Classifications

• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
  • A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
  • A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
  • A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
  • A01N43/14—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings
  • A01N43/16—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings with oxygen as the ring hetero atom
• • A—HUMAN NECESSITIES
  • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
  • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
  • A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
  • A01N65/40—Liliopsida [monocotyledons]

Definitions

• oligoapiogalacturonans and its derivatives for the stimulation of the defense and resistance reactions of plants against biotic and abiotic stresses.
• the present invention relates to the use of oligoapiogalacturonan compounds as a stimulating agent for the defense and resistance reactions of a plant, in particular of an ornamental plant or of agronomic interest, against biotic and / or abiotic stresses.
• the invention relates to the use of oligoapiogalacturonans extracted from at least one aquatic plant of the genus Zostera or Lemna, preferentially from the species Zostera marina, Zostera noltii and Lemna minor, even more preferably from the species Zostera marina and Zostera noltii as an agent for stimulating the defense and resistance reactions of a plant against biotic and / or abiotic stress, for example resulting from a pathogen attack.
• Plants are, in their environment, continually subjected to stress, or constraints, caused by external factors likely to affect their metabolism and to induce defensive reactions.
• stress may be of biotic or abiotic origin.
• Stress is said to be of biotic origin when it comes from living organisms, for example, pathogenic microorganisms such as fungi, bacteria, viruses, but also nematodes, insects or parasitic plants.
• Stress is of abiotic origin when it results from the non-living, such as water stress, oxidative stress, chemical stress or heat stress. Biotic and / or abiotic stresses are the cause of significant economic losses for farmers.
• the defense reaction is most effective. Indeed, the interaction between the product of a resistance gene R carried by the plant and the product of a corresponding Avr avirulence gene (specific elicitor) carried by the pathogen, leads to the rapid activation of specific and effective defenses responses.
• the result of this interaction is manifested in particular by the rapid necrosis of the cells of the plant on the site of penetration of the pathogen, so-called hypersensitivity reaction, which aims to confine it on this site and stop its progression in the plant.
• the plant and / or the pathogen do not respectively carry the resistance gene R and the corresponding avirulence gene Avr.
• the stimulation of the plant's defense reactions may be caused by the effects of the lesions caused by the attack of the pathogen.
• the oligosaccharides produced on the one hand under the action of fungal enzymes on the cell walls of the host plant and, on the other hand, by the action of the hydrolases of the plant on the parietal polysaccharides of the pathogen can act as nonspecific or general eliciters and activate defense responses in response to pathogen attack.
• the defense reactions implemented are then late and / or weaker than the defense reactions during an incompatible plant-pathogenic interaction and are nonspecific. Symptoms and yield losses remain moderate in a tolerant plant, while infestation will be severe or even fatal in a plant susceptible to this attack.
• the most commonly induced or stimulated defense responses of the plant during a pathogenic attack are the programmed cell death of the cells at the site of pathogen penetration (hypersensitive response), a strengthening of the cell walls of the plant due to production.
• compounds such as lignin, callose or crosslinking proteins, the accumulation of antimicrobial compounds such as phytoalexins, the synthesis of PR (pathogenesis related) proteins such as chitinases, glucanases, peroxidases or inhibitors of enzymes acting against the hydrolytic enzymes of the pathogen.
• RSA distributed systemic resistance
• a possible strategy is to induce defense reactions of the plant before it is attacked by a pathogen, by the plant. exogenous application of eliciting compounds.
• eliciting compounds are preferentially sought for their natural origin, this in particular to limit the use of pollutants.
• molecules are known that can stimulate plant defense mechanisms in response to stress and thus increase the level of plant resistance.
• These molecules can be of various chemical nature, for example proteins, peptides, glycoproteins, lipids or oligosaccharides and can be of various origin, for example of bacterial origin, such as harpin, of plant origin, such as oligogalacturonic acid, of algal origin, such as laminarine, or of synthetic origin, such as nicotonic acid derivatives.
• bacterial origin such as harpin
• plant origin such as oligogalacturonic acid
• algal origin such as laminarine
• synthetic origin such as nicotonic acid derivatives.
