WO2018167347A2 - Métodos y composiciones para mejorar la salud de plantas y/o rendimiento de plantas - Google Patents
Métodos y composiciones para mejorar la salud de plantas y/o rendimiento de plantas Download PDFInfo
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- WO2018167347A2 WO2018167347A2 PCT/ES2018/070193 ES2018070193W WO2018167347A2 WO 2018167347 A2 WO2018167347 A2 WO 2018167347A2 ES 2018070193 W ES2018070193 W ES 2018070193W WO 2018167347 A2 WO2018167347 A2 WO 2018167347A2
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/34—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom
- A01N43/40—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom six-membered rings
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
- A01N63/22—Bacillus
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
- A01N63/27—Pseudomonas
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
- A01N63/28—Streptomyces
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
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- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/065—Azotobacter
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
- C12R2001/125—Bacillus subtilis ; Hay bacillus; Grass bacillus
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/38—Pseudomonas
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/41—Rhizobium
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/465—Streptomyces
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/885—Trichoderma
Definitions
- the present invention relates to methods and compositions for increasing a plant's growth characteristic, increasing the efficiency of the use of nutrients from a plant, or improving the ability of a plant to overcome biotic or abiotic aggression comprising applying a composition comprising a fungal mycelial extract comprising piperidine and / or an analogue thereof, and / or, a salt thereof, or any combination thereof to a plant, part of a plant, or a plant propagation material.
- Plants are constantly challenged by biotic and abiotic aggressions including microbial pathogens, such as bacteria, fungi, and viruses, and / or changing climatic factors. Therefore, plants need to regulate and adapt their cellular metabolism to optimize the distribution of resources between growth, storage, or production of defense compounds.
- microbial pathogens such as bacteria, fungi, and viruses
- Plants have developed a mutifhasic and complex network of defense responses, including preformed physical barriers (for example, cortex and cuticle) and inducible perception systems, both on the cell surface and intracellularly, with the ability to distinguish between "own” and “not own”, “damaged own” or “modified own”, which is the basis of immunity and is evolutionarily conserved.
- the main triggers of plant immunity are the so-called molecular patterns associated with microbes or pathogens (MAMP / PAMP) or patterns associated with endogenous danger (DAMP) that they induce de novo production of antimicrobial defense proteins and metabolites, including phenolic compounds, terpenes, alkaloids and non-protein amino acids.
- PAMP / MAMP / DAMP The immunity triggered by PAMP / MAMP / DAMP protects plants against most pathogens and reflects baseline resistance.
- PAMP / MAMP / DAMP are derivatives of major structural polysaccharides of the bacterial, fungal or plant cell wall, for example, lipopolysaccharides (LPS) of gram negative bacteria and peptidoglycan (PGN) of gram + bacteria, chitin and fungal glucans, or pectin from plants.
- LPS lipopolysaccharides
- PPN peptidoglycan
- proteins of bacterial or fungal origin trigger an immune response in plant cells. The most prominent examples are the flagelin bacterial proteins (the main constituent of the bacterial scourge), thermosetting elongation factor (EF-Tu) and cold shock (CSP).
- the present invention overcomes defects in the art by providing novel methods to increase resistance to biotic and abiotic aggressions.
- Figure 1 shows photographs of strawberries and onions collected after treatment with a fungal mycelial extract (PRBT) (upper panel) versus strawberries and control onions (untreated) (lower panel).
- PRBT fungal mycelial extract
- Figure 2 shows photographs of soybean plants inoculated with Sclerotinia sclerotiorum and treated with PRBT (right panel) compared to control plants inoculated with Sclerotinia sclerotiorum (middle panel) and inoculated control treated plants of reference (left panel).
- Figure 3 shows photographs of zucchini plants infected with New Delhi tomato curly virus treated with PRBT (panel B) compared to control (untreated) plants (panel A).
- Immunodetection membranes of 34 zucchini plants treated with PRBT (panel D) compared to control (untreated) plants (panel C) are shown where plants infected with the New Delhi tomato leaf curl virus are highlighted with a rectangle continuous line with impressions of stem tissue (left) and leaf (right) while negative (left) and positive (right) controls are indicated with a dashed rectangle.
- compositions for increasing a growth characteristic of a plant that comprises applying a composition comprising a fungal mycelial extract comprising piperidine and / or an analogue thereof (eg, 6-oxopiperidine-2- acid carboxylic), a salt thereof, or any combination thereof (referred to herein as "PRBT") to a plant or part thereof.
- a fungal mycelial extract comprises 6-oxopiperidine-2-carboxylic acid and / or an analog and / or a salt thereof (for example, 6-oxopiperidine-2-carboxylate).
- a further embodiment discloses increasing the efficiency of nutrient use of a plant comprising applying a composition comprising a fungal mycelial extract comprising piperidine and / or an analogue thereof (for example, 6-oxopiperidine-2-carboxylic acid ), a salt thereof, or any combination thereof to a plant or part thereof.
- a fungal mycelial extract comprises 6-oxopiperidine-2-carboxylic acid and / or an analog and / or a salt thereof (for example, 6-oxopiperidine-2-carboxylate).
- a further embodiment discloses increasing tolerance or resistance to biotic and / or abiotic aggressions in a plant or part thereof which comprises applying a composition comprising a fungal mycelial extract comprising piperidine and / or an analogue thereof (by for example, 6- oxopiperidine-2-carboxylic acid), a salt thereof, or any combination thereof to a plant or part thereof.
- a fungal mycelial extract comprises 6-oxopiperidine-2-carboxylic acid and / or an analog and / or a salt thereof (for example, 6-oxopiperidine-2-carboxylate).
- the fungal mycelial extract may further comprise peptides, proteins, carbohydrates and / or sugars.
- a composition of the invention may further comprise a surfactant, a humectant, an adjuvant, an antioxidant, a preservative, a plant macronutrient, a plant micronutrient, a plant growth regulator, a pesticide, a fungicide, an antiviral, an antibacterial, and / or a herbicide.
- a measurable value such as an amount or concentration or the like
- a measurable value such as an amount or concentration or the like
- approximately X where X is the measurable value, is intended to include X, as well as variations of ⁇ 10%, ⁇ 5%, ⁇ 1%, ⁇ 0.5%, or even ⁇ 0.1% of X.
- An interval provided herein for a measurable value may include any other range and / or individual value therein.
- phrases such as “between X and Y” and “between approximately X and Y” should be construed to include X and Y.
- the term “understand”, “comprises” and “comprising” as used herein, specifies the presence of features, integers, stages, operations, elements and / or components expressed, but does not exclude the presence or addition of one or more other characteristics, integers, stages, operations, elements and / or components and / or groups thereof.
- the transitional phrase “consist essentially of” means that the scope of a claim should be construed to cover the specified materials or stages listed in the claim and those that do not materially affect the characteristic (s). ) basic (s) and novel (s) of the claimed invention. Therefore, the term “consist essentially of” when used in a claim of this invention is not intended to be construed as being equivalent to "understanding.”
- the terms “increase”, “increase”, “increase”, “increase”, “increase”, “increase”, “increase”, “increase”, and “increase” describe a elevation of at least about 25%, 50%, 75%, 100%, 150%, 200%, 300%, 400%, 500% or more compared to a control.
- the terms “reduce”, “reduced”, “that reduce”, “reduction”, “decrease”, and “decrease” describe, for example, a decrease of at least about 5%, 10%, 15%, 20%, 25%, 35%, 50%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% compared to a control.
- the reduction may produce no or essentially no (ie, an insignificant amount, for example, less than about 10%, or even 5%) activity or detectable amount.
- growth characteristic refers to any plant trait associated with growth, for example, biomass, yield, size / weight of inflorescence, fruit yield, fruit quality, fruit size, production of seed, leaf tissue weight, number of nodulations, nodulation mass, nodulation activity, number of seed heads, number of shoots, number of flowers, numbers of tubers, mass of tubers, bulb mass, number of seeds, total mass of seeds, leaf rate, emergence, sprout emergence rate, seedling emergence rate and combinations thereof. Therefore, in some aspects, an increased growth characteristic may be increased fruit production, increased inflorescence production, increased fruit quality and / or increased biomass compared to a control plant or part thereof to which the compositions of the invention have not been applied.
- fertilizer use efficiency refers to the ability of a plant to use available nutrients.
- use efficiency of nutrients can be defined in terms of total absorption of nutrients (nutrient concentration in plant tissue ⁇ total biomass) and / or yield per unit of nutrient applied.
- abiotic aggression refers to external, non-living factors that may cause harmful effects to plants.
- abiotic aggression includes, but is not limited to, cold temperature that produces freezing, cold or cold temperature, heat or high temperatures, drought, high light intensity, low light intensity, salinity, flood (excess water / waterlogging), ozone, and / or combinations thereof.
- the parameters for aggression factors abiotic are species specific and even variety specific and therefore vary widely according to the species / variety exposed to abiotic aggression. As one species may be severely affected by a high temperature of 23 ° C, another species may not be affected until at least 30 ° C, and the like.
- compositions comprising a fungal mycelial extract comprising piperidine and / or an analogue thereof (eg, 6- oxopiperidine-2-carboxylic acid), a salt thereof (for example, 6-oxopiperidine-2-carboxylate) or any combination thereof (also referred to herein as "PBRT") may increase a growth characteristic, nutrient use efficiency, and / or tolerance / resistance to abiotic and / or biotic aggression of the plant or part thereof.
- a fungal mycelial extract comprising piperidine and / or an analogue thereof (eg, 6- oxopiperidine-2-carboxylic acid), a salt thereof (for example, 6-oxopiperidine-2-carboxylate) or any combination thereof (also referred to herein as "PBRT”)
- PBRT any combination thereof
- the present invention provides a composition to increase a growth characteristic of a plant or part thereof, to increase the efficiency of nutrient use of a plant or part thereof, and / or to increase the tolerance to abiotic aggression and / or biotic aggression of a plant or part thereof, the composition comprises an effective amount of a fungal mycelial extract comprising piperidine and / or an analogue thereof (for example, acid 6- oxopiperidine-2-carboxylic acid), a salt thereof (for example, 6-oxopiperidine-2-carboxylate) or any combination thereof.
