WO1996017069A2 - Plantes transgeniques presentant une production de biomasse amelioree - Google Patents

Plantes transgeniques presentant une production de biomasse amelioree Download PDF

Info

Publication number
WO1996017069A2
WO1996017069A2 PCT/EP1995/004705 EP9504705W WO9617069A2 WO 1996017069 A2 WO1996017069 A2 WO 1996017069A2 EP 9504705 W EP9504705 W EP 9504705W WO 9617069 A2 WO9617069 A2 WO 9617069A2
Authority
WO
WIPO (PCT)
Prior art keywords
phosphate
transgenic plant
plants
dna
plant
Prior art date
Application number
PCT/EP1995/004705
Other languages
German (de)
English (en)
Other versions
WO1996017069A3 (fr
Inventor
Jens Kossmann
Franziska Springer
Original Assignee
Hoechst Schering Agrevo Gmbh
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Hoechst Schering Agrevo Gmbh filed Critical Hoechst Schering Agrevo Gmbh
Priority to JP8518189A priority Critical patent/JPH10510162A/ja
Priority to EP95940259A priority patent/EP0797673A2/fr
Priority to AU41770/96A priority patent/AU715002B2/en
Priority to HU9800264V priority patent/HU221515B/hu
Publication of WO1996017069A2 publication Critical patent/WO1996017069A2/fr
Publication of WO1996017069A3 publication Critical patent/WO1996017069A3/fr

Links

Classifications

    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • C12N9/1217—Phosphotransferases with a carboxyl group as acceptor (2.7.2)
    • 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
    • C12N15/8214—Plastid 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)
    • C12N15/8241—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
    • C12N15/827—Flower development or morphology, e.g. flowering promoting factor [FPF]
    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • C12N9/1229—Phosphotransferases with a phosphate group as acceptor (2.7.4)
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
    • Y02A40/146—Genetically Modified [GMO] plants, e.g. transgenic plants

