular the gene MtATPG2 having the nucleotide sequence of SEQ ID NO: 1, into the plant .
  • the "transformed plant” is obtained by crossing with a transformed plant as well as when the gene is introduced and transformed into a plant cell, plant tissue, or plant seed capable of developing and growing into a mature plant. Genome modified plants, plant seeds derived from the plants, plant cells, plant tissues.
  • the MtATPG2 protein and its gene, MtATPG2 which have a productivity enhancing function and a stress resistance function of the plant, and a delaying function of aging. Since the gene provides a function of increasing productivity and stress resistance and delaying aging of the plant, when the plant is transformed with the gene, the gene may not only increase productivity but also produce a plant having a stress-resistant and aging delaying function of the plant. Can be.
  • Figure 1 shows the structure (schematic) of the pCSEN-MtATPG2 recombinant vector in which the MtATPG2 gene having productivity enhancement function, stress resistance function, and aging delay function was introduced in the sense direction.
  • Figure 2 is a photograph of the Arabidopsis grown 50 days and 70 days after germination of the Arabidopsis T 2 line transformed with the pCSEN-MtATPG2 recombinant vector of FIG. DAG, day after germination.
  • MtATPG2-7 Arabidopsis T 2 line transformed with pCSEN-MtATPG2 recombinant vector
  • MtATPG2-8 Arabidopsis T 2 line transformed with pCSEN-MtATPG2 recombinant vector
  • MtATPG2-15 Arabidopsis T 2 line transformed with pCSEN-MtATPG2 recombinant vector
  • MtATPG2-16 Arabidopsis T 2 line transformed with pCSEN-MtATPG2 recombinant vector
  • MtATPG2-17 Arabidopsis T 2 line transformed with pCSEN-MtATPG2 recombinant vector
  • Figure 3 shows the results of analyzing the expression of the MtATPG2 gene of Arabidopsis thalass for 25 days after cotyledon generation of the Arabidopsis T 2 line transformed with the pCSEN-MtATPG2 recombinant vector of Figure 1 through qRT-PCR TUB was used as a PCR positive control.
  • MtATPG2-7 Arabidopsis T 2 line transformed with pCSEN-MtATPG2 recombinant vector
  • MtATPG2-8 Arabidopsis T 2 line transformed with pCSEN-MtATPG2 recombinant vector
  • MtATPG2-15 Arabidopsis T 2 line transformed with pCSEN-MtATPG2 recombinant vector
  • MtATPG2-16 Arabidopsis T 2 line transformed with pCSEN-MtATPG2 recombinant vector
  • MtATPG2-17 Arabidopsis T 2 line transformed with pCSEN-MtATPG2 recombinant vector
  • FIG. 4 is a diagram for increasing productivity and seed size of the Arabidopsis line of FIG. 3.
  • MtATPG2-7 Arabidopsis T 2 line transformed with pCSEN-MtATPG2 recombinant vector
  • MtATPG2-8 Arabidopsis T 2 line transformed with pCSEN-MtATPG2 recombinant vector
  • MtATPG2-15 Arabidopsis T 2 line transformed with pCSEN-MtATPG2 recombinant vector
  • MtATPG2-16 Arabidopsis T 2 line transformed with pCSEN-MtATPG2 recombinant vector
  • MtATPG2-17 Arabidopsis T 2 line transformed with pCSEN-MtATPG2 recombinant vector
  • Figure 6 is a photograph of the Arabidopsis grown 50 days after germination of the Arabidopsis T 3 or T 4 homo line transformed with the pCSEN-MtATPG2 recombinant vector of FIG. DAG, day after germination.
  • MtATPG2 -7-15-11 Arabidopsis T 4 homo line transformed with pCSEN-MtATPG2 recombinant vector
  • MtATPG2 -7-15-12 Arabidopsis T 4 homo line transformed with pCSEN-MtATPG2 recombinant vector
  • MtATPG2 -8-7 pCSEN-MtATPG2 Transgenic Arabidopsis T 3 homo line with a recombinant vector
  • MtATPG2 -8-8 pCSEN-MtATPG2 Transgenic Arabidopsis T 3 homo line with a recombinant vector
  • MtATPG2 -15-3 pCSEN-MtATPG2 Transgenic Arabidopsis T 3 homo line with a recombinant vector
  • MtATPG2 -15-6 pCSEN-MtATPG2 Transgenic Arabidopsis T 3 homo line with a recombinant vector
  • Figure 7 is a photograph of the Arabidopsis grown 70 days after germination of the Arabidopsis T 3 or T 4 homo line transformed with the pCSEN-MtATPG2 recombinant vector of FIG. DAG, day after germination.