- a fungal mycelial extract comprising piperidine and / or an analogue thereof (for example, acid 6- oxopiperidine-2-carboxylic acid), a salt thereof (for example, 6-oxopiperidine-2-carboxylate) or any combination thereof.
- the invention provides a composition comprising a fungal mycelial extract comprising piperidine and / or an analog thereof (for example, 6-oxopiperidine-2-carboxylic acid), a salt thereof (for example , 6- oxopiperidine-2-carboxylate) or any combination thereof.
- a fungal mycelial extract comprises 6- oxopiperidine-2-carboxylic acid and / or an analogue and / or a salt thereof (for example, 6- oxopiperidine-2-carboxylate).
- a composition may comprise an amount of piperidine and / or an analog thereof (for example, 6-oxopiperidine-2-carboxylic acid), a salt thereof (for example, 6-oxopiperidine-2- carboxylate) in a range from about 0.1 grams (g) per liter (I) to about 50 g / l of the composition.
- piperidine and / or an analog thereof for example, 6-oxopiperidine-2-carboxylic acid
- a salt thereof for example, 6-oxopiperidine-2- carboxylate
- a composition of the invention may comprise piperidine and / or an analog thereof and / or a salt thereof in a range from about 0.1 g / l to about 1 g / l , from approximately 0.1 g / l to about 5 g / l, from about 0.1 g / l to about 10 g / l, from about 0.1 g / l to about 15 g / l, from about 0.1 g / l l to about 20 g / l, from about 0.1 g / l to about 30 g / l, from about 0.1 g / l to about 40 g / l, from about 0.5 g / l to about 1 g / l, from about 0.5 g / l to about 5 g l, from about 0.5 g / l to about 10 g / l, from about 0.5 g / l to about 20 g
- a composition of the invention may comprise piperidine and / or an analog thereof or a salt thereof. in an amount of about 0.01, 0.05, 0.1, 0.5, 1, 1, 5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5 , 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 1 1, 1 1, 5, 12, 12.5, 13, 13.5 , 14, 14.5, 15, 15.5, 16, 16.5, 17, 17.5, 18, 18.5, 19, 19.5, 20, 21, 22, 23, 24, 25, 26 , 27, 28, 29, 30, 31, 32, 33, 34, 35, 40, 45, or 50 g per liter of the composition, or any interval or value therein.
- the amount of piperidine and / or analog thereof, or salt thereof in a composition of the invention may be in a range of about 0.1 g / l to about 5 g / l (per example, about 0.01, 0.05, 0.1, 0.5, 1, 1, 5, 2, 2.5, 3, 3.5, 4, 4.5, 5 g / l of the composition or any interval or value in it).
- a composition of the invention may comprise a fungal mycelial extract (eg, "PBRT") in an amount from about 0.01% to about 100% w / w of the composition. Therefore, in some embodiments, a composition of the invention may comprise a fungal mycelial extract in an amount from about 0.01% to about 0.1%, from about 0.01% to about 1% , from about 0.01% to about 3%, from about 0.01% to about 5%, from about 0.01% to about 10%, from about 0.01% to about 15%, from about 0.01% to about 20%, from about 0.01% to about 25%, from about 0.01% to about 30%, from about 0.01% to about 35%, from about 0.01% to about 40%, from about 0.01% to about 45%, from about 0.01% to about 50%, from about 0.01 % until approximately 60%, from about 0.01% to about 70%, from about 0.01% to about 80%, from about 0.01% to about 90%, from about 0.01 % to about 95%, from about 0.1%
- a composition of the invention may comprise a fungal mycelial extract in an amount of about 0.01%, 0.05%, 0.1%, 0.5%, 1%, 1.5% , 2%, 2.5%, 3, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8% , 8.5%, 9%, 9.5%, 10%, 1 1%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21% , 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, or 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% w / w of the composition, or any interval or value therein.
- an effective amount of a fungal mycelial extract is an amount sufficient to increase a growth characteristic of a plant or part thereof, increase the efficiency in the use of nutrients of a plant or part thereof, and / or increase tolerance / resistance to abiotic aggression and / or biotic aggression of a plant or part thereof.
- an effective amount of a fungal mycelial extract in a composition may be from about 0.005 g per liter to about 150 g per liter of composition.
- an effective amount of a fungal mycelial extract may be from about 0.005 g / l to about 1 g / l, from about 0.005 g / l to about 5 g / l, from about 0.005 g / l to about 10 g / l, from about 0.005 g / l to about 15 g / l, from about 0.005 g / l to about 20 g / l, from about 0.005 g / l to about 30 g / l, from about 0.01 g / l to about 1 g / l, from about 0.01 g / l to about 5 g / l, from about 0.01 g / l to about 10 g / l, from about 0.01 g / l to about 15 g / l, from about 0.01 g / l to about 20 g / l, from about 0.01 g / / /
- an effective amount of a fungal mycelial extract in a composition may be about 0.005, 0.01, 0.05, 0.1, 0.5, 1, 1, 5, 2, 2 , 5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5 , 1 1, 1 1, 5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16, 16.5, 17, 17.5, 18, 18.5 , 19, 19.5, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 40, 45, 50, 55, 60, 65 , 70, 75, 80, 85, 90, 95, 100, 105, 1 10, 1 15, 120, 125, 130, 135, 140, 145, or 150 g per liter of the composition, or any interval or value in the same.
- An extract of the invention may comprise an analog and / or piperidine salt (for example, -oxypiperidine-2-carboxylic acid, 6-oxopiperidine-2-carboxylate).
- an "analog" of a respective compound may include, but is not limited to, a tautomer and / or an isomer (for example, a stereoisomer and / or a structural isomer) of the respective compound.
- An “analog" of a respective compound may also include a compound having a similar or equal central structure (eg, a central structure comprising piperidine) than the respective compound and optionally one or more substituent (s) that may be different than a substituent of the respective compound and / or in a different position purchased with a substituent of the respective compound.
- a similar or equal central structure eg, a central structure comprising piperidine
- substituents that may be different than a substituent of the respective compound and / or in a different position purchased with a substituent of the respective compound.
- an analog of a compound of the present invention may have a structure represented by formula I:
- R 1 is selected from the group consisting of H and -COOH
- R 1 is -COOH
- R 3 is selected from the group consisting of H, NH 2 and CC 4 alkyl.
- 6-oxopiperidine-2-carboxylic acid is in the form of a salt
- 6-oxopiperidine-2-carboxylate is referred to herein.
- An IUPAC name for 6-oxopiperidine-2-carboxylate includes 6-hydroxy acid.
- 6-oxopiperidine-2-carboxylate is also known as 6-oxo » pipendin-2-carboxylic acid, adipo - ⁇ .
- E-iactam cyclic alpha-aminoadipic acid, 8-Gxopiperidine -2-carboxylate, cyclic a-aminoadipate, cyclic a-aminoadipic acid, cyclic ai-aminoadipate, cyclic a-aminoadipate, cyclic a-aminoadipic acid, or 6 ⁇ oxo-pipecoic acid.
- An extract of the invention may comprise an analogue and / or piperidine salt (for example, -oxypiperidine-2-carboxylic acid, 6-oxopiperidine-2-carboxylate).
- piperidine salt for example, -oxypiperidine-2-carboxylic acid, 6-oxopiperidine-2-carboxylate.
- a piperidine analog can include, but is not limited to 6-oxopiperidine-2-carboxylic acid, 6- hydroxypiperidine-2-carboxylic acid; 6-oxopiperidine-3-carboxylic acid; 6- hydroxypiperidine-3-carboxylic acid; 3-hydroxypiperidine-2-carboxylic acid; 4- hydroxypiperidine-2-carboxylic acid; 5-oxopiperidine-2-carboxylic acid; 5- hydroxypiperidine-2-carboxylic acid; 5-oxopiperidine-3-carboxylic acid; 5- hydroxypiperidine-3-carboxylic acid; 4-oxopiperidine-2-carboxylic acid; 4- hydroxypiperidine-2-carboxylic acid; 4-oxopiperidine-3-carboxylic acid; 4- hydroxypyridine-3-carboxylic acid; 3-oxopiperidine-4-carboxylic acid; 3- oxopiperidine-2-carboxylic acid; 2-oxopiperidine-4-carboxylic acid;
- a 6-oxopiperidine-2-carboxylate analog can be, for example, 6-hydroxypiperidine-2-carboxylic acid, 4-hydroxypiperidine-2-carboxylic acid and / or 3-hydroxypiperidine-2-carboxylic acid.
- a salt useful with this invention includes, but is not limited to, sodium, potassium, ammonium, copper, magnesium, calcium, zinc, molybdenum, iron, aluminum, lead, cadmium, chromium, nickel, mercury, and / or arsenic.
- a fungal mycelial extract may further comprise a peptide, a protein, a sugar, and / or a carbohydrate. In some embodiments, a fungal mycelial extract may comprise peptides and / or proteins in amounts from about 0.1% to about 10% w / w of the extract.
- a mycelial extract fungal may comprise peptides and / or proteins in an amount from about 0.1% to about 1%, from about 0.1% to about 3%, from about 0.1% to about 5%, from about 0.1% to about 7%, from about 0.5% to about 1%, from about 0.5% to about 3%, from about 0.5% to about 5% %, from about 0.5% to about 7%, from about 0.5% to about 10%, from about 1% to about 3%, from about 1% to about 5%, from about 1% to about 7%, from about 1% to about 10%, from about 3% to about 5%, from about 3% to about 7%, from about 3% to approximate me from 10%, from about 5% to about 7%, from about 5% to about 10%, or from about 7% to about 10%, or any range or value therein of the composition .