Definitions

  • the present invention relates to transgenic plant cells in which the introduction and expression of certain DNA molecules leads to the formation of easily mobilizable phosphate pools outside the vacuole.
  • Transgenic plants which contain such plant cells show an increased biomass production and / or a changed flowering behavior in comparison to non-transformed plants.
  • the invention further relates to methods for increasing yields or changing the flowering behavior in plants, in which plants are changed in such a way that easily mobilizable phosphate pools outside the vacuole are formed in the cells.
  • sucrose transporter Riesmeier et al., EMBO J. 13 (1994), 1-7) to influence the sucrose transport is also considered (see also WO 94/00574).
  • the present invention is therefore based on the object of making available further genetic engineering methods which can generally be used in plants to increase the biomass production or the yield.
  • the invention thus relates to transgenic plant cells in which at least one easily mobilizable phosphate pool is formed outside the vacuole due to the introduction and expression of a DNA molecule which codes for a protein which is involved in the synthesis of a phos - is involved in a molecule containing phosphate.
  • Increasing the biomass production in plants is understood in the context of this invention to mean an increase in the biomass of the entire plant (measured as dry weight) and / or individual parts in comparison with wild-type plants, preferably an increase of at least 5% and in particular an increase of more than 10%.
  • the increase in biomass production thus also includes the increase in the yield of agriculturally usable parts of plants, for example storage organs, such as potato tubers, beets, seeds, fruits, or of leaves, stems, etc., in plants in which phosphate pools outside the vacuole were produced compared to wild-type plants, preferably by at least 5%, and in particular by more than 10%.
  • storage organs such as potato tubers, beets, seeds, fruits, or of leaves, stems, etc.
  • a change in the flowering behavior or premature flower formation means that transformed plants compared to non-transformed plants at least a few days, preferably one to several weeks, in particular 1-2 weeks early bloom here.
  • the term “phosphate pool” is understood to mean a class of phosphate-containing molecules which contain phosphate covalently bound and from which phosphate is easily mobilized, ie released or onto other molecules, by generally reversible enzymatic reactions can be transferred. This increases the availability of phosphate in the cells for various phosphate-dependent reactions.
  • phosphate mobilisable is understood to mean that the phosphate is available more quickly in the cell's cytosol than is usually possible by transporting phosphate from the vacuole into the cytosol.
  • the release of phosphate from the vacuum into the cytosol in the event of a phosphate deficiency in the cytosol generally takes place in the region of several hours (Woodrow et al., Planta 161 (1984), 525-530).
  • the phosphate pools are preferably pools of phosphate-containing molecules which do not impair the cytosolic homeostasis necessary for metabolic processes with regard to the phosphate concentration, but from which phosphate can be easily released or transferred to other molecules, i.e. which ensure an increased availability of phosphate.
  • phosphate-containing compounds which do not normally occur in higher plants. These include polyphosphate (Wood, Ann. Rev. Biochem. 57 (1988), 235-260), a compound which is synthesized by most organisms, except for higher plants, as a storage substance for phosphate, and e.g. also acetyl phosphate, which is used in bacteria for ATP regeneration (see e.g. Matsuyama et al., J. Bacteriol. 171 (1989), 577-580).
  • the phosphate pools can also consist of trehalose-6-phosphate, phosphoenolpyruvate or fructose-1-phosphate.
  • Such phosphate pools outside the vacuum are preferably produced by the fact that in vegetable Cells DNA molecules are introduced and expressed, which encode proteins whose enzymatic activity leads to the generation of the respective phosphate pool in transgenic plant cells.
  • the DNA molecules introduced into the plant cells code for proteins with the enzymatic property of a polyphosphate kinase, an acetate kinase, a phosphotransacetylase, a trehalose-6-phosphate synthase, a phosphoenolpyruvate mutase or a ketohexokinase.
  • ATP polyphosphate phosphotransferase
  • the product of this reversible reaction is a linear polymer of orthophosphate residues.
  • the length of the polymers can range from 3 to over 1000 phosphate residues.
  • the enzyme has already been described in various organisms, including E. coli (Ahn and Kornberg, J. Biol. Chem. 265 (1990), 11734-11739; Akiyama et al., J. Biol. Chem. 267 (1992), 22556-22561), Klebsiella aerogenes (Kato et al ., Gene 137 (1993), 237-242), S. cerevisiae (Felter and Stahl, Biochimie 55 (1973), 245-251), Propionibacterium shermanii (Robinson and Wood, J. Biol.
  • Acetate kinases (ATP: acetate phosphotransferase; E.C. 2.7.2.1.) Catalyze the reaction:
  • Acetate + ATP s acetyl phosphate + ADP The enzyme has been identified in a number of microorganisms and DNA sequences encoding acetate kinases have also been isolated, e.g. the ack gene from E. coli K-12 (Matsuyama et al., J. Bacteriol. 171 ( 1989), 577-580) and the ac ⁇ gene from Methanosarcina thermophila (Latimer and Ferry, J. Bacteriol. 175 (1993), 6822-6829).
  • Phosphotransacetylases (acetyl-CoA: orthophosphate acetyl transferase; E.C. 2.3.1.8.) Catalyze the following reaction:
  • Acetyl Coenzyme A + Orthophosphate Acetyl Phosphate + Coenzyme A
  • DNA sequences which code for this enzyme are also described, e.g. the pta gene from Methanosarcina thermophila (Latimer and Ferry, J. Bacteriol. 175 (1993), 6822-6829)
  • Trehalose-6-phosphate synthase catalyzes the synthesis of trehalose-6-phosphate.
  • Such an enzyme from yeast is described, for example, in US Pat. No. 5,422,254.
  • Phosphoenolpyruvate mutase catalyzes the synthesis of phosphoenolpyruvate.
  • Such an enzyme from Tetrahymena pyriformis is described, for example, in Seidel et al. (Biochemistry 31 (1995), 2598-2608).
  • Ketohexokinase catalyzes the synthesis of fructose-1-phosphate.
  • Such rat and human enzymes are described, for example, in Donaldson et al. (Biochem. J. 291 (1993), 179-186) or in Bonthron et al. (Hum. Mol. Genet. 3 (1994), 1627-1631).
  • the DNA molecules which code for proteins whose enzymatic activity leads to the formation of phosphate pools outside the vacuole in particular proteins with the enzymatic see property of a polyphosphate kinase, an acetate kinase, a phosphotransacetylase, a trehalose-6-phosphate synthase, a phosphoenolpyruvate mutase or a ketohexokinase
  • it can be genomic or cDNA molecules from any organism, preferably around DNA molecules from prokaryotic, in particular bacterial organisms.
  • the DNA molecules can be isolated from cells with the aid of common molecular biological methods or produced synthetically.
  • the molecules can be modified according to methods known to those skilled in the art in such a way that they contain plant-specific codons in order to improve expression in plant cells.
  • DNA molecules are preferably used for introduction and expression in plant cells which encode proteins which have the enzymatic property of a polyphosphate kinase and which have a low K - value for ADP and a high 1 ⁇ value for ATP.
  • the activity of these enzymes is usually influenced by the ratio of ATP to ADP. This means that a phosphate pool is only formed if there is an excess of ATP compared to ADP. This leads to a depletion of the cytosolic phosphate concentration under conditions which prefer the formation of ATP and to a constant reflux of phosphate from the vacuole. These newly formed pools have the advantage over the vacuolar phosphate pool that they are easier to mobilize.
  • the release of phosphate from the vacuole into the cytosol in the event of a phosphate deficiency in the cytosol generally takes place in the region of several hours (Woodrow et al., Planta 161 (1984), 525-530).
