EP0906438A2 - Plants de pomme de terre a activite reduite de la phosphorylase cytosolique de l'amidon et a comportement en germination modifie - Google Patents

Plants de pomme de terre a activite reduite de la phosphorylase cytosolique de l'amidon et a comportement en germination modifie

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Publication number
EP0906438A2
EP0906438A2 EP97923093A EP97923093A EP0906438A2 EP 0906438 A2 EP0906438 A2 EP 0906438A2 EP 97923093 A EP97923093 A EP 97923093A EP 97923093 A EP97923093 A EP 97923093A EP 0906438 A2 EP0906438 A2 EP 0906438A2
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EP
European Patent Office
Prior art keywords
tubers
plants
cytosolic
wild
transgenic potato
Prior art date
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Withdrawn
Application number
EP97923093A
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German (de)
English (en)
Inventor
Jens Kossmann
Martin Steup
Elke Duwenig
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Max Planck Gesellschaft zur Foerderung der Wissenschaften eV
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Max Planck Gesellschaft zur Foerderung der Wissenschaften eV
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Publication of EP0906438A2 publication Critical patent/EP0906438A2/fr
Withdrawn legal-status Critical Current

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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10Transferases (2.)
    • C12N9/1048Glycosyltransferases (2.4)
    • C12N9/1051Hexosyltransferases (2.4.1)
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8242Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
    • C12N15/8243Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine
    • C12N15/8245Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine involving modified carbohydrate or sugar alcohol metabolism, e.g. starch biosynthesis
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8262Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield involving plant development
    • C12N15/8267Seed dormancy, germination or sprouting