- a fungal mycelial extract may comprise peptides and / or proteins in an amount of about 0.1, 0.25, 0.5, 0.75, 1, 1, 5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, or 10% p / po any interval or value in it of the composition.
- a fungal mycelial extract may comprise sugars and / or carbohydrates in an amount from about 1% to about 35% w / w of the extract. In some embodiments, a fungal mycelial extract may comprise sugars and / or carbohydrates in an amount from about 1% to about 5%, from about 1% to about 10%, from about 1% to about 15%, from about 1% to about 20%, from about 1% to about 25%, from about 1% to about 30%, from about 5% to about 10%, from about 5% to about 15%, from about 5% to about 20%, from about 5% to about 25%, from about 5% to about 30%, from about 5% to about 35%, from about 10% to about 15%, from about 10% to about 20%, from about 10% to about 25%, from about 10% to about 30%, from about 10% to about 35%, from about 15% to about 20%, from about 15% to about 25%, from about 15% to about 30%, from about 15% to about 35%, from about 20% to about 25%, from about 20% to about 30%, from about 20% to approximately 35%,
- the fungal mycelial extract may comprise sugars and / or carbohydrates in an amount of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35% p / p of the extract or any interval or value in it.
- a sugar and / or a carbohydrate that may be comprised in the fungal mycelial extract may include, but is not limited to glucose, mannose, galactose, glucan oligosaccharides, low-branched glucose-derived polysaccharides, glycogen, morning oligosaccharides, low-branched polysaccharides derived from mannose, galactans and / or galactomannans.
- a fungal mycelial extract may comprise piperidine and / or an analog thereof, a salt thereof, and / or any combination thereof in an amount from about 1% to about 40% p / p of the extract.
- a fungal mycelial extract may comprise piperidine and / or an analog thereof, a salt thereof, and / or any combination thereof in an amount from about 1% to about 5%, from about 1% to about 10%, from about 1% to about 15%, from about 1% to about 20%, from about 1% to about 25%, from about 1% to about 30%, from about 1% to about 35%, from about 5% to about 10%, from about 5% to about 15%, from about 5% to about 20%, from about 5% to about 25%, from about 5% to about 30%, from about 5% to about 35%, from about 5% to about 40%, from about 10% to about 15%, from about 10% to about 20 %, from about 10% to about 25%, from about 10% to about 30%, from about 10% to about 35%, from about
- a fungal mycelial extract may comprise piperidine and / or an analog thereof, a salt thereof and / or any combination thereof in an amount of approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28 , 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40% w / w of the extract or any interval or value in it.
- a composition of the invention may further comprise additional components including, but not limited to, a surfactant, a humectant, an adjuvant, an antioxidant, a preservative, a plant macronutrient, a plant micronutrient, a growth regulator vegetable, a pesticide, a fungicide, an antiviral, an antibacterial, a herbicide, or any combination thereof.
- additional components including, but not limited to, a surfactant, a humectant, an adjuvant, an antioxidant, a preservative, a plant macronutrient, a plant micronutrient, a growth regulator vegetable, a pesticide, a fungicide, an antiviral, an antibacterial, a herbicide, or any combination thereof.
- surfactants may include, but are not limited to, alkali metal salts, alkaline earth metals and lignosulfonic acid, naphthalenesulfonic acid, phenylsulfonic acid, dibutylnaphthalenesulfonic acid, aquylarylsulfonates, sodium dodecyl sulfate, alkyl sulfates, alkyl sulphonates, sulfate sulfates, alcohoate sulfates fatty, glycol ethers of fatty acids and sulfated fatty alcohols, sulphonated condensate of naphthalene and naphthalene derivatives with formaldehyde, condensates of naphthalene or of naphthalenesulfonic acid with phenol and formaldehyde, polyoxyethylene octyl- phenyl ether, ethoxylated isooctyl
- a surfactant may be present in a composition in an amount from about 10% to about 40% w / w of the composition. In some embodiments, a surfactant may be present in a composition in an amount from about 10% to about 15%, from about 10% to about 20%, from about 10% to about 25%, from about 10% to about 30%, from about 10% to about 35%, from about 15% to about 20%, from about 15% to about 25%, from about 15% to approximately 30%, from approximately 15% to approximately 35%, from approximately 15% to approximately 40%, from approximately 20% to approximately 25%, from approximately 20% to approximately 30%, from approximately 20% up about 35%, from about 20% to about 40%, from about 25% to about 30%, from about 25% to about 35%, from about 25% to about 40%, from about 30% to about 35%, from about 30% to about 40%, or from about 35% to about 40%, w / w of the composition or any range or value therein.
- the surfactant may be present in a composition in an amount of approximately 10, 1, 1, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40% w / w of the composition or any interval or value therein .
- Example humectants may include, but are not limited to, glycerol, sorbitol, xylitol, maltitol, glyceryl triacetate, sodium lactate, urea formaldehyde, propylene glycol, ethylene glycol and / or fatty acids.
- An example antioxidant may include, but is not limited to, ascorbic acid, tocopherols, propyl gallate, tertiary butyl hydroquinone, butylated hydroxyanisole, and / or butylated hydroxytoluene.
- An example preservative may include, but is not limited to, sorbic acid, sodium sorbate, sorbates, benzoic acid, sodium benzoate, benzoates, hydroxybenzoate and derivatives, sulfur dioxide and sulphites, nitrite, nitrate, lactic acid, propionic acid and sodium propionate, tocopherol, vegetable extract, hops, salt, sugar, vinegar, alcohol (for example, methanol and ethanol), diatomaceous earth and castor oil, citric acid, ascorbic acid, sodium ascorbate, phenol derivatives ( butylated hydroxytoluene, butylated hydroxyanisole, BHA, BHT, TBHQ and propyl gallate), gallic acid, sodium gallate, sulfur dioxide, sulphites, tocopherols, and / or methylchloroisothiazolinone, 1, 2-benzothiazolin-3-one (BIT), hexahydro-1, 3,5-tris-hydroxyethyl-s-tri
- a preservative may be present in a composition in a range from about 0.001% to about 5% w / w or any range or value therein. In some embodiments, a composition may comprise a preservative in an amount ranging from about 0.001% to about
- the preservative may be present in the composition in an amount of about 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1, 1%, 1, 2%, 1, 3%, 1, 4%, 1, 5%, 1, 6%, 1, 7%, 1, 8%, 1, 9%, 2%, 2.5%, 3%, 3 , 5%, 4%, 4.5%, 5% w / w of the composition or any range or value therein.
- Example plant macronutrients include, but are not limited to, nitrogen, potassium, calcium, magnesium, phosphorus, and / or sulfur.
- Example plant micronutrients may include, but are not limited to, iron, manganese, boron, molybdenum, copper, zinc, chlorine and / or cobalt.
- Example plant growth regulators include, but are not limited to, auxin (including, but not limited to naphthalene acetic acid (NAA), and / or indole-3-butyric acid (IBA) and / or indole-3-acetic acid (IAA, 3-IAA)), cytokinin, abscisic acid, gibberellin, ethylene, salicylic acid, jasmonic acid, brasinosteriode (for example, brazed), or any combination thereof.
- auxin including, but not limited to naphthalene acetic acid (NAA), and / or indole-3-butyric acid (IBA) and / or indole-3-acetic acid (IAA, 3-IAA)
- cytokinin including, but not limited to naphthalene acetic acid (NAA), and / or indole-3-butyric acid (IBA) and / or indole-3-acetic acid (IAA, 3-IAA)
- Example pesticides include, but are not limited to, malation, paration, methyl paration, chlorpyrifos, diazinon, dichlorvos, fosmet, fenitrotion, tetrachlorvinfos, azamethylphos, fenvalerate, ciflutrin, lambda-cyhalotrine, zeta-cypermethrin, permethrin pyrobutyl, butoxide , imidacloprid, acetamiprid, clotianidin, nitenpyram, nitiacin, thiacloprid, thiamethoxam, ryanodol, 9,21-didehydrorianodol, chlorantraniliprol, flubendiamide, and / or ciantraniliprol
- Example fungicides include, but are not limited to, protioconazole trifloxistrobin, azoxistrobin, propiconazole, and / or pyraclostrobin.
- Exemplary antibacterials include, but are not limited to, methylisothiazolinone, chloromethylisothiazolinone, benzisothiazolinone, octylisothiazolinone, dichloroctylisothiazolinone, and / or butylbenzothiazolinone.
- Example herbicides may include, but are not limited to, glyphosate, 2,4-dichlorophenoxyacetic acid, atracine, S-metholachlor, and / or 3,6-dichloro-2-methoxybenzoic acid.
- a composition of the invention may further comprise an antifoaming agent. Any antifoam agent may be used for use with agricultural and / or food products.
- Example antifoaming agents include, but are not limited to, long chain unsaturated fatty acids including, but not limited to C12 to C14, C18: 1 and C18: 2 unsaturated fatty acids, and / or synthetic polysiloxanes (silicones) including , but not limited to, polydimethylsiloxane and / or hydrophobic silica.
- a composition of the invention may comprise an amount of antifoam agent in a range from about 0.0001% to about 0.05% w / w of the composition or any range or value therein.
- the antifoam agent may be present in the composition in an amount of approximately 0.0001%, 0.0002%, 0.0003%, 0.0004%, 0.0005%, 0 , 0006%, 0.0007%, 0.0008%, 0.0009%, 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.01% , 0.02%, 0.03%, 0.04%, or 0.05% w / w of the composition or any range or value therein.
- a composition of the invention may further comprise a beneficial microbe.
- the beneficial microbe may be Bacillus subtilis, Pseudomonas spp, Azotobacter spp, Azospirillum spp, Rhizobium spp, Azorhizobium spp, Chaetomium spp, Streptomyces spp. Trichoderma spp., And / or mycorrhizal fungi.
- a composition of the invention may be in the form of an aqueous solution, a non-aqueous solution, a suspension, a gel, a foam, a paste, a powder, a powder, a solid and / or an emulsion .
