EP4153318A1 - Use of polyamines, tyramine and/or a plant extract containing same to stabilise microorganisms - Google Patents

Use of polyamines, tyramine and/or a plant extract containing same to stabilise microorganisms

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Publication number
EP4153318A1
EP4153318A1 EP21732467.2A EP21732467A EP4153318A1 EP 4153318 A1 EP4153318 A1 EP 4153318A1 EP 21732467 A EP21732467 A EP 21732467A EP 4153318 A1 EP4153318 A1 EP 4153318A1
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EP
European Patent Office
Prior art keywords
bacteria
composition
extract
microorganisms
spp
Prior art date
Legal status (The legal status 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 status listed.)
Pending
Application number
EP21732467.2A
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German (de)
French (fr)
Inventor
Xavier Briand
Fabrice Houdusse
Morgane COMBY
Matthieu C BEZIAT
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Agro Innovation International SAS
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Agro Innovation International SAS
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Application filed by Agro Innovation International SAS filed Critical Agro Innovation International SAS
Publication of EP4153318A1 publication Critical patent/EP4153318A1/en
Pending legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F11/00—Other organic fertilisers
    • C05F11/02—Other organic fertilisers from peat, brown coal, and similar vegetable deposits
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05C—NITROGENOUS FERTILISERS
    • C05C11/00—Other nitrogenous fertilisers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F11/00—Other organic fertilisers
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F5/00—Fertilisers from distillery wastes, molasses, vinasses, sugar plant or similar wastes or residues, e.g. from waste originating from industrial processing of raw material of agricultural origin or derived products thereof
    • C05F5/004—Liquid waste from mechanical processing of material, e.g. wash-water, milling fluid, filtrate

Definitions

  • the invention relates to the use of an extract of algae and / or plants, said extract containing polyamines and / or tyramine, for stabilizing microorganisms in the face of environmental stress.
  • microorganisms contained in the probiotics must remain functional up to their site of action.
  • these microorganisms are fragile because they are very sensitive to environmental stresses, for example variations in pH.
  • silage namely the conservation of fodder by lactic fermentation
  • the microorganisms are also subjected to a certain number of environmental stresses such as oxidative stress, osmotic or saline stress, thermal stress, acid stress. - basic and / or stress linked to competing microorganisms.
  • the microorganisms used in animal feed are therefore exposed to the same types of stress as those encountered in agriculture, which therefore prevents their growth, their stability and / or their effects.
  • the present invention which finds application in the agro-ecological and agricultural field, aims to provide the use of polyamines, tyramine and / or a plant extract containing it to stabilize microorganisms in the face of environmental stress, such as oxidative stress, osmotic stress or salt stress, heat stress, acid-base stress and / or stress linked to competing microorganisms.
  • environmental stress such as oxidative stress, osmotic stress or salt stress, heat stress, acid-base stress and / or stress linked to competing microorganisms.
  • the invention relates to the use of an extract of algae and / or plants, said extract containing polyamines and / or tyramine, for stabilizing microorganisms in the face of stress. environmental.
  • the invention relates to a composition
  • a composition comprising:
  • microorganisms chosen from: (i) bacteria fixing atmospheric nitrogen, such as Azotobacter or Azospirillum (ii) rhizobacteria promoting plant growth (PGPR or Plant Growth Promoting Rhizobacteria), (iii) phosphorus solubilizing bacteria such as BaciHus amyloliquefaciens, (iv) root protection bacteria (PGPR) capable of opposing the activity of pathogens such as BaciHus subti / is or Pseudomonas spp., (v) bacteria producing phytohormones such as BaciHus amyloliquefaciens or BaciHus radicoia, (vi) bacteria involved in the process of mineralization of organic matter such as LactobaciHus rhamnosus or LactobaciHus faciminis, (vii) iron solubilizing bacteria such as Pseudomonas spp., (viii) silica solubilizing bacteria, (ix) sulfur oxidizing bacteria, (
  • the invention relates to a method for fertilizing a plant characterized in that it comprises supplying said plant with a composition according to the invention.
  • Tyramine denotes a chemical compound of monoamine type, the CAS number of which is 51-67-2. Tyramine has the following formula:
  • polyamines denotes organic compounds containing at least two amine functions. Polyamines are found in almost all plants, animals and microorganisms. Polyamines are involved in a variety of cellular processes such as the expression of certain genes, cell growth, cell survival and cell proliferation.
  • the polyamines can be chosen from cadaverine, spermidine, spermine and / or putrescine.
  • the plant extract used in the context of the invention can therefore comprise one, two, three or all four polyamines chosen from cadaverine, spermidine, spermine and putrescine.
  • Cadaverine or pentamethylenediamine
  • the CAS number of which is 462-94-2 has the following formula:
  • Putrescine (or tetramethylene diamine), the CAS number of which is 110-60-1, has the following formula:
  • plant extract denotes the product resulting from an extraction of the contents of the cells of a plant.
  • the plant extract contains an effective amount of polyamines and / or tyramine.
  • the plant extract can be an extract of algae and / or plants.
  • the term "extract of algae and / or plants” denotes the product resulting from an extraction of the contents of the cells of algae and / or plants respectively.
  • the algae extract can be an extract of brown algae, such as an extract of brown algae of the Fucaceae family.
  • the extract of brown algae can be an extract of Fucus vesicu / osus or Ascophyllum noc / osum.
  • the plant extract can be an extract of beet molasses vinasse.
  • the preparation of a plant extract containing an effective amount of polyamines and / or tyramine does not present any particular difficulty, many extraction processes are described in the literature.
  • the extraction process is not limited to any particular process, and conventional processes can be used to prepare such an extract.
  • it can be obtained by a process comprising the following steps: washing, grinding, extraction with a solvent (eg water), solid / liquid separation and optionally fractionation and / or concentration.
  • a solvent eg water
  • the plant extract obtained can be more or less concentrated depending on the use envisaged, for example it is possible to concentrate the extract by an evaporation technique. Total dehydration of this extract allowing presentation in water-soluble powder form can be obtained, for example, by tumble dryer or by atomization.
  • the extraction conditions and the nature of the plant will be chosen such that the extract obtained exhibits the quantity of polyamines and / or tyramine desired in the envisaged application.
  • the extract when it is an extract of beet molasses vinasse, it can be obtained by a process comprising the following steps: washing of the beets, grinding of the beets, extraction of the crystallizable sugar from the beet, obtaining two by-products: molasses and beet pulp, recovery of molasses, fermentation of molasses to obtain molasses stillage and concentration [2] [3]. It is also possible to enrich the plant extract with polyamines and / or tyramine, for example by an ultrafiltration technique.
  • the term "effective amount of polyamines and / or tyramine” denotes the amount of polyamines and / or tyramine sufficient to stabilize the microorganisms in the face of environmental stress.
  • the plant extract preferably contains at least 0.01% by mass of polyamines and / or tyramine relative to the total dry mass of the extract (% w / w), for example at least 0.05% w / w, at least 0.5% w / w, at least 0.2% w / w, for example between 0.01% w / w and 1% w / w, between 0.05% w / w and 0, 5% w / w.
  • the assay of the amount of polyamines and tyramines in the extract does not present any particular difficulty, assay methods being described in the literature, such as for example the HPLC method [1].
  • the plant extract can be more or less concentrated in polyamines and / or in tyramine depending on the use envisaged.
  • microorganisms designates microscopic organisms such as bacteria, microscopic fungi, for example filamentous microscopic fungi, yeasts.
  • the microorganisms can be chosen from (i) bacteria fixing atmospheric nitrogen, such as Azotobacter or Azospirillum (ii) rhizobacteria promoting plant growth (PGPR or Plant Growth
  • Rhizobacteria Promoting Rhizobacteria
  • phosphorus solubilizing bacteria such as BaciHus amyloliquefaciens
  • plant protection bacteria PGPR
  • bacteria producing phytohormones such as BaciHus amyloliquefaciens or BaciHus radicoia
  • bacteria involved in the mineralization process of organic matter such as LactobaciHus rhamnosus or LactobaciHus faciminis
  • iron solubilizing bacteria such as Pseudomonas spp.
  • silica solubilizing bacteria sulfur oxidizing bacteria
  • lactic acid bacteria such as LactobaciHus spp., Lactococcus spp.
  • lactic acid bacteria such as LactobaciHus spp., Lactococcus spp.
  • Bifidobacterium spp. (Xi) bacteria of the genus Enterococcus spp., (Xii) bacteria of the genus Pediococcus spp., (Xiii) bacteria of the genus BaciHus Hcheniformis, (xiv) mycorrhizal fungi such as Rhizophagus irregularis, (xv) yeasts of the genus Saccharomyces cerevisiae and (xvi) a mixture of at least two microorganisms chosen from (i) to (xv).
  • composition denotes a mixture of one or more substances distinct from one another.
  • a composition may be a fertilizing composition, a composition intended for animal feed or a composition intended for the preservation of silage.
  • a “fertilizing composition” denotes a composition comprising a fertilizing substance, or a mixture of fertilizing substances, natural or of synthetic origin, used in agriculture, horticulture and forestry.
  • fertilizer means a fertilizer and / or an amendment.
  • fertilizer designates fertilizing materials whose main function is to provide the plants with elements directly useful for their nutrition (major fertilizing elements, secondary fertilizing elements and trace elements).
  • amendment denotes a substance intended to improve the quality of soils, and in particular intended to improve the pH of the soils.
  • the amendment is chosen from basic mineral amendments of limestone and / or limestone and magnesium type; humus amendments such as composts or manure.
  • composition intended for animal feed can comprise substances intended to improve the performance and / or the health of the animals.
  • a composition intended for animal feed can, for example, comprise probiotic foods, mineral-vitamin supplements, etc.
  • composition intended for the preservation of silage can comprise substances intended to improve the preservation and the quality of the silages.
  • a composition intended for silage preservation can, for example, include lactose, xylanase or beta-glucanase enzymes, etc.
  • plant is intended in the present application to denote the plant considered as a whole, including its root system, its vegetative system, seeds, seeds and fruits.
  • the present invention results from the surprising advantages demonstrated by the inventors of the effect of polyamines, of tyramine and / or of a plant extract containing it to stabilize microorganisms.
  • the present application relates to the use of polyamines, tyramine and / or a plant extract containing it to stabilize microorganisms in the face of environmental stress, such as oxidative stress, stress. osmotic or saline stress, thermal stress, acid-base stress and / or stress linked to competing microorganisms.
  • the first object of the invention relates to the use of an extract of algae and / or plants, said extract containing polyamines and / or tyramine, for stabilizing microorganisms in the face of environmental stress. , such as oxidative stress, osmotic stress or salt stress, heat stress, acid-base stress and / or stress linked to competing microorganisms.
  • the plant extract contains an effective amount of polyamines and / or tyramine.
  • the plant extract is an extract of brown algae, for example an extract of brown algae of the Fucaceae family, and / or an extract of beet molasses vinasse. It is understood that the plant used for the preparation of the plant extract naturally comprises polyamines and / or tyramine. Consequently, the polyamines and / or the tyramine contained in the extract of algae and / or plants come directly from the algae and / or plants used to prepare said extract.
  • Oxidative stress corresponds to an attack on microorganisms by free radicals, also called "reactive oxygen species" (ROS).
  • ROS reactive oxygen species
  • An acid-base stress corresponds to a stress linked to the pH. Indeed, it is known that microorganisms are more or less stable at certain pHs, for example a pH ranging from 1 to 12, for example ranging from 3 to 9, for example a pH 5 or for example a pH 9.
  • Osmotic stress corresponds to stress linked to osmolarity, ie to the salt content in the medium. In fact, it is known that microorganisms are more or less stable at certain osmolarities, for example at a salt concentration greater than 0.3 M, microorganisms are generally less stable.
  • Water stress corresponds to stress linked to the absence of water, ie to the water content in the medium.
  • Thermal stress corresponds to stress induced by cold or by heat. Depending on the microorganism considered, the temperature inducing thermal stress varies. Stress linked to competing microorganisms corresponds to stress linked to the presence of microorganisms other than microorganisms stabilized with the plant extract. It may be, for example, micro ⁇ organisms naturally present in the soil that will compete for space in the ground and for access to nutrients with microorganisms added to the soil.
  • the competing microorganisms are bacteria of the genus Agrobacterium spp., Of the genus Erwinia spp., Or of the genus Xanthomonas spp., Or fungi of the genus Fusarium spp.
  • stabilization consists in increasing the survival rate of microorganisms, in particular measured after 24 hours, 48 hours or 72 hours of growth, under conditions of environmental stress compared to the same conditions without polyamines, tyramine and / or plant extract containing it.
  • the survival rate of microorganisms with polyamines, tyramine and / or the plant extract containing them, and subjected to environmental stress is advantageously at least 1 log, at least 2 log, at least 3 log, at least 4 log, at least 5 log greater than the survival rate of the same microorganisms subjected to the same environmental stress but without polyamines, tyramine and / or plant extract containing it.
  • the survival rate is measured by comparing the quantity of bacteria (for example in CFU) after a given time, for example 24 hours, 48 hours or 72 hours.
  • the microorganisms are contained in a composition, such as a fertilizing composition, a composition intended for animal feed or a composition intended for the preservation of silage.
  • the microorganisms may be contained in said composition in an amount ranging from 10 2 to 10 50 CFU per tonne of composition, preferably ranging from 10 5 to 10 20 CFU per tonne of composition, preferably approximately 10 11 CFU per tonne of composition.
  • Polyamines, tyramine and / or the plant extract containing it can therefore be used as a complement in a composition, for example a fertilizing composition, a composition intended for animal feed or a composition intended for the preservation of silage, to stabilize the microorganisms contained in these compositions.
  • the present application relates to a composition
  • a composition comprising:
  • microorganisms chosen from: (i) bacteria fixing atmospheric nitrogen, such as Azotobacter or Azospirillum (ii) rhizobacteria promoting plant growth (PGPR or Plant Growth Promoting Rhizobacteria), (iii) phosphorus solubilizing bacteria such as BaciHus amyloliquefaciens, (iv) plant protection bacteria (PGPR) capable of opposing the activity of pathogens such as BaciHus subti / is or Pseudomonas spp., (v) producing bacteria phytohormones such as BaciHus amyloliquefaciens or BaciHus radicoia, (vi) bacteria involved in the mineralization process of organic matter such as LactobaciHus rhamnosus or LactobaciHus faciminis, (vii) iron solubilizing bacteria such as Pseudomonas spp., (viii) silica solubilizing bacteria, (ix) sulfur oxidizing bacteria,
  • a second object of the invention relates to a composition
  • a composition comprising: (a) an extract of algae and / or plants, said extract containing polyamines and / or tyramine, and (b) selected microorganisms among: (i) atmospheric nitrogen fixing bacteria, such as Azotobacter or Azospirillum (ii) rhizobacteria promoting plant growth (PGPR or Plant Growth Promoting Rhizobacteria), (iii) phosphorus solubilizing bacteria such as BaciHus amyloliquefaciens , (iv) plant protection bacteria of the roots (PGPR) capable of opposing the activity of pathogenic agents such as BaciHus subti / is or Pseudomonas spp., (v) bacteria producing phytohormones such as BaciHus amyloliquefaciens or BaciHus radicoia, (vi) bacteria involved in the mineralization process of organic matter such as LactobaciHus rhamnosus or
  • yeasts of the genus Saccharomyces cerevisiae and (xvi) a mixture of at least two microorganisms chosen from (i) to (xv).
