Plant growth promoter composition, processes for preparing the same and uses thereof

Abstract

To provide a composition to effectively promote plant growth by supplying a sufficient quantity of nutrients to a plant without causing any damage on foliage, and especially protecting the human and animal health, crops and environment.SOLUTION: Provided is a composition comprising a fungus of Trichoderma genus and a lignin fraction, wherein: the fungus is selected from Trichoderma species, their protoplast fusants, and mixtures thereof, the lignin fraction comprises fragments having a weight average molecular weight up to 20,000 Daltons, as measured by Size-Exclusion Chromatography, the fragments comprising up to 111 phenylpropane units on weight average, and wherein the fungus is in a concentration up to 1×107 spores / ml (composition), and the lignin fraction in a concentration up to 5 wt.%, based on the weight of the composition.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a composition comprising a fungus of the genus Trichoderma and a lignin fraction, as well as a process for its preparation and use as a plant growth promoter and fruit production promoter in agriculture. [Background technology]

[0002] Fertilizers consist essentially of various nutrients for plant growth and are widely used throughout the world to improve agricultural yields. Fertilizers may generally be in the form of pure liquids, suspensions, or solids. They can be introduced to plants through soil feeding or by foliar application (e.g., spraying, irrigation, etc.) to plants. In recent years, foliar fertilizers are gradually replacing the use of common soil fertilizers in agricultural fields, as they have less adverse environmental impacts. Studies have shown that traditional fertilization methods through soil feeding contribute to the pollution of surface and groundwater.

[0003] This is mainly due to leaching of soluble nutrients from the fertilizer, such as nitrogen, into the aquifer. In this situation, the transport of nutrients to the plant cells may be insufficient. It has therefore been found to be more desirable to supply nutrients directly to the plant through the plant's leaves. Foliar fertilization appears to overcome the shortcomings of soil-fed methods, but improper application of this foliar fertilization to the plant (e.g., when high concentrations of nutrients are applied directly to the leaves) may lead to foliar damage in the form of leaf necrosis or leaf burn, thus causing a reduction in crop yield. Foliar fertilization with reduced amounts of nutrients is believed to be able to prevent this foliar damage.

[0004] However, applying foliar fertilizers with low nutrient content to plants requires laborious work and is not practical. As an example, Patent Document 1 discloses a foliar fertilizer composition that promotes plant growth by foliar fertilization. The composition is an aqueous solution of at least one coenzyme, further at least one carbohydrate source, a complexing agent and a preservative, the coenzyme being preferably vitamin B, more preferably folic acid and / or pyridoxine. The subject fertilizer can significantly improve nutrient uptake by plants by increasing the metabolic activity of the plants, but the availability of nutrients taken up from soil or foliar fertilizer into plant cells remains an open question.

[0005] Therefore, a need is felt to effectively promote plant growth by providing sufficient amounts of nutrients to plants without damaging the leaves, particularly protecting human and animal health, crops and the environment. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] U.S. Patent No. 6,475,258 Summary of the Invention

[0007] The above object is achieved by a composition comprising a fungus of the genus Trichoderma and a lignin fraction as claimed in claim 1.

[0008] In another aspect, the present invention relates to the use of said compositions as plant growth promoters and fruit production promoters in agriculture.

[0009] In a further aspect, the present invention relates to an agricultural chemical product comprising said composition and an agricultural chemical additive.

[0010] In a further aspect, the present invention relates to a method of promoting plant growth and fruit production, said method comprising the step of applying the composition or agrochemical product to the plant or to the soil of the plant.

[0011] The term "plants" refers to plants that can be grown and harvested for profit or subsistence, and includes crops, grains, vegetables, fruits, and flowers, as well as those grown and harvested for horticultural or personal use.

[0012] The term "soil of a plant" refers to the soil in which the plant is growing or in which the plant is sown or in which the plant is to be sown, and thus includes ground, land, and soilless media such as water culture and hydroponics.