- Example preparation of a fungal mycelial extract (PRBT) for example, a fungal mycelial extract comprising piperidine and / or an analog thereof, a salt thereof, or any combination thereof).
- a mycelial extract PRBT
- approximately 150 g of dry mycelium of Penicillium spp. they can be added to 1 liter (I) of water and stir while heating at about 90 ° C for about 3 hours.
- the mixture can be centrifuged and the resulting supernatant can contain from about 25 to about 30 g 1 of dry matter (after removal of water by evaporation or lyophilization).
- PRBT can be used directly in the methods of the present invention or PRBT can be formulated to comprise additional components. Therefore, as an example, the supernatant obtained from the preparation of a fungal mycelial extract (as an example, see above) can be mixed with an antifoaming agent to produce a mixture.
- an antifoaming agent may be present in a range from about 0.001 g 1 to about 0.5 g 1 . In some embodiments, the amount of antifoaming agent may be present in about 0.0001% to about 0.05% w / w of the composition.
- a surfactant may be added to the supernatant (eg, fungal mycelial extract) in a ratio of about 2: 1.5 v / v to about 2: 0.5 v / v.
- the supernatant can be mixed in a ratio of 2: 1 v / v with a surfactant.
- a surfactant can be added to a composition of the invention in a range from about 10% to about 40% w / w of the composition.
- a biocide can be added to the supernatant.
- a biocide may be present from about 0.5 g 1 to about 20 g 1 .
- a final mixture may contain from about 15 to about 20 g 1 of dry matter (from fungal mycelial extract) when mixed in a 2: 1 v / v ratio with a surfactant.
- a fungal extract (PBRT) of the invention can be concentrated using any method including, but not limited to, lyophilization (eg, freezing and drying).
- PRBT can be used directly, in a concentrated form, or before use, the PRBT can be diluted to concentrations ranging from, for example, about 0.005 g 1 to about 20 g 1 or more.
- the supernatant can be mixed with a surfactant alone (for example, without antifoaming agent or biocide) in a ratio of about 2: 1 v / p.
- any amount of dry mycelium can be used, for example, from about 0.01 g to about 300 g.
- the temperature for preparing a fungal mycelial extract may vary from about 40 ° C to about 70 ° C, from about 40 ° C to about 90 ° C, about 40 ° C to about 1 10 ° C, from about 50 ° C to about 70 ° C, from about 50 ° C to about 90 ° C, about 50 ° C to about 120 ° C, from about 70 ° C to about 90 ° C, from about 70 ° C to about 100 ° C, from about 70 ° C to about 120 ° C, from about 80 ° C to about 100 ° C, from about 80 ° C to about 1 10 ° C, from about 80 ° C to about 120 ° C, from about 90 ° C to about 100 ° C, from about 90 ° C to about 100 ° C, from about 90 ° C to about 100 ° C, from about 90 ° C to about 100 ° C, from about 90 ° C to about 100 ° C, from about 90 ° C to about 100 ° C, from about 90
- the temperature for preparing the mycelial extract may be approximately 40 ° C, 45 ° C, 50 ° C, 55 ° C, 60 ° C, 65 ° C, 70 ° C, 75 ° C, 80 ° C, 85 ° C, 90 ° C, 95 ° C, 100 ° C, 105 ° C, 1 10 ° C, 1 15 ° C, or 120 ° C or any interval or value therein.
- a biocide can be added in an amount, for example, ranging from about 0.5 g to about 2 g per liter (for example, approximately, 0.6, 0.7, 0.8, 0.9, 1, 1, 1, 1, 2, 1, 3, 1, 4, 1, 5, 1, 6, 1, 7, 1, 8, 1, 9, or 2 g per liter).
- the above composition may further comprise micronutrients, such as magnesium (Mg), boron (B), copper (Cu), iron (Fe), manganese (Mn), molybdenum (Mo) and zinc (Zn), fungicides, herbicides, insecticides, macronutrients, such as nitrogen (N), phosphorus (P) and potassium (K), and adjuvants.
- micronutrients such as magnesium (Mg), boron (B), copper (Cu), iron (Fe), manganese (Mn), molybdenum (Mo) and zinc (Zn
- fungicides such as magnesium (Mg), boron (B), copper (Cu), iron (Fe), manganese (Mn), molybdenum (Mo) and zinc (Zn
- fungicides herbicides, insecticides, macronutrients, such as nitrogen (N), phosphorus (P) and potassium (K), and adjuvants.
- a composition of the invention can be applied to plants or parts thereof to, for
- the present invention provides a method of increasing a growth characteristic of a plant or part thereof, the method comprises applying a composition comprising an effective amount of a fungal mycelial extract to a plant or part thereof, wherein the extract comprises 6-oxopiperidine-2-carboxylate and / or piperidine, and / or an analogue thereof, or a salt thereof, thereby increasing the growth characteristic of the plant or part thereof compared to a control plant or part thereof (for example, a plant or part thereof to which the composition or extract of the invention has not been applied).
- the method comprises applying a composition at least once (for example, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12 or more times).
- the method comprises applying the composition at least twice (for example, approximately 2, 3, 4, 5, 6, 7, 8, 9, 10, 1, 12 or more times).
- a "prior application” refers to any application of a composition of the invention to a plant or part of a plant that is followed by another application (eg, a subsequent application) of the composition.
- a method is provided to increase the nutrient use efficiency of a plant or part thereof, the method comprises applying a composition comprising an effective amount of a fungal mycelial extract to a plant or part of the plant.
- the method comprises applying a composition at least once (for example, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12 or more times). In some embodiments, the method comprises applying the composition at least twice (for example, approximately 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or more times).
- the present invention provides a method for increasing the disease tolerance of a plant or part thereof, the method comprises applying a composition comprising an effective amount of a fungal mycelial extract to a plant or part of the plant. same, wherein the extract comprises 6-oxopiperidine-2-carboxylate and / or piperidine, and / or an analog thereof, or a salt thereof, or any combination thereof, thereby increasing disease tolerance of the plant or part thereof compared to a control plant or part thereof (for example, a plant or part thereof to which the composition or extract of this invention has not been applied).
- the method comprises applying a composition at least once (for example, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12 or more times).
- the method comprises applying the composition at least twice (for example, approximately 2, 3, 4, 5, 6, 7, 8, 9, 10, 1, 12 or more times). In some embodiments, the method comprises applying the composition at least three times. In some embodiments, the method comprises applying the composition at least four times.
- a subsequent application for example, any application that follows a previous application; for example, a second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth application and so on successively
- a subsequent application can be any time from about 1 week to about six months after the previous application, therefore, for example, a subsequent application can be applied about 1, 2, 3, 4, 5, 6 , 7, 8, 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or 26 weeks after a previous application or approximately 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6 months after a previous application.
- applying a composition of the invention to a plant or part thereof may increase tolerance or resistance to a viral pathogen disease including, but not limited to, a virus from the family of Caulimoviridae, Potyviridae, Sequiviridae , Rheoviridae, Capillovirus, Geminiviridae, Bromoviridae, Closteroviridae, Comoviridae Tombusviridae, Rhabdoviridae, Bunyaviridae, Partitiviridae, Carlavirus, Enamovirus, Furovirus, Hordeivirus, Idaeovirus, Luteovirus, Marafivirus, Potexvirus, Sobemovirus, Tenuivirus, Tobamovirus, Tobravirus, Trichovirus, Tymovirus and / or Umbravirus.
- a viral pathogen disease including, but not limited to, a virus from the family of Caulimoviridae, Potyviridae, Sequiviridae , Rheoviridae,
- applying a composition of the invention to a plant or part thereof may increase resistance to a virus including, but not limited to, turnip mosaic virus, papaya ring spot virus, virus. of the wilting of the yolks, bean pod speckling virus, lettuce mosaic virus, corn mosaic virus, cauliflower mosaic virus, tobacco mosaic virus, soy mosaic virus, African cassava mosaic virus, tomato mosaic virus, cucumber mosaic virus, yellow zucchini mosaic virus, sharka virus, branched tomato dwarf virus, tomato tan virus, curly virus yellow tomato leaf, tattered rice dwarf virus, bacilliform rice tungro virus, spherical rice tungro virus, yellow rice mottle virus, cucumber mosaic virus, cebadilla mosaic virus, virus d the yellow wheat mosaic, yellow dwarf barley virus, sugar cane mosaic virus, beet yellowing virus, lettuce yellowing virus, dwarf corn mosaic virus, corn scratch virus, virus of peanut dwarfism, citrus sadness virus, potato leaf curl virus, potato X virus, potato virus Y, sweet potato feathery chlorotic mottled poti
- applying a composition of the invention to a plant or part thereof may increase resistance to a fungal pathogen including, but not limited to, a fungus of the family of Physodermataceae, Synchytriaceae, Olpidiaceae, Choanephoraceae, Gilbertellaceae , Mucoraceae, Dipodascaceae, Eremotheciaceae, Taphrinaceae, Botryosphaeriaceae, Capnodiaceae, Phaeosphaeriaceae, Leptosphaeriaceae, Cucurbitariaceae, Didymellaceae Davidiellaceae, Mycosphaerellaceae, Schizothyriaceae, Dothideaceae, Dothioraceae, Lahomaceae, Lahomaceae, Lahomaceae, Lahomaceae, Lahomaceae, Lahomaceae Pleosporaceae, Venturiaceae, Trichochomaceae, Erysipha
- Hemiphacidiaceae Hyaloscyphaceae, Phacidiaceae, Sclerotiniaceae, Ascodichaenaceae, Mediolariaceae, Rhytismataceae, Meliolaceae,
- Caloscyphaceae Sarcosomataceae, Cryphonectriaceae, Diaporthaceae, Gnomoniaceae, Valsaceae, Glomerellaceae, Plectosphaerellaceae, Bionectriaceae, Clavicipitaceae, Hypocreaceae, Nectriaceae, Magnaporthaceae, Pyriculariaceae, Ceratocystideae, Ophiostomataceae, Phyllachoraceae, Chaetomiaceae, Amphisphaeriaceae, Diatrypaceae, Xylariaceae, psathyrellaceae, Marasmiaceae, mycenaceae, Schizophyllaceae, Typhulaceae.Thelephoraceae, Atheliaceae, Atheliaceae, Stereaceae, Echinodontiaceae, Corticiaceae, Ganodermataceae, Hymenochaetace
- Bipolaris maydis, Bipolaris oryzae, Bipolaris sacchari, Bipolaris victoriae, Curvularia spp., Leptosphaerulina trifolii, Venturia inaequalis, Aspergillus, Piprillus spp.