  • the DNA molecules which encode a polyphosphate kinase and which are introduced into the plant cells are DNA molecules from E. coli, Klebsiella aerogenes, Neisseria meningi tidis, or Synechocystia sp. , It is particularly preferably the pp gene from E. coli (Akiyama et al., J. Biol. Chem.
  • DNA molecules are introduced which code for an acetate kinase and originate from Methanosarcina thermophila, E. coli, Haemophilus influenza, Bacillus subtilis or Mycoplasma geni taliu. It is preferably the ac gene from E. coli K-12 (Matsuyama et al., J. Bacteriol. 171 (1989), 577-580), the ack gene from Methanosarcina thermophila, which is described in Fleischmann et al. (Science 269 (1995), 496-512) published sequence from Haemophilus influenza to the in Grundy et al. ⁇ J. Bacteriol.
  • DNA molecules that encode a phosphotransacetylase preferably originate from Methanosarcina thermophila, Escherichia coli or Mycoplasma geni talium. It is particularly preferably the pta gene from Methanosarcina thermophila
  • DNA molecules that encode a trehalose-6-phosphate synthase ⁇ preferably come from yeast.
  • the DNA molecule described in US Pat. No. 5,422,254 is preferably used.
  • DNA molecules that encode a phosphoenolpyruvate mutase are preferably derived from Tetrahymena pyriformis, such as that in Seidel et al. (see above) described molecule.
  • DNA molecules that encode a ketohexokinase are preferably from humans or rats, such as those in Bonthron et al. (see above) or Donaldson et al. (see above) molecules described.
  • DNA molecules which encode a protein which leads to the synthesis of polyphosphate DNA molecules which encode a protein with the enzymatic activity of an acetate kinase and a DNA sequence coding for a protein with the enzymatic activity of a phosphotransacetylase.
  • the enzymatic activities of acetate kinase and phosphotransacetylase lead to the conversion of acetyl-coenzyme A and ADP to acetate and ATP.
  • the resulting ATP can then be used by the polyphosphate kinase for the synthesis of polyphosphate.
  • the mobilization of the phosphate pools formed by the expressed proteins in the cells outside the vacuole, in particular polyphosphate can also be carried out by other enzymes in addition to the enzymes which catalyze the reversible reactions described above.
  • Enzymes using polyphosphate as a substrate are known. DNA sequences which encode such enzymes must then also be introduced into the plant cells and expressed.
  • polyphosphate glucokinase which has already been described in a number of microorganisms, is known to for example in Mycobacterium phlei (Szymona, Bull. Acad. Pol. Sei. Ser. Sei. Biol. 5 (1956), 379-381; Szymona and Ostrowski, Biochim. Biophys.
  • 1,3-diphosphoglycerate polyphosphate phosphotransferase (Kulaev et al., Biokhimiya 33 (1968), 419-434; Wood and Goss, Proc. Natl. Acad. Sci. USA 82 (1985), 312-315), the polyphosphate-dependent NAD kinase (Murata et al., Agric. Biol. Chem. 44 (1980), 61-68), exopolyphosphatase (see e.g. Akiyama et al., J. Biol. Chem.
  • the protein expressed in the transgenic plant cell which due to its enzymatic activity causes the formation of a phosphate pool outside the vacuole, in particular polyphosphate or acetylphosphate, is localized in any compartment of the plant cell can.
  • the coding region In order to achieve localization in a specific compartment, the coding region must be linked to DNA sequences which ensure localization in the respective compartment.
  • the protein to be expressed will preferably be located in the cytosol, the plastids or the mitochondria. No special signal sequence is required for localization in the cytosol. When using prokaryotic DNA molecules, care must be taken that the coding regions have no signal sequences which cause secretion of the protein to be expressed.
  • Plant cells into which the DNA molecules described above are introduced can generally originate from any plant species, in particular from any monocotyledonous or dicotyledonous plants.
  • Plant cells of agricultural useful plants are preferably used, in particular of cereals (such as barley, rye, oats, millet, rice, corn, wheat etc.), types of fruit, potatoes, rape, sugar beet, soybeans, vegetables (such as peas, Bean, tomato etc.) or from ornamental plants.
  • Photosynthetically active cells are particularly preferred.
  • the invention also relates to transgenic plants which contain transgenic plant cells according to the invention. These can be obtained, for example, by regeneration from the transgenic plant cells described, using methods known to the person skilled in the art.
  • the invention also relates to propagation material of a plant according to the invention.
  • the present invention relates to methods for increasing the biomass product or the yield and / or changing the flowering behavior in plants, characterized in that easily mobilizable phosphate pools are generated in plant cells outside the vacuole, which guarantee an increased availability of phosphate in the cells.
  • Such a procedure usually comprises the following steps:
  • any promoter which is functional in plants is suitable for the promoter mentioned under (i) in the processes according to the invention.
  • the expression of the said DNA sequences can generally take place in any tissue of a transformed plant and at any time.
  • the 35S promoter of the Cauliflower mosaic virus (Odell et al., Nature 313 (1985), 810-812) is suitable, which ensures constitutive expression in all tissues of a plant.
  • promoters can also be used which lead to an expression of subsequent sequences in a certain tissue of the plant, preferably in photosynthetically active tissue (see, for example, Stockhaus et al., EMBO J. 8 (1989), 2245- 2251) or only at a point in time determined by external influences (see for example WO / 9307279).
  • the termination sequence serves to correctly terminate the transcription and to add a poly-A tail to the transcript, which is believed to have a function in stabilizing the transcripts.
  • Such elements are described in the literature (cf. Gielen et al., EMBO J. 8 (1989), 23-29) and can be interchanged as desired.
  • the expression cassette produced in the course of the method according to the invention is also a subject of the invention. It is preferably located on a plasmid, in particular on the plasmids p35S-PPK (FIG. 1) or p35S-ACK (FIG. 3), and is preferably used using a plasmid which is suitable for the transformation of plant cells is introduced into plant cells.
  • the binary vector pBinAR Höfgen and Willmitzer, Plant Sei. 66 (1990), 221-230
  • This vector is a derivative of the binary vector pBin19 (Bevan, Nucl. Acids Res.
  • Cells of agricultural crops are preferably used in the process, in particular cereals (such as barley, rye, oats, millet, rice, corn, wheat etc.), fruit varieties, potatoes, rapeseed, sugar beet, soybeans, Vegetable types (such as peas, beans, tomatoes etc.) or cells from ornamental plants.
  • cereals such as barley, rye, oats, millet, rice, corn, wheat etc.
  • fruit varieties such as barley, rye, oats, millet, rice, corn, wheat etc.
  • Fruit varieties such as barley, rye, oats, millet, rice, corn, wheat etc.
  • fruit varieties such as barley, rye, oats, millet, rice, corn, wheat etc.
  • rapeseed such as sugar beet
  • soybeans such as Vegetable types (such as peas, beans, tomatoes etc.) or cells from ornamental plants.
  • Vegetable types such as
  • the invention also relates to the transgenic plant cells and transgenic plants resulting from the processes according to the invention.
  • the transgenic plants are characterized in that in cells of these plants, due to the introduction and stable integration of an expression cassette constructed according to the method according to the invention into the genome, there is expression of a protein which, owing to its enzymatic activity, forms polyphosphate, Acetyl phosphate, trehalose-6-phosphate, phosphoenol pyruvate or fructose-1-phosphate outside the vacuole in the transgenic cells.