• oligosaccharides such as laminarine, ulvans, fucans, carrageenans, are known for their ability to stimulate certain defenses in the plants to which they are applied.
• the identified compounds can be used as a preventive treatment to stimulate or potentiate defense mechanisms, for example according to the concentration used, or in curative treatment.
• the compounds identified by the applicant company also have the advantage of being slowly degraded in the terrestrial environment, and in particular in the soil, thus making it possible to apply lesser amounts to the plants compared to to the compounds cited. They also have the advantage of being particularly suitable for application on the root system, for example, via nutrient solutions during an aboveground culture.
• the present invention thus relates to the use of at least one oligoapiogalacturonan composed of 3 to 300 monosaccharide units, in particular extracted from aquatic plants of the genus Lemna or Zostera, as an agent for stimulating the defense and resistance reactions of a plant, or part of a plant, in particular of an ornamental plant or of agronomic interest, against biotic and / or abiotic stresses.
• Oligoapiogalacturonan allows the activation of intracellular signaling pathways leading to the stimulation of defense and resistance genes and the synthesis of compounds able to fight against the stress generated by the plant.
• Eelgrass are marine phanerogams that develop in sandy and sandy-sandy sediments in the intertidal and subtidal zones in which they form sometimes dense underwater meadows.
• the eelgrass are organized according to a more or less subterranean creeping stem called rhizome, on which bands of ribbon-shaped leaves are inserted.
• a biochemical characteristic of aquatic plants of the genus Zostera is that their cell walls contain particular polysaccharides, pectic substances, or pectins, called apiogalacturonans. The developed chemical formula of such a compound is shown. Apiogalacturonans of plants of the genus Zostera are known as zosterines or zosteranes.
• Lemna aquatic plants, or duckweeds also contain apiogalacturonans in their cell walls.
• Apiogalacturonans of plants of the genus Lemna are known as lemnans.
• pectic substances are macromolecules of a carbohydrate nature composed essentially of galacturonic acid.
• the skeleton of these molecules is formed by the linear sequence of an identical monomeric unit, ⁇ -D-galacturonic acid.
• the motifs are linked together by ⁇ [1-4] bonds.
• the particularity of these pectins lies in the substitution of certain groups of the main chain of ⁇ -D-galacturonic acids by monomeric or dimeric units of D-apiosis.
• Apiose (3-C- (hydroxymethyl) -D-glycero-tetrose), a rare sugar, is a branched chain pentose.
• the main saccharide chain is also substituted by galactose, rhamnose, xylose and arabinose.
• oligoapiogalacturonans increased resistance to pectinolytic enzymes secreted by microorganisms and thus a prolonged life in the terrestrial environment.
• oligoapiogalacturonan compounds thus durably protected from degradations by their apiose motifs, makes it possible to optimize their effectiveness of stimulating the defense and resistance reactions of a plant against biotic and / or abiotic stress.
• the presence of the apiose motifs thus confers on the oligoapiogalacturonans a biological activity enhanced and adapted to an application on plants in order to stimulate its defense and resistance reactions.
• the increased resistance of these compounds to biodegradation makes it possible to limit the quantities of products to be applied and thereby optimize the efficiency of the treatment of the plants.
• Such compounds are particularly suitable for application to the root system of the target plants.
• said one or more oligoapiogalacturonans are obtained from at least one aquatic plant of the genus Zostera or Lemna, preferentially from the species Zostera marina, Zostera noltii and Lemna minor, even more preferably from species Zostera marina and Zostera noltii.
• the compounds according to the invention can be obtained by an extraction process comprising the following steps: a step of washing the leaves and / or rhizomes of the plant, a grinding step, an aqueous extraction step of solid-type, liquid, which can be followed by a fractionation step and concentration of the extract obtained.
• said oligoapiogalacturonans are obtained by enzymatic and / or chemical hydrolysis of an apiogalacturonan compound.
• said oligoapiogalacturonans are composed of 3 to 100 and even more preferentially of 3 to 30 monosaccharide units.