- Botrytis cinerea Monilinia spp., Monilinia fructicola, Sclerotinia sclerotiorum, Amphilogia gyrosa, Cryphonectria parasitica, Diaporthe citri, Diaporthe helianthi, Diaporthe phaseolorum, Cytospora leucostoma, Colletotrichum spp., Colletotletumumumumumumumumumumumumumumumumumumum, Colletotrichumumumumumumumumumumumus, Colletotrichumumumumumumumumus, Colletotrichumumumumumumumumus, Colletotumumumumumumumus, Colletotumumumumumumumus, Colletotrichumumumumumumumus, Colletotrichumumumumumumus, Colletotrichum coccodes, Colletotumumumumumumumumus, cucumerina, Verti
- applying a composition of the invention to a plant or part thereof may increase resistance to a bacterial pathogen including, but not limited to, a bacterial pathogen of the Enterobacteriaceae, Pseudomonadaceae, Rhizobiaceae, Microbacteriaceae family, Xanthomonadaceae, Rhizobiaceae, Corynebacteriaceae, Acetobacteraceae, Comamonadaceae, Bacillaceae, Burkholderiaceae, Micrococcaceae, Ralstoniaceae, Xanthomonadaceae, Spiroplasmataceae, Sphingomonadaceae, Acholeplasmataceae, Corynebacteriaceae, and / or Streceae
- applying a composition of the invention to a plant or part thereof may increase resistance to a bacterial pathogen including, but not limited to, a bacterial pathogen of the genus of Erwinia spp., P
- applying a composition of the invention to a plant or part thereof may increase resistance to a bacterial pathogen including, but not limited to, Erwinia amylovora, Ea carotovora var. chrysanthemi. Pseudomonas tabaci, P. angulate, P. phaseolicola, P. lachrymans, P. pisi, P. fluorescens, P. glycinea, P. vesicatoria, P. savastanoi, P. syringae, P. solanacearum, Xanthamonas phaseoli, X. malvacearum, X. oryzae, X translucens, X.
- a bacterial pathogen including, but not limited to, Erwinia amylovora, Ea carotovora var. chrysanthemi. Pseudomonas tabaci, P. angulate, P. phaseolicola, P. lachryman
- disease resistance or “disease tolerance” are used interchangeably and refer to a decrease in disease symptoms and / or a decrease in growth and reproduction of the pathogen of a plant or plant part.
- the percentage (%) of increase in disease resistance / tolerance compared to a control may be in a range from about 0.1% to about 100%.
- the percentage increase in resistance / disease tolerance may be an increase in a range from about 0.1% to about 10%, from about 0.1% to about 30%, from about 0.1% to about 50%, from about 0.1% to about 80%, from about 0.1% to about 90%, from about 0.1% to about 95% , from about 1% to about 10%, from about 1% to about 20%, from about 1% to about 40%, from about 1% to about 50%, from about 1% to about 75%, from about 1% to about 95%, from about 1% to about 100%, from about 10% to about 20%, from about 10% to about 40%, from about 10% to about 50%, from about 10% to about 70%, from about 10% to about 80%, from about 10% to about 90 %, from about 10% to about 100%, from about 20% to about 40%, from about 20% to about 75%, from about 20% to about 90%, from about 20 % until approximately 95%, from approximately 20% to approximately 100%, from approximately 25% to approximately 50%, from approximately 50% to approximately 75%, from approximately 50% to approximately 95%, from approximately 50% to approximately 100%, from approximately 7
- the% increase in resistance / disease tolerance may be approximately 0.1%, 0.5%, 1%, 1.5%, 2%, 2.5%, 3, 3, 5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5% , 10%, 1 1%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, or 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90 %, 95%, 96%, 97%, 98%, 99% or 100% or any value or interval in it, compared to a control.
- a method is provided to increase the tolerance to abiotic aggression of a plant or part thereof, the method is to apply a composition comprising an effective amount of a fungal mycelial extract to a plant or part of the plant. same, wherein the extract comprises 6-oxopiperidine-2-carboxylate and / or piperidine and / or an analogue thereof, a salt thereof, or any combination thereof, thereby increasing tolerance to abiotic aggression. of the plant or part thereof compared to a control plant or part thereof (for example, a plant or part thereof to which the composition or extract of the invention has not been applied).
- the method comprises applying the composition at least once (for example, approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or more times).
- the method comprises applying the composition at least twice (for example, approximately 2, 3, 4, 5, 6, 7, 8, 9, 10, 1, 12 or more times).
- abiotic aggression is drought.
- the abiotic aggression is salinity.
- an "increased tolerance to abiotic aggression” or “increased resistance to abiotic aggression” is used interchangeably and refers to the capacity of a plant or part thereof exposed to abiotic aggression and put into contact with a composition of the invention to resist a determined abiotic aggression better than a control plant or part thereof (i.e., a plant or part thereof that has been exposed to the same abiotic aggression, but which has not been placed in contact with a composition comprising a fungal mycelial extract comprising piperidine and / or an analogue and / or salt thereof).
- Increased tolerance to abiotic aggression can be measured using a variety of parameters including, but not limited to, the size and number of plants or parts thereof, and the like (eg, number and size of fruits), level or quantity of cell division, the amount of floral abortion, the amount of sunburn damage, crop yield, and the like. Therefore, in some embodiments of this invention, a plant or part thereof that has been contacted with a composition of the present invention, and which has increased tolerance to abiotic aggression, for example, would have number and / or increased weight of fruit / seed compared to a plant or part thereof exposed to the same aggression, but without having contacted said composition.
- the percentage increase in resistance / tolerance to abiotic aggression compared to a control may be an increase in a range from about 0.1% to about 100%.
- the percentage increase in disease resistance / tolerance may be in a range from about 0.1% to about 10%, from about 0.1% to about 30%, from about 0.1% to about 50%, from about 0.1% to about 80%, from about 0.1% to about 90%, from about 0.1% to about 95% , from about 1% to about 10%, from about 1% to about 20%, from about 1% to about 40%, from about 1% to about 50%, from about 1% up to about 75%, from about 1% to about 95%, from about 1% to about 100%, from about 10% to about 20%, from about 10% to about 40%, from about 10% to about 50%, from about 10% to about 70%, from about 10% to about 80%, from about 10% to about 90%, from about 10% to about 100%, from about 20% to about 40%, from about 20% to about 75%, from about 20% to about 90%, from about 20% to about 95%, from about 20% to about
- the% increase in resistance / tolerance to abiotic aggression may be approximately 0.01%, 0.05%, 0.1%, 0.5%, 1%, 1.5%, 2 %, 2.5%, 3, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8 , 5%, 9%, 9.5%, 10%, 1 1%, 12%, 13%, 14%, 15%, 16%, 17, 5, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, or 50%, 55%, 60%, 65%, 70 %, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% or any value or interval in it, compared to a control.
- an extract or a composition of the invention can be applied to any plant or part thereof. Therefore, an extract or a composition of the invention can be applied to a variety of plants in various forms or sites, such as, for example, foliage, buds, flowers, fruits, ears or spikes, seeds, bulbs, stem tubers , roots, and seedlings.
- bulb means a discoidal stem, rhizomes, root tubers and rhizophores.
- an extract or composition of the invention can also be applied to cuttings (eg, cuttings of the sugar cane stem).
- "one plant” means any monocot and dicot plant, and any annual and perennial dicot plant and monocot.
- plants include, but are not limited to, those of the genera Glycine, Vitis, Asparagus, Populus, Pennisetum, Lolium, Oryza, Zea, Avena, Hordeum, Sécale, Triticum, Helia nthus, Gossypium, Medicago, Pisum, Acer , Actinidia, Abelmoschus, Agropyron, Allium, Amaranthus, Apium, Arachis, Asparagus, Beta, Brassica, Camellia, Canna, Capsicum, Carex, Carica papaya, Carya, Castanea, Cinnamomum, Citrullus, Citrus, coconuts, Coffea, Colocasia, Cola, Coriandrum, Corylus, Crataegus, Crocus, Cucurbita, Cucumis, Cynara, Daucus, Desmodium, Dimocarpus, Dioscorea, Diospyros, Echinochloa, Ela
- a plant or part thereof is of the genera Glycine, Vitis, Asparagus, Populus, Pennisetum, Lolium, Oryza, Zea, Avena, Hordeum, Sécale, Triticum, Sorghum, Saccharum and Lycopersicum, or class Liliaceae
- a "plant” includes mature plants, seeds, shoots and seedlings, parts of plants, propagation material, plant organs, tissue, protoplasts, corns and other crops, for example, cell cultures derived from the above, and all other types of plant cell associations that give functional or structural units.
- a “mature plant” refers to a plant at any stage of development beyond the seedling stage and includes, but is not limited to, an adult or mature plant, a plant with buds, a flowering plant, and / or a fruit plant
- “Seedling” refers to a young, immature plant in an early stage of development.