  • (a) is a plant extract containing an effective amount of polyamines and / or tyramine.
  • the plant extract is an extract of algae and / or plants, preferably an extract of brown algae, for example an extract of brown algae of the Fucaceae family, and / or an extract of beet molasses stillage.
  • the plant used for the preparation of (a) the plant extract naturally comprises polyamines and / or tyramine. Consequently, the polyamines and / or the tyramine contained in (a) the extract of algae and / or plants come directly from the algae and / or plants used to prepare said extract.
  • the composition according to the invention may in particular comprise from 10 2 to 10 50 CFU of microorganisms (b) per tonne of composition, preferably from 10 5 to 10 20 CFU of microorganisms (b) per tonne of composition, preferably 10 11 CFU of microorganisms (b) per tonne of the composition.
  • the composition may in particular comprise from 0.1 to 100 grams of polyamines and / or tyramine per tonne of composition, preferably from 0.4 to 10 grams of polyamines and / or tyramine per tonne of composition.
  • the composition comprises: - from 0.1 to 100 grams of polyamines and / or tyramine per tonne of composition, preferably from 0.4 to 10 grams of polyamines and / or tyramine per tonne of composition, and
  • composition according to the invention can also comprise (c) a fertilizer and / or an amendment.
  • the fertilizer can be one or more active substances chosen from nitrogen, phosphorus, potassium, urea, ammonium sulfate, ammonium nitrate, phosphate, potassium chloride, ammonium sulfate, magnesium nitrate, manganese nitrate, zinc nitrate, copper nitrate, phosphoric acid, potassium nitrate, boric acid and their mixtures, preferably a mixture of nitrogen , potassium and phosphorus or a mixture of phosphorus and potassium.
  • the amendment can be one or several active substances chosen from basic mineral amendments of the limestone type, basic mineral amendments of the magnesium type, humus amendments of the composts type and / or humus amendments of the manure type, preferably fertilizers and mineral amendments.
  • Polyamines, tyramine and / or the plant extract containing them preferably the plant extract (a) containing an effective amount of polyamines and / or tyramine, for example the algae extract and / or of plants (a), preferably the extract of algae and / or plants (a) containing an effective amount of polyamines and / or tyramine, stabilize the microorganisms (b) in the face of environmental stress, such as oxidative stress, osmotic stress or salt stress, heat stress, acid-base stress and / or stress linked to competing microorganisms.
  • environmental stress such as oxidative stress, osmotic stress or salt stress, heat stress, acid-base stress and / or stress linked to competing microorganisms.
  • the microorganisms (b) can be sensitive and / or unstable in the presence of a fertilizer, at a pH ranging from 1 to 12, preferably at a pH ranging from 3 to 9, for example at a pH of '' about 5 or at a pH of about 9 and / or at a salinity going beyond the threshold of sensitivity / stability to the salt of a microorganism in question, for example at a salt concentration greater than 0.3 M for bacteria of the genus Azobacter chroococcum.
  • the composition further comprises a fertilizer and has a pH ranging from 1 to 12, preferably a pH ranging from 3 to 9, for example a pH of about 5 or a pH of approximately 9 and / or a salinity going beyond the threshold of sensitivity / stability to the salt of a microorganism in question, for example at a salt concentration greater than 0.3 M for bacteria of the genus Azobacter chroococcum.
  • the composition according to the invention can be in solid or liquid form, preferably in solid form.
  • composition according to the invention can also comprise other substances, such as biostimulant substances, phytohormones, polyphenols, amino acids, etc.
  • the composition according to the invention may be a fertilizing composition, a composition intended for animal feed or a composition intended for the preservation of silage. The other substances will therefore be chosen according to the intended use.
  • compositions comprising limestone amendments, organic amendments and growing media, root fertilizers of NP, PK type,
  • NPK foliar fertilizers and / or biostimulants or root nutrient solutions.
  • compositions intended for animal feed according to the invention mention will be made of compositions comprising, for example, sodium bicarbonate, magnesium oxide, nutritional yeast.
  • compositions intended for the preservation of silage according to the invention, mention will be made of compositions comprising, for example, lactose, enzymes of xylanase, beta-glucanase, etc. type.
  • a third object of the invention relates to a method for fertilizing a plant characterized in that it comprises supplying said plant with a composition according to the invention.
  • the method according to the invention makes it possible to stimulate the development of a plant and / or to stimulate the yield of a plant.
  • the composition can be added to said plant in an amount ranging from 0.5 to 1500 kg / ha, preferably ranging from 1 to 1000 kg / ha kg / ha.
  • the composition can be provided in solid or liquid form. According to the process of the invention, the composition can advantageously be a fertilizing composition, such as that described above.
  • the application to said plant will be carried out by foliar route or by root route, preferably by root route.
  • the present invention finds application in the treatment of a very wide variety of plants.
  • - field crops such as cereals (wheat, corn, sugar cane, etc.), - protein crops (peas),
  • the plant belongs to the order of monocotyledons, preferably to the Poaceae family, preferably the plant is chosen from wheat, rice, barley, l oats, rye, meadow, sugar cane or corn.
  • Example 1 Preparation of an extract of alaues and / or plants
  • Example 1 a) Preparation of an extract of brown algae of the Fucaceae family of the genus Ascophyllum nodosum 200 g of fresh algae of the genus Ascophyllum nodosum were ground to obtain fragments of fresh algae having a size of about 5 mm.
  • the aqueous extraction of fresh algae was carried out using 200 g of Ascophyllum nodosum per liter of water.
  • the extraction was carried out in a 0.5N solution of sulfuric acid with stirring which lasts 2 hours at room temperature.
  • the extract was then filtered through a membrane (80 ⁇ m porosity) and then concentrated on a falling flow evaporator.
  • the analysis of one kilo of extract was carried out by High Performance Liquid Chromatography.
  • the analytical method used for the identification and Quantification of polyamines is based on the method described by HB Papenfus (2011).
  • Table 1 One kilogram of dry extract (Table 1) contains 3300 mg of polyamines in total, ie 480 mg of putrescine, 1470 mg of cadaverine, 840 mg of spermidine and 510 mg of spermine.
  • the aqueous extraction of fresh algae was carried out using 200 g of Fucus vesicu / osus per liter of water.
  • the extraction was carried out in a 1N solution of sulfuric acid with stirring which lasts 2 hours at room temperature.
  • the extract was then filtered through a membrane (80 ⁇ m porosity) and then concentrated on a falling flow evaporator.
  • the analysis of one kilo of extract was carried out by High Performance Liquid Chromatography.
  • the analytical method used for the identification and quantification of polyamines is based on the method described by H. B. Papenfus (2011).
  • Table 2 One kilo of dry extract (Table 2) contains 7170 mg of polyamines in total: 1140 mg of putrescine, 150 mg of cadaverine, 1830 mg of spermidine and 4050 mg of spermine. Table 2
  • An extract of beet molasses vinasse suitable for the implementation of the invention is available under the following trade names: Proteinal, Sirional, Betainex, Viprotal, Vinasse FS, Citrocol and Prodyn [3].
  • One kilogram of dry extract (Table 3) contains 477 mg of polyamines in total: 344 mg of tyramine, 73 mg of putrescine, 48 mg of cadaverine and 12 mg of spermidine.
  • the starting inoculum for each of the strains was calibrated by measuring the OD (620 nm) in order to reach an initial concentration around 10 3 -10 4 CFU / mL.
  • the strains were incubated for 24 to 72 h.
  • Counts were carried out in TSA (Tryptone Soya Agar) agar at T0 and after 24 h, 48 h or 72 h of incubation at 30 ° C (+/- 1 ° C), depending on the strains studied.
  • survival rates in the presence or absence of the extract and / or stress were calculated relative to the bacterial concentration obtained at the end of kinetics in the modality containing the strain alone under stress conditions without addition of the extract.
  • Example 2 b Effect of the extract of Fucus vesicu / osus prepared according to example 1 a] on the growth of Bacillus Hcheniformis under acid stress condition
  • the addition of 0.4 M NaCl in the culture medium made it possible to reach a limiting level of salt stress.
  • the addition of the Ascophyllum nodosum extract made it possible to increase the bacterial concentration by 1.5 logs after 72 h of incubation.
  • the bacterial concentration reached is 2.6 logs higher than that of the strain control alone but remains lower than that of the salt-free condition with the addition of the Ascophyllum nodosum extract.
  • Example 2 d Effect of the extract ô'Ascophyllum nodosum prepared according to example la] on the growth of ô'Azospirillum brasilense under heat stress conditions
  • the Azospirillum brasilense strain was tested under heat stress conditions, that is to say by increasing the temperature of the culture medium from
  • the incubation of A brasiiense 11 h at 45 ° C made it possible to reproduce the conditions of thermal stress with a loss of 5.9 logs of the bacterial concentration after 72 h of incubation compared to the condition at 30 ° C. Under these conditions, the addition of the Ascophyllum nodosum extract allowed an increase of 5.5 logs in the bacterial concentration after 72 h of incubation.
  • the bacterial concentration reached is globally similar to that of the control at 30 ° C, around 5.10 6 CFU / mL.
  • Example 2 fi Effect of the extract of beet molasses stillage prepared according to example 1c on the growth of AzosDirillum brasilense under conditions of basic stress
  • Example 2 cO Effect of the extract of beet molasses vinasse prepared according to example 1c on the growth of a complex of nitrogen-fixing rhizobacteria (Azotobacter diazotrophicus. Paenibacillus azotofixans.
  • Glycine betaine known as an osmoprotector, was tested in comparison with the extract ô'Ascophyllum nodosum prepared according to example la).
  • Polyamines make it possible to stabilize Azotobacter chroococcum in the face of osmotic stress, whereas no stabilization is observed with glycine betaine under conditions of osmotic stress.
  • Example 3 Effect of a biofertilizer composition based on an extract of Ascoohyllum nodosum on the growth of tomatoes grown in the open field
  • the test was carried out on a crop of tomatoes of the species So / anum lycopersicum.
  • the experimental set-up includes modalities with 5 repetitions.
  • the Ascophyllum nodosum extract was incorporated in an amount of lg / T in a solution of PK (5-30), which was enriched with the strain Bacillus Hcheniformis.
  • the following treatments are applied at the 3 leaf stage of tomato plants: - a control not including fertilizer,
  • the PK 5-30 + B. Hcheniformis solution slightly improved the yield, ie 5.2% compared to the control.
  • the use of the extract in combination with the PK 5-30 + bacteria solution allowed an improvement in the yield compared to the control of more than 14.8%. Compared to the solution
  • Example 4 Biofertilizing compositions or compositions intended for animal feed according to the invention [0127]
  • Example 4 c root fertilizers
  • Table 16 below describes the composition of an N P type root fertilizer.
  • Table 17 describes the composition of a root fertilizer of NPK and magnesium oxide type.
  • Table 18 below describes the composition of a solution of NPK and magnesium type.
  • Table 19 describes the composition of a solution of N, Calcium, Magnesium type.
  • Example 4 e Compositions intended for animal feed
  • Table 20 below describes the composition of a probiotic.
  • Table 20 Table 21 below describes the composition of a silage preservative.

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Abstract

The invention relates to the use of polyamines, tyramine and/or a plant extract containing same to stabilise microorganisms under environmental stress, such as oxidative stress, osmotic stress or saline stress, heat stress, acid-base stress and/or stress linked to competitor microorganisms; and a composition comprising (a) polyamines, tyramine and/or a plant extract containing same, and (b) microorganisms.

Description

Utilisation de polyamines, de tyramine et/ou d'un extrait végétal en contenant pour stabiliser des micro-organismesUse of polyamines, tyramine and / or a plant extract containing them to stabilize microorganisms
Domaine Technique Technical area
[0001] L'invention concerne l'utilisation d'un extrait d'algues et/ou de plantes, ledit extrait contenant des polyamines et/ou de la tyramine, pour stabiliser des micro-organismes face à un stress environnemental. The invention relates to the use of an extract of algae and / or plants, said extract containing polyamines and / or tyramine, for stabilizing microorganisms in the face of environmental stress.
Technique antérieure Prior art
[0002] Dans le domaine agricole, les sols sont des systèmes dynamiques qui contiennent une grande variété de micro-organismes. Cependant de nombreux facteurs, comme les techniques agricoles utilisées ces dernières décennies, ainsi que les changements climatiques ont bouleversé l'ensemble des équilibres préexistants. Ainsi, l'utilisation de grandes quantités d’intrants chimiques, les travaux culturaux etc. ont provoqué une raréfaction, voire une élimination de certains micro-organismes de la plupart des sols cultivés, ce qui contribue à une perte de productivité des sols. [0002] In the agricultural field, soils are dynamic systems which contain a wide variety of microorganisms. However, many factors, such as the agricultural techniques used in recent decades, as well as climate change have upset all the pre-existing balances. Thus, the use of large amounts of chemical inputs, cultural work etc. have caused a depletion or even elimination of certain microorganisms from most cultivated soils, which contributes to a loss of soil productivity.
[0003] De même, dans le domaine de l'alimentation animale, pour exercer leurs effets, les micro-organismes contenus dans les probiotiques doivent rester fonctionnels jusqu'à leur site d'action. Cependant, ces micro-organismes sont fragiles car ils sont très sensibles aux stress environnementaux, par exemple les variations de pH. Dans le cas des ensilages, à savoir la conservation de fourrage par fermentation lactique, les micro-organismes sont aussi soumis à un certain nombre de stress environnementaux tel qu'un stress oxydatif, un stress osmotique ou salin, un stress thermique, un stress acido-basique et/ou un stress lié aux micro-organismes compétiteurs. Les micro-organismes utilisés en alimentation animale sont donc exposés aux mêmes types de stress que ceux rencontrés dans le domaine agricole, ce qui empêche donc leur croissance, leur stabilité et/ou altère leurs effets. Likewise, in the field of animal nutrition, in order to exert their effects, the microorganisms contained in the probiotics must remain functional up to their site of action. However, these microorganisms are fragile because they are very sensitive to environmental stresses, for example variations in pH. In the case of silage, namely the conservation of fodder by lactic fermentation, the microorganisms are also subjected to a certain number of environmental stresses such as oxidative stress, osmotic or saline stress, thermal stress, acid stress. - basic and / or stress linked to competing microorganisms. The microorganisms used in animal feed are therefore exposed to the same types of stress as those encountered in agriculture, which therefore prevents their growth, their stability and / or their effects.
[0004] Le faible taux de survie des micro-organismes dans les préparations ou après leur incorporation dans le sol constitue l'un des facteurs limitant majeurs de leurs efficacités. Il est donc nécessaire de stabiliser les micro-organismes face aux stress environnementaux. [0004] The low survival rate of microorganisms in preparations or after their incorporation into the soil constitutes one of the major limiting factors of their efficiencies. It is therefore necessary to stabilize microorganisms in the face of environmental stresses.