Definitions

  • the present invention relates to transgenic potato plants which contain cells with a reduced activity of the cytosolic starch phosphorylase compared to wild-type plants.
  • the tubers of such potato plants show a different germination behavior compared to tubers of wild-type plants, which leads to the formation of an increased number of shoot ends and consequently to an increased number of stolens and tubers. Such plants also have an increased yield.
  • the yield from the agricultural cultivation of potatoes is primarily determined by the number of shoot ends that are formed per tuber. Normally, a germinating potato tuber forms only one, sometimes 2 to 3, shoot ends, the growth of further potentially existing side shoots being suppressed due to the apikaidominance of these shoot ends. Starting from the shoot ends, stolons are formed during the further growth of the potato plants, on which the tubers are later formed. Since the crop yield correlates with the number of stolons formed by the developed shoots, there is an effort to manipulate the germination of potato plants in such a way that the largest possible number of "eyes" germinate and develop into shoot ends.
  • One method is to break off the first sprout of a tuber, which suppresses the growth of further sprouts due to its apica dominance. This leads to the growth of further shoots.
  • Another method is to pre-germinate the potatoes to be laid out fine under regulated conditions in special containers (see e.g. Bouman, kannbau 47 (1996), 18-21; van de Waart, etcbau 44 (1993), 18-20).
  • Bouman, istbau 47 (1996), 18-21; van de Waart, cauliflower 44 (1993), 18-20 are very cost-intensive since, in addition to the special containers, they also require storage rooms in which both the light and the temperature conditions can be regulated.
  • a genetic engineering manipulation of potato plants was already proposed in DE-A1 42 13 444 in such a way that enzymes involved in starch metabolism are inhibited. Appropriate approaches for increased sprout formation of the potato tubers are not yet known. There is therefore a need for potato plants or processes in which the labor-intensive or cost-intensive steps described above are unnecessary and which lead to an increased number
  • the present invention is therefore based on the object of providing potato plants whose tubers form a large number of shoot ends when germinated.
  • the invention relates to transgenic potato plants that contain cells with a compared to wild-type plants, i.e. corresponding non-transformed plants, contain at least 60% reduced activity of the cytosolic starch phosphoryl.
  • the activity of the cytosolic starch phosphorylase is reduced by at least 80% and particularly preferably by at least 95% compared to wild-type potato plants.
  • cytosolic starch phosphorylase is understood to mean the isoform of starch phosphorylase (EC 2.4.1.1) localized in the cytoplasm of plant cells, which also is known as isoform H or I. This differs from the second, the plastid isoform, in that it shows, for example, a much higher affinity for highly branched glucans (Shimomura et al., J. Biochem. (Tokyo) 91 (1982), 703-717; Yang and Steup, Plant Physiol. 94, 960-969) and a low affinity for oligoglucans.
  • the enzyme catalyzes the reversible phosphorolysis of ⁇ -1, 4-glucans.
  • the activity of the starch phosphorylase can be determined, for example, as described in Parvin and Smith (Anal. Biochem. 27 (1969), 65-72), Conrads et al. (Biochim. Biophys. Acta 882 (1986), 452-463), Steup and Latzko (Planta 145 (1979), 69-75), Steup (In. Methods in Plant Biochemistry 3; Academic Press Limited (1990), 103- 128) or Sonnewald et al. (Plant Mol. Biol. 27: 567-576 (1995)).
  • the activity of the cytosolic starch phosphorylase is preferably reduced in all or in almost all cells of the plant. At least, however, in the bulbs and the shoot ends that develop from them.
  • tubers of potato plants which have such a reduced activity of the cytosolic starch phosphorylase show a drastically changed germination behavior compared to tubers of wild-type plants. Germination here means the growth of shoot ends from tubers.
  • tubers of wild-type plants usually form 1, at most 2 to 3, shoot ends during normal germination. To increase the number of shoots, either the elimination of the apikai shoot or a pre-germination under special conditions is necessary.
  • tubers of the potato plants according to the invention which contain cells with a reduced cytosolic starch phosphorylase, show a drastically increased number of shoot ends when germinated, in particular after storage at 20 ° C. in the dark. This leads to the formation of more stolons and consequently more tubers per plant. Overall, the tuber yield per plant increases. This concerns both the number of tubers per plant and the total fresh weight of tubers per plant.
  • the average number of shoots which are formed per tuber when germination takes place after storage for 5 months at 20 ° C. in the dark is at least doubled in comparison to wild-type plants ⁇ zen, ie corresponding non-transformed plants.
  • the number of shoot ends which are formed per budding eye when the germination takes place after storage for 5 months at 20 ° C. in the dark is at least doubled in comparison to tubers of wild-type Plants, ie corresponding non-transformed plants.
  • the plants according to the invention also have an increased yield with regard to the number of tubers and the weight of the tubers.
  • the number of tubers per plant is preferably at least 20%, preferably at least 50% and particularly preferably at least 100% higher than in corresponding ones non-transformed plants under the same growth conditions.
  • the fresh tuber weight of all tubers per plant is preferably at least 10%, preferably at least 15% and particularly preferably at least 20% higher than in corresponding non-transformed plants under the same growth conditions.