- plant part includes, but is not limited to, reproductive tissues (eg, petals, sepals, stamens, pistils, receptacles, anthers, pollen, flowers, fruits, buds of flowers, ovules, seeds, embryos, nuts, pips, ears, ears, pods); vegetative tissues (eg, petioles, stems, roots, root hairs, root tips, albedo, coleoptyls, reeds, buds, branches, bark, apical meristem, auxiliary bud, cotyledon, hypocotyls, and leaves); vascular tissues (for example, phloem and xylem); specialized cells such as epidermal cells, parenchyma cells, colenchyme cells, sclerenchyma cells, stomata, guard cells, cuticle, mesophilic cells, callus tissue; and cuttings.
- reproductive tissues eg, petals, sepals, stamens, pistils,
- plant part also includes plant cells, including plant cells that are intact in plants and / or parts of plants, plant protoplasts, plant tissues, plant organs, plant tissue cultures, plant corns, plant agglomerations, and the like
- flower refers to parts above ground including leaves and stems.
- tissue culture encompasses tissue cultures, cells, protoplasts and calluses.
- plant propagation material or “plant propagating material” refers to any plant material from which a plant or part of the plant can be derived.
- the plant propagation material includes, but is not limited to, seeds, seedlings, young plants, cuttings, cell suspensions, protoplasts, callus culture, tissue culture, protoorms, explants, germplasm, bulbs and / or tubers, or any combination thereof.
- a variety of seeds or bulbs may be used in the methods described herein including, but not limited to, plants in the Solanaceae and Cucurbitaceae families, as well as selected Calibrachoa plants, Capsicum, Nicotiana, Nierembergia, Petunia, Solanum, Brassica, Cuc ⁇ rbita, Cucumis, Citrullus, Glycine, such as Glycine max (Soy), Calibrachoa ⁇ hybrida, Capsicum annuum (pepper), Nicotiana tabacum (tobacco), Nierenbergia scoparia (tobacco) Petunia, Solanumlycopersicum (tomato), Solanum tuberosum (potato), Solanum melongena (eggplant), Maximum Cucurbita (pumpkin), Cucurbita pepo (pumpkin, zucchini), Cucumis metuliferus (African cucumber) Cucumis melon (melon), Cucumis sativus (cucumber) and Citrullus
- Several monocotyledonous planytas in particular those belonging to the Poaceae family, can be used with the methods described herein, including, but not limited to, Hordeum, Avena, Sécale, Triticum, Sorghum, Zea, Saccharum, Oryza, Hordeum vulgare (barley), Triticum aestivum (wheat), Triticum aestivum subsp. spelta (spelled), Triticale, Avena sativa (oatmeal), Sécale cereale (rye), Sorghum bicolor (sorghum), Zea mays (corn), Saccharum officinarum (sugarcane) and Oryza sativa (rice).
- plants for which health and / or yield can be improved using the methods described herein include the following crops: rice, corn, cane, soy, wheat, buckwheat, beet, rapeseed, sunflower, cane of sugar, tobacco and pea, etc .; Vegetables: Solanaceae vegetables such as red pepper or potato; cucurbitaceous vegetables; cruciferous vegetables such as Japanese radish, white turnip, horseradish, kohlrabi, Chinese cabbage, cabbage, leaf mustard, broccoli, and cauliflower, asteric vegetables such as burdock, chrysanthemum, artichoke, and lettuce; liliaceous vegetables such as scallions, onions, garlic, and asparagus; anmiaceae vegetables such as carrots, parsley, celery and parsnips; chenopodiaceous vegetables such as spinach, chard; Laminaceous vegetables such as Perilla frutescens, mint, basil; strawberry, sweet potato, Dioscorea japan, colocasia;
- An embodiment of the present disclosure also provides plants that have increased production of inflorescences compared to plants where the composition of the present disclosure has not been applied.
- An embodiment of the present disclosure also provides plants that have increased fruit quality compared to plants where the composition of the present disclosure has not been applied.
- An embodiment of the present disclosure further provides plants that have increased fungal tolerance compared to plants where the composition of the present disclosure has not been applied.
- An embodiment of the present disclosure further provides plants that have increased tolerance to white mold (Sclerotinia sclerotiorum) compared to plants where the composition of the present disclosure has not been applied.
- white mold Sclerotinia sclerotiorum
- An embodiment of the present disclosure further provides plants that have increased tolerance to Botrytis cinerea compared to plants where the composition of the present disclosure has not been applied.
- An embodiment of the present disclosure further provides plants that have increased disease tolerance when treated with the composition of the present disclosure in combination with beneficial microbes when compared to untreated plants.
- An embodiment of the present disclosure further provides plants that have increased fungal tolerance when treated with the composition of the present disclosure in combination with beneficial microbes when compared to untreated plants.
- An embodiment of the present disclosure further provides plants that have increased tolerance to white mold (Sclerotinia sclerotiorum) when treated with the composition of the present disclosure in combination with Bacillus subtilis when compared to untreated plants.
- white mold Sclerotinia sclerotiorum
- An embodiment of the present disclosure also provides plants that have increased tolerance to plant viruses when compared to plants where the composition of the present disclosure has not been applied.
- An embodiment of the present disclosure further provides plants that have decreased viral load when compared to plants where the composition of the present disclosure has not been applied.
- An embodiment of the present disclosure further provides plants that have increased tolerance to the New Delhi tomato leaf curl virus (ToLCNDV) compared to plants where the composition of the present disclosure has not been applied.
- ToLCNDV New Delhi tomato leaf curl virus
- a fungal mycelial extract can be applied as a soil treatment in the form of a solid or a liquid. Therefore, in some embodiments, the composition can be applied as a spray on land, incorporation to land, and / or perfusion of a chemical liquid in the soil (chemical liquid irrigation, injection into the earth and dripping of chemical liquid ).
- the placement of PRBT during the treatment of the land includes, but is not limited to, planting hole, groove, around a planting hole, around a groove, entire area of farmland, the parts between the land and the plant, area between the roots, area under the trunk, main groove, growth box, seedling cultivation tray and seedbed, seedling cultivation.
- the treatment of the soil with PRBT can be before sowing, at the time of sowing, immediately after sowing, cultivation period, before the settled plantation, in the time of the settled plantation and / or the period of growth after the settled plantation.
- an irrigation liquid may be mixed with PRBT in advance and, for example, used for treatment by an appropriate irrigation method including the irrigation method mentioned above and the other methods such as sprinkling and flooding.
- PRBT can also be applied by wrapping a crop with a resin formulation processed in a sheet or a rope, putting a string of the resin formulation around the crop so that the crop is surrounded by the string, and / or casting the sheet of the resin formulation on the surface of the earth near the root of a crop.
- PRBT can be used to treat seeds or bulbs, as well as a spray treatment of PRBT for seeds in which a suspension of PRBT is atomized and sprayed on a seed surface or bulb surface.
- a smearing treatment can also be used, for example, where a wettable powder, an emulsion or a PRBT fluid agent is applied to seeds or bulbs with a small amount of water added or applied as it is without dilution.
- an immersion treatment can be used in which seeds are immersed in a PRBT solution for a certain period of time, film coating treatment, and pellet coating treatment.
- PRBT can be used for the treatment of seedlings, including spray treatment composed of spraying the whole seedlings with a dilution having an appropriate concentration of the active ingredients prepared by diluting PRBT with water.
- spray treatment composed of spraying the whole seedlings with a dilution having an appropriate concentration of the active ingredients prepared by diluting PRBT with water.
- the soil can be treated / contacted with PRBT before and / or after sowing seedlings including spraying a dilution that has an appropriate concentration of the active ingredients prepared by diluting PRBT with water and applying the mixture to seedlings or the soil around the seedlings. after sowing the seedlings.
- a PRBT spray treatment can also be used. formulated in a solid formulation such as a granule to land around the seedlings when planting the seedlings.
- PRBT When PRBT can be applied as a treatment to foliage, floral organs, or ears or spikes of plants, such as foliage spraying; seed treatment, such as seed sterilization, seed immersion or seed coating; seedling treatment; bulb treatment; and treatment of plant farmland, such as soil treatment.
- PRBT can be applied only to specific plant sites, such as a floral organ in the flowering season including before flowering, during flowering and after flowering, and the cob or spike in the cob-forming station, or You can apply to whole plants.
- Example 1 Results of PRBT in increased metabolites in Arabidopsis and tomato
- PRBT fungal mycelial extract
- Samples were prepared using the Hamilton Company automated MicroLab STAR system. Several recovery standards were added before the first stage in the extraction process for QC purposes. To remove protein, dissociated small molecules bound to protein or trapped in the precipitated protein matrix, and to recover chemically diverse metabolites, the proteins were precipitated with methanol with vigorous stirring for 2 minutes (Glen Mills GenoGrinder 2000) followed by centrifugation.
- the resulting extract was divided into four fractions: two were analyzed by two separate reverse phase (RP) / UPLC-MS / MS methods with electrospray ionization (ESI) in positive ion mode, one by RP / UPLC-MS / MS with ESI in negative ion mode, and one by HILIC / UPLC-MS / MS with ESI in negative ion mode.
- RP reverse phase
- UPLC-MS / MS electrospray ionization
- HILIC / UPLC-MS / MS with ESI in negative ion mode A total of 373 biochemical substances of known identity were determined.
- ANOVA contrasts were used to identify biochemical substances that differed significantly between the control plants and those treated with PRBT.
- Table 1 shows biochemical substances (column 1) that differed significantly (P ⁇ 0.05) between control and PRBT-treated plants that were common between Arabidopsis and tomato.
- the PubChem identification number of the compound is shown in column 2 for each biochemical substance.
- the change in times of the plants treated with PRBT versus those treated control (column 4), the p-value of ANOVA (column 5), as well as the average values for each biochemical substance (columns 6 and 7) are indicated.
- the application of PRBT increases the levels of 6-oxopiperidine-2-carboxylate, staquidrin and the mannitol / sorbitol ratio.
- RNA sequencing experiment was performed on 4 samples. Before further analysis, a quality control was performed on the raw sequencing data using FastQC, then the low quality parts of the readings were removed with BBDuk.