• said oligapiogalacturonans are obtained by grafting apiose units on oligogalacturonan compounds.
• each oligoapiogalacturonans comprises at least 5% of apiose units, preferably between 10 and 80%, more preferably between 20 and 70% of apiose units.
• said oligoapiogalacturonans or said oligoapiogalacturonans are supplied to said plant, or part of a plant, in an amount of between 0.01 and 100 g / l in a liquid form via the foliar system, the root system, the seed of said plant or the environment of said plant, in particular in a nutrient solution for a hydroponic-type culture, or in an amount of between 0.01 and 100 g / kg in a solid form in the environment of said plant , before or after planting or harvesting said plant.
• the supply to the plants can thus be achieved by means of a solid composition, such as a stick or a granule that can, for example, be placed in the soil in which the plant is planted.
• the feed can also be carried out by spraying on the plant or on part of the plant, or else by watering the foot of the plant with a solution comprising the compounds of interest.
• the contribution can also be made on the plant after harvest.
• the feed can be carried out via a nutrient solution provided for this purpose.
• the term "plant” designates any plant, in particular any plant of the gymnosperm or monocotyledonous or dicotyledonous angiosperm type, in particular any plant of ornamental or agronomic interest, whatever its stage of development.
• plant part is meant any fragment of a plant containing at least one plant cell, for example a plant organ, such as a leaf, a bud, a flower, a root, a fruit, or a seed , a plant tissue, such as a meristem, a plant callus or a plant cell.
• oligoapiogalacturonans resides in the stimulation of the defense reactions against pathogens.
• the invention also relates to a method for stimulating reactions of the natural defenses and resistance of a plant, or part of a plant, against biotic and / or abiotic stress, which is characterized in that it comprises the application on said plant, or said part of a plant or in the plant environment of at least one oligoapiogalacturonans, having from 3 to 300 monosaccharide units, in liquid form or in solid form.
• the amount of oligoapiogalacturonans is between 0.01 and 100 g / L in a liquid composition and between 0.01 and 100 g / kg in a solid composition.
• the invention also relates to a product for stimulating reactions of the natural defenses and resistance of a plant, or part of a plant, against biotic and / or abiotic stress, which is characterized in that it is composed of: minus one oligoapiogalacturonans having from 3 to 300 monosaccharide units.
• said one or more oligoapiogalacturonans are extracted from at least one aquatic plant of the genus Zostera and / or Lemna, preferentially from the species Zostera marina, Zostera noltii, Lemna minor, even more preferably from the species Zostera marina and Zostera noltii.
• such a product further comprises p-sulfoxy cinnamic acid, or zosteric acid.
• the product according to the invention may be in the form of a solution, an emulsion, a powder, a granule or a coating solution, in particular for coating seeds.
• the product according to the invention comprises between 0.01 and 100 g / l of at least one oligoapiogalacturonan, when said product is in a liquid form or between 0.01 and 100 g / kg of at least one compound.
• apiogalacturonan and / or derivatives of apiogalacturonan compound when said product is in a solid form.
• the product according to the invention further comprises at least one fertilizing material.
• the product according to the invention further comprises at least one phytosanitary product and, preferably, at least one elicitor product.
• Fertilizers that can be used in a product according to the invention may be chosen from urea, ammonium sulphate, ammonium nitrate, natural phosphate, potassium chloride, ammonium sulphate, magnesium nitrate, manganese nitrate, zinc nitrate, copper nitrate, phosphoric acid, boric acid, NP, PK, NPK type fertilizers.
• Eelgrass leaching In a 250-mL flask, previously cleaned eelgrass (8 g) is added to 72 mL of distilled water. The pH is adjusted to 4 by the addition of a 3N hydrochloric acid solution. The medium is stirred for 3 minutes. hours at a temperature of 50 ° C. After returning to ambient temperature, the reaction medium is filtered on sintered glass (porosity of 15 to 50 ⁇ m). The filtrate is then dialyzed overnight using a cellulose membrane to remove chloride anions for analysis. The retentate is directly used in the next step.