  • the invention relates to transgenic plants containing at least one DNA sequence which contains a protein with the enzymatic activity of a polyphosphate kinase, an acetate kinase, a phosphotransacetylase, a trehalose-6-phosphate synthase, a phosphoenolpyruvate mutase, or a ketohexokinase encoded, this DNA sequence being linked to regulatory DNA regions for transcription and translation in plant cells, being stably integrated into the genome and, owing to the expression of the said DNA sequence, forming polyphosphate, acetylphosphate, trehalose-6 Phosphate, phosphoenolpyruvate or fructose-1-phosphate comes outside the vacuole in plant cells.
  • Another object of the invention is the use of DNA molecules which encode proteins which, owing to their enzymatic activity, form phosphate pools which can be easily mobilized, in particular polyphosphate, acetylphosphate, trehalose-6-phosphate, fructose-1 Phosphate or phosphoenol pyruvate, outside the vacuole in plant cells, lead to the transformation of plant cells and to expression in plant cells, in particular to improve the phenotype of these cells.
  • the invention relates to the use of DNA molecules, the proteins with the enzymatic activity of a polyphosphate kinase, an acetate kinase, a phosphotransacetylase, a trehalose-6-phosphate synthase.
  • DNA molecules include in particular the DNA molecules described above, which encode proteins with the enzymatic activity of a polyphosphate kinase, an acetate kinase, a phosphotransacetylase, a trehalose-6-phosphate synthase, a phosphoenolpyruvate mutase or a ketohexokinase.
  • cloning vectors which contain a replication signal for E. coli and a marker gene for the selection of transformed bacterial cells.
  • examples of such vectors are pBR322, pUC series, M13mp series, pACYC184 etc.
  • the desired sequence can be introduced into the vector at a suitable restriction site.
  • the plasmid obtained is used for the transformation of E. coli cells.
  • Transformed E. coli cells are grown in a suitable medium, then harvested and lysed.
  • the plasmid is recovered. Restriction analyzes are generally used as the analysis method for characterizing the plasmid DNA obtained, Gel electrophoresis and other biochemical-molecular biological methods are used.
  • the plasmid DNA can be cleaved and DNA fragments obtained can be linked to other DNA sequences.
  • Each plasmid DNA sequence can be cloned into the same or different plasmids.
  • a variety of techniques are available for introducing DNA into a plant host cell. These techniques include the transformation of plant cells with T-DNA using Agrobacterium tumefaciens or Agrobacterium rhizogenes as the transformation agent, the fusion of protoplasts, the injection, the electroporation of DNA, the introduction of DNA using the biolistic method and More options.
  • DNA When DNA is injected and electroporated into plant cells, there are no special requirements for the plasmids used. Simple plasmids such as e.g. pUC derivatives can be used. However, if whole plants are to be regenerated from such transformed cells, the presence of a selectable marker gene is necessary. Depending on the method of introducing desired genes into the plant cell, further DNA sequences may be required. E.g. If the Ti or Ri plasmid is used for the transformation of the plant cell, at least the right boundary, but often the right and left boundary of the Ti and Ri plasmid T-DNA as the flank region, must be linked to the genes to be introduced .
  • Simple plasmids such as e.g. pUC derivatives can be used. However, if whole plants are to be regenerated from such transformed cells, the presence of a selectable marker gene is necessary. Depending on the method of introducing desired genes into the plant cell, further DNA sequences may be required. E.g. If the Ti or Ri plasm
  • the DNA to be introduced must be cloned into special plasmids, either in an intermediate vector or in a binary vector.
  • the intermediate vectors can be integrated into the Ti or Ri plasmid of the agrobacteria on the basis of sequences which are homologous to sequences in the T-DNA by homologous recombination. This also contains the vir region necessary for the transfer of the T-DNA. Intermediate vectors cannot replicate in agrobacteria. Using a helper plasmid, the intermediate vector can be transferred to Agrobacterium tumefaciens (conjugation). Binary vectors can replicate both in E. coli and in agrobacteria.
  • the agrobacterium serving as the host cell is said to contain a plasmid which carries a vir region. The vir region is necessary for the transfer of the T-DNA into the plant cell. Additional T-DNA may be present.
  • the agrobacterium transformed in this way is used to transform plant cells.
  • T-DNA for the transformation of plant cells has been intensively investigated and is sufficient in EP 120516; Hoekema, In: The Binary Plant Vector System Offsetdrukkerij Kanters BV, Alblasserdam, Chapter V; Fraley et al., Crit. Rev. Plant. Sci., 4 (1985), 1-46 and An et al. , EMBO J. 4 (1985), 277-287.
  • plant explants can expediently be cultivated with Agrobacterium tumefaciens or Agrobacterium rhizogenes.
  • Whole plants can then be regenerated from the infected plant material (for example leaf pieces, stem segments, roots, but also protoplasts or suspension-cultivated plant cells) in a suitable medium, which can contain antibiotics or biocides for the selection of transformed cells .
  • the plants thus obtained can then be examined for the presence of the introduced DNA.
  • the introduced DNA is integrated in the genome of the plant cell, it is generally stable there and remains in the offspring of the originally transformed cell. It normally contains a selection marker which gives the transformed plant cells resistance to a biocide or an antibiotic such as kanamycin, G 418, bleomycin, hygromycin or phosphinotricin and others.
  • the individually selected marker should therefore allow the selection of transformed cells over cells which lack the introduced DNA.
  • the transformed cells grow within the plant in the usual way (see also McCormick et al., Plant Cell Reports 5 (1986), 81-84).
  • the resulting plants can be grown normally and crossed with plants which have the same transformed genetic makeup or other genetic makeup.
  • the resulting hybrid individuals have the corresponding phenotypic properties. Two or more generations should be grown to ensure that the phenotypic trait is stably maintained and inherited. Seeds should also be harvested to ensure that the appropriate phenotype or other characteristics have been preserved.
  • NSEB buffer 0.25 M sodium phosphate buffer pH 7.2
  • Fragment A CaMV 35S promoter, nt 6909-7437 (Franck et al., Cell 21 (1980), 285-294)
  • fragment B DNA fragment from Escherichia coli coding for polyphosphate kinase
  • the arrow indicates the direction of transcription of the ppk gene
  • FIG. 2 shows the plasmid pACK
  • the thin line corresponds to the plasmid pUC19
  • the strong line represents a DNA insert which comprises the coding region of the ac gene from Methanosarcina thermophila (Latimer and Ferry, J. Bacteriol. 175 (1993), 6822-6829).
  • the arrow indicates the direction of transcription of the ac gene.
  • FIG. 3 shows the plasmid p35S-ACK
  • A Fragment A: CaMV 35S promoter, nt 6909-7437 (Franck et al., Cell 21 (1980), 285-294)
  • B fragment B: DNA fragment from Methanosarcina thermophila coding for acetate kinase; Nucleotides 1314-2748 of the acetate kinase (ack) gene (Latimer and Ferry, J. Bacteriol. 175 (1993), 6822-6829);
  • the arrow indicates the direction of transcription of the ack gene
  • the vector pUC19 was used for cloning in E. coli.
  • the gene constructions were cloned into the binary vector pBinAR (Höfgen and Willmitzer, Plant Sei. 66 (1990), 221-230).
  • E. coli strain DH5 ⁇ (Bethesda Research Laboratories, Gaithersburgh, USA) was used for the pUC19 vector and for the pBinAR constructs.
  • the transformation of the plasmids into the potato plants was carried out using the Agrobacterium tumefaciens-Sta es C58C1 pGV2260 (Deblaere et al., Nucl. Acids Res. 13 (1985), 4777-4788).
  • the DNA was transferred by direct transformation according to the method of Höfgen & Willmitzer (Nucleic Acids Res. 16 (1988), 9877).