[0005] Il existe donc un besoin de trouver de nouvelles stratégies pour stabiliser les micro-organismes face à un stress environnemental. [0006] C'est dans ce contexte, le demandeur a mis en évidence, et ceci constitue le fondement de la présente invention, que l'utilisation de polyamines, de tyramine et/ou d'un extrait végétal en contenant permet de stabiliser des micro-organismes face à un stress environnemental, tel qu'un stress oxydatif, un stress osmotique ou un stress salin, un stress thermique, un stress acido- basique et/ou un stress lié à des micro-organismes compétiteurs. [0005] There is therefore a need to find new strategies for stabilizing microorganisms in the face of environmental stress. It is in this context, the applicant has demonstrated, and this constitutes the basis of the present invention, that the use of polyamines, tyramine and / or a plant extract containing it makes it possible to stabilize microorganisms in the face of environmental stress, such as oxidative stress, osmotic stress or salt stress, heat stress, acid-base stress and / or stress linked to competing microorganisms.
Résumé de l'invention Summary of the invention
[0007] Ainsi, la présente invention, qui trouve application dans le domaine agro-écologique et agricole, vise à proposer l'utilisation de polyamines, de tyramine et/ou d'un extrait végétal en contenant pour stabiliser des micro- organismes face à un stress environnemental, tel qu'un stress oxydatif, un stress osmotique ou un stress salin, un stress thermique, un stress acido- basique et/ou un stress lié à des micro-organismes compétiteurs. [0007] Thus, the present invention, which finds application in the agro-ecological and agricultural field, aims to provide the use of polyamines, tyramine and / or a plant extract containing it to stabilize microorganisms in the face of environmental stress, such as oxidative stress, osmotic stress or salt stress, heat stress, acid-base stress and / or stress linked to competing microorganisms.
[0008] Selon un premier aspect, l'invention concerne l'utilisation d'un extrait d'algues et/ou de plantes, ledit extrait contenant des polyamines et/ou de la tyramine, pour stabiliser des micro-organismes face à un stress environnemental. [0008] According to a first aspect, the invention relates to the use of an extract of algae and / or plants, said extract containing polyamines and / or tyramine, for stabilizing microorganisms in the face of stress. environmental.
[0009] Selon un second aspect, l'invention concerne une composition comprenant : According to a second aspect, the invention relates to a composition comprising:
(a) un extrait d'algues et/ou de plantes, ledit extrait contenant des polyamines et/ou de la tyramine, et (a) an extract of algae and / or plants, said extract containing polyamines and / or tyramine, and
(b) des micro-organismes choisis parmi : (i) les bactéries fixatrices de l'azote atmosphérique, telles que Azotobacter ou Azospirillum (ii) les rhizobactéries favorisant la croissance des plantes (PGPR ou Plant Growth Promoting Rhizobacteria), (iii) les bactéries solubilisatrices du phosphore telles que BaciHus amyloliquefaciens, (iv) les bactéries phytoprotectrices des racines (PGPR) capables de s'opposer à l'activité d'agents pathogènes telles que BaciHus subti/is ou Pseudomonas spp., (v) les bactéries productrices de phytohormones telles que BaciHus amyloliquefaciens ou BaciHus radicoia, (vi) les bactéries impliquées dans le processus de minéralisation de la matière organique telles que LactobaciHus rhamnosus ou LactobaciHus faciminis, (vii) les bactéries solubilisatrices de fer telles que Pseudomonas spp., (viii) les bactéries solubilisatrices de la silice, (ix) les bactéries oxydatrices du soufre, (x) les bactéries lactiques telles que LactobaciHus spp., Lactococcus spp., Bifidobacterium spp., (xi) les bactéries du genre Enterococcus spp., (xii) les bactéries du genre Pediococcus spp., (xiii) les bactéries du genre BaciHus iicheniformis, (xiv) les champignons mycorhizes tel que Rhizophagus irreguiaris, (xv) les levures du genre Saccharomyces cerevisiae et (xvi) un mélange d'au moins deux micro-organismes choisis parmi (i) à (xv). (b) microorganisms chosen from: (i) bacteria fixing atmospheric nitrogen, such as Azotobacter or Azospirillum (ii) rhizobacteria promoting plant growth (PGPR or Plant Growth Promoting Rhizobacteria), (iii) phosphorus solubilizing bacteria such as BaciHus amyloliquefaciens, (iv) root protection bacteria (PGPR) capable of opposing the activity of pathogens such as BaciHus subti / is or Pseudomonas spp., (v) bacteria producing phytohormones such as BaciHus amyloliquefaciens or BaciHus radicoia, (vi) bacteria involved in the process of mineralization of organic matter such as LactobaciHus rhamnosus or LactobaciHus faciminis, (vii) iron solubilizing bacteria such as Pseudomonas spp., (viii) silica solubilizing bacteria, (ix) sulfur oxidizing bacteria, (x) lactic acid bacteria such as LactobaciHus spp., Lactococcus spp., Bifidobacterium spp., (xi) bacteria of the genus Enterococcus spp., (xii) bacteria of the genus Pediococcus spp., (xiii) bacteria of the genus BaciHus iicheniformis, (xiv ) mycorrhizal fungi such as Rhizophagus irreguiaris, (xv) yeasts of the genus Saccharomyces cerevisiae and (xvi) a mixture of at least two microorganisms chosen from (i) to (xv).
[0010] Selon un troisième aspect, l'invention concerne un procédé pour fertiliser une plante caractérisé en ce qu'il comprend l'apport à ladite plante d'une composition selon l'invention. According to a third aspect, the invention relates to a method for fertilizing a plant characterized in that it comprises supplying said plant with a composition according to the invention.
Description détaillée detailed description
[0011] Définitions [0012] Le terme « tyramine » désigne un composé chimique de type monoamine, dont le numéro CAS est 51-67-2. La tyramine a la formule suivante : Definitions The term "tyramine" denotes a chemical compound of monoamine type, the CAS number of which is 51-67-2. Tyramine has the following formula:
Formule 1 [0013] Le terme « polyamines » désigne des composés organiques contenant au moins deux fonctions amine. Les polyamines sont présentes dans presque toutes les plantes, les animaux et les micro-organismes. Les polyamines sont impliquées dans une variété de processus cellulaires tels que l’expression de certains gènes, la croissance cellulaire, la survie cellulaire et la prolifération cellulaire. Dans le cadre de la présente invention, les polyamines peuvent être choisies parmi la cadavérine, la spermidine, la spermine et/ou la putrescine. L'extrait végétal utilisé dans le cadre de l'invention peut donc comprendre un, deux, trois ou les quatre polyamines choisies parmi la cadavérine, la spermidine, la spermine et la putrescine. Formula 1 The term "polyamines" denotes organic compounds containing at least two amine functions. Polyamines are found in almost all plants, animals and microorganisms. Polyamines are involved in a variety of cellular processes such as the expression of certain genes, cell growth, cell survival and cell proliferation. In the context of the present invention, the polyamines can be chosen from cadaverine, spermidine, spermine and / or putrescine. The plant extract used in the context of the invention can therefore comprise one, two, three or all four polyamines chosen from cadaverine, spermidine, spermine and putrescine.
[0014] La cadavérine (ou pentaméthylènediamine), dont le numéro CAS est 462-94-2, a la formule suivante : Cadaverine (or pentamethylenediamine), the CAS number of which is 462-94-2, has the following formula:
Formule 2 Formula 2
[0015] La spermidine (ou 1,5,10-triazadécane), dont le numéro CAS est 124- 20-9, a la formule suivante : [0015] Spermidine (or 1,5,10-triazadecane), the CAS number of which is 124-20-9, has the following formula:
Formule 3 [0016] La spermine (également appelée gérontine, musculamine ou neuridine), dont le numéro CAS est 71-44-3, a la formule suivante : Formula 3 [0016] Spermine (also called gerontine, musculamine or neuridine), the CAS number of which is 71-44-3, has the following formula:
Formule 4 Formula 4
[0017] La putrescine (ou tétraméthylène diamine), dont le numéro CAS est 110-60-1, a la formule suivante : [0017] Putrescine (or tetramethylene diamine), the CAS number of which is 110-60-1, has the following formula:
Formule 5 Formula 5
[0018] Le terme « extrait végétal » désigne le produit résultant d'une extraction du contenu des cellules d'un végétal. Dans le cadre de la présente invention, l'extrait végétal contient une quantité efficace de polyamines et/ou de tyramine. Dans le cadre de la présente description, l'extrait végétal peut être un extrait d'algues et/ou de plantes. The term "plant extract" denotes the product resulting from an extraction of the contents of the cells of a plant. As part of this invention, the plant extract contains an effective amount of polyamines and / or tyramine. In the context of the present description, the plant extract can be an extract of algae and / or plants.
[0019] Le terme « extrait d'algues et/ou de plantes » désigne le produit résultant d'une extraction du contenu des cellules d'algues et/ou de plantes respectivement. Dans le cadre de la présente invention, l'extrait d'algues peut être un extrait d'algues brunes, tel qu'un extrait d'algues brunes de la famille des Fucacées. Par exemple, l'extrait d'algues brunes peut être un extrait de Fucus vesicu/osus ou ù'Ascophyllum noc/osum. Dans le cadre de la présente invention, l'extrait de plantes peut être un extrait de vinasse de mélasse de betteraves. The term "extract of algae and / or plants" denotes the product resulting from an extraction of the contents of the cells of algae and / or plants respectively. In the context of the present invention, the algae extract can be an extract of brown algae, such as an extract of brown algae of the Fucaceae family. For example, the extract of brown algae can be an extract of Fucus vesicu / osus or Ascophyllum noc / osum. In the context of the present invention, the plant extract can be an extract of beet molasses vinasse.
[0020] La préparation d'un extrait végétal contenant une quantité efficace de polyamines et/ou de tyramine ne présente aucune difficulté particulière, de nombreux procédés d'extraction sont décrits dans la littérature. Le procédé d'extraction n'est pas limité à un procédé particulier, et les procédés classiques peuvent être mis en œuvre pour préparer un tel extrait. Par exemple, il peut être obtenu par un procédé comportant les étapes suivantes : lavage, broyage, extraction avec un solvant (ex. de l'eau), séparation solide/liquide et éventuellement fractionnement et/ou concentration. L'extrait végétal obtenu peut être plus ou moins concentré selon l'utilisation envisagée, par exemple il est possible de concentrer l'extrait par une technique d'évaporation. Une déshydratation totale de cet extrait permettant une présentation sous forme pulvérulente hydrosoluble peut être obtenue, par exemple, par sécheur à tambour ou par atomisation. Les conditions d'extraction et la nature du végétal seront choisies de telle façon que l'extrait obtenu présente la quantité de polyamines et/ou de tyramine souhaitée dans l’application envisagée. Selon un exemple particulier, lorsque l'extrait est un extrait de vinasse de mélasse de betteraves, il peut être obtenu par un procédé comportant les étapes suivantes : lavage des betteraves, broyage des betteraves, extraction du sucre cristallisable de la betterave, obtention de deux co-produits : la mélasse et la pulpe de betteraves, récupération de la mélasse, fermentation de la mélasse pour obtenir une vinasse de mélasse et concentration [2] [3]. Il est également possible d'enrichir l'extrait végétal en polyamines et/ou en tyramine, par exemple par une technique d'ultrafiltration. The preparation of a plant extract containing an effective amount of polyamines and / or tyramine does not present any particular difficulty, many extraction processes are described in the literature. The extraction process is not limited to any particular process, and conventional processes can be used to prepare such an extract. For example, it can be obtained by a process comprising the following steps: washing, grinding, extraction with a solvent (eg water), solid / liquid separation and optionally fractionation and / or concentration. The plant extract obtained can be more or less concentrated depending on the use envisaged, for example it is possible to concentrate the extract by an evaporation technique. Total dehydration of this extract allowing presentation in water-soluble powder form can be obtained, for example, by tumble dryer or by atomization. The extraction conditions and the nature of the plant will be chosen such that the extract obtained exhibits the quantity of polyamines and / or tyramine desired in the envisaged application. According to a particular example, when the extract is an extract of beet molasses vinasse, it can be obtained by a process comprising the following steps: washing of the beets, grinding of the beets, extraction of the crystallizable sugar from the beet, obtaining two by-products: molasses and beet pulp, recovery of molasses, fermentation of molasses to obtain molasses stillage and concentration [2] [3]. It is also possible to enrich the plant extract with polyamines and / or tyramine, for example by an ultrafiltration technique.
[0021] Le terme « quantité efficace de polyamines et/ou de tyramine » désigne la quantité de polyamines et/ou de tyramine suffisante pour stabiliser les micro-organismes face au stress environnemental. L'extrait végétal contient de préférence au moins 0,01% en masse de polyamines et/ou de tyramine par rapport à la masse sèche totale de l'extrait (% w/w), par exemple au moins 0,05% w/w, au moins 0,5% w/w, au moins 0,2% w/w, par exemple entre 0,01% w/w et 1% w/w, entre 0,05% w/w et 0,5% w/w. Le dosage de la quantité de polyamines et de tyramines dans l'extrait ne présente aucune difficulté particulière, des méthodes de dosage étant décrites dans la littérature, comme par exemple la méthode HPLC [1]. L'extrait végétal peut être plus ou moins concentré en polyamines et/ou en tyramine selon l'utilisation envisagée. The term "effective amount of polyamines and / or tyramine" denotes the amount of polyamines and / or tyramine sufficient to stabilize the microorganisms in the face of environmental stress. The plant extract preferably contains at least 0.01% by mass of polyamines and / or tyramine relative to the total dry mass of the extract (% w / w), for example at least 0.05% w / w, at least 0.5% w / w, at least 0.2% w / w, for example between 0.01% w / w and 1% w / w, between 0.05% w / w and 0, 5% w / w. The assay of the amount of polyamines and tyramines in the extract does not present any particular difficulty, assay methods being described in the literature, such as for example the HPLC method [1]. The plant extract can be more or less concentrated in polyamines and / or in tyramine depending on the use envisaged.