  • MtATPG2 -7-15-11 Arabidopsis T 4 homo line transformed with pCSEN-MtATPG2 recombinant vector
  • MtATPG2 -7-15-12 Arabidopsis T 4 homo line transformed with pCSEN-MtATPG2 recombinant vector
  • MtATPG2 -8-7 pCSEN-MtATPG2 Transgenic Arabidopsis T 3 homo line with a recombinant vector
  • MtATPG2 -8-8 pCSEN-MtATPG2 Transgenic Arabidopsis T 3 homo line with a recombinant vector
  • MtATPG2 -15-3 pCSEN-MtATPG2 Transgenic Arabidopsis T 3 homo line with a recombinant vector
  • MtATPG2 -15-6 pCSEN-MtATPG2 Transgenic Arabidopsis T 3 homo line with a recombinant vector
  • FIG. 8 shows the expression patterns of the MtATPG2 gene of Arabidopsis thaliana grown in 25 days after cotyledon generation using T 3 or T 4 homoline transformed with the pCSEN-MtATPG2 recombinant vector of FIG. 1 through qRT-PCR.
  • FIG. One result is shown and TUB was used as a PCR positive control.
  • FIG. 9 is generated after the cotyledons from 16 wild-type Arabidopsis thaliana (Con) and three progress with T 3 or T 4 3-4 times left lobe (rosette leaf) of homo-line observing the phenotype of the leaves to 56 days every 4 days
  • Figure 10 is a figure of the investigation of the chlorophyll content of the leaves for this. DAE, day after emersion.
  • MtATPG2 -7-15-11 Arabidopsis T 4 homo line transformed with pCSEN-MtATPG2 recombinant vector
  • MtATPG2 -7-15-12 Arabidopsis T 4 homo line transformed with pCSEN-MtATPG2 recombinant vector
  • MtATPG2 -8-7 pCSEN-MtATPG2 Transgenic Arabidopsis T 3 homo line with a recombinant vector
  • MtATPG2 -8-8 pCSEN-MtATPG2 Transgenic Arabidopsis T 3 homo line with a recombinant vector
  • MtATPG2 -15-3 pCSEN-MtATPG2 Transgenic Arabidopsis T 3 homo line with a recombinant vector
  • MtATPG2 -15-6 pCSEN-MtATPG2 Transgenic Arabidopsis T 3 homo line with a recombinant vector
  • FIG. 11 shows the phenotype of leaves up to 6 days every 2 days after detaching the left lobe 3-4 of the Arabidopsis wild-type (Con) and generation-advanced T 3 or T 4 homo lines after germination to maintain cancer status.
  • FIG. 12 is a diagram illustrating the chlorophyll content of leaves. DAT, day after treatment.
  • MtATPG2 -7-15-11 Arabidopsis T 4 homo line transformed with pCSEN-MtATPG2 recombinant vector
  • MtATPG2 -7-15-12 Arabidopsis T 4 homo line transformed with pCSEN-MtATPG2 recombinant vector
  • MtATPG2 -8-7 pCSEN-MtATPG2 Transgenic Arabidopsis T 3 homo line with a recombinant vector
  • MtATPG2 -8-8 pCSEN-MtATPG2 Transgenic Arabidopsis T 3 homo line with a recombinant vector
  • MtATPG2 -15-3 pCSEN-MtATPG2 Transgenic Arabidopsis T 3 homo line with a recombinant vector
  • MtATPG2 -15-6 pCSEN-MtATPG2 Transgenic Arabidopsis T 3 homo line with a recombinant vector
  • FIG. 13 shows the leaves treated with 4 mM H 2 O 2 in 3 mM MES solution by detaching the left lobe No. 3-4 of the Arabidopsis wild-type (Con) and generation-advanced T 3 or T 4 homo lines 25 days after germination.