  • the plasmid DNA of transformed agrobacteria was isolated by the method of Birnboim & Doly (Nucleic Acids Res. 7 (1979), 1513-1523) and analyzed by gel electrophoresis after a suitable restriction cleavage.
  • the leaves for callus induction were then on MS medium with 1.6% glucose, 5 mg / 1 naphthylacetic acid, 0.2 mg / 1 benzylaminopurine, 250 mg / 1 claforan, 50 mg / 1 kanamycin or 1 mg / 1 hygromycin B, and 0.80% Bacto agar. After a week's incubation at 25 ° C.
  • the leaves were sprouted for induction on MS medium with 1.6% glucose, 1.4 mg / 1 zeatin ribose, 20 mg / 1 naphthylacetic acid, 20 mg / 1 Giberellic acid, 250 mg / 1 claforan, 50 mg / 1 kanamycin or 3 mg / 1 hygromycin B, and 0.80% Bacto agar. 5. Radioactive labeling of DNA fragments
  • the radiocative labeling of DNA fragments was carried out using a DNA random primer labeling kit from Boehringer (Germany) according to the manufacturer's instructions.
  • the membrane was prehybridized in NSEB buffer for 2 h at 68 ° C. and then hybridized in NSEB buffer overnight at 68 ° C. in the presence of the radioactively labeled sample.
  • a DNA fragment which codes for the polyphosphate kinase from E. coli was first amplified using the PCR technique ("polymerase chain reaction"). Genomic DNA from E. coli cells of the DH5 ⁇ strain were isolated by standard methods (see, for example, Sambrook et al., Molecular Cloning, A Laboratory Manual, 2nd ed. (1989), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY). Using the two oligonucleotides
  • Oligo 1 5 '-AAGGATCCAGGAACCCGGGCCATGGGTCAGGAAAAG-3' (Seq ID No. 1) and
  • Oligo 2 5 '-GGGGATCCCGGGCCATGGGTTATTCAGGTTG-3' (Seq ID No. 2)
  • nucleotides 187 to 2253 of the type described in Akiyama et al. (J. Biol. Chem. 267 (1992), 22558) comprises the DNA sequence (ppk gene) which encodes polyphosphate kinase from E. coli.
  • ppk gene polyphosphate kinase from E. coli.
  • restriction sites for the restriction endonucleases BamH I and an Sma I are introduced at the 5' end of the amplified DNA fragment.
  • Oligonucleotide 2 which is partially complementary to the 3 'end of the ppk gene, introduces interfaces for BamH I, Sma I and Nco I at the 3' end of the fragment.
  • the DNA fragment resulting from the PCR reaction was cut with BamH I and ligated into the binary vector pBinAR cut with BamH I (H ⁇ fgen and Willmitzer, Plant Sei. 66 (1990), 221-230). This is a derivative of the binary vector pBin19 (Bevan, Nucl. Acids Res. 12 (1984), 8711-8721).
  • pBinAR was constructed as follows:
  • a 529 bp fragment comprising nucleotides 6909-7437 of the 35S promoter of the Cauliflowermosaic virus was selected as the EcoRI / Kpn I fragment the plasmid pDH51 (Pietrzak et al., Nucl. Acids Res. 14, 5857-5868) isolated and between the EcoR I and the Kpn J sites of the polylinker from pBinl9 were ligated.
  • the plasmid pBin19-A was formed.
  • a 192 bp fragment was isolated from the plasmid pAGV40 (Herrera-Estrella et al., Nature 303, 209-213) with the aid of the restriction endonucleases Pvu II and Hind III which contains the polyadenylation signal of gene 3 of the T-DNA of the Ti plasmid pTiACH5 (Gielen et al., EMBO J. 3, 835-846) comprises (nucleotides 11749-11939). After addition of Sph I linkers to the Pvu J site, the fragment was ligated between the Sph I and Hind HI sites pBin19-A. This created pBinAR.
  • the insertion of the amplified DNA fragment creates an expression cassette which is composed of fragments A, B and C as follows:
  • Fragment A contains the 35S promoter of the cauliflower mosaic virus (CaMV).
  • the fragment comprises nucleotides 6909 to 7437 of the CaMV (Franck et al., 1980, Cell 21 (285-294)).
  • Fragment B contains the coding region for polyphosphate kinase from E. coli.
  • the fragment comprises nucleotides 187-2253 of the polyphosphate kinase gene (Akiyama et al., J. Biol. Chem. 267 (1992), 22556-22561). This coding region was ligated to the 35S promoter in sense orientation.
  • Fragment C (192 bp) contains the polyadenylation signal of gene 3 of the T-DNA of the Ti plasmid pTiACH5 (Gielen et al., EMBO J. 3 (1984), 835-846).
  • the size of the plasmid p35S-PPK is approximately 13 kb. Since the coding region of the gene for polyphosphate kinase from E. coli used did not comprise a signal sequence, the expressed protein should be present in the cytoplasm of transformed cells.
  • the plasmid was transferred into cells of potato plants using Agrobacterium-mediated transformation as described above. Whole plants were regenerated from the transformed cells. In this way, 40 lines of transformed plants were produced, of which 7 lines, in particular lines JP1 16, JP1 26, JP1 29, JP1 31, JP1 33, JP1 34 and JP1 35, were analyzed in more detail.
  • transgenic potato plants showed the expression of the gene for the polyphosphate kinase from E. coli.
  • the expression was verified with the help of Northern blot analyzes.
  • RNA was isolated from tissue from transgenic plants, separated by gel electrophoresis, transferred to a nylon membrane and hybridized with the radioactively labeled coding region of the ppk gene from E. coli.
  • 7 transgenic potato lines mentioned above one plant was examined with regard to the expression of the ppk gene. All showed a clear expression of the ppk gene from E. coli. In contrast, no transcripts of this gene could be detected in wild-type plants.
  • the starch content of the tubers of transformed plants corresponded to the starch content of tubers of wild-type plants (measurement by determining the density of the tubers). Furthermore, the transformed plants showed premature flowering compared to wild-type plants, as shown in the following tables: Table II n% of plants
  • Tables II and III illustrate the premature flower formation of plants which were transformed with the plasmid p35S-PPK in comparison with non-transformed plants (wild type).
  • Transformed potato plants of the lines JP1 16, JP1 26, JP1 29, JP1 31, JP1 33, JP1 34 and JP1 35 as well as wild-type plants of the potato variety Desiree were cultivated under greenhouse conditions.
  • Table II shows the percentage of the plants examined in each line or of the wild-type plants which, after 80 days after the exposure of the Plants bloomed.
  • Table III shows the percentage of plants examined in each line or of the wild-type plants which bloomed after 84 days after the plants had been exposed.
  • n number of plants analyzed
  • % Plants percentage of plants that bloom
  • p35S-ACK For the construction of the plasmid p35S-ACK, a DNA fragment was first isolated which encodes the acetate kinase from Methanosarcina thermophila (ack gene). The gene is inserted in the Sma J site of a pUC19 plasmid. The construction of this pUC plasmid is described in detail in Latimer and Ferry (J. Bacteriol. 175 (1993), 6822-6829; see page 6823, right column). The plasmid used, which is called pACK in the context of this invention, is shown in FIG. 2.
  • Fragment A contains the 35S promoter of the cauliflower mosaic virus (CaMV).
  • the fragment includes the Nucleotides 6909 to 7437 of CaMV (Franck et al., 1980, Cell 21, 285-294).
  • Fragment B contains the coding region for acetate kinase from Methanosarcina thermophila.
  • the fragment comprises nucleotides 1314-2748 of the sequence shown in Latimer and Ferry (J. Bacteriol. 175 (1993), 6822-6829) (acetylkinase iack) gene). This coding region was ligated to the 35S promoter in sense orientation.
  • Fragment C (192 bp) contains the polyadenylation signal of gene 3 of the T-DNA of the Ti plasmid pTiACH5 (Gielen et al., EMBO J. 3 (1984), 835-846).
  • the size of the plasmid p35S-ACK is approximately 12.5 kb.
  • the plasmid was transferred into cells of potato plants using agrobacterium-mediated transformation. Whole plants were regenerated from the transformed cells.
  • transgenic potato plants showed the expression of the acetate kinase from Methanosarcina thermophila in the cytosol of the cells.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Zoology (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Cell Biology (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • Medicinal Chemistry (AREA)
  • Physiology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Compounds Of Unknown Constitution (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)