[0022] Le terme « micro-organismes » désigne des organismes microscopiques tels que les bactéries, les champignons microscopiques, par exemple les champignons microscopiques filamenteux, les levures. Le terme « spp. » signifie « plusieurs espèces », du latin species plurimae. Dans le cadre de la présente invention, les micro-organismes peuvent être choisis parmi (i) les bactéries fixatrices de l'azote atmosphérique, telles que Azotobacter ou Azospirillum (ii) les rhizobactéries favorisant la croissance des plantes (PGPR ou Plant GrowthThe term "microorganisms" designates microscopic organisms such as bacteria, microscopic fungi, for example filamentous microscopic fungi, yeasts. The term "spp. "Means" several species ", from the Latin species plurimae. In the context of the present invention, the microorganisms can be chosen from (i) bacteria fixing atmospheric nitrogen, such as Azotobacter or Azospirillum (ii) rhizobacteria promoting plant growth (PGPR or Plant Growth
Promoting Rhizobacteria), (iii) les bactéries solubilisatrices du phosphore telles que BaciHus amyloliquefaciens, (iv) les bactéries phytoprotectrices des racines (PGPR) capables de s'opposer à l'activité d'agents pathogènes telles que BaciHus subti/is ou Pseudomonas spp., (v) les bactéries productrices de phytohormones telles que BaciHus amyloliquefaciens ou BaciHus radicoia, (vi) les bactéries impliquées dans le processus de minéralisation de la matière organique telles que LactobaciHus rhamnosus ou LactobaciHus faciminis, (vii) les bactéries solubilisatrices de fer telles que Pseudomonas spp., (viii) les bactéries solubilisatrices de la silice, (ix) les bactéries oxydatrices du soufre, (x) les bactéries lactiques telles que LactobaciHus spp., Lactococcus spp.,Promoting Rhizobacteria), (iii) phosphorus solubilizing bacteria such as BaciHus amyloliquefaciens, (iv) plant protection bacteria (PGPR) capable of opposing the activity of pathogens such as BaciHus subti / is or Pseudomonas spp ., (v) bacteria producing phytohormones such as BaciHus amyloliquefaciens or BaciHus radicoia, (vi) bacteria involved in the mineralization process of organic matter such as LactobaciHus rhamnosus or LactobaciHus faciminis, (vii) iron solubilizing bacteria such as Pseudomonas spp., (viii) silica solubilizing bacteria, (ix) sulfur oxidizing bacteria, (x) lactic acid bacteria such as LactobaciHus spp., Lactococcus spp.,
Bifidobacterium spp., (xi) les bactéries du genre Enterococcus spp., (xii) les bactéries du genre Pediococcus spp., (xiii) les bactéries du genre BaciHus Hcheniformis, (xiv) les champignons mycorhizes tel que Rhizophagus irregularis, (xv) les levures du genre Saccharomyces cerevisiae et (xvi) un mélange d'au moins deux micro-organismes choisis parmi (i) à (xv). Bifidobacterium spp., (Xi) bacteria of the genus Enterococcus spp., (Xii) bacteria of the genus Pediococcus spp., (Xiii) bacteria of the genus BaciHus Hcheniformis, (xiv) mycorrhizal fungi such as Rhizophagus irregularis, (xv) yeasts of the genus Saccharomyces cerevisiae and (xvi) a mixture of at least two microorganisms chosen from (i) to (xv).
[0023] Le terme « composition » désigne un mélange d'une ou plusieurs substances distinctes l'une de l'autre. Dans le cadre de l'invention, une composition peut être une composition fertilisante, une composition destinée à l'alimentation animale ou une composition destinée à la conservation d'ensilage. The term "composition" denotes a mixture of one or more substances distinct from one another. In the context of the invention, a composition may be a fertilizing composition, a composition intended for animal feed or a composition intended for the preservation of silage.
[0024] Une « composition fertilisante » désigne une composition comprenant une substance fertilisante, ou un mélange de substances fertilisantes, naturelle ou d’origine synthétique, utilisée en agriculture, en horticulture et sylviculture. A "fertilizing composition" denotes a composition comprising a fertilizing substance, or a mixture of fertilizing substances, natural or of synthetic origin, used in agriculture, horticulture and forestry.
[0025] On entend par « substance fertilisante » un engrais et/ou un amendement. The term "fertilizing substance" means a fertilizer and / or an amendment.
[0026] Le terme « engrais » désigne des matières fertilisantes dont la fonction principale est d'apporter aux plantes des éléments directement utiles à leur nutrition (éléments fertilisants majeurs, éléments fertilisants secondaires et oligo-éléments). The term "fertilizer" designates fertilizing materials whose main function is to provide the plants with elements directly useful for their nutrition (major fertilizing elements, secondary fertilizing elements and trace elements).
[0027] Le terme « amendement » désigne une substance destinée à améliorer la qualité des sols, et notamment destinée à améliorer le pH des sols. Avantageusement, l'amendement est choisi parmi les amendements minéraux basiques de type calcaire et/ou calcaires et magnésiens ; les amendements humifères de type composts ou les fumiers. The term "amendment" denotes a substance intended to improve the quality of soils, and in particular intended to improve the pH of the soils. Advantageously, the amendment is chosen from basic mineral amendments of limestone and / or limestone and magnesium type; humus amendments such as composts or manure.
[0028] Au sens de l'invention, une « composition destinée à l'alimentation animale » peut comprendre des substances destinées à améliorer la performance et/ou la santé des animaux. Une composition destinée à l'alimentation animale peut, par exemple, comprendre des aliments probiotiques, des compléments minéraux-vitaminiques, etc. [0028] Within the meaning of the invention, a "composition intended for animal feed" can comprise substances intended to improve the performance and / or the health of the animals. A composition intended for animal feed can, for example, comprise probiotic foods, mineral-vitamin supplements, etc.
[0029] Au sens de l'invention, une « composition destinée à la conservation d'ensilage » peut comprendre des substances destinées à améliorer la conservation et la qualité des ensilages. Une composition destinée à la conservation d'ensilage peut, par exemple, comprendre du lactose, des enzymes xylanases ou beta-glucanases, etc. [0029] Within the meaning of the invention, a "composition intended for the preservation of silage" can comprise substances intended to improve the preservation and the quality of the silages. A composition intended for silage preservation can, for example, include lactose, xylanase or beta-glucanase enzymes, etc.
[0030] Par l’expression « plante » on entend désigner dans la présente demande la plante considérée dans son ensemble, incluant son appareil racinaire, son appareil végétatif, les graines, semences et fruits. The expression "plant" is intended in the present application to denote the plant considered as a whole, including its root system, its vegetative system, seeds, seeds and fruits.
[0031] La présente invention découle des avantages surprenants mis en évidence par les inventeurs de l'effet de polyamines, de tyramine et/ou d'un extrait végétal en contenant pour stabiliser des micro-organismes. The present invention results from the surprising advantages demonstrated by the inventors of the effect of polyamines, of tyramine and / or of a plant extract containing it to stabilize microorganisms.
[0032] Utilisation [0033] La présente demande concerne l'utilisation de polyamines, de tyramine et/ou d'un extrait végétal en contenant pour stabiliser des micro-organismes face à un stress environnemental, tel qu'un stress oxydatif, un stress osmotique ou un stress salin, un stress thermique, un stress acido-basique et/ou un stress lié à des micro-organismes compétiteurs. [0034] Le premier objet de l'invention concerne l'utilisation d'un extrait d'algues et/ou de plantes, ledit extrait contenant des polyamines et/ou de la tyramine, pour stabiliser des micro-organismes face à un stress environnemental, tel qu'un stress oxydatif, un stress osmotique ou un stress salin, un stress thermique, un stress acido-basique et/ou un stress lié à des micro-organismes compétiteurs. En particulier, l'extrait végétal contient une quantité efficace de polyamines et/ou de tyramine. De préférence, l'extrait végétal est un extrait d'algues brunes, par exemple un extrait d'algues brunes de la famille des Fucacées, et/ou un extrait de vinasse de mélasse de betteraves. Il est entendu que le végétal utilisé pour la préparation de l'extrait végétal comprend naturellement des polyamines et/ou de la tyramine. Par conséquence, les polyamines et/ou la tyramine contenus dans l'extrait d'algues et/ou de plantes proviennent directement des algues et/ou des plantes utilisées pour préparer ledit extrait. [0032] Use [0033] The present application relates to the use of polyamines, tyramine and / or a plant extract containing it to stabilize microorganisms in the face of environmental stress, such as oxidative stress, stress. osmotic or saline stress, thermal stress, acid-base stress and / or stress linked to competing microorganisms. The first object of the invention relates to the use of an extract of algae and / or plants, said extract containing polyamines and / or tyramine, for stabilizing microorganisms in the face of environmental stress. , such as oxidative stress, osmotic stress or salt stress, heat stress, acid-base stress and / or stress linked to competing microorganisms. In particular, the plant extract contains an effective amount of polyamines and / or tyramine. Preferably, the plant extract is an extract of brown algae, for example an extract of brown algae of the Fucaceae family, and / or an extract of beet molasses vinasse. It is understood that the plant used for the preparation of the plant extract naturally comprises polyamines and / or tyramine. Consequently, the polyamines and / or the tyramine contained in the extract of algae and / or plants come directly from the algae and / or plants used to prepare said extract.
[0035] Un stress oxydatif correspond à une agression des micro-organismes par des radicaux libres, aussi appelés « espèces réactives de l’oxygène » (ERO). [0036] Un stress acido-basique correspond à un stress lié au pH. En effet, il est connu que les micro-organismes sont plus ou moins stables à certains pH, par exemple un pH allant de 1 à 12, par exemple allant de 3 à 9, par exemple un pH 5 ou par exemple un pH 9. [0037] Un stress osmotique correspond à un stress lié à la l'osmolarité, i.e. à la teneur en sel dans le milieu. En effet, il est connu que les micro-organismes sont plus ou moins stables à certaines osmolarités, par exemple à une concentration saline supérieure à 0,3 M les micro-organismes sont généralement moins stables. [0038] Un stress hydrique correspond à un stress lié à l'absence d'eau, i.e. à la teneur en eau dans le milieu. Oxidative stress corresponds to an attack on microorganisms by free radicals, also called "reactive oxygen species" (ROS). An acid-base stress corresponds to a stress linked to the pH. Indeed, it is known that microorganisms are more or less stable at certain pHs, for example a pH ranging from 1 to 12, for example ranging from 3 to 9, for example a pH 5 or for example a pH 9. Osmotic stress corresponds to stress linked to osmolarity, ie to the salt content in the medium. In fact, it is known that microorganisms are more or less stable at certain osmolarities, for example at a salt concentration greater than 0.3 M, microorganisms are generally less stable. Water stress corresponds to stress linked to the absence of water, ie to the water content in the medium.
[0039] Un stress thermique correspond à un stress induit par le froid ou par la chaleur. En fonction du micro-organisme considéré, la température induisant un stress thermique varie. [0040] Un stress lié à des micro-organismes compétiteurs correspond à un stress lié à la présence de micro-organismes autres que les micro-organismes stabilisés avec l'extrait végétal. Il peut s'agir, par exemple, des micro¬ organismes naturellement présents dans le sol qui vont entrer en compétition pour l'espace dans le sol et pour l'accès aux nutriments avec les micro- organismes rajoutés au sol. Par exemple, les micro-organismes compétiteurs sont les bactéries du genre Agrobacterium spp., du genre Erwinia spp., ou du genre Xanthomonas spp., ou les champignons du genre Fusarium spp. Thermal stress corresponds to stress induced by cold or by heat. Depending on the microorganism considered, the temperature inducing thermal stress varies. Stress linked to competing microorganisms corresponds to stress linked to the presence of microorganisms other than microorganisms stabilized with the plant extract. It may be, for example, micro ¬ organisms naturally present in the soil that will compete for space in the ground and for access to nutrients with microorganisms added to the soil. For example, the competing microorganisms are bacteria of the genus Agrobacterium spp., Of the genus Erwinia spp., Or of the genus Xanthomonas spp., Or fungi of the genus Fusarium spp.
[0041] En particulier, la stabilisation consiste à augmenter le taux de survie des micro-organismes, notamment mesuré après 24 heures, 48 heures ou 72 heures de croissance, en condition de stress environnemental par rapport aux mêmes conditions sans polyamines, tyramine et/ou extrait végétal en contenant. Le taux de survie de micro-organismes avec les polyamines, la tyramine et/ou l'extrait végétal en contenant, et soumis à un stress environnemental, est avantageusement d'au moins 1 log, au moins 2 log, au moins 3 log, au moins 4 log, au moins 5 log supérieur au taux de survie des mêmes micro-organismes soumis au même stress environnemental mais sans polyamines, tyramine et/ou extrait végétal en contenant. Le taux de survie se mesure en comparant la quantité de bactéries (par exemple en UFC) après un temps donné, par exemple 24 heures, 48 heures ou 72 heures. In particular, stabilization consists in increasing the survival rate of microorganisms, in particular measured after 24 hours, 48 hours or 72 hours of growth, under conditions of environmental stress compared to the same conditions without polyamines, tyramine and / or plant extract containing it. The survival rate of microorganisms with polyamines, tyramine and / or the plant extract containing them, and subjected to environmental stress, is advantageously at least 1 log, at least 2 log, at least 3 log, at least 4 log, at least 5 log greater than the survival rate of the same microorganisms subjected to the same environmental stress but without polyamines, tyramine and / or plant extract containing it. The survival rate is measured by comparing the quantity of bacteria (for example in CFU) after a given time, for example 24 hours, 48 hours or 72 hours.
[0042] Avantageusement, les micro-organismes sont contenus dans une composition, telle qu'une composition fertilisante, une composition destinée à l'alimentation animale ou une composition destinée à la conservation d'ensilage. Les micro-organismes peuvent être contenus dans ladite composition en une quantité allant de 102 à 1050 UFC par tonne de composition, de préférence allant de 105 à 1020 UFC par tonne de composition, de préférence environ 1011 UFC par tonne de composition. Advantageously, the microorganisms are contained in a composition, such as a fertilizing composition, a composition intended for animal feed or a composition intended for the preservation of silage. The microorganisms may be contained in said composition in an amount ranging from 10 2 to 10 50 CFU per tonne of composition, preferably ranging from 10 5 to 10 20 CFU per tonne of composition, preferably approximately 10 11 CFU per tonne of composition.
[0043] Les polyamines, la tyramine et/ou l'extrait végétal en contenant peuvent donc être utilisé en complément dans une composition, par exemple une composition fertilisante, une composition destinée à l'alimentation animale ou une composition destinée à la conservation d'ensilage, pour stabiliser les micro-organismes contenus dans ces compositions. Polyamines, tyramine and / or the plant extract containing it can therefore be used as a complement in a composition, for example a fertilizing composition, a composition intended for animal feed or a composition intended for the preservation of silage, to stabilize the microorganisms contained in these compositions.