  • Fig. 13A shows a phenotypic change in Fig. 13A
  • Fig. 13A shows a phenotypic change in leaves treated with 150 mM NaCl in a 3 mM MES solution for 6 days in a sample under the same conditions (Fig. 13B). DAT, day after treatment.
  • MtATPG2 -7-15-11 Arabidopsis T 4 homo line transformed with pCSEN-MtATPG2 recombinant vector
  • MtATPG2 -7-15-12 Arabidopsis T 4 homo line transformed with pCSEN-MtATPG2 recombinant vector
  • MtATPG2 -8-7 pCSEN-MtATPG2 Transgenic Arabidopsis T 3 homo line with a recombinant vector
  • MtATPG2 -8-8 pCSEN-MtATPG2 Transgenic Arabidopsis T 3 homo line with a recombinant vector
  • MtATPG2 -15-3 pCSEN-MtATPG2 Transgenic Arabidopsis T 3 homo line with a recombinant vector
  • MtATPG2 -15-6 pCSEN-MtATPG2 Transgenic Arabidopsis T 3 homo line with a recombinant vector
  • 14 and 15 shows the amino acid sequence of the base sequence of each protein and MtATPG2 MtATPG2 gene.
  • Example 1 Medicago truncatula from In addition to increasing productivity and stress tolerance of plants Providing delayed aging MtATPG2 Isolation of genes
  • Medicago uses the MtATPG2 gene, which has increased productivity and stress resistance in plants and also delays aging. To separate from truncatula , the following process was performed.
  • Example 1-1 Medicago planting and cultivation of truncatula
  • Medicago truncatula Jemalong A17 was cultivated in a growth chamber controlled in a soil containing pots at a temperature of 22 ° C. and a cycle of 16/8 hours.
  • Example 1-3 Isolation of MtATPG2 Gene Having Increased Productivity, Stress Tolerance, and Delay in Aging of Plants
  • a forward primer (BglII / MTR8g 098390-F, represented by SEQ ID NO: 3 and containing the sequence of restriction enzyme BglII)
  • a reverse primer (BstEII / MTR8g 098390, 5GAC CTC ATC CTC TTG TCA AGT CAA AGC-3) represented by SEQ ID NO: 4 and containing the sequence of restriction enzyme BstEII were synthesized.
  • the primer was used to amplify and isolate full-length cDNA from M. truncatula cDNA using polymerase chain reaction (PCR).
  • the isolated cDNA As a result of analysis of the isolated cDNA, it has a 1,056 bp transcriptional translation frame (ORF) encoding 351 amino acids having a molecular weight of about 35.9 kDa, it was confirmed that it consists of one exon, AT -hook has a motif I and named it as MtATPG2 (M edicago t runcatula AT -hook p rotein of enomine G 2).
  • ORF transcriptional translation frame
  • the isoelectric point of the MtATPG2 protein encoded by the gene was found to be 10.28 (hereinafter, the gene is called “ MtATPG2 " or " MtATPG2 gene” using italics, and the protein is called “MtATPG2” or “MtATPG2 protein”).
  • a transgenic Arabidopsis in which the MtATPG2 gene was introduced in the sense direction was prepared to change the expression of the MtATPG2 transcript.
  • MtATPG2 cDNA using PCR from cDNA of M. truncatula using a forward primer represented by SEQ ID NO: 3 and containing a sequence of restriction enzyme Bgl II and a reverse primer represented by SEQ ID NO: 4 and a sequence of restriction enzyme BstE II was amplified.
  • the DNA was digested with restriction enzymes Bgl II and BstE II, cloned in the sense direction into a pCSEN vector designed to be controlled by the SEN1 promoter, an inducible promoter, and the pCSEN-MtATPG2 recombinant vector, a sense construct for the MtATPG2 gene.
  • the SEN1 promoter has specificity for the gene expressed according to the growth stage of the plant.
  • FIG. 1 is a diagram illustrating a pCSEN-MtATPG2 recombinant vector in which a MtATPG2 gene is introduced in a sense direction into a pCSEN vector.
  • BAR refers to a BAR gene (phosphinothricin acetyltransferase gene) that confers resistance to Basta herbicide, RB to the right border, LB to the left border, p35S to the CaMV 35S promoter, and T35S to CaMV 35S RNA polyA
  • pSEN refers to the SEN1 promoter
  • NOA-polyA refers to the polyA of the nopaline synthase gene.