Abstract

L'invention concerne des cellules végétales transgéniques dans lesquelles l'introduction et l'expression de molécules particulières d'ADN entraînent la formation de groupes de molécules renfermant du phosphate, facilement mobilisables, à l'extérieur de la vacuole. Les plantes transgéniques qui contiennent ces cellules présentent une production de biomasse améliorée et/ou une floraison modifiée. L'invention concerne en outre un procédé entraînant une augmentation du rendement et/ou une modification de la floraison des plantes.
PCT/EP1995/004705 1994-11-29 1995-11-29 Plantes transgeniques presentant une production de biomasse amelioree WO1996017069A2 (fr)

Priority Applications (4)

Application Number Priority Date Filing Date Title
JP8518189A JPH10510162A (ja) 1994-11-29 1995-11-29 バイオマス生産が改良されたトランスジェニック植物
EP95940259A EP0797673A2 (fr) 1994-11-29 1995-11-29 Plantes transgeniques presentant une production de biomasse amelioree
AU41770/96A AU715002B2 (en) 1994-11-29 1995-11-29 Transgenic plants with improved biomass production
HU9800264V HU221515B (en) 1994-11-29 1995-11-29 Transgenic plants with improved biomass production

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DEP4444460.5 1994-11-29
DE4444460A DE4444460A1 (de) 1994-11-29 1994-11-29 Verfahren zur Steigerung des Ertrags sowie zur Veränderung des Blühverhaltens bei Pflanzen

Publications (2)

Publication Number Publication Date
WO1996017069A2 true WO1996017069A2 (fr) 1996-06-06
WO1996017069A3 WO1996017069A3 (fr) 1996-08-29

Family

ID=6535750

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP1995/004705 WO1996017069A2 (fr) 1994-11-29 1995-11-29 Plantes transgeniques presentant une production de biomasse amelioree

Country Status (7)

Country Link
EP (1) EP0797673A2 (fr)
JP (1) JPH10510162A (fr)
AU (1) AU715002B2 (fr)
CA (1) CA2205849A1 (fr)
DE (1) DE4444460A1 (fr)
HU (1) HU221515B (fr)
WO (1) WO1996017069A2 (fr)