- the minimum length of the readings after trimming was adjusted to 35 bp and the minimum base quality score to 25.
- ATCG00280 genes are related to photosynthesis (ATCG00280 genes; ATCG00680, ATCG00550, ATCG00070, ATCG00080, ATCG00580, ATCG00270, AT1G29930, AT2G34430, ATCG00020, AT1G29910, ATCG00350, ATCG00340, ATCG00280, ATCG00680, AT1G29920, ATCG00730, ATCG00280, AT2G01918, ATCG00680, ATCG00580), response to saline aggression (AT5G58580), transport and nutrient binding (AT4G33000, AT1G21840, AT2G47400, AT1G76560, AT4G09640, AT4G 13800, AT3G23870, AT3G26740, AT5G57345, AT5G27290 cell phosphide response (a5G27290) AT5G20150), hormone transport (AT1G17140, AT5G64770, AT4G09
- Table 1 below shows a list of genes that are augmented in Arabidopsis plants treated with PRBT compared to untreated plants.
- Column 1 shows the ontological identification of the gene (GO ID)
- columns 2 and 3 list the type and description of the genes, respectively
- column 4 shows the p-value
- column 5 shows the enrichment score (enrichment times of the corresponding GO term in the list of genes differentially expressed with respect to the reference genome)
- column 7 shows the genetic loci.
- AT2G34430 ATCG00020; AT1 G29910; ATCG00350; ATCG00340; ATCG00280. ATCG00680; AT1 G29920
- AT2G34430 ATCG00020; AT1 G29910; AT2G39470; AT3G55330; AT2G30790; AT2G28605; ATCG00550; ATCG00280; AT2G01918; AT1 G77090; ATCG00710; ATCG00070; ATCG00680; AT1 G29920; AT4G19100; ATCG00080; ATCG00580
- AT2G34430 AT1 G29910; ATCG00520; ATCG00350; ATCG00340; ATCG00630; AT1 G29920
- AT3G62750 AT1 G60270; AT2G44480; AT1 G60260
- AT3G61060 AT2G42900 AT1 G74670 AT2G47400 AT2G 17880 AT3G 13450 AT4G23870
- AT2G42530 AT2G23430 AT4G39260 AT5G37260 AT5G53160 AT4G00310 AT3G 17510 AT1 G22640 AT5G59220 AT1 G08810 AT4G 15910 AT5G15970 AT3G22060 AT5G39610 AT4G27410 AT4G34000 AT1 G1682902 ATG2802 AT2G26980;
- Example 3 - PRBT increases yield in a wide variety of crops
- plants of strawberry variety 'Sabrina', variety of garlic 'white', variety of lettuce 'Iceberg', kohlrabi, onion, fennel, and FAO700 corn were grown under conditions of standard production in 2013 and 120 plants of each variety were analyzed per treatment (where the treatment was a control that included standard irrigation and 0.054 g 1 of sprayed PRBT (once for garlic in combination with 0.000006 g I "1 of B , 0.000012 g I "1 of chelated Cu, 0.00003 g I "1 of chelated Fe and 0.000012 g 1 and 0.00003 g I " 1 of chelated Mn; four times for lettuce, kohlrabi, onion and fennel and once for corn in combination with 0.012 g I "1 of N, 0.003 g I " 1 of P 2 0 5, 0.003 g I "1 of K 2 0 , 0.024 g I "1 of P 2 0 3 ,
- the plants were located in four different positions for each group of 30 plants of the same treatment. Fruits, leaves or roots of individual plants were collected and the total weight for each plant was determined.
- Sabrina strawberry plants were grown under standard production conditions (from October 2013 to April 2014) and 120 plants were analyzed per treatment (control sprayed with the adjuvant or plants with PRBT ( 0.054 g 1 ) (with 2: 1, 5 v / p surfactant) were sprayed a total of 3 times, once every four weeks for three months.
- the plants were located in four different positions for each group of 30 plants of the same treatment Fruits were picked from individual plants and the total weight was determined for each plant.
- broccoli plants when treated with PRBT, showed a 12% increase in inflorescence production in conditions without aggression, and a 22% increase in high salinity conditions, compared to plants without treating.
- broccoli plants showed a 12% increase in plant weight under high salinity conditions compared to untreated plants.
- broccoli plants showed an increase in plant quality under normal and high salinity conditions compared to untreated plants.
- Example 5 Tomato plants treated with PRBT have an increased fruit production
- PRBT applied exogenously increases the production of tomato plants. Var. Tomato plants were sown. Mayoral in a greenhouse on August 31, 2014. A total of 2244 plants, 1122 control plants and 1 122 plants treated with PRBT distributed in randomized blocks were used. Each pair of lines are separated by 1.5 meters. In a line, the plants are separated by 50 cm. The plants treated with PRBT were sprayed once a month, from September 2014 to March 2015 (7 treatments) with PRBT (0.054 g I "1 ) (with surfactant 2: 1, 5 v / p) at 0.36 ml / plant / month Plants were collected 10 times and production data was collected for each plant per harvest day.
- tomato plants showed an increase in total fruit production weight of up to 42% when PRBT was applied compared to untreated plants.
- the plants flourished before increasing the yield of the first day of harvest.
- Example 6 Watermelon plants treated with PRBT have an increased production and fruit quality.
- PRBT applied exogenously increases the production of watermelon plants. Watermelon plants were planted var. Motril in 2014 and Boston in 2016 in a greenhouse on December 19, 2014 and 2016. A total of 1600 plants, 800 control plants and 800 PRBT treated plants distributed in randomized blocks were used.
- Plants treated with PRBT (with 2: 1, 5 v / p surfactant in 2014; and with 2: 1 v / p surfactant, antifoaming agent and biocide in 2016) were sprayed four times once a month, from January to April with PRBT (0.054 g 1 ) at 0.36 ml / plant / month and with spinosad in all treatments, as well as mancozeb in the last treatment.
- the control plants received the treatments with spinosad and mancozeb without the PRBT.
- the plants were collected once in 2014 and twice in 2016 in April and production data were collected per harvest day according to the category, category 1 (CAT 1) being the best and category 2 (CAT2) being the lowest quality.
- Example 7 Pepper plants treated with PRBT have an increased production and fruit quality
- PRBT applied exogenously increases the production of pepper plants. California or Guepard variety pepper plants were planted in three independent greenhouses and the fruits were collected in 10 crops of 64 plants per randomly distributed greenhouse. The plants grew between January 20, 2014 and October 23, 2014. A total of 512 plants were collected per greenhouse, 256 control plants and 256 plants treated with PRBT (with 2: 1, 5 v / p surfactant) distributed in random blocks. The plants treated with PRBT were sprayed nine times, once a month from January to April with PRBT (0.054 g 1 ) at 0.36 ml / pl anta / month.
- PRBT 1 138.38 ⁇ 79.76
- pepper plants treated with PRBT had an increase in total number of fruits per plant between 12.9% and 21.9% compared to untreated plants.
- Table 12 Weight per pepper fruit in grams for plants treated with PRBT or control spray
- Example 8 Soybeans treated with PRBT have an increased tolerance against white mold (Sclerotinia sclerotiorum)
- Soybean plants were sown on September 16, 2016 and distributed in randomized blocks of 9 plants with 4 replicates (36 plants per treatment) and grown in a standard greenhouse. Plants treated with PRBT (with 2: 1 v / p surfactant, antifoam and biocide) were sprayed once on October 25 with PRBT (0.054 g 1 ) at 6 ml / plant. Two days later the plants were inoculated with 10 ml / plant of a mixed mycelium foliar spray of Sclerotinia sclerotiorum strain CH 109 that had been grown at an optical density at 600 nm of 1.5 (in C1 inoculated plants) or 2 , 0 in the inoculated plants C2).
- the plants were maintained at a relative humidity of 100% during the rest of the experiment.
- the plants were evaluated after 7, 13 and 18 days post-inoculation (dpi) according to a score of the disease severity index where 0 corresponds to no symptoms and 5 corresponds to a dead plant.
- the area under the disease assessment curve (AUCPC) was calculated as a quantitative summary of the disease intensity over time.
- FIG. 2 shows photographs of soybean plants inoculated with Sclerotinia sclerotiorum (C1) and treated with PRBT (right panel) compared to control plants inoculated with Sclerotinia sclerotiorum (C1) (middle panel) and inoculated control plants of reference (left panel).
- Example 9 - Tomato plants treated with PRBT have an increased tolerance against white mold (Sclerotinia sclerotiorum) and increased fruit production under conditions of infection
- Var. Tomato plants were sown. Royesta on January 28, 2014 and were distributed in randomized blocks of 5 plants with 6 replicates (30 plants per treatment) and grown in a standard production greenhouse. Plants treated with PRBT were sprayed twice on February 18 and on March 1 with PRBT (0.054 g 1 ) (without surfactant) at 6 ml / plant or with stemicol (4.5 g 1 ) at 6 ml / plant . Two days later the plants were inoculated with 20 ml / plant of a mixed mycelium foliar spray of Sclerotinia sclerotiorum strain CH109 that had been grown at an optical density at 600 nm of 1, 7.
- the plants were maintained at a relative humidity of 100% for ten days and then the humidity was decreased to 80% during the rest of the experiment.
- the plants were evaluated after 1 1, 18, 41 and 55 days post-inoculation (dpi) according to a score of the disease severity index where 0 corresponds to no symptoms and 5 corresponds to a dead plant. Tomatoes were collected at 90 dpi and the total weight per plant was recorded.
- Table 14 disease index and total production per plant in kilograms for plants treated with PRBT spray, stemicol or control inoculated with white mold and treated with PRBT
- TREATMENT 1 1 dpi 18dpi 41 dpi 55 dpi
- the disease rate in tomato plants treated with PRBT was significantly lower than in untreated tomato plants and equivalent to plants treated with stemicol that had been inoculated with Sclerotinia sclerotiorum.