  • the pCSEN-MtATPG2 recombinant vector was introduced into Agrobacterium tumefaciens using an electroporation method.
  • the transformed Agrobacterium cultures were incubated at 28 until the OD600 value was 1.0, and the cells were harvested by centrifugation at 5,000 rpm for 10 minutes at 25 ° C.
  • Harvested cells were suspended in Infiltration Medium (IM; 1X MS SALTS, 1X B5 vitamin, 5% sucrose, 0.005% Silwet L-77, Lehle Seed, USA) medium until the final OD600 value was 2.0.
  • IM Infiltration Medium
  • 1X MS SALTS 1X B5 vitamin, 5% sucrose, 0.005% Silwet L-77, Lehle Seed, USA
  • the Arabidopsis was placed in a polyethylene bag for 24 hours. Thereafter, the transformed Arabidopsis cultivars continued to grow to harvest seeds (T 1 ).
  • T 1 a non-transformed wild type Arabidopsis or a Arabidopsis transformed with only a vector (pCSEN vector) containing no MtATPG2 gene was used.
  • Seeds harvested from the transformed Arabidopsis larvae as in ⁇ Example 2-1> were selected by immersing and incubating for 30 minutes in a 0.1% Bassta herbicide (light, South Korea) solution. Thereafter, during the growth of the transformed Arabidopsis, the pollen was treated 5 times with the Basta herbicide, and the transformed Arabidopsis was selected from each pollen.
  • the T 1 Arabidopsis transformed with the pCSEN-MtATPG2 vector compared with the phenotype transformed only with a control (a vector without the MtATPG2 gene (pCSEN vector) or their wild-type Arabidopsis), surprisingly the variants were distinct. Multiple traits and biomass enhancing traits were shown.
  • T 2 transgenic Arabidopsis In order to more accurately identify the phenotypic changes of the transgenic Arabidopsis, the phenotypes of these lines were examined by receiving T 2 transgenic seeds from the T 1 transgenic Arabidopsis. First, T 2 transformed seed that had been cold-treated (4 ° C.) for 3 days was cultivated in a pollen, and then T 2 transgenic Arabidopsis was selected through the treatment of Basta herbicide. Phenotyping of selected Arabidopsis T 2 transformation lines was performed 50 days and 70 days after germination (FIG. 2).
  • RNA was extracted from the control and leaf of the mutant line 25 days after cotyledon formation using the RNasey Plant Mini Kit (QIAGEN, Germany), respectively. . 1 ⁇ g of RNA each as a template and 5 minutes at 65 ° C. using Superscript III Reverse Tanscriptase (INVITROGEN, USA); 60 minutes at 50 ° C; And cDNA was synthesized at 70 ° C. for 15 minutes. Thereafter, the synthesized cDNA was used as a template, and PCR was performed using the primers specific to the following Table 1 for the MtATPG2 gene and the TUB gene used as a PCR positive control.
  • MtATPG2 can be used as an excellent gene source for the development of high-productivity crops, as it is easy to manufacture plants that increase productivity with the same harvesting season as Arabidopsis wild type through the regulation of gene expression.
  • MtATPG2 It is shown in Figure 2 that the plant obtained through the introduction of the gene has an expression trait for multiplicity. In order to analyze the phenotypic characteristics of these multiple traits more accurately, productivity indicators such as seed yields were investigated in five transformation lines and compared with Arabidopsis control.
  • productivity indicators applied are plant height, silique number (NTS), biomass (FW), biomass (DW), total seed weight (TSW), and 1,000 seed weights (1,000 SW). Is the average value of 20 objects per line.
  • MtATPG2 gene mutant lines MtATPG2 -7, MtATPG2 -8, MtATPG2 -15, MtATPG2 -16 and All of MtATPG2 -17 increased seed production more than 1.5 times compared to Arabidopsis control, especially MtATPG2 -16 and MtATPG2 -17 more than doubled seed production compared to the control.
  • all variant lines had a long and numbered pattern of increase.
  • the seed weight is 1,000 or almost've found that similar to the control group, MtATPG2 - 7 as compared to the control group It weighed about 1.4 times or more.