  • the activity of the cytosolic starch phosphorylase in the cells of the potato plants according to the invention can be reduced in principle by various methods known to the person skilled in the art.
  • the activity of the cytosolic starch phosphorylase is reduced by inhibiting the expression of endogenous genes which code for this enzyme.
  • Molecular biological techniques based on an antisense, ribozyme or a cosuppression effect are preferred.
  • a corresponding RNA is expressed in the antisense orientation. This preferably has a length of at least 30 nucleotides, preferably of at least 50 nucleotides and particularly preferably of at least 100 nucleotides.
  • the expressed antisense-RNA should have a high homology to the transcripts expressed endogenously in the plant, which encode cytosolic starch phosphorylase.
  • the homology is preferably at least 90%, preferably at least 95% and particularly preferably at least 99%.
  • an RNA is expressed which can specifically cleave transcripts of cytosolic starch phosphorylase.
  • the expression of ribozymes for reducing the activity of certain enzymes in cells is also known to the person skilled in the art and is described, for example, in EP-Bl 0 321 201.
  • the expression of ribozymes in plant cells was described, for example, in Feyter et al. (Mol. Gen. Genet. 250 (1996), 329-338).
  • the cosuppression effect is based on the expression of a sense RNA which expresses the expression suppressed by endogenous starch phosphorylase mRNA.
  • the implementation of these techniques is known to the person skilled in the art.
  • the method of cosuppression is described, for example, in Jorgensen (Trends Biotechnol. 8 (1990), 340-344), Niebel et al. (Curr. Top. Microbiol. Immuno. 197 (1995), 91-103), Flavell et al. (Curr. Top Microbiol. Immuno. 197 (1995), 43-46), Palaqui and Vaucheret (Plant. Mol. Biol. 29 (1995), 149-159), Vaucheret et al. (Mol. Gen. Genet. 248 (1995), 311-317), de Borne et al. (Mol. Gen. Genet. 243 (1994), 613-621) and other sources.
  • the reduction in the activity of a cytosolic starch phosphorylase in the cells is achieved by producing transgenic potato plants which contain a recombinant DNA molecule which is stably integrated into the genome and comprises the following elements:
  • the promoter of the patatin gene B33 from potato is suitable for expression in the tubers of the potato plants (Rocha-Sosa et al., EMBO J.8 (1989), 23-29).
  • the 35S promoter of the CaMV (Franck et al., Cell 21 (1980), 285-292) is suitable, for example, for constitutive expression.
  • DNA sequences which encode a potato cytosolic starch phosphorylase have already been described (see, for example, Mori et al., J. Biol. Chem. 266 (1991), 18446-18453). With the help of these DNA sequences it is possible for the person skilled in the art to isolate further sequences using standard methods which cyto- encode potato starch phosphorylase, if necessary.
  • the activity of the cytosolic starch phosphorylase can also be reduced by inactivating the endogenously present genes which code for this enzyme. Techniques for this are, for example, transposon mutagenesis or gene tagging. Alternatively, there is also the possibility of expressing antibodies in the cells which specifically recognize cytosolic starch phosphorylase.
  • the present invention relates to propagation material of the potato plants according to the invention, in particular seeds and particularly preferably potato tubers. These contain cells with a reduced activity of the cytosolic starch phosphorylase in comparison with tubers of wild-type plants and an altered germination behavior as described above.
  • the invention also relates to the use of nucleic acid molecules which encode a cytosolic starch phosphorylase or parts thereof for the production of transgenic potato plants with a reduced activity of the cytosolic starch phosphorylase, in particular by at least 60% in comparison to corresponding non-transfor ⁇ mated plants, preferably by at least 80% and particularly preferably by at least 95% and a changed germination behavior.
  • Figure 1 shows schematically the plasmid pBin-Anti-STPI K ⁇ r ⁇ .
  • cSTP 1.7 approx. 1.7 kb long DNA fragment which comprises part of the coding region for cytosolic starch phosphorylase from potato and is linked in antisense orientation to the 35S promoter.
  • FIG. 2 shows two polyacrylamide gels for detecting the activity of the cytosolic starch phosphorylase in leaf (A) and tuber tissue (B) transgenic potato plants which have been transformed with the plasmid pBin-Anti-STPI K ⁇ n .
  • Raw protein extracts from leaf and tuber tissue were separated in a non-denaturing polyacrylamide gel (discontinuous system; 12% and 4% (w / v) acrylamide).
  • the separating gel contained 2.4% (w / v) glycogen.
  • Approx. 20 ⁇ g protein was applied per lane.
  • the electrophoresis was carried out at 100 volts for 4 h. The direction of migration is from the top (cathode) to the bottom (anode).
  • the gels were incubated overnight in 20 mM glucose-1-phosphate / 100 mM citrate, pH 6.0 at room temperature. Protein bands with starch-synthesizing activity are visible as blue bands.
  • the cytosolic starch phosphorase (STPI) is strongly inhibited in its mobility by the immobilized polysaccharide.
  • the plastidic starch phosphorylase (STPII) is not so badly affected in its rate of migration.
  • FIG. 3 shows the germination behavior of tubers of wild-type plants (S. tuberosum L. cv Desiree; right) in comparison to tubers of transformed line cSTP 9 (left) after storage for 5 months at 20 ° C in the dark.
  • FIG. 4 shows the germination behavior of tubers from wild-type plants (center) in comparison to tubers of the transformed lines cSTP 6 (left) and cSTP 7 (right) after storage for 10 months at 20 ° C. in the dark .