- plants inoculated and treated with PRBT had a significantly lower disease rate (1, 6 c) compared to plants treated with stemicol (1, 9 b) and untreated (2.4 a).
- Example 10 Pepper plants treated with PRBT have an increased tolerance to Botrytis cinerea and increased fruit production under conditions of infection
- Var. Pepper plants were planted. Ferrari on November 26, 2014 and were distributed in randomized blocks of 5 plants with 6 replicates (30 plants per treatment) and grown in a standard production greenhouse. Plants treated with PRBT (with 2: 1 v / p surfactant) were sprayed on January 13, 2015 with PRBT (0.054 g I "1 ) at 20 ml / plant or with stemicol (4.5 g I " 1 ) a 20 ml / plant One day later the plants were inoculated with 20 ml / plant of a foliar spray of 10 6 conidia / ml of Botrytis cinérea strain CH98.
- the plants were maintained at a relative humidity of 100% for ten days and then the humidity was decreased to 80% during the rest of the experiment.
- the plants were evaluated after 22, 49 and 58 days post-inoculation (dpi) according to a score of the disease severity index where 0 corresponds to no symptoms and 5 corresponds to a dead plant.
- the peppers were collected at 122, 135, 150, 170 and 200 dpi and the total weight per plant was recorded.
- Stemicol inoculated 1, 65 to 1, 6 be 1, 7 to 1, 6 to 51, 6
- PRBT inoculated 1, 35 to 1, 35 be 1, 45 b 2,7b 87, 1
- the disease rate in pepper plants treated with PRBT was significantly lower than in untreated pepper plants and equivalent to plants treated with stemicol that had been inoculated with Botrytis cinerea.
- plants inoculated and treated with PRBT had a significantly lower disease rate (1, 35 be) compared to untreated inoculated plants (2.0 a).
- Example 11 - Tomato plants treated with PRBT and Bacillus subtilis have increased tolerance against white mold (Sclerotinia sclerotiorum)
- Var. Tomato plants were sown. Ventero on April 5, 2016 and distributed in randomized blocks of 9 plants with 7 replicates (63 plants per treatment) and grown in a standard greenhouse.
- PRBT (with 2: 1, 5 v / p and antifoam surfactant), Bacillus subtilis or a combination of both was sprayed three times before inoculation on June 20, July 7 and July 20 with PRBT (0.038 g 1 ) (C1 ), or PRBT (0.054 g I "1 ) (C2) and / or Bacillus subtilis at 4 l / ha (C3), or Bacillus subtilis at 6 l / ha (C4).
- C2 PRBT 0.054 g ⁇ 1 of the PRBT with surfactant and antifoam
- C3 Bacillus subtilis Bacillus subtilis at 4 l / ha
- C4 Bacillus subtilis Bacillus subtilis at 6 l / ha
- the disease index and AUDPC in tomato plants treated with two concentrations of PRBT or with two concentrations of Bacillus subtilis were significantly lower than in untreated tomato plants inoculated with Sclerotinia sclerotiorum.
- the disease rate and AUDPC in tomato plants treated with a combination of two concentrations of PRBT and Bacillus subtilis were significantly lower than in untreated tomato plants inoculated with Sclerotinia sclerotiorum.
- Example 12 Soybeans treated with PRBT and Bacillus subtilis have increased tolerance against white mold (Sclerotinia sclerotiorum)
- Soybean plants were sown and distributed in randomized blocks of 9 plants with 7 replicates (63 plants per treatment) and grown in a standard greenhouse.
- PRBT (with 2: 1, 5 v / p and antifoam surfactant), Bacillus subtilis or a combination of both was sprayed once before inoculation with PRBT (0.038 g I "1 ) (C1), or PRBT (0.054 g I " 1 ) (C2) and / or Bacillus subtilis at 4 l / ha (C3), or Bacillus subtilis at 6 l / ha (C4).
- the plants were inoculated with 10 ml / plant of a mixed mycelium foliar spray of Sclerotinia sclerotiorum strain CH109 that had been grown at an optical density at 600 nm of 1, 3.
- the plants were maintained at a relative humidity of 100% for ten days and then the humidity was lowered to 80% during the rest of the experiment.
- the plants were evaluated after 6, 9 and 14 days post-inoculation (dpi) according to a score of the disease severity index where 0 corresponds to no symptoms and 5 corresponds to a dead plant.
- the area under the disease assessment curve was calculated as a quantitative summary of the intensity of the disease over time, as well as the effectiveness of protection.
- Inoculated C1 PRBT 2.5 be 2.9 b 2.9 b 22.7 b 17.7 c
- Example 13 - Zucchini plants treated with PRBT have an increased tolerance against the New Delhi tomato leaf curl virus (ToLCNDV)
- the digoxigenin - labeled probe was obtained by PCR amplification of the partial AV1 gene DNA-A ToLCNDV using primer pairs Tonda-580F: 5'-TCACACATCGCGTAGGCAAG-3 '(SEQ ID NO: 1) and Tonda-935R: 5 ' -TGCCGGCCTCTTGTTGATTG-3 ' (SEQ ID NO: 2) with the PCR DIG mapping mixture (Roche Diagnostics, Switzerland) and according to the manufacturer's instructions.
- Immunodetection was performed with anti-digoxigenin antibody conjugated with alkaline phosphatase (Roche Diagnostics, Switzerland) and chemiluminescence with CSPD (Roche Diagnostics, Switzerland) as a substrate, according to the manufacturer's instructions and different exposure times (15 min-overnight) to a movie Lumi (Amersham Bioscience, RU).
- the virus disease index was calculated as: number of symptomatic ToLCNDV plants / total number of plants for 2014 and 2015 or number of plants positive for ToLCNDV / 180 total number of plants sampled for 2016.
- FIG. 3 shows photographs of zucchini plants infected with ToLCNDV treated with PRBT (panel B) compared to control (untreated) plants (panel A). Immunodetection membranes of 34 zucchini plants treated with PRBT (panel D) compared to control (untreated) plants (panel C) are shown where plants infected with the New Delhi tomato leaf curl virus are marked with a Continuous line rectangle with tissue impressions of the stem (left) and leaf (right) while the negative (left) and positive (right) controls are indicated with a dashed rectangle. Control (untreated) plants infected with ToLCNDV (panel C) they show a higher virus load on both stems and leaves than plants treated with PRBT (panel D).
- Table 19 below shows the total zucchini fruit production per plant in treated and untreated plants.
- the treated plants were sprayed every four weeks with 0.054 grams per liter of PRBT or every two weeks with 0.038 grams per liter of PRBT.
- Column 1 shows the season
- column 2 shows the group
- column 3 shows the average collected production for each plant in kilograms
- column 4 shows the number of plants for each group
- column 5 shows the percentage of profit with respect to to the control values
- column 6 shows the P value corresponding to T2 statistical analysis for daily production by plant and treatment data.
- the plants were grown in a conventional greenhouse by an experienced farmer during the last 3 seasons in Almeria, Spain. The percentage is shown with respect to the control values for harvest data for each year.
- Table 19 Total production of zucchini fruits per plant (kg) sprayed every four weeks with PRBT (0.054 g per liter) once a month or every two weeks with PRBT (0.038 g per liter) twice a month and untreated plants (control)
- Example 14 - PRBT contains 6-oxo-piperidine-2-carboxylic acid and not 6- hydroxypiperidine-2-carboxylic acid
- the content of 6-hydroxypyridine-2-carboxylic acid and 6-oxo-piperidine-2-carboxylic acid in PRBT was determined by quantitative MRM (LC-QQQ-MS).
- a total of 75 ⁇ of triplicate PRBT samples were dissolved in 100 ml of water, filtered and analyzed by LC-MS with an injection volume of 20 ⁇ at a flow rate of 0.4 ml / min and run time of 22 min (excluding 1 1 min of washing between samples).
- MRM transitions were: 6-hydroxypyridine-2-carboxylic acid, quantifier (m / z): 138, 10> 93.95 CE: +13, qualifier (m / z): 138, 10> 40, 10 CE: + 36, 6-oxo-piperidinyl acid, quantifier (m / z): 144.1> 97.95 CE: -15, qualifier (m / z): 144, 1> 55.05 EC: -26.
- the analytical column was a ZORBAX RX-SIL 5u 1 10 A 150 x 2, 1 mm.
- PRBT does not contain 6- hydroxypiperidine-2-carboxylic acid, while it contains an average of 2022 ⁇ 15.8 mg / l of 6-oxopiperidine-2-carboxylic acid.
- Example 15 Tomato plants treated with 6-oxopiperidine-2-carboxylic acid or 6-hydroxypiperidine-2-carboxylic acid have increased tolerance against white mold (Sclerotinia sclerotiorum) Var. Tomato plants were sown. Money Maker on December 13, 2016, distributed in randomized blocks of 9 plants with 4 replicates (36 plants per treatment) and grown in a standard greenhouse.
- the treated plants were sprayed on March 6, 2017 with 25 ml / plant at 3 ml / l (0.054 g 1 ) of PRBT (with 2: 1 surfactant, v / p, antifoam and biocide) or with 25 ml (0, 1 g 1 ) 6- hydroxypiperidine-2-carboxylic acid (6-HP-2CA) or with 25 ml (0.1 g 1 ) of 6- oxopiperidine-2-carboxylic acid (6-OXO-2CA).
- Table 21 disease index for plants treated with PRBT spray, 6-hydroxypiperidine-2-carboxylic acid and 6-oxopiperidine-2-carboxylic acid and untreated plants inoculated with white mold
- Example 15 Pepper plants treated with 6-hydroxypiperidine-2-carboxylic acid have an increased tolerance against white mold (Sclerotinia sclerotiorum). Pep pepper plants were seeded var. Murano on October 20, 2016 and were distributed in random blocks of 3 plants with 4 replicates (12 plants per treatment) and grown in a standard greenhouse.
- the plants were maintained at a relative humidity of 100% for four days and then the humidity was lowered to 80% during the rest of the experiment.
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