  • MtATPG2-7 increased seed size compared to Arabidopsis control and other transformation lines (FIG. 4). Part of the seed size increased MtATPG2 -7 variants are expected to continue to proceed with further research. Overall, the expression of the MtATPG2 gene does not seem to have a significant effect on the seed size, but it is thought to induce an increase in the long shell length and the number, leading to an increase in seed yield. There was no significant difference in the size of the plant between the mutant and the control, but there was a marked increase in both the biomass and the dry weight, especially in the biodry weight.
  • CLV1 gene involved in the separated MtATPG2 in M. truncatula increase productivity by controlling the cytokinin signaling in Arabidopsis thaliana or / and CLV1 / WUS pathway of cytokinin signaling pathway in order to ensure that provide phenotypic features of senescence delay (receptor protein MtATPG2 the histidine kinase 4, AT2G01830) expression of the gene -: kinase CLAVATA1, AT1G75820) and histidine kinase HK2 (histidine kinase 2, AT5G35750) of the genes having a function, HK3 (histidin kinase3, AT1G27320) and CRE1 (HK4 as cytokinin receptor 7, MtATPG2 -8, and It was aimed at MtATPG2 -15 (Fig.
  • RT-PCR primer information for the applied gene and MtATPG2 and the positive control tubulin are shown in Table 2.
  • Table 2 RT-PCR primer information for the applied gene and MtATPG2 and the positive control tubulin.
  • CLV1 in the cytokinin signaling pathway Primer sequence and sequence number for RT-PCR for the gene, histidine kinase (HK) gene group , MtATPG2 , and positive control TUB gene gene Forward / Reverse Primer (SEQ ID NO) CLV1 CLV1-RT-F: 5'-ACT TAC CTC TGT CTC CCT CA-3 '(SEQ ID NO: 9) / CLV1-RT-R: 5'-GAC CAC CTT TAG ATC CAT GC-3 '(SEQ ID NO: 10) HK3 HK3-RT-F: 5'-CAA CAA CCA GCC CAT ATT CTC-3 '(SEQ ID NO: 11) / HK3-RT-R: 5'-TTC CAA TAC CCA ATC CCC TC-3 '(Dial No.
  • AHP1 from the phosphotransfer proteins (HP) gene family (AT3G21510) , AHP2 (AT3G29350) , AHP3 (AT5G39340), AHP4 (AT3G16360) , AHP5 (AT1G03430) , and AHP6 Expression patterns of the (AT1G80100) gene were also examined (FIG. 5). The expression pattern of the gene was analyzed by RT-PCR, primer information used is shown in Table 3.
  • AHP1, AHP2, and in the case of AHP3 showed a high expression in MtATPG2 MtATPG2 -7 and -15 mutants compared to the control, in the case of AHP4 exhibited high expression in MtATPG2 MtATPG2 -7 and -8 variant compared to the control.
  • AHP1 (AT3G21510)
  • AHP1-RT-F 5'-ATGGATTTGGTTCAGAAGCAGAA-3 '(SEQ ID NO: 17)
  • AHP1-RT-R 5'-TCAAAATCCGAGTTCGACGGCC-3'
  • AHP2 (AT3G29350)
  • AHP2I-RT-F 5'-ATGGACGCTCTCATTGCTCAGC-3 '(SEQ ID NO: 19)
  • AHP2I-RT-R 5'-TTA GTT AAT ATC CAC TTG AGG AAC-3' (SEQ ID NO: 20)
  • AHP3 (AT5G39340)
  • AHP3-RT-F 5'-GGACACACTCATTGCTCAGT-3 '(
  • MtATPG2 is not significantly involved in the CLV1 / WUS pathway in the cytokinin pathway.
  • MtATPG2 regulates the expression of the cytokinin receptor gene family and histidine phosphotransfer proteins (HP) gene family to regulate cytokinin signaling to increase plant yield.
  • Productivity traits such as increased biomass and delayed aging traits, and 3) these phenotypic characteristics are regulated according to the expression level of the MtATPG2 gene. That is, normal or relatively low expression of the MtATPG2 gene provides a phenotype of increased productivity, and a relatively high gene expression provides a phenotype of extended melting with a productivity-enhancing trait.