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997007221A1 (fr) * 1995-08-11 1997-02-27 Planttec Biotechnologie Gmbh Cellules vegetales transgeniques et plantes a taux de glycolyse augmente
WO1997042326A2 (fr) * 1996-05-03 1997-11-13 Mogen International N.V. Regulation du metabolisme par modification du taux de trehalose-6-phosphate
WO1998036084A2 (fr) * 1997-02-14 1998-08-20 Agricola Technologies, Inc. Amelioration de la croissance des vegetaux a l'aide de genes codant pour une anhydrase carbonique, une proteine fixant le calcium, une proteine fixant un metal, ou une proteine de biomineralisation
WO1998050561A1 (fr) * 1997-05-02 1998-11-12 Mogen International N.V. Metabolisme regulateur modifiant le niveau de trehalose-6-phosphate par inhibition des niveaux de trehalase endogene
WO1999023225A1 (fr) * 1997-10-30 1999-05-14 Mogen International N.V. Nouveaux micro-organismes a fermentation elevee
WO1999023234A1 (fr) * 1997-10-30 1999-05-14 Mogen International N.V. Inhibition de la remobilisation des composes stockes avant et apres recolte
WO1999024558A2 (fr) * 1997-10-30 1999-05-20 Mogen International N.V. Micro-organismes pour fermentation haute
WO2002034925A1 (fr) * 2000-10-20 2002-05-02 University Of Kentucky Research Foundation Utilisation d'acetate kinase bacterienne et des genes codant de celle-ci de façon a proteger des plantes contre differents agents pathogenes
US6476293B1 (en) 1999-10-01 2002-11-05 University Of Kentucky Research Foundation Use of bacterial acetate kinase and their genes for protection of plants against different pathogens
US8847006B2 (en) 2007-03-28 2014-09-30 Monsanto Technology Llc Utility of SNP markers associated with major soybean plant maturity and growth habit genomic regions

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE19619917A1 (de) * 1996-05-17 1997-11-20 Max Planck Gesellschaft Kartoffelpflanzen mit einer verringerten Aktivität der cytosolischen Stärkephosphorylase und einem veränderten Keimungsverhalten
WO1998004725A1 (fr) * 1996-07-31 1998-02-05 Yale University Procedes de modification de la vitesse de croissance de plantes et plantes ainsi obtenues
CN101815433B (zh) 2007-09-11 2013-06-05 孟山都技术公司 α-优化的β-伴大豆球蛋白增高的大豆
KR101370283B1 (ko) * 2012-09-19 2014-03-06 전남대학교산학협력단 애기장대의 유래의 ＡｔＲＢＰ1 단백질을 암호화하는 유전자로 형질전환된 생육촉진 및 종자수량이 증진된 식물체의 제조방법
CN109609542A (zh) * 2018-12-28 2019-04-12 博域环保技术研究院(南京)有限公司 多聚磷酸盐激酶基因ppk1在水稻中的基因工程应用

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0438904A1 (fr) * 1989-12-21 1991-07-31 Advanced Technologies (Cambridge) Limited Modification du métabolisme végétal
WO1993017093A2 (fr) * 1992-02-14 1993-09-02 Oy Alko Ab Augmentation de la teneur en trehalose des organismes par transformation a l'aide de combinaisons de genes de structure pour la trehalose-synthase
WO1994000563A1 (fr) * 1992-06-24 1994-01-06 Institut Für Genbiologische Forschung Berlin Gmbh Sequences d'adn et plasmides servant a preparer des plantes a concentration de sucrose modifiee
WO1995001446A1 (fr) * 1993-06-30 1995-01-12 Mogen International N.V. Production de trehalose dans des plantes

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0438904A1 (fr) * 1989-12-21 1991-07-31 Advanced Technologies (Cambridge) Limited Modification du métabolisme végétal
WO1993017093A2 (fr) * 1992-02-14 1993-09-02 Oy Alko Ab Augmentation de la teneur en trehalose des organismes par transformation a l'aide de combinaisons de genes de structure pour la trehalose-synthase
WO1994000563A1 (fr) * 1992-06-24 1994-01-06 Institut Für Genbiologische Forschung Berlin Gmbh Sequences d'adn et plasmides servant a preparer des plantes a concentration de sucrose modifiee
WO1995001446A1 (fr) * 1993-06-30 1995-01-12 Mogen International N.V. Production de trehalose dans des plantes

Cited By (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU719452B2 (en) * 1995-08-11 2000-05-11 Planttec Biotechnologie Gmbh Transgenic plant cells and plants with an increased glycolysis rate
WO1997007221A1 (fr) * 1995-08-11 1997-02-27 Planttec Biotechnologie Gmbh Cellules vegetales transgeniques et plantes a taux de glycolyse augmente
WO1997042326A2 (fr) * 1996-05-03 1997-11-13 Mogen International N.V. Regulation du metabolisme par modification du taux de trehalose-6-phosphate
WO1997042326A3 (fr) * 1996-05-03 1998-03-12 Mogen Int Regulation du metabolisme par modification du taux de trehalose-6-phosphate
US8124840B2 (en) 1996-05-03 2012-02-28 Syngenta Mogen B.V. Regulating metabolism by modifying the level of trehalose-6-phosphate
US7247770B2 (en) 1996-05-03 2007-07-24 Syngenta Mogen B.V. Regulating metabolism by modifying the level of trehalose-6-phosphate
US6833490B1 (en) 1996-05-03 2004-12-21 Mogen International N.V. Regulating metabolism by modifying the level of trehalose-6-phosphate
WO1998036084A2 (fr) * 1997-02-14 1998-08-20 Agricola Technologies, Inc. Amelioration de la croissance des vegetaux a l'aide de genes codant pour une anhydrase carbonique, une proteine fixant le calcium, une proteine fixant un metal, ou une proteine de biomineralisation
WO1998036084A3 (fr) * 1997-02-14 1998-12-17 Agricola Tech Inc Amelioration de la croissance des vegetaux a l'aide de genes codant pour une anhydrase carbonique, une proteine fixant le calcium, une proteine fixant un metal, ou une proteine de biomineralisation
WO1998050561A1 (fr) * 1997-05-02 1998-11-12 Mogen International N.V. Metabolisme regulateur modifiant le niveau de trehalose-6-phosphate par inhibition des niveaux de trehalase endogene
WO1999024558A2 (fr) * 1997-10-30 1999-05-20 Mogen International N.V. Micro-organismes pour fermentation haute
US6559364B1 (en) 1997-10-30 2003-05-06 Mogen International N.V. Pre- and postharvest inhibition of remobilisation of storage compounds
WO1999024558A3 (fr) * 1997-10-30 1999-07-15 Mogen Int Micro-organismes pour fermentation haute
WO1999023234A1 (fr) * 1997-10-30 1999-05-14 Mogen International N.V. Inhibition de la remobilisation des composes stockes avant et apres recolte
WO1999023225A1 (fr) * 1997-10-30 1999-05-14 Mogen International N.V. Nouveaux micro-organismes a fermentation elevee
US6476293B1 (en) 1999-10-01 2002-11-05 University Of Kentucky Research Foundation Use of bacterial acetate kinase and their genes for protection of plants against different pathogens
WO2002034925A1 (fr) * 2000-10-20 2002-05-02 University Of Kentucky Research Foundation Utilisation d'acetate kinase bacterienne et des genes codant de celle-ci de façon a proteger des plantes contre differents agents pathogenes
AU2001213383B2 (en) * 2000-10-20 2007-08-16 University Of Kentucky Research Foundation Use of bacterial acetate kinase and their genes for protection of plants against different pathogens
US8847006B2 (en) 2007-03-28 2014-09-30 Monsanto Technology Llc Utility of SNP markers associated with major soybean plant maturity and growth habit genomic regions