  • FIG. 5 shows a statistical overview of the number of shoot ends formed on average per tuber or per 25 tuber in tubers of wild-type plants (cDesi) or the transformed lines (cSTP-6, -7, -9, -14, - 15, -16 and -18) after storage for 5 months at 20 ° C in the dark.
  • FIG. 6 shows a statistical overview of the average number of shoot ends formed per "eye" in tubers from wild-type plants (cDesi) and tubers in transformed lines (cSTP-6, -7, -9, -14, -15, -16 and -18).
  • the E.coli strain DH5 ⁇ (Bethesda Research Laboratories, Gaithersburgh, USA) was used for cloning.
  • the DNA was transferred by direct transformation using the Höfgen & Willmitzer method (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 membrane was prehybridized in NSEB buffer for 2 h at 68 ° C. and then hybridized in NSEB buffer overnight at 6B ° C. in the presence of the radioactively labeled sample.
  • Potato plants are kept in the greenhouse under the following conditions:
  • the plants are kept in individual pots (200 cm 2 , 15 cm deep) and watered daily.
  • the tubers are harvested 4 months after the transfer of the tissue culture plants into the greenhouse.
  • Tubers with a fresh weight of 8-16 g are used for biochemical analyzes. The fresh weight is determined immediately after harvesting.
  • the harvested tubers are washed and stored in boxes at 20 ° C for 5 to 10 months in the dark. Examples
  • an antisense construct which encodes an anti-sense RNA to transcripts which encode cytosolic starch phosphorylase from potato
  • part of the coding region described in Mori et al. (loc. cit.) described cDNA amplified by means of PCR from a ⁇ ZAP cDNA library from tuber tissue.
  • a 1.7 kb Asp718 / Smal fragment was inserted with smooth ends into the Smal interface of the binary plant transformation vector pBIN19 (Bevan, Nucl. Acids Res. 12 (1984), 8711-8721). This contains the 35S promoter of the CaMV and the polyadenylation signal of the octopine synthase gene. Restriction digestion ensured that the coding region was arranged in an antisense orientation to the promoter.
  • the resulting construct was designated pBin-Anti-STPI Km (see FIG. 1).
  • the vector pBin-Anti-STP Km was introduced into the Agrobacterium tumefaciens strain C58C1: pGV2260 (Höfgen and Willmitzer, Nucl. Acid Res. 16
  • RNA from leaf or tuber material was isolated from various independent lines of the transgenic potato plants produced according to Example 2 and analyzed by means of Northern blot analysis for the expression of mRNA which encodes cytosolic starch phosphorylase.
  • mRNA which encodes cytosolic starch phosphorylase.
  • samples from wild-type plants Solanum tuberosum L. cv. Desiree
  • the detection of the activity of the cytosolic starch phosphorylase in tissues of the transformed potato plants was carried out by the method of Steup (In: Methods in Plant Biochemistry 3, Academic Press Limited (1990), 103-128).
  • crude protein extracts were first obtained from the tissues to be examined by freezing tissue in extraction buffer (100 mM HEPES-NaOH, pH 7.5; 1 mM EDTA, 10% (vol. / Vol.) Glycerol; 5 mM DTT , 200 mg Na 2 S0 3 ; 150 mg Na 2 S 2 0 5 ) homogenized. After centrifugation, the clear supernatant was separated in a polyacrylamide gel.
  • Electrophoresis was carried out as described in Steup and Latzko (Planta 145 (1979), 69-759). Here a non-de- natural discontinuous system used (12% or 4% (w / v) acrylamide).
  • the separating gel contained 2.4% (w / v) glycogen.
  • the gel was run after the gel run in a solution of 20 mM glucose-1-phosphate / 100 mM citrate; pH 6.0 incubated overnight at room temperature. The gel was then incubated in a Lugolian solution for 5 min. Decolorization was carried out by extensive washing with water over a longer period of time. Blue color indicates the presence of starch-synthesizing enzyme activities.
  • the activity of the cytosolic starch phosphorylase can subsequently be determined, for example, by densitometric analysis of the corresponding protein bands.
  • FIG. 2 shows a polyacrylamide gel in which seven independent transgenic potato lines produced according to Example 2 were tested for their activity on cytosolic starch phosphorylase.
  • tubers of the plants were stored at 20 ° C. and the germination behavior of the tubers was examined after various periods.
  • Figure 3 shows a comparison of tubers of Solanum tuberosum L. cv. Desiree (wild type) and tubers of the line cSTPI-9 transformed with the plasmid pBin-Anti-STPI Km , each of which had been stored at 20 ° C. for 5 months.
  • FIG. 4 shows a comparison of wild-type tubers and tubers from two other transformed lines (cSTPI-6 and cSTPI-7) which have been stored at 20 ° C. for 10 months. It is clear from the two figures that the tubers of the transformed lines, which have a reduced activity of the cytosolic starch phosphorylase, have a drastically changed germination behavior.
  • the bulbs of the transformed lines form on the one hand significantly more shoot ends per bulb and also more shoot ends per out of no eye. Furthermore, more eyes usually germinate in the tubers of the transformed plants with reduced activity of the cytosolic starch phosphorylase.
  • a statistical evaluation according to - is shown in FIGS. 5 and 6.
  • the potato plants growing from the tubers of the transformed plants furthermore show an increased yield (in fresh bulb weight / plant). This is shown in the following table using the example of the transformed lines cStP6, cSTP7, cSTP9, CSTP14, CSTP15, CSTP16 and cSTP18.
  • the table shows that plants with a reduced activity of the cytosolic starch phosphorylase on the one hand form more tubers per plant than Wild-type plants and, moreover, the fresh tuber weight per plant is significantly higher than in wild-type plants.
  • the transformed potato plants do not differ from wild-type plants with regard to the starch formed in the tubers.