  • the homologous lines were selected by generation advancement, and then the six phenotypes were identified whether their phenotypes were the same as the previous generation phenotypes. .
  • most of the transformant T 3 or T 4 lines at 50 and 70 days after germination showed distinct multiplicity and biomass enhancement traits as in the previous results.
  • MtATPG2 -7-15-11 and MtATPG2 -7-15-12 lines in the transformation lines exhibited some aging delay traits as well as productivity enhancing traits. Investigation of the gene expression of the transformants showed that the aging delay traits appearing with this multiplicity were due to the increased expression level of the present gene (FIG. 8). This phenotypic characteristic of the transformant may be slightly different from the previous generation, which is believed to be due to changes in gene expression as the homo line is selected from the hetero line.
  • MtATPG2 Gene expression is expected to provide many advantages for stable generation of productivity-enhancing traits when applied to crops in that the expression of genes maintains the stabilization of phenotypic traits for productivity-enhancing traits.
  • MtATPG2 Expression T 3 or T 4 are transformed variants of the melt extending from the gene group that appears coding for the Arabidopsis thaliana are AT-hook protein did not appear very strong, but still in the mutant lines such as gene expression levels are high MtATPG2 -7 variants with the trait of delayed senescence I could see that.
  • MtATPG2-8 and MtATPG2 -15 mutant lines age of wild - I've found similar age-dependent phenomenon MtATPG2 -7 mutant lines were found to have the traits of aging delay compared to the control (Fig. 9).
  • chlorophyll content was measured during age-dependent aging. Chlorophyll content was measured according to the method of Lichtenthaler and Wellburn ( Biochemical Society Transduction 603: 591 ⁇ 592, 1983) using extinction coefficients of 663.2 nm and 664.8 nm.
  • a second study of aging delayed trait analysis examined the characteristics of cancer-induced aging.
  • MtATPG2 -8 and MtATPG2 -15 variant of the control lines are cancer-induced aging and appear nateu or MtATPG2 -7 mutant lines were found to have similar traits of aging delay by keeping the degree to melt even after 6 days (Fig. 11).
  • chlorophyll content was investigated during cancer-induced aging, and the chlorophyll content decrease pattern was proportional to the phenotypic features (FIG. 12).
  • MtATPG2 Since gene expression maintains the stabilization of phenotypic traits against pluripotent traits and aging delayed traits even in generational development, it may be possible to develop multiply crops through stable generation of agricultural traits when applied to crops.

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Abstract

La présente invention concerne une protéine MtATPG2 à fonction d'augmentation de productivité, améliorant la résistance au stress et retardant la sénescence d'une plante, un gène MtATPG2 associé, et l'utilisation de ladite protéine et dudit gène. Une plante dans laquelle le gène est introduit et exprimé présente la caractéristique d'une résistance anti-stress et d'un retard de sénescence en plus d'une augmentation de productivité.
PCT/KR2017/010859 2016-10-07 2017-09-28 Protéine mtatpg2 à fonction d'augmentation de productivité, améliorant la résistance au stress et retardant la sénescence d'une plante, gène associé, et utilisation de ladite protéine et dudit gène WO2018066898A2 (fr)

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KR101850003B1 (ko) * 2010-02-08 2018-06-05 제노마인(주) 식물의 노화 지연 기능 및 스트레스 내성 기능을 갖는 ａｔｈｇ１ 단백질과 그 유전자 및 이들의 용도
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KR101855135B1 (ko) * 2012-02-20 2018-05-09 제노마인(주) 식물의 생산성 증대 기능, 노화 지연 기능 및 스트레스 내성 기능을 갖는 ａｔｐｇ３ 단백질과 그 유전자 및 이들의 용도
KR20150003099A (ko) * 2013-06-28 2015-01-08 제노마인(주) 식물의 생산성 증대 기능, 스트레스 내성 기능 및 노화 지연 기능을 갖는 atpg6 단백질과 그 유전자 및 이들의 용도
KR20150001926A (ko) * 2013-06-28 2015-01-07 제노마인(주) 식물의 생산성 증대 기능, 스트레스 내성 기능 및 노화 지연 기능을 갖는 atpg10 단백질과 그 유전자 및 이들의 용도

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