Also Published As

Publication number Publication date
JPH10510162A (ja) 1998-10-06
AU715002B2 (en) 2000-01-13
HUT77465A (hu) 1998-05-28
EP0797673A2 (fr) 1997-10-01
HU221515B (en) 2002-10-28
AU4177096A (en) 1996-06-19
WO1996017069A3 (fr) 1996-08-29
CA2205849A1 (fr) 1996-06-06
DE4444460A1 (de) 1996-05-30

Similar Documents

Publication Publication Date Title
DE69733945T2 (de) Metabolismusregulierung durch verändern des gehaltes an trehalose-6-phosphat
EP0973921B1 (fr) Sequences d'adn codant pour une 2-desoxyglucose-6-phosphate (2-dog-6-p) phosphatase et servant de marqueur de selection chez les vegetaux
DE69534467T2 (de) Modifikation des starke-gehalts in pflanzen
DE4447387A1 (de) Debranching-Enzyme aus Pflanzen und DNA-Sequenzen kodierend diese Enzyme
DE19509695A1 (de) Verfahren zur Herstellung einer modifizieren Stärke in Pflanzen, sowie die aus den Pflanzen isolierbare modifizierte Stärke
WO1996017069A2 (fr) Plantes transgeniques presentant une production de biomasse amelioree
DE112010000876T5 (de) Steuerung Kälte-induzierten Süßens und Verringerung von Acrylamid-Niveaus in Kartoffeln oder Süßkartoffeln
EP0479180A2 (fr) Plantes résistantes aux virus, procédé pour leur production
DE4035756A1 (de) Neue plasmide zur herstellung von im habitus und ertrag veraenderten transgenen pflanzen
DE3933384A1 (de) Multifunktionelle rna mit selbstprozessierungsaktivitaet, ihre herstellung und ihre verwendung
DE19502053A1 (de) Verfahren und DNA-Moleküle zur Steigerung der Photosyntheserate in Pflanzen, sowie Pflanzenzellen und Pflanzen mit gesteigerter Photosyntheserate
DE69434173T2 (de) Herstellung von trehalose in pflanzen
EP0513054A1 (fr) Genes de resistance aux virus/herbicides, leur procede de production et leur utilisation
KR102272557B1 (ko) CRISPR/Cas9 시스템을 이용한 갈변 억제 감자 식물체의 제조방법
EP0977878B1 (fr) Sequences d'adn codant pour la sous-unite chld de chelatases vegetales de magnesium pour determiner leurs effets
EP0428881B1 (fr) ARNs à activité endonucléase et antisens, leurs production et utilisation
WO1996013595A1 (fr) Procede permettant de modifier la floraison de plantes
WO2001059135A1 (fr) Procede permettant d'influencer la production de pollen par modification du metabolisme du saccharose
EP0918849A1 (fr) Cellules vegetales et plantes transgeniques a formation acetyle-coa modifiee
DE60008962T2 (de) Klonierung einer N-Methyltransferase, die an der Koffeinbiosynthese beteiligt ist
DE19732926C2 (de) DNA-Sequenzen, kodierend einen Glukose-6-phosphat-Phosphat-Translokator, sowie Plasmide, Bakterien, Hefen und Pflanzen, enthaltend diesen Transporter
WO2003014364A1 (fr) Procede pour influencer l'absorption de substances minerales chez des plantes transgeniques
CN115335528A (zh) 孢囊线虫属病原体抗性基因
BG100268A (bg) Продуциране на трехалоза в растения
WO1997025346A1 (fr) Sequence d'adn codant pour un translocateur de phosphoenolpyruvate-phosphate, plasmides, bacteries, levures et plantes contenant ce transporteur

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A2

Designated state(s): AU CA HU JP SI US

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LU MC NL PT SE

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
AK Designated states

Kind code of ref document: A3

Designated state(s): AU CA HU JP SI US

AL Designated countries for regional patents

Kind code of ref document: A3

Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LU MC NL PT SE

121 Ep: the epo has been informed by wipo that ep was designated in this application
ENP Entry into the national phase

Ref document number: 2205849

Country of ref document: CA

Ref country code: CA

Ref document number: 2205849

Kind code of ref document: A

Format of ref document f/p: F

WWE Wipo information: entry into national phase

Ref document number: 1995940259

Country of ref document: EP

ENP Entry into the national phase

Ref country code: US

Ref document number: 1997 849384

Date of ref document: 19970820

Kind code of ref document: A

Format of ref document f/p: F

WWP Wipo information: published in national office

Ref document number: 1995940259

Country of ref document: EP

WWR Wipo information: refused in national office

Ref document number: 1995940259

Country of ref document: EP

WWW Wipo information: withdrawn in national office

Ref document number: 1995940259

Country of ref document: EP