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Abstract

L'invention concerne des pommes de terre transgéniques qui contiennent des cellules à activité réduite de la phosphorylase cytosolique de l'amidon, comparativement aux plantes de type sauvage. Les tubercules de ce type de pommes de terre présentent, comparativement aux tubercules de plantes de type sauvage, un comportement en germination radicalement changé, qui entraîne la formation d'un nombre élevé d'extrémités de pousses et par conséquent un nombre élevé de stolons et de tubercules. Les plantes issues de ce type de tubercules permettent d'obtenir des rendements plus élevés.
EP97923093A 1996-05-17 1997-05-15 Plants de pomme de terre a activite reduite de la phosphorylase cytosolique de l'amidon et a comportement en germination modifie Withdrawn EP0906438A2 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
DE19619917A DE19619917A1 (de) 1996-05-17 1996-05-17 Kartoffelpflanzen mit einer verringerten Aktivität der cytosolischen Stärkephosphorylase und einem veränderten Keimungsverhalten
DE19619917 1996-05-17
PCT/EP1997/002513 WO1997044471A2 (fr) 1996-05-17 1997-05-15 Plants de pomme de terre a activite reduite de la phosphorylase cytosolique de l'amidon et a comportement en germination modifie

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EP0906438A2 true EP0906438A2 (fr) 1999-04-07

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EP (1) EP0906438A2 (fr)
AU (1) AU2899297A (fr)
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DE19709775A1 (de) 1997-03-10 1998-09-17 Planttec Biotechnologie Gmbh Nucleinsäuremoleküle codierend Stärkephosphorylase aus Mais
AU6221599A (en) * 1998-10-16 2000-05-08 Scottish Crop Research Institute Tissue-specific promoters for gene expression
FR2868080B1 (fr) * 2004-03-29 2007-11-16 Genoplante Valor Soc Par Actio Procede d'amelioration des plantes
CN101115840A (zh) * 2004-11-27 2008-01-30 梅坦诺米克斯有限公司 通过减少基因表达增加产量
WO2010046423A2 (fr) 2008-10-22 2010-04-29 Basf Se Utilisation d'herbicides sulfonylurées sur des plantes cultivées
WO2010046422A2 (fr) 2008-10-22 2010-04-29 Basf Se Utilisation d'herbicides de type auxine sur des plantes cultivées
MX2015004175A (es) 2012-10-01 2015-06-10 Basf Se Uso de compuestos de n-tio-antranilamida en plantas cultivadas.
WO2014079820A1 (fr) 2012-11-22 2014-05-30 Basf Se Utilisation de composés d'anthranilamides pour réduire les infections virales véhiculées par les insectes
EP3028573A1 (fr) 2014-12-05 2016-06-08 Basf Se Utilisation d'un triazole fongicide sur des plantes transgéniques
WO2016091674A1 (fr) 2014-12-12 2016-06-16 Basf Se Utilisation de cyclaniliprole sur des plantes cultivées
US11064696B2 (en) 2015-04-07 2021-07-20 Basf Agrochemical Products B.V. Use of an insecticidal carboxamide compound against pests on cultivated plants
EP3338552A1 (fr) 2016-12-21 2018-06-27 Basf Se Utilisation d'un fongicide tetrazolinone sur des plantes transgéniques
CN114085838B (zh) * 2021-12-02 2023-08-15 甘肃农业大学 马铃薯stu-miRn220及其应用

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GB9117159D0 (en) * 1991-08-08 1991-09-25 Cambridge Advanced Tech Modification of sucrose accumulation
DE4213444A1 (de) * 1992-04-18 1993-10-28 Inst Genbiologische Forschung Verfahren zur Herstellung von Kartoffelpflanzen, deren Knollensprossung unterdrückt ist
US6103893A (en) * 1994-03-25 2000-08-15 National Starch And Chemical Investment Holding Corporation High amylose starch from transgenic potato plants
DE4444460A1 (de) * 1994-11-29 1996-05-30 Inst Genbiologische Forschung Verfahren zur Steigerung des Ertrags sowie zur Veränderung des Blühverhaltens bei Pflanzen
US5716837A (en) * 1995-02-10 1998-02-10 Monsanto Company Expression of sucrose phosphorylase in plants

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WO1997044471A2 (fr) 1997-11-27
WO1997044471A3 (fr) 1997-12-24
DE19619917A1 (de) 1997-11-20

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