[0013] Features and advantages of the present invention will become apparent from the following detailed description, the examples provided by way of example, and the accompanying drawings. [Brief description of the drawings]

[0014] [Figure 1] Figure 1 shows the effect of the composition of the present invention on tomato yield. The bar graph shows the average yield per plant in grams of tomato fruit produced per fruiting branch. The different letters on the bars indicate significant differences according to ANOVA and LSD test (p < 0.05) according to Example 3. [Diagram 2] Figure 2 shows the effect of the composition of the invention on tomato yield. The bar graph shows the average yield per plant as the number of tomato fruits produced per plant. The different letters on the bars indicate significant differences according to ANOVA and LSD test (p < 0.05) according to Example 3. [Diagram 3] Figure 3 shows the effect of the composition of the present invention on tomato yield. The bar graph shows the average yield per plant in grams of tomato fruit produced per plant. Different letters on the bars indicate significant differences according to ANOVA and LSD test (p < 0.05) according to Example 3. [Figure 4] Figure 4 shows the effect of the composition of the present invention on tomato yield. The bar graph shows the average yield per plant as the number of tomato fruits produced per plant. The different letters on the bars indicate significant differences according to ANOVA and LSD test (p < 0.05) according to Example 3. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] [Detailed Description of the Invention] The subject of the present invention is therefore a composition comprising a fungus of the genus Trichoderma and a lignin fraction, which comprises: - the fungus is selected from Trichoderma species, protoplast fusions thereof, and mixtures thereof; - the lignin fraction comprises fragments having a weight average molecular weight of at most 20,000 Daltons as measured by size exclusion chromatography, the fragments comprising a weight average of at most 111 phenylpropane units; and Fungi have a maximum of 1×10 7 The concentration is in spores / ml of composition and the lignin fraction is at a concentration of up to 5 wt% based on the weight of the composition.

[0016] Preferably, the composition is in the form of an aqueous solution, dispersion, or suspension.

[0017] Preferably, the species of Trichoderma is selected from the group consisting of Trichoderma aggressivum, Trichoderma asperellum, Trichoderma atroviride, Trichoderma citrinoviride, Trichoderma cremeum, Trichoderma harzianum, Trichoderma koningii, Trichoderma longibrachiatum, Trichoderma reesei, Trichoderma virens, Trichoderma viride, Trichoderma spp. viride, and Trichoderma viridescens.

[0018] Fungi belonging to the genus Trichoderma as defined above are able to colonize various crevices, antagonize and control phytopathogenic microorganisms and establish direct beneficial interactions with the plant, thus improving growth, nutrient uptake and systemic resistance to diseases. In particular, improved plant growth is generally associated with an increase in seed germination, root system, plant weight and leaf area, as well as the size and / or number of seeds, flowers and / or fruits, resulting in an increase in yield and often an increase in the content of important nutritional factors.

[0019] Lignin is known to have antibacterial activity against both fungi and bacteria. Therefore, it was expected that the treatment of Trichoderma, a known soil fungus, would be directly inhibited at lignin concentrations that are already toxic to many other fungi tested. Therefore, a reasonable expectation was that Trichoderma would be affected at typical lignin concentrations considered for use in agricultural applications. Surprisingly and unexpectedly, the above Trichoderma species were not only unaffected by certain concentrations of lignin fractions, but also enhanced their activity in terms of plant productivity and plant growth, as shown in the following examples.

[0020] In particular, the plant growth promoting effect was observed to be increased by up to 25% in some cases compared to the application of a single component.

[0021] In addition to the above, it should be recognized that Trichoderma species and lignin fractions function in different ways and use different mechanisms: this makes the effect of the resulting composition more stable and suitable for a variety of conditions, while reducing the occurrence of resistance mechanisms (for biocontrol).

[0022] Moreover, advantageously, the compositions are easy and cost-effective to prepare as the respective concentrations are very low, which also means that the compositions can be prepared in concentrated form that can be easily diluted to the above concentrations with water prior to use.

[0023] The term "protoplast fusion" is meant to include hybrid strains of Trichoderma species obtained by protoplast fusion.

[0024] Protoplasts are cells from which the cell wall has been removed, and the cytoplasmic membrane is the outermost layer of the cell. Protoplasts can be obtained by certain lytic enzymes that remove the cell wall. Protoplast fusion is a physical phenomenon during which two or more protoplasts come into contact and attach to each other, either spontaneously or in the presence of a fusogenic agent. Protoplast fusion makes it possible to transfer some genes into another species. Protoplast fusion is an important tool for strain improvement to effect genetic modification and development of hybrid strains in filamentous fungi. The improvement can include, for example, higher yields in cellulase production.