[0044] Composition [0044] Composition
[0045] La présente demande concerne une composition comprenant : The present application relates to a composition comprising:
(a) des polyamines, de la tyramine et/ou un extrait végétal en contenant, et(a) polyamines, tyramine and / or a plant extract containing them, and
(b) des micro-organismes choisis parmi : (i) les bactéries fixatrices de l'azote atmosphérique, telles que Azotobacter ou Azospirillum (ii) les rhizobactéries favorisant la croissance des plantes (PGPR ou Plant Growth Promoting Rhizobacteria), (iii) les bactéries solubilisatrices du phosphore telles que BaciHus amyloliquefaciens, (iv) les bactéries phytoprotectrices des racines (PGPR) capables de s'opposer à l'activité d'agents pathogènes telles que BaciHus subti/is ou Pseudomonas spp., (v) les bactéries productrices de phytohormones telles que BaciHus amyloliquefaciens ou BaciHus radicoia, (vi) les bactéries impliquées dans le processus de minéralisation de la matière organique telles que LactobaciHus rhamnosus ou LactobaciHus faciminis, (vii) les bactéries solubilisatrices de fer telles que Pseudomonas spp., (viii) les bactéries solubilisatrices de la silice, (ix) les bactéries oxydatrices du soufre, (x) les bactéries lactiques telles que LactobaciHus spp., Lactococcus spp., Bifidobacterium spp., (xi) les bactéries du genre Enterococcus spp., (xii) les bactéries du genre Pediococcus spp., (xiii) les bactéries du genre BaciHus iicheniformis, (xiv) les champignons mycorhizes tel que Rhizophagus irreguiaris, (xv) les levures du genre Saccharomyces cerevisiae et (xvi) un mélange d'au moins deux micro-organismes choisis parmi (i) à (xv). (b) microorganisms chosen from: (i) bacteria fixing atmospheric nitrogen, such as Azotobacter or Azospirillum (ii) rhizobacteria promoting plant growth (PGPR or Plant Growth Promoting Rhizobacteria), (iii) phosphorus solubilizing bacteria such as BaciHus amyloliquefaciens, (iv) plant protection bacteria (PGPR) capable of opposing the activity of pathogens such as BaciHus subti / is or Pseudomonas spp., (v) producing bacteria phytohormones such as BaciHus amyloliquefaciens or BaciHus radicoia, (vi) bacteria involved in the mineralization process of organic matter such as LactobaciHus rhamnosus or LactobaciHus faciminis, (vii) iron solubilizing bacteria such as Pseudomonas spp., (viii) silica solubilizing bacteria, (ix) sulfur oxidizing bacteria, (x) lactic acid bacteria such as LactobaciHus spp., Lactococcus spp., Bifidobacterium spp., (Xi) bacteria of the genus Enterococcus spp., (Xii) bacteria of the genus Pediococcus spp., (Xiii) bacteria of the genus BaciHus iicheniformis, (xiv) mycorrhizal fungi such as Rhizophagus irreguiaris, (xv) yeasts of the genus Saccharomyces cerevisiae and (xvi) a mixture of at least two microorganisms chosen from (i) to (xv).
[0046] Un second objet de l'invention concerne une composition comprenant : (a) un extrait d'algues et/ou de plantes, ledit extrait contenant des polyamines et/ou de la tyramine, et (b) des micro-organismes choisis parmi : (i) les bactéries fixatrices de l'azote atmosphérique, telles que Azotobacter ou Azospirillum (ii) les rhizobactéries favorisant la croissance des plantes (PGPR ou Plant Growth Promoting Rhizobacteria), (iii) les bactéries solubilisatrices du phosphore telles que BaciHus amyloliquefaciens, (iv) les bactéries phytoprotectrices des racines (PGPR) capables de s'opposer à l'activité d'agents pathogènes telles que BaciHus subti/is ou Pseudomonas spp., (v) les bactéries productrices de phytohormones telles que BaciHus amyloliquefaciens ou BaciHus radicoia, (vi) les bactéries impliquées dans le processus de minéralisation de la matière organique telles que LactobaciHus rhamnosus ou LactobaciHus faciminis, (vii) les bactéries solubilisatrices de fer telles que Pseudomonas spp., (viii) les bactéries solubilisatrices de la silice, (ix) les bactéries oxydatrices du soufre, (x) les bactéries lactiques telles que LactobaciHus spp., Lactococcus spp., Bifidobacterium spp., (xi) les bactéries du genre Enterococcus spp., (xii) les bactéries du genre Pediococcus spp., (xiii) les bactéries du genre BaciHus iicheniformis, (xiv) les champignons mycorhizes tel que Rhizophagus irreguiaris,A second object of the invention relates to a composition comprising: (a) an extract of algae and / or plants, said extract containing polyamines and / or tyramine, and (b) selected microorganisms among: (i) atmospheric nitrogen fixing bacteria, such as Azotobacter or Azospirillum (ii) rhizobacteria promoting plant growth (PGPR or Plant Growth Promoting Rhizobacteria), (iii) phosphorus solubilizing bacteria such as BaciHus amyloliquefaciens , (iv) plant protection bacteria of the roots (PGPR) capable of opposing the activity of pathogenic agents such as BaciHus subti / is or Pseudomonas spp., (v) bacteria producing phytohormones such as BaciHus amyloliquefaciens or BaciHus radicoia, (vi) bacteria involved in the mineralization process of organic matter such as LactobaciHus rhamnosus or LactobaciHus faciminis, (vii) iron solubilizing bacteria such as Pseudomonas sp p., (viii) silica solubilizing bacteria, (ix) sulfur oxidizing bacteria, (x) lactic acid bacteria such as LactobaciHus spp., Lactococcus spp., Bifidobacterium spp., (xi) bacteria of the genus Enterococcus spp., (xii) bacteria of the genus Pediococcus spp., (xiii) bacteria of the genus BaciHus iicheniformis, (xiv) mycorrhizal fungi such as Rhizophagus irreguiaris,
(xv) les levures du genre Saccharomyces cerevisiae et (xvi) un mélange d'au moins deux micro-organismes choisis parmi (i) à (xv). (xv) yeasts of the genus Saccharomyces cerevisiae and (xvi) a mixture of at least two microorganisms chosen from (i) to (xv).
[0047] En particulier, (a) est un extrait végétal contenant une quantité efficace de polyamines et/ou de tyramine. Avantageusement, l'extrait végétal est un extrait d'algues et/ou de plantes, de préférence un extrait d'algues brunes, par exemple un extrait d'algues brunes de la famille des Fucacées, et/ou un extrait de vinasse de mélasse de betteraves. Il est entendu que le végétal utilisé pour la préparation de (a) l'extrait végétal comprend naturellement des polyamines et/ou de la tyramine. Par conséquence, les polyamines et/ou la tyramine contenus dans (a) l'extrait d'algues et/ou de plantes proviennent directement des algues et/ou des plantes utilisées pour préparer ledit extrait. In particular, (a) is a plant extract containing an effective amount of polyamines and / or tyramine. Advantageously, the plant extract is an extract of algae and / or plants, preferably an extract of brown algae, for example an extract of brown algae of the Fucaceae family, and / or an extract of beet molasses stillage. It is understood that the plant used for the preparation of (a) the plant extract naturally comprises polyamines and / or tyramine. Consequently, the polyamines and / or the tyramine contained in (a) the extract of algae and / or plants come directly from the algae and / or plants used to prepare said extract.
[0048] La composition selon l'invention peut notamment comprendre de 102 à 1050 UFC de micro-organismes (b) par tonne de composition, de préférence de 105 à 1020 UFC de micro-organismes (b) par tonne de composition, de préférence 1011 UFC de micro-organismes (b) par tonne de la composition. [0049] La composition peut notamment comprendre de 0,1 à 100 grammes de polyamines et/ou de tyramine par tonne de composition, préférentiellement de 0,4 à 10 grammes de polyamines et/ou de tyramine par tonne de composition. The composition according to the invention may in particular comprise from 10 2 to 10 50 CFU of microorganisms (b) per tonne of composition, preferably from 10 5 to 10 20 CFU of microorganisms (b) per tonne of composition, preferably 10 11 CFU of microorganisms (b) per tonne of the composition. The composition may in particular comprise from 0.1 to 100 grams of polyamines and / or tyramine per tonne of composition, preferably from 0.4 to 10 grams of polyamines and / or tyramine per tonne of composition.
[0050] Ainsi, dans un mode particulier de l'invention, la composition comprend : - de 0,1 à 100 grammes de polyamines et/ou de tyramine par tonne de composition, préférentiellement de 0,4 à 10 grammes de polyamines et/ou de tyramine par tonne de composition, et Thus, in a particular embodiment of the invention, the composition comprises: - from 0.1 to 100 grams of polyamines and / or tyramine per tonne of composition, preferably from 0.4 to 10 grams of polyamines and / or tyramine per tonne of composition, and
- de 102 à 1050 UFC de micro-organismes (b) par tonne de composition, de préférence de 105 à 1020 UFC de micro-organismes (b) par tonne de composition, de préférence 1011 UFC de micro-organismes (b) par tonne de composition. - from 10 2 to 10 50 CFU of microorganisms (b) per tonne of composition, preferably from 10 5 to 10 20 CFU of microorganisms (b) per tonne of composition, preferably 10 11 CFU of microorganisms (b) per tonne of composition.
[0051] La composition selon l'invention peut comprendre en outre (c) un engrais et / ou un amendement. The composition according to the invention can also comprise (c) a fertilizer and / or an amendment.
[0052] L'engrais peut être une ou plusieurs substances actives choisie parmi l'azote, le phosphore, le potassium, l’urée, le sulfate d’ammonium, le nitrate d'ammonium, le phosphate, le chlorure de potassium, du sulfate d’ammonium, le nitrate de magnésium, le nitrate de manganèse, le nitrate de zinc, le nitrate de cuivre, l’acide phosphorique, le nitrate de potassium, l’acide borique et leurs mélanges, de préférence un mélange d'azote, de potassium et de phosphore ou un mélange de phosphore et de potassium. L'amendement peut être une ou plusieurs substances actives choisie parmi les amendements minéraux basiques de type calcaire, les amendements minéraux basiques de type magnésien, les amendements humifères de type composts et/ou les amendements humifères de type fumiers, de préférence engrais et amendements minéraux. [0053] Les polyamines, la tyramine et/ou l'extrait végétal en contenant, de préférence l'extrait végétal (a) contenant une quantité efficace de polyamines et/ou de tyramine, par exemple l'extrait d'algues et/ou de plantes (a), de préférence l'extrait d'algues et/ou de plantes (a) contenant une quantité efficace de polyamines et/ou de tyramine, stabilisent les micro-organismes (b) face à un stress environnemental, tel qu'un stress oxydatif, un stress osmotique ou un stress salin, un stress thermique, un stress acido-basique et/ou un stress lié à des micro-organismes compétiteurs. En effet, les micro-organismes (b) peuvent être sensibles et/ou instables en présence d'un engrais, à un pH allant de 1 à 12, de préférence à un pH allant de 3 à 9, par exemple à un pH d'environ 5 ou à un pH d'environ 9 et/ou à une salinité allant au-delà du seuil de sensibilité/stabilité au sel d'un micro-organisme considéré, par exemple à une concentration saline supérieure à 0,3 M pour les bactéries du genre Azobacter chroococcum. The fertilizer can be one or more active substances chosen from nitrogen, phosphorus, potassium, urea, ammonium sulfate, ammonium nitrate, phosphate, potassium chloride, ammonium sulfate, magnesium nitrate, manganese nitrate, zinc nitrate, copper nitrate, phosphoric acid, potassium nitrate, boric acid and their mixtures, preferably a mixture of nitrogen , potassium and phosphorus or a mixture of phosphorus and potassium. The amendment can be one or several active substances chosen from basic mineral amendments of the limestone type, basic mineral amendments of the magnesium type, humus amendments of the composts type and / or humus amendments of the manure type, preferably fertilizers and mineral amendments. Polyamines, tyramine and / or the plant extract containing them, preferably the plant extract (a) containing an effective amount of polyamines and / or tyramine, for example the algae extract and / or of plants (a), preferably the extract of algae and / or plants (a) containing an effective amount of polyamines and / or tyramine, stabilize the microorganisms (b) in the face of environmental stress, such as oxidative stress, osmotic stress or salt stress, heat stress, acid-base stress and / or stress linked to competing microorganisms. Indeed, the microorganisms (b) can be sensitive and / or unstable in the presence of a fertilizer, at a pH ranging from 1 to 12, preferably at a pH ranging from 3 to 9, for example at a pH of '' about 5 or at a pH of about 9 and / or at a salinity going beyond the threshold of sensitivity / stability to the salt of a microorganism in question, for example at a salt concentration greater than 0.3 M for bacteria of the genus Azobacter chroococcum.
[0054] Dans un mode de réalisation particulier, la composition comprend en outre un engrais et présente un pH allant de 1 à 12, de préférence un pH allant de 3 à 9, par exemple un pH d'environ 5 ou un pH d'environ 9 et/ou une salinité allant au-delà du seuil de sensibilité/stabilité au sel d'un micro-organisme considéré, par exemple à une concentration saline supérieure à 0,3 M pour les bactéries du genre Azobacter chroococcum. [0055] La composition selon l'invention peut être sous forme solide ou liquide, préférentiellement sous forme solide. In a particular embodiment, the composition further comprises a fertilizer and has a pH ranging from 1 to 12, preferably a pH ranging from 3 to 9, for example a pH of about 5 or a pH of approximately 9 and / or a salinity going beyond the threshold of sensitivity / stability to the salt of a microorganism in question, for example at a salt concentration greater than 0.3 M for bacteria of the genus Azobacter chroococcum. The composition according to the invention can be in solid or liquid form, preferably in solid form.
[0056] La composition selon l'invention peut également comprendre d'autres substances, telles que des substances biostimulantes, phytohormones, polyphénols, acides aminés, etc. [0057] La composition selon l'invention pourra être une composition fertilisante, une composition destinée à l'alimentation animale ou une composition destinée à la conservation d'ensilage. Les autres substances seront donc choisies en fonction de l'utilisation envisagée. The composition according to the invention can also comprise other substances, such as biostimulant substances, phytohormones, polyphenols, amino acids, etc. The composition according to the invention may be a fertilizing composition, a composition intended for animal feed or a composition intended for the preservation of silage. The other substances will therefore be chosen according to the intended use.
[0058] A titre d’exemples de composition fertilisante selon l’invention, on citera les compositions comprenant les amendements calcaires, les amendements organiques et les supports de culture, les engrais racinaires de type NP, PK,As examples of a fertilizing composition according to the invention, mention will be made of compositions comprising limestone amendments, organic amendments and growing media, root fertilizers of NP, PK type,
NPK, etc., les engrais et/ou biostimulants foliaires ou encore les solutions nutritives racinaires. NPK, etc., foliar fertilizers and / or biostimulants or root nutrient solutions.
[0059] A titre d’exemples de composition destinée à l'alimentation animale selon l’invention, on citera les compositions comprenant par exemple du bicarbonate de sodium, de l'oxyde de magnésium, de la levure alimentaire. As examples of a composition intended for animal feed according to the invention, mention will be made of compositions comprising, for example, sodium bicarbonate, magnesium oxide, nutritional yeast.
[0060] A titre d'exemples de composition destinée à la conservation d'ensilage selon l'invention, on citera les compositions comprenant par exemple du lactose, des enzymes de type xylanase, beta-glucanase, etc. By way of examples of a composition intended for the preservation of silage according to the invention, mention will be made of compositions comprising, for example, lactose, enzymes of xylanase, beta-glucanase, etc. type.
[0061] Procédé [0062] Un troisième objet de l'invention concerne un procédé pour fertiliser une plante caractérisé en ce qu'il comprend l'apport à ladite plante d'une composition selon l'invention. Le procédé selon l'invention permet de stimuler le développement d'une plante et/ou de stimuler le rendement d'une plante. A third object of the invention relates to a method for fertilizing a plant characterized in that it comprises supplying said plant with a composition according to the invention. The method according to the invention makes it possible to stimulate the development of a plant and / or to stimulate the yield of a plant.