• the medium is then transferred into a 500 ml beaker in which the zosterine is precipitated by adding 200 ml (3 volumes) of ethanol.
• Zosterine is recovered after centrifugation (20 min, 15 ° C, 3000 g).
• Zosterine is then solubilized in water, dialyzed on a 10 KD membrane and then lyophilized. 1.2 g of zosterine are thus isolated, which corresponds to an extraction yield of 15% with respect to the eelgrass extracted.
• This method makes it possible to obtain zosterine called LJl. It is brown because it contains 2.7% polyphenols.
• the zosterine LJ1 can then be purified by the use of a hydrophobic resin, such as the hydrophobic resin Amberlite TM XAD TM 16 (marketed by Rohm and Haas) with a yield of 63%. This produces zosterine LJ 2 white, with a monosaccharide content of 31%. This zosterine contains only 0.5% polyphenols.
• a hydrophobic resin such as the hydrophobic resin Amberlite TM XAD TM 16 (marketed by Rohm and Haas) with a yield of 63%.
• This zosterine contains only 0.5% polyphenols.
• zosterine 400 mg is solubilized in 74 mL of water at 50 ° C. Once solubilized, Amberlite TM XAD TM 16 resin (30 g) is added in the middle. This is then stirred and maintained at a temperature of 50 ° C. overnight. The resin is removed by filtration on sintered glass under atmospheric pressure. The filtrate is concentrated until the beginning of the precipitation. Zosterine precipitates by adding three volumes of ethanol. The zosterine is then filtered on Buchner, dialyzed on a membrane of 10 KD, and freeze-dried.
• the neutral ose composition of the two zosterines LJ1 and LJ2 is given in FIG. 1 illustrating the neutral ose composition of zosterines LJl and LJ2 extracted from Zostera noltii expressed as a percentage.
• Rha Rhamnose; Fuc: fucose; Ara: arabinose; ; Api: apiose; Xyl: xylose; Man: manose; GaI: galactose; GIc: glucose;
• the zosterines LJ1 and LJ2 are hydolyzed enzymatically so as to obtain oligoapiogalacturonans comprising between 3 and 300 saccharide units, respectively LJ '1 and LJ'2.
• apiogalacturonan extracted according to the preceding example are solubilized in 4 g of demineralized water.
• the pH is adjusted to 5 with sodium acetate / acetic acid buffer.
• the reaction mass is then boiled to inactivate the enzyme. Once denatured, the enzyme precipitates and is separated by centrifugation. The centrifugation supernatant is collected and desalted by dialysis on a 1000 Da cut-off membrane.
• the dialysis retentate is frozen and
• the average degree of polymerization by number of Poligoapiogalacturonan obtained is 27.
• the extract used is purified on resin and corresponds to
• LJ'l The use of LJ'l is not excluded.
• Liquid compositions were prepared by mixing an extract obtained by a process as described above and zosteric acid.
• the liquid compositions comprise between 0.2 and 2 g / l of apiogalacturonan compounds and 10 g / l of zosteric acid. Such solutions can be sprayed directly onto the leaves of plants.
• liquid compositions were prepared by adding an extract obtained by a process as described above to a NPK fertilizer solution.
• Solutions include 0.2 and 2 g / L of apiogalacturonan compounds. Such solutions can be made to the root systems of plants by watering.
• Plant seed coating compositions were prepared by incorporating 2 g / L of an extract obtained by a process as described above into a coating composition known to those skilled in the art, for example a composition of coating based on hemicellulose. Plant seeds can thus be coated with such a composition.

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• Engineering & Computer Science (AREA)
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• Agronomy & Crop Science (AREA)
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• Fertilizers (AREA)

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• 2008
  • 2008-08-12 FR FR0855529A patent/FR2934943B1/fr not_active Expired - Fee Related
• 2009
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  • 2009-08-11 EP EP09777803A patent/EP2317848A2/de not_active Withdrawn
  • 2009-08-11 US US13/058,431 patent/US20110172102A1/en not_active Abandoned
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