[0063] Selon le procédé de l'invention, la composition peut être apportée à ladite plante dans une quantité allant de 0,5 à 1500 kg/ha, de préférence allant de 1 à 1000kg/ha kg/ha. According to the process of the invention, the composition can be added to said plant in an amount ranging from 0.5 to 1500 kg / ha, preferably ranging from 1 to 1000 kg / ha kg / ha.
[0064] La composition peut être apportée sous forme solide ou liquide. Selon le procédé de l'invention, la composition peut avantageusement être une composition fertilisante, telle que celle décrite ci-dessus. Avantageusement, l’application à ladite plante sera réalisée par voie foliaire ou par voie racinaire, préférentiellement par voie racinaire. The composition can be provided in solid or liquid form. According to the process of the invention, the composition can advantageously be a fertilizing composition, such as that described above. Advantageously, the application to said plant will be carried out by foliar route or by root route, preferably by root route.
[0065] La présente invention trouve application dans le traitement d’une très grande variété de plantes. Parmi celles-ci, on citera en particulier : - les plantes de grande culture telles que les céréales (blé, maïs, canne à sucre,...), - les protéagineux (pois), The present invention finds application in the treatment of a very wide variety of plants. Among these, we can cite in particular: - field crops such as cereals (wheat, corn, sugar cane, etc.), - protein crops (peas),
- les oléagineux (soja, tournesol), - oilseeds (soya, sunflower),
- la vigne, - Vine,
- les plantes prairiales utiles pour l'alimentation animale, - les cultures spécialisées telles qu'en particulier le maraîchage (laitue, épinards, tomate, melon), la vigne, l'arboriculture (oranger, poire, pomme, nectarine), ou l'horticulture (rosiers), - grassland plants useful for animal feed, - specialized crops such as market gardening (lettuce, spinach, tomato, melon), vines, arboriculture (orange tree, pear, apple, nectarine), or 'horticulture (roses),
- les cultures industrielles (pomme de terre, betterave, ...) - industrial crops (potatoes, beets, etc.)
[0066] Dans un mode de réalisation préféré selon l'invention, la plante appartient à l'ordre des monocotylédones, de préférence à la famille des Poacées, de préférence la plante est choisie parmi le blé, le riz, l'orge, l'avoine, le seigle, la prairie, la canne à sucre ou le maïs. In a preferred embodiment according to the invention, the plant belongs to the order of monocotyledons, preferably to the Poaceae family, preferably the plant is chosen from wheat, rice, barley, l oats, rye, meadow, sugar cane or corn.
[0067] La présente invention est illustrée par les exemples non limitatifs suivants. The present invention is illustrated by the following non-limiting examples.
EXEMPLES EXAMPLES
[0068] Exemple 1 : Préparation d'un extrait d'alaues et/ou de plantesExample 1: Preparation of an extract of alaues and / or plants
[0069] Exemple 1 a) : Préparation d'un extrait d'alaues brunes de la famille des Fucacées du genre Ascophyllum nodosum [0070] 200 g d'algues fraîches du genre Ascophyllum nodosum ont été broyées pour obtenir des fragments d'algues fraîches ayant une taille d'environ 5 mm. Example 1 a) Preparation of an extract of brown algae of the Fucaceae family of the genus Ascophyllum nodosum 200 g of fresh algae of the genus Ascophyllum nodosum were ground to obtain fragments of fresh algae having a size of about 5 mm.
[0071] L'extraction aqueuse d'algues fraîches a été réalisée en utilisant 200 g d' Ascophyllum nodosum par litre d'eau. [0072] L'extraction a été réalisée dans une solution 0,5 N d'acide sulfurique sous agitation qui dure 2h à température ambiante. L'extrait a ensuite été filtré sur une membrane (80 pm de porosité) puis concentré sur évaporateur à flux tombant. L'analyse d'un kilo d'extrait a été réalisée par Chromatographie Liquide haute Performance. La méthode analytique utilisée pour l'identification et la quantification des polyamines est basée sur la méthode décrite par H. B. Papenfus (2011). The aqueous extraction of fresh algae was carried out using 200 g of Ascophyllum nodosum per liter of water. The extraction was carried out in a 0.5N solution of sulfuric acid with stirring which lasts 2 hours at room temperature. The extract was then filtered through a membrane (80 μm porosity) and then concentrated on a falling flow evaporator. The analysis of one kilo of extract was carried out by High Performance Liquid Chromatography. The analytical method used for the identification and Quantification of polyamines is based on the method described by HB Papenfus (2011).
[0073] Un kilo d'extrait sec (tableau 1) contient 3300 mg de polyamines au total, soit 480 mg de putrescine, 1470 mg de cadaverine, 840 mg de spermidine et 510 mg de spermine. One kilogram of dry extract (Table 1) contains 3300 mg of polyamines in total, ie 480 mg of putrescine, 1470 mg of cadaverine, 840 mg of spermidine and 510 mg of spermine.
Tableau 1 Table 1
[0074] Exemple 1 b) : Préparation d'un extrait d'algues brunes de la famille des Fucacées du genre Fucus vesicu/osus [0075] 200 g d'algues fraîches du genre Fucus vesicu/osus ont été broyées pour obtenir des fragments d'algues fraîches ayant une taille d'environ 5 mm. Example 1 b) Preparation of an extract of brown algae of the Fucaceae family of the genus Fucus vesicu / osus 200 g of fresh algae of the genus Fucus vesicu / osus were crushed to obtain fragments fresh seaweed having a size of about 5 mm.
[0076] L'extraction aqueuse d'algues fraîches a été réalisée en utilisant 200 g de Fucus vesicu/osus par litre d'eau. The aqueous extraction of fresh algae was carried out using 200 g of Fucus vesicu / osus per liter of water.
[0077] L'extraction a été réalisée dans une solution 1 N d'acide sulfurique sous agitation qui dure 2h à température ambiante. L'extrait a ensuite été filtré sur une membrane (80 pm de porosité) puis concentré sur évaporateur à flux tombant. L'analyse d'un kilo d'extrait a été réalisée par Chromatographie Liquide haute Performance. La méthode analytique utilisée pour l'identification et la quantification des polyamines est basée sur la méthode décrite par H. B. Papenfus (2011). The extraction was carried out in a 1N solution of sulfuric acid with stirring which lasts 2 hours at room temperature. The extract was then filtered through a membrane (80 μm porosity) and then concentrated on a falling flow evaporator. The analysis of one kilo of extract was carried out by High Performance Liquid Chromatography. The analytical method used for the identification and quantification of polyamines is based on the method described by H. B. Papenfus (2011).
[0078] Un kilo d'extrait sec (tableau 2) contient 7170 mg de polyamines au total: 1140 mg de putrescine, 150 mg de cadaverine, 1830 mg de spermidine et 4050 mg de spermine. Tableau 2 One kilo of dry extract (Table 2) contains 7170 mg of polyamines in total: 1140 mg of putrescine, 150 mg of cadaverine, 1830 mg of spermidine and 4050 mg of spermine. Table 2
[0079] Exemple 1 c) : Analyse d'un extrait de vinasse de mélasse de betteraves. [0080] Un extrait de vinasse de mélasse de betteraves convenant à la mise en œuvre de l'invention est disponible sous les noms commerciaux suivants : Protéinal, Sirional, Betainex, Viprotal, Vinasse FS, Citrocol et Prodyn [3]. Example 1 c): Analysis of an extract of beet molasses vinasse. An extract of beet molasses vinasse suitable for the implementation of the invention is available under the following trade names: Proteinal, Sirional, Betainex, Viprotal, Vinasse FS, Citrocol and Prodyn [3].
[0081] L'analyse d'un kilo d'extrait liquide (environ 60% de masse sèche) a été réalisée par chromatographie Liquide haute Performance. La méthode analytique utilisée pour l'identification et la quantification des polyamines est basée sur la méthode décrite par H. B. Papenfus (2011) [1]. The analysis of a kilo of liquid extract (approximately 60% of dry mass) was carried out by high performance liquid chromatography. The analytical method used for the identification and quantification of polyamines is based on the method described by H. B. Papenfus (2011) [1].
[0082] Un kilo d'extrait sec (tableau 3) contient 477 mg de polyamines au total: 344 mg de tyramine, 73 mg de putrescine, 48 mg de cadavérine et 12 mg de spermidine. One kilogram of dry extract (Table 3) contains 477 mg of polyamines in total: 344 mg of tyramine, 73 mg of putrescine, 48 mg of cadaverine and 12 mg of spermidine.
Tableau 3 [0083] Exemple 2 : Effet d'un extrait d'alaues ou de plantes sur la croissance de micro-organismes Table 3 Example 2: Effect of an extract of alaues or plants on the growth of microorganisms
[0084] Méthode [0084] Method
[0085] L'effet de chacun des extraits préparés selon l'exemple la), lb) ou le) a été étudié sur la croissance de différentes souches bactériennes, en condition ou non de stress thermique, de stress osmotique ou de stress acido-basique. Les extraits obtenus aux exemples la), lb) ou le) sont ajoutés dans un milieu de culture en différentes quantités, afin d'obtenir les quantités de polyamines et/ou de tyramine souhaités. The effect of each of the extracts prepared according to example la), lb) or le) was studied on the growth of different bacterial strains, under condition or not of heat stress, osmotic stress or acid stress. basic. The extracts obtained in Examples la), lb) or le) are added to a culture medium in different amounts, in order to obtain the desired amounts of polyamines and / or tyramine.
[0086] L'étude a été réalisée en bouillon de culture SMM Spizizen Minimal Medium ((NH4)2S0 : 2 g/L, K2HP0 : 14 g/L, KH2P0 : 6 g/L, MgS0 : 0.2 g/L, C6H5Na307 2H20 : 1 g/L, D-glucose : 5 g/L). The study was carried out in SMM Spizizen Minimal Medium culture broth ((NH 4 ) 2 S0: 2 g / L, K 2 HP0: 14 g / L, KH 2 P0: 6 g / L, MgS0: 0.2 g / L, C6H 5 Na 3 07 2H 2 0: 1 g / L, D-glucose: 5 g / L).
[0087] L'inoculum de départ pour chacune de souches a été calibré par mesure de DO (620 nm) afin d'atteindre une concentration initiale autour de 103-104 UFC/mL. Les souches ont été mises à incuber pendant 24 à 72 h. Des dénombrements ont été réalisés en gélose TSA (Tryptone Soya Agar) à T0 et après 24 h, 48 h ou 72 h d'incubation à 30°C (+/- 1°C), selon les souches étudiées. The starting inoculum for each of the strains was calibrated by measuring the OD (620 nm) in order to reach an initial concentration around 10 3 -10 4 CFU / mL. The strains were incubated for 24 to 72 h. Counts were carried out in TSA (Tryptone Soya Agar) agar at T0 and after 24 h, 48 h or 72 h of incubation at 30 ° C (+/- 1 ° C), depending on the strains studied.
[0088] Pour chacune des souches, des taux de survie en présence de l'extrait et/ou de stress ont été calculés par rapport à la concentration bactérienne obtenue en fin de cinétique (à T= 24h, 48h ou 72h) dans la modalité témoin contenant la souche seule en condition non stressante sans ajout de l'extrait. De la même manière, des taux de survie en présence ou absence de l'extrait et/ou de stress ont été calculés par rapport à la concentration bactérienne obtenue en fin de cinétique dans la modalité contenant la souche seule en condition de stress sans ajout de l'extrait. For each of the strains, survival rates in the presence of the extract and / or stress were calculated relative to the bacterial concentration obtained at the end of kinetics (at T = 24h, 48h or 72h) in the modality control containing the strain alone in non-stressful condition without addition of the extract. Likewise, survival rates in the presence or absence of the extract and / or stress were calculated relative to the bacterial concentration obtained at the end of kinetics in the modality containing the strain alone under stress conditions without addition of the extract.
[0089] Exemple 2 a) : Effet de l'extrait de Fucus vesicu/osus préparé selon l'exemple 1 a) sur la croissance de BaciHus SD. en condition de stress osmotique [0090] La souche BaciHus sp. a été testée en condition de stress osmotique, c'est-à-dire en ajoutant 0,3 M de NaCI dans le milieu de culture à un instant T=0. Example 2 a): Effect of the extract of Fucus vesicu / osus prepared according to example 1 a) on the growth of BaciHus SD. under conditions of osmotic stress The BaciHus sp. was tested under conditions of osmotic stress, that is to say by adding 0.3 M of NaCl to the culture medium at a time T = 0.
[0091] Les résultats, présentés au tableau 4, indiquent que l'ajout de l'extrait de Fucus vesicu/osus préparé selon l'exemple 1 a), en condition non stressante a permis de stimuler la croissance de BaciHus sp. en augmentant la concentration bactérienne de 2,8 logs après 24 h d'incubation. L'ajout de NaCI à 0,3 M dans le milieu de culture a bien permis de reproduire les conditions d'un stress salin avec une perte de 2,2 logs de la concentration bactérienne après 24 h d'incubation par rapport à la condition sans stress salin. En condition de stress salin, l'ajout de l'extrait de Fucus vesicu/osus a permis une augmentation de 4,9 logs de la concentration bactérienne après 24 h d'incubation. La concentration bactérienne obtenue est globalement similaire à celle observée dans le milieu sans sel avec ajout de l'extrait de Fucus vesicu/osus, autour de 107 UFC/mL. [0092] Tableau 4 [0093] Exemple 2 b : Effet de l'extrait de Fucus vesicu/osus préparé selon l'exemple 1 a] sur la croissance de Bacillus Hcheniformis en condition de stress acide The results, presented in Table 4, indicate that the addition of the extract of Fucus vesicu / osus prepared according to Example 1 a), under non-stressful conditions made it possible to stimulate the growth of BaciHus sp. by increasing the bacterial concentration by 2.8 logs after 24 h of incubation. The addition of 0.3 M NaCl in the culture medium made it possible to reproduce the conditions of a salt stress with a loss of 2.2 logs of the bacterial concentration after 24 h of incubation compared to the condition. without salt stress. Under conditions of salt stress, the addition of the extract of Fucus vesicu / osus allowed an increase of 4.9 logs in the bacterial concentration after 24 h of incubation. The bacterial concentration obtained is generally similar to that observed in the medium without salt with the addition of the extract of Fucus vesicu / osus, around 10 7 CFU / mL. Table 4 Example 2 b: Effect of the extract of Fucus vesicu / osus prepared according to example 1 a] on the growth of Bacillus Hcheniformis under acid stress condition
[0094] La souche Bacillus Hcheniformis a été testée en condition de stress acide, c'est-à-dire en ajoutant du HCl pour atteindre un pH de 4,6 dans le milieu de culture à un instant T = 0. The Bacillus Hcheniformis strain was tested under acid stress conditions, that is to say by adding HCl to reach a pH of 4.6 in the culture medium at a time T = 0.
[0095] Les résultats sont présentés au tableau 5 : l'incubation de B. Hcheniformis 72 h à pH 4,6 a bien permis de reproduire les conditions d'un stress acide avec une perte de 3,7 logs de la concentration bactérienne après 72 h d'incubation par rapport à la condition à pH 7. The results are presented in Table 5: the incubation of B. Hcheniformis 72 h at pH 4.6 made it possible to reproduce the conditions of an acid stress with a loss of 3.7 logs of the bacterial concentration after 72 h of incubation relative to the condition at pH 7.
[0096] En condition de stress acide, l'ajout de l'extrait de Fucus vesicu/osus a permis une augmentation de 1,9 logs de la concentration bactérienne après 72 h d'incubation. Under acid stress conditions, the addition of the extract of Fucus vesicu / osus allowed an increase of 1.9 logs in the bacterial concentration after 72 h of incubation.
[0097] Tableau 5 [0098] Exemple 2 c) : Effet de l'extrait ô'Ascoobv/lum nodosum préparé selon l'exemple la] sur la croissance ô' Azospirillum brasilense en condition de stress osmotique Table 5 Example 2 c): Effect of the extract ô'Ascoobv / lum nodosum prepared according to example la] on the growth of ô'Azospirillum brasilense under conditions of osmotic stress
[0099] La souche Azospirillum brasilense a été testée en condition de stress osmotique, c'est-à-dire en ajoutant 0,4 M de NaCI dans le milieu de culture à un instant T = 0. The Azospirillum brasilense strain was tested under conditions of osmotic stress, that is to say by adding 0.4 M of NaCl in the culture medium at a time T = 0.
[0100] Les résultats, présentés au tableau 6, indiquent que l'ajout de l'extrait ô'Ascophyllum nodosum préparé selon l'exemple la) en condition non stressante a permis de stimuler la croissance û'A. brasilense en augmentant la concentration bactérienne de 4,6 logs après 72 h d'incubation. The results, presented in Table 6, indicate that the addition of the extract ô'Ascophyllum nodosum prepared according to example la) in non-stressful condition made it possible to stimulate the û'A growth. brasilense by increasing the bacterial concentration by 4.6 logs after 72 h of incubation.
[0101] L'ajout de NaCI à 0,4 M dans le milieu de culture a permis d'atteindre un niveau limite de stress salin. Dans ces conditions l'ajout de l'extrait ô'Ascophyllum nodosum a permis une augmentation de 1,5 logs de la concentration bactérienne après 72 h d'incubation. La concentration bactérienne atteinte est supérieure de 2,6 logs à celle du témoin souche seule mais reste inférieure à celle de la condition sans sel avec ajout l'extrait ô'Ascophyllum nodosum. The addition of 0.4 M NaCl in the culture medium made it possible to reach a limiting level of salt stress. Under these conditions, the addition of the Ascophyllum nodosum extract made it possible to increase the bacterial concentration by 1.5 logs after 72 h of incubation. The bacterial concentration reached is 2.6 logs higher than that of the strain control alone but remains lower than that of the salt-free condition with the addition of the Ascophyllum nodosum extract.
Tableau 6 [0102] Exemple 2 d : Effet de l'extrait ô'Ascophyllum nodosum préparé selon l'exemple la] sur la croissance ô' Azospirillum brasilense en condition de stress thermique Table 6 [0102] Example 2 d: Effect of the extract ô'Ascophyllum nodosum prepared according to example la] on the growth of ô'Azospirillum brasilense under heat stress conditions
[0103] La souche Azospirillum brasilense a été testée en condition de stress thermique, c'est-à-dire en augmentant la température du milieu de culture deThe Azospirillum brasilense strain was tested under heat stress conditions, that is to say by increasing the temperature of the culture medium from
30°C à 45°C à un instant T = 0. 30 ° C to 45 ° C at a time T = 0.
[0104] Les résultats, présentés au tableau 7, indiquent que l'ajout de l'extrait ô'Ascophyllum nodosum, préparé selon l'exemple la), en condition non stressante a permis une légère stimulation de la croissance d'A brasilense en augmentant la concentration bactérienne de 0,5 log après 72 h d'incubation. The results, presented in Table 7, indicate that the addition of the extract ô'Ascophyllum nodosum, prepared according to example la), in non-stressful conditions allowed a slight stimulation of the growth of A. brasilense in increasing the bacterial concentration by 0.5 log after 72 h of incubation.
[0105] L'incubation d'A brasiiense ll h à 45°C a bien permis de reproduire les conditions d'un stress thermique avec une perte de 5,9 logs de la concentration bactérienne après 72 h d'incubation par rapport à la condition à 30°C. Dans ces conditions l'ajout de l'extrait ô'Ascophyllum nodosum a permis une augmentation de 5,5 logs de la concentration bactérienne après 72 h d'incubation. La concentration bactérienne atteinte est globalement similaire à celle du témoin à 30°C, autour de 5.106 UFC/mL. The incubation of A brasiiense 11 h at 45 ° C made it possible to reproduce the conditions of thermal stress with a loss of 5.9 logs of the bacterial concentration after 72 h of incubation compared to the condition at 30 ° C. Under these conditions, the addition of the Ascophyllum nodosum extract allowed an increase of 5.5 logs in the bacterial concentration after 72 h of incubation. The bacterial concentration reached is globally similar to that of the control at 30 ° C, around 5.10 6 CFU / mL.
Tableau 7 Table 7
[0106] Exemple 2 fi : Effet de l'extrait de vinasse de mélasse de betterave préparé selon l'exemple le] sur la croissance ù'AzosDirillum brasilense en condition de stress basique [0107] La souche Azospirillum brasilense a été testée en condition de stress basique, c'est-à-dire en ajoutant NaOH dans le milieu de culture à un instant T = 0. [0106] Example 2 fi: Effect of the extract of beet molasses stillage prepared according to example 1c on the growth of AzosDirillum brasilense under conditions of basic stress [0107] The Azospirillum brasilense strain was tested under conditions of basic stress, that is to say by adding NaOH to the culture medium at a time T = 0.
[0108] Les résultats, présentés au tableau 8, indiquent que l'ajout de l'extrait de vinasse de mélasse de betterave, préparé selon l'exemple le), en condition non stressante a permis une légère stimulation de la croissance d'A brasilense en augmentant la concentration bactérienne de 0,6 log après 72 h d'incubation. The results, presented in Table 8, indicate that the addition of the extract of beet molasses stillage, prepared according to example 1c), under non-stressful conditions allowed a slight stimulation of the growth of A. brasilense by increasing the bacterial concentration by 0.6 log after 72 h of incubation.
[0109] L'incubation d'A brasiiense ll h à pH 10 a permis d'atteindre un niveau limite de stress basique. Dans ces conditions l'ajout de l'extrait de vinasse de mélasse de betterave a permis une augmentation de 1,1 logs de la concentration bactérienne après 72 h d'incubation. La concentration bactérienne atteinte est supérieure de 1,3 log à celle du témoin souche seule à pH 7. Tableau 8 [0109] The incubation of A brasiiense 11 h at pH 10 made it possible to reach a limiting level of basic stress. Under these conditions, the addition of the extract of beet molasses vinasse allowed an increase of 1.1 logs in the bacterial concentration after 72 h of incubation. The bacterial concentration reached is 1.3 log higher than that of the strain control alone at pH 7. Table 8
[0110] Exemple 2 cO Effet de l'extrait de vinasse de mélasse de betterave préparé selon l'exemple le) sur la croissance d'un complexe de rhizobactéries fixatrices d'azote ( Azotobacter diazotrophicus. Paenibacillus azotofixans.[0110] Example 2 cO Effect of the extract of beet molasses vinasse prepared according to example 1c on the growth of a complex of nitrogen-fixing rhizobacteria (Azotobacter diazotrophicus. Paenibacillus azotofixans.
Azotobacter chroococcurri ) en condition de stress osmotique Azotobacter chroococcurri) in conditions of osmotic stress
[0111] Le complexe de rhizobactéries fixatrices d'azote a été testé en condition de stress osmotique, c'est-à-dire en ajoutant 0,8 M de NaCI dans le milieu de culture à un instant T = 0. [0112] Les résultats, présentés au tableau 9, indiquent que l'ajout de l'extrait de vinasse de mélasse de betterave préparé selon l'exemple le) en condition non stressante a permis une légère stimulation de la croissance des bactéries fixatrices d'azote en augmentant la concentration bactérienne de 0,5 logs après 48 h d'incubation. [0111] The complex of rhizobacteria fixing nitrogen was tested under conditions of osmotic stress, that is to say by adding 0.8 M of NaCl to the culture medium at a time T = 0. [0112] The results, presented in Table 9, indicate that the addition of the extract of beet molasses stillage prepared according to example 1c) under condition non-stressful allowed a slight stimulation of the growth of nitrogen-fixing bacteria by increasing the bacterial concentration by 0.5 logs after 48 h of incubation.
[0113] L'ajout de NaCI à 0,8 M dans le milieu de culture a bien permis de reproduire les conditions d'un stress salin avec une perte de 5,4 logs de la concentration bactérienne après 48 h d'incubation par rapport à la condition sans sel. The addition of 0.8 M NaCl in the culture medium made it possible to reproduce the conditions of a salt stress with a loss of 5.4 logs of the bacterial concentration after 48 h of incubation compared to under the condition without salt.
[0114] En condition de stress salin, l'ajout de de l'extrait de vinasse de mélasse de betterave a permis une augmentation de 5,6 logs de la concentration bactérienne après 48 h d'incubation. La concentration bactérienne obtenue est proche de celle observée dans le milieu sans sel avec ajout de de l'extrait de vinasse de mélasse de betterave, autour de 108 UFC/mL. [0114] Under conditions of salt stress, the addition of extract of beet molasses vinasse allowed an increase of 5.6 logs in the bacterial concentration after 48 h of incubation. The bacterial concentration obtained is close to that observed in the medium without salt with the addition of extract of beet molasses stillage, around 10 8 CFU / mL.
Tableau 9 [0115] Exemple 2) h) : Effet de la glycine betaïne en comparaison avec l'effet de polvamines issues d'un extrait dA' scophyllum Nodosum Table 9 Example 2) h): Effect of glycine betaine in comparison with the effect of polvamines obtained from an extract of scophyllum Nodosum
[0116] La souche Azotobacter chroococcum a été testée en condition de stress osmotique, c'est-à-dire en ajoutant 0,3 M NaCI dans le milieu de culture à un instant T =0. The Azotobacter chroococcum strain was tested under conditions of osmotic stress, that is to say by adding 0.3 M NaCl to the culture medium at a time T = 0.
[0117] La glycine bétaïne, connue comme osmo-protecteur, a été testée en comparaison avec l'extrait ô'Ascophyllum nodosum préparé selon l'exemple la). Glycine betaine, known as an osmoprotector, was tested in comparison with the extract ô'Ascophyllum nodosum prepared according to example la).
[0118] Les résultats, présentés au tableau 10, indiquent que l'ajout de l'extrait ô'Ascophyllum nodosum préparé selon l'exemple la) en condition non stressante a permis une légère stimulation de la croissance des bactéries en augmentant la concentration bactérienne de 0,2 log après 72 h d'incubation. The results, presented in Table 10, indicate that the addition of the extract ô'Ascophyllum nodosum prepared according to example la) in non-stressful condition allowed a slight stimulation of the growth of bacteria by increasing the bacterial concentration. 0.2 log after 72 h of incubation.
[0119] L'ajout de NaCI à 0,3 M dans le milieu de culture sans glycine bétaïne ni extrait, a induit une perte de 4,8 logs de la concentration bactérienne après 72 h d'incubation par rapport à la condition sans sel. [0120] En condition de stress salin, l'ajout de de l'extrait de ô'Ascophyllum nodosum a permis une augmentation de 2,9 logs de la concentration bactérienne après 72 h d'incubation par rapport à la condition stressante, alors que l'ajout de glycine bétaïne en condition stressante provoque une perte de 0,8 logs de la concentration bactérienne après 72h d'incubation par rapport à la condition stressante. The addition of 0.3 M NaCl in the culture medium without glycine betaine or extract, induced a loss of 4.8 logs of the bacterial concentration after 72 h of incubation compared to the condition without salt. . [0120] Under conditions of saline stress, the addition of extract of ô'Ascophyllum nodosum allowed an increase of 2.9 logs of the bacterial concentration after 72 h of incubation compared to the stressful condition, whereas the addition of glycine betaine under stressful conditions causes a loss of 0.8 logs of the bacterial concentration after 72 hours of incubation compared to the stressful condition.
[0121] Les polyamines permettent de stabiliser Azotobacter chroococcum face à un stress osmotique, alors qu'aucune stabilisation n'est observée avec la glycine bétaïne en condition de stress osmotique. Tl Polyamines make it possible to stabilize Azotobacter chroococcum in the face of osmotic stress, whereas no stabilization is observed with glycine betaine under conditions of osmotic stress. Tl
Tableau 10 Table 10
[0122] Exemple 3 : Effet d'une composition biofertilisante à base d'un extrait à'Ascoohyllum nodosum sur la croissance tomates cultivées en plein champ [0122] Example 3: Effect of a biofertilizer composition based on an extract of Ascoohyllum nodosum on the growth of tomatoes grown in the open field
[0123] L'essai a été réalisé sur une culture de tomates de l'espèce So/anum lycopersicum. Le dispositif expérimental comprend des modalités à 5 répétitions. L'extrait ô'Ascophyllum nodosum a été incorporé à raison de lg/T dans une solution de PK (5-30), qui a été enrichie avec la souche Bacillus Hcheniformis. Les traitements suivants sont appliqués au stade 3 feuilles des plants de tomates: - un témoin ne comprenant pas de fertilisant, The test was carried out on a crop of tomatoes of the species So / anum lycopersicum. The experimental set-up includes modalities with 5 repetitions. The Ascophyllum nodosum extract was incorporated in an amount of lg / T in a solution of PK (5-30), which was enriched with the strain Bacillus Hcheniformis. The following treatments are applied at the 3 leaf stage of tomato plants: - a control not including fertilizer,
- la solution de PK 5 - 30, appliquée à raison de 500 kg/ha, - the solution of PK 5 - 30, applied at a rate of 500 kg / ha,
- la solution de PK 5 - 30 avec la souche Bacillus Hcheniformis, appliquée à raison de 500 kg/ha correspondant à 5.108 UFC/ha, ou - la solution de PK 5 - 30 avec la souche Bacillus Hcheniformis, et avec l'extrait ù'Ascophyiium nodosum, 500 Kg/ha correspondant à 5.108 UFC/ha. - the solution of PK 5 - 30 with the strain Bacillus Hcheniformis, applied at a rate of 500 kg / ha corresponding to 5.10 8 CFU / ha, or - the solution of PK 5 - 30 with the strain Bacillus Hcheniformis, and with the extract ù'Ascophyiium nodosum, 500 Kg / ha corresponding to 5.10 8 CFU / ha.
[0124] Le rendement en T/ha a été mesuré. Les résultats sont présentés au tableau 11. The yield in T / ha was measured. The results are shown in Table 11.
Tableau 11 Table 11
[0125] La solution PK 5-30 seule a amélioré le rendement en tomates de 4,2% par rapport au témoin. La solution PK 5-30 + B. Hcheniformis a peu amélioré le rendement, soit 5,2% par rapport au témoin. Par contre l'utilisation de l'extrait en combinaison avec la solution PK 5-30 + bactéries a permis une amélioration du rendement par rapport au témoin de plus de 14,8%. Par rapport à la solution[0125] The PK 5-30 solution alone improved the tomato yield by 4.2% compared to the control. The PK 5-30 + B. Hcheniformis solution slightly improved the yield, ie 5.2% compared to the control. On the other hand, the use of the extract in combination with the PK 5-30 + bacteria solution allowed an improvement in the yield compared to the control of more than 14.8%. Compared to the solution
PK 5-30 seule, l'incorporation de l'extrait a permis d'augmenter le rendement tomates de près de 17,5 T/ha, soit une augmentation de 9,2%. PK 5-30 alone, the incorporation of the extract made it possible to increase the tomato yield by nearly 17.5 T / ha, ie an increase of 9.2%.
[0126] Exemple 4 : Compositions biofertilisantes ou compositions destinées à l'alimentation animale selon l'invention [0127] Exemple 4 a) : Amendement calcaire Tableau 12 [0126] Example 4: Biofertilizing compositions or compositions intended for animal feed according to the invention [0127] Example 4 a): Limestone amendment Table 12
Tableau 13 Tableau 14 Table 13 Table 14
G01281 Exemple 4 bl : Amendement organique et supports de cultureG01281 Example 4 bl: Organic amendment and growing media
Tableau 15 [0129] Exemple 4 c : engrais racinaires Table 15 [0129] Example 4 c: root fertilizers
[0130] Le tableau 16 ci-après décrit la composition d'un engrais racinaire de type N P. Table 16 below describes the composition of an N P type root fertilizer.
Tableau 16 Table 16
[0131] Le tableau 17 ci-après décrit la composition d'un engrais racinaire de type NPK et Oxyde de magnésium. Table 17 below describes the composition of a root fertilizer of NPK and magnesium oxide type.
Tableau 17 Table 17
[0132] Exemple 4 d) : solutions nutritives racinaires [0132] Example 4 d): root nutrient solutions
[0133] Le tableau 18 ci-après décrit la composition d'une solution de type NPK et magnésium. Tableau 18 Table 18 below describes the composition of a solution of NPK and magnesium type. Table 18
[0134] Le tableau 19 ci-après décrit la composition d'une solution de type N, Calcium, Magnésium. Tableau 19 Table 19 below describes the composition of a solution of N, Calcium, Magnesium type. Table 19
[0135] Exemple 4 e) : Compositions destinées à l'alimentation animale[0135] Example 4 e): Compositions intended for animal feed
[0136] Le tableau 20 ci-après décrit la composition d'un probiotique. [0136] Table 20 below describes the composition of a probiotic.
Tableau 20 [0137] Le tableau 21 ci-après décrit la composition d'un conservateur d'ensilage. Table 20 Table 21 below describes the composition of a silage preservative.
Tableau 21 Table 21
BIBLIOGRAPHIE BIBLIOGRAPHY
[1] "Po/yamines in Ecklonia maxima and their effects on plant growtii', Heino Benoni Papenfus, the Research Centre for Plant Growth and Development School of Biological and Conservation Sciences University of KwaZulu-Natal Pietermaritzburg Decem ber 2011. [1] "Po / yamines in Ecklonia maxima and their effects on plant growtii ', Heino Benoni Papenfus, the Research Center for Plant Growth and Development School of Biological and Conservation Sciences University of KwaZulu-Natal Pietermaritzburg Decem ber 2011.
[2] « Vinasse de mélasse de betterave », Fiche n° 10 du Comité National des Coproduits. [2] “Beet molasses vinasse”, Sheet n ° 10 of the National Co-products Committee.
[3] « La vinasse de mélasse de betteraves pour les ruminants », INRA Prod.Anim., 1989, 2 (4), 245-248, J.L. TROCCON, C. DEMARQUILLY. [3] "Beet molasses stillage for ruminants", INRA Prod.Anim., 1989, 2 (4), 245-248, J.L. TROCCON, C. DEMARQUILLY.

Claims

REVENDICATIONS
1. Utilisation d'un extrait d'algues et/ou de plantes, ledit extrait contenant des polyamines et/ou de la tyramine, pour stabiliser des micro-organismes face à un stress environnemental. 1. Use of an extract of algae and / or plants, said extract containing polyamines and / or tyramine, for stabilizing microorganisms in the face of environmental stress.
2. Utilisation selon la revendication 1, dans laquelle l'extrait d'algues et/ou de plantes est un extrait d'algues brunes, de préférence un extrait d'algues brunes de la famille des Fucacées, et/ou un extrait de vinasse de mélasse de betteraves. 2. Use according to claim 1, wherein the algae and / or plant extract is an extract of brown algae, preferably an extract of brown algae of the Fucaceae family, and / or an extract of vinasse. of beet molasses.
3. Utilisation selon l'une quelconque des revendications 1 ou 2, dans laquelle les micro-organismes sont contenus dans une composition, telle qu'une composition fertilisante, une composition destinée à l'alimentation animale ou une composition destinée à la conservation d'ensilage. 3. Use according to any one of claims 1 or 2, wherein the microorganisms are contained in a composition, such as a fertilizing composition, a composition intended for animal feed or a composition intended for the preservation of. silage.
4. Utilisation selon la revendication 3, dans laquelle ladite composition comprend en outre un engrais et/ou un amendement. 4. Use according to claim 3, wherein said composition further comprises a fertilizer and / or an amendment.
5. Utilisation selon l'une quelconque des revendications 1 à 4, dans laquelle les micro-organismes sont choisis parmi (i) les bactéries fixatrices de l'azote atmosphérique, telles que Azotobacter ou Azospirillum (ii) les rhizobactéries favorisant la croissance des plantes (PGPR ou Plant Growth Promoting Rhizobacteria), (iii) les bactéries solubilisatrices du phosphore telles que BaciHus amyloliquefaciens, (iv) les bactéries phytoprotectrices des racines (PGPR) capables de s'opposer à l'activité d'agents pathogènes telles que BaciHus subti/is ou Pseudomonas spp., (v) les bactéries productrices de phytohormones telles que BaciHus amyloliquefaciens ou BaciHus radicoia, (vi) les bactéries impliquées dans le processus de minéralisation de la matière organique telles que LactobaciHus rhamnosus ou LactobaciHus faciminis, (vii) les bactéries solubilisatrices de fer telles que Pseudomonas spp., (viii) les bactéries solubilisatrices de la silice, (ix) les bactéries oxydatrices du soufre, (x) les bactéries lactiques telles que LactobaciHus spp., Lactococcus spp., Bifidobacterium spp., (xi) les bactéries du genre Enterococcus spp., (xii) les bactéries du genre Pediococcus spp., (xiii) les bactéries du genre BaciHus Hcheniformis, (xiv) les champignons mycorhizes tel que Rhizophagus irregularis, (xv) les levures du genre Saccharomyces cerevisiae et (xvi) un mélange d'au moins deux micro-organismes choisis parmi (i) à (xv). 5. Use according to any one of claims 1 to 4, wherein the microorganisms are chosen from (i) bacteria fixing atmospheric nitrogen, such as Azotobacter or Azospirillum (ii) rhizobacteria promoting plant growth. (PGPR or Plant Growth Promoting Rhizobacteria), (iii) phosphorus solubilizing bacteria such as BaciHus amyloliquefaciens, (iv) root protection bacteria (PGPR) capable of opposing the activity of pathogens such as BaciHus subti / is or Pseudomonas spp., (v) bacteria producing phytohormones such as BaciHus amyloliquefaciens or BaciHus radicoia, (vi) bacteria involved in the mineralization process of organic matter such as LactobaciHus rhamnosus or LactobaciHus faciminis, (vii) iron solubilizing bacteria such as Pseudomonas spp., (viii) silica solubilizing bacteria, (ix) sulfur oxidizing bacteria, (x) lactic acid bacteria such as LactobaciHus spp., Lactococcus spp., Bifidobacterium spp., (Xi) bacteria of the genus Enterococcus spp., (Xii) bacteria of the genus Pediococcus spp., (Xiii) bacteria of the genus BaciHus Hcheniformis, (xiv) mycorrhizal fungi such as Rhizophagus irregularis, (xv) yeasts of the genus Saccharomyces cerevisiae and (xvi) a mixture of at least two microorganisms chosen from (i) to (xv).
6. Utilisation selon l'une quelconque des revendications 3 à 5, dans laquelle les micro-organismes sont contenus dans la composition en une quantité allant de 102 à 1050 UFC par tonne de composition, de préférence allant de 105 à 1020 UFC par tonne de composition, de préférence environ 1011 UFC par tonne de composition. 6. Use according to any one of claims 3 to 5, in which the microorganisms are contained in the composition in an amount ranging from 10 2 to 10 50 CFU per tonne of composition, preferably ranging from 10 5 to 10 20. CFU per tonne of composition, preferably about 10 11 CFU per tonne of composition.
7. Utilisation selon l'une quelconque des revendications 1 à 6, dans laquelle les polyamines sont choisies parmi la cadavérine, la spermidine, la spermine et/ou la putrescine. 7. Use according to any one of claims 1 to 6, in which the polyamines are chosen from cadaverine, spermidine, spermine and / or putrescine.
8. Composition comprenant : 8. Composition comprising:
(a) un extrait d'algues et/ou de plantes, ledit extrait contenant des polyamines et/ou de la tyramine, et (b) des micro-organismes choisis parmi : (i) les bactéries fixatrices de l'azote atmosphérique, telles que Azotobacter ou Azospirillum (ii) les rhizobactéries favorisant la croissance des plantes (PGPR ou Plant Growth Promoting Rhizobacteria), (iii) les bactéries solubilisatrices du phosphore telles que BaciHus amyloliquefaciens, (iv) les bactéries phytoprotectrices des racines (PGPR) capables de s'opposer à l'activité d'agents pathogènes telles que BaciHus subti/is ou Pseudomonas spp., (v) les bactéries productrices de phytohormones telles que BaciHus amyloliquefaciens ou BaciHus radicoia, (vi) les bactéries impliquées dans le processus de minéralisation de la matière organique telles que LactobaciHus rhamnosus ou LactobaciHus faciminis, (vii) les bactéries solubilisatrices de fer telles que Pseudomonas spp., (viii) les bactéries solubilisatrices de la silice, (ix) les bactéries oxydatrices du soufre, (x) les bactéries lactiques telles que LactobaciHus spp., Lactococcus spp., Bifidobacterium spp., (xi) les bactéries du genre Enterococcus spp., (xii) les bactéries du genre Pediococcus spp., (xiii) les bactéries du genre BaciHus Hcheniformis, (xiv) les champignons mycorhizes tel que Rhizophagus irregularis, (xv) les levures du genre Saccharomyces cerevisiae et (xvi) un mélange d'au moins deux micro-organismes choisis parmi (i) à (xv). (a) an extract of algae and / or plants, said extract containing polyamines and / or tyramine, and (b) microorganisms chosen from: (i) bacteria fixing atmospheric nitrogen, such as Azotobacter or Azospirillum (ii) rhizobacteria promoting plant growth (PGPR or Plant Growth Promoting Rhizobacteria), (iii) phosphorus solubilizing bacteria such as BaciHus amyloliquefaciens, (iv) phytoprotective bacteria of the roots (PGPR) capable of s '' oppose the activity of pathogens such as BaciHus subti / is or Pseudomonas spp., (v) bacteria producing phytohormones such as BaciHus amyloliquefaciens or BaciHus radicoia, (vi) bacteria involved in the mineralization process of the organic matter such as LactobaciHus rhamnosus or LactobaciHus faciminis, (vii) iron solubilizing bacteria such as Pseudomonas spp., (viii) silica solubilizing bacteria, (ix) bacteria o sulfur xidators, (x) lactic acid bacteria such as LactobaciHus spp., Lactococcus spp., Bifidobacterium spp., (Xi) bacteria of the genus Enterococcus spp., (Xii) bacteria of the genus Pediococcus spp., (Xiii) bacteria of the genus BaciHus Hcheniformis, (xiv) mycorrhizal fungi such as Rhizophagus irregularis, (xv) yeasts of the genus Saccharomyces cerevisiae and (xvi) a mixture of at least two microorganisms chosen from (i) to (xv).
9. Composition selon la revendication 8, comprenant de 102 à 1050 UFC de micro¬ organismes (b) par tonne de composition, de préférence de 105 à 1020 UFC de micro-organismes (b) par tonne de composition, de préférence environ 1011 UFC de micro-organismes (b) par tonne de composition. 9. The composition of claim 8, comprising 10 2 to 10 50 CFU of micro ¬ organisms (b) per ton of composition, preferably October 5 to October 20 CFU of microorganisms (b) per ton of composition, preferably about 10 11 CFU of microorganisms (b) per tonne of composition.
10. Composition selon l'une quelconque des revendications 8 ou 9, comprenant de 0,1 à 100 grammes de polyamines et/ou de tyramine (a) par tonne de composition (g/T), de préférence de 0,4 à 10 g/T de polyamines et/ou de tyramine (a) par tonne de composition (g/T). 10. Composition according to any one of claims 8 or 9, comprising from 0.1 to 100 grams of polyamines and / or tyramine (a) per tonne of composition (g / T), preferably from 0.4 to 10 g / T of polyamines and / or tyramine (a) per tonne of composition (g / T).
11. Composition selon l'une quelconque des revendications 8 à 10, comprenant en outre (c) un engrais et / ou un amendement. 11. A composition according to any one of claims 8 to 10, further comprising (c) a fertilizer and / or an amendment.
12. Composition selon l'une quelconque des revendications 8 à 11, dans laquelle l'extrait d'algues et/ou de plantes est un extrait d'algues brunes, par exemple un extrait d'algues brunes de la famille des Fucacées, et/ou un extrait de vinasse de mélasse de betteraves. 12. Composition according to any one of claims 8 to 11, in which the algae and / or plant extract is an extract of brown algae, for example an extract of brown algae of the Fucaceae family, and / or an extract of beet molasses vinasse.
13. Composition selon l'une quelconque des revendications 8 à 12, dans laquelle les polyamines sont choisies parmi la tyramine, la cadaverine, la spermidine, la spermine et/ou la putrescine. 13. Composition according to any one of claims 8 to 12, in which the polyamines are chosen from tyramine, cadaverine, spermidine, spermine and / or putrescine.
14. Procédé pour fertiliser une plante caractérisé en ce qu'il comprend l'apport à ladite plante d'une composition selon l'une quelconque des revendications 8 à 13. 14. A method for fertilizing a plant characterized in that it comprises supplying said plant with a composition according to any one of claims 8 to 13.
15. Procédé selon la revendication 14, dans lequel la composition est apportée à ladite plante en une quantité allant de 0,5 à 1500 kg /ha, de préférence allant de 1 à 1000 kg/ha. 15. The method of claim 14, wherein the composition is supplied to said plant in an amount ranging from 0.5 to 1500 kg / ha, preferably ranging from 1 to 1000 kg / ha.
EP21732467.2A 2020-05-19 2021-05-18 Use of polyamines, tyramine and/or a plant extract containing same to stabilise microorganisms Pending EP4153318A1 (en)

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