[0025] Protoplast fusions for the purposes of the present invention can be obtained according to techniques known in the art (e.g., Hassan MM (2014); Influence of protoplast fusion between two Trichoderma spp. on extracellular enzymes production and antagonistic activity; Biotechnology & Biotechnological Equipment, 28:6, 1014-1023).

[0026] In a preferred embodiment of the composition of the present invention, the fungus of the genus Trichoderma is selected from T. harzianum, T. atroviride, and T. virens, and mixtures thereof.

[0027] In some embodiments, the composition comprises a mixture of Trichoderma species.

[0028] In a more preferred embodiment, the fungus is selected from T. harzianum HK2, T. atroviride HK4 and T. virens GV41, and mixtures thereof, with "HK2", "HK4" and "GV41" being preferred strains, respectively.

[0029] In some embodiments, the composition comprises a mixture of Trichoderma strains.

[0030] When a mixture is present in the composition, each species or strain is in the same or approximately the same concentration.

[0031] In a preferred embodiment, the composition comprises two Trichoderma species or two Trichoderma strains in a concentration ratio of 2:1 to 1:2, preferably 1:1.

[0032] Preferably, the fungus is at a concentration of 1×10 6 ~8×10 6 Concentration of spores / ml of composition.

[0033] More preferably, the fungus is at least 2×10 6 ~6×10 6 In a preferred embodiment, the fungus is at a concentration of 3×10 6 ~5×10 6 Concentration of spores / ml of composition.

[0034] Lignin is a type of complex organic polymer that forms the important structure of the support tissues of some algae, vascular plants including their bark, and herbaceous plants, e.g. trees (i.e. conifers and broad-leaved trees), all cereal straw, sugarcane bagasse, grasses, hemp, jute, hemp or cotton. Lignin can also be of mineral origin, e.g. peat, leonardite, coal.

[0035] In its natural form, lignin is chemically a highly irregular, randomly cross-linked polymer of phenylpropane units linked in many different linkages, with a weight average molecular weight of over 20,000 Daltons. A representative illustrative lignin fragment (I) containing the most important linkage patterns is shown below:

[0036] [ka]

[0037] The polymers are the result of the enzymatically mediated dehydrogenative polymerization of three phenylpropanoid monomer precursors:

[0038] [ka]

[0039] The parts are as follows:

[0040] [ka]

[0041] Coniferyl alcohol is present in all species and is the major monomer in conifers (conifers). Deciduous tree (broadleaf) species contain up to 40% sinapyl alcohol units, while grasses and crops may also contain coumaryl alcohol units.

[0042] Lignin is classified into softwood lignin and hardwood lignin according to the origin of its raw biomass.

[0043] The raw biomass sources that may be suitable starting materials for obtaining suitable lignin fractions are any lignin including substantially pure lignin, kraft lignin, biomass-derived lignin, lignin from an alkaline pulping process, lignin from a soda process, lignin from organosolv pulping, lignin from an enzymatic process, lignin from a steam explosion process, and combinations thereof.

[0044] The expression "substantially pure lignin" is to be understood as at least 80% pure lignin, preferably at least 90% pure lignin, more preferably at least 95% pure lignin, based on the dry raw biomass, the remainder being extractives, carbohydrates such as hemicellulose, and inorganic matter.

[0045] The expression "kraft lignin" is understood to mean lignin derived from kraft black liquor. Black liquor is an alkaline aqueous solution of lignin residues, hemicelluloses and inorganic chemicals used in the kraft pulping process. The black liquor of the pulping process contains components derived from different softwood and hardwood species in different proportions. Lignin can be separated from the black liquor by different methods, including for example precipitation and filtration. Lignin usually starts to precipitate at pH values ​​below 11-12. Different pH values ​​can be used to precipitate lignin fractions with different properties.

[0046] These lignin fractions are characterized by their molecular weight distribution (e.g., M w and M n ), polydispersity, hemicellulose and extractive content, inorganic content, etc. The precipitated lignin can be purified from inorganic impurities, hemicellulose and wood extractives using an acid washing process. It can also be further purified by filtration.

[0047] Alternatively, lignin can be separated from the pure biomass. The separation process can begin with liquefying the biomass with strong alkali, followed by a neutralization process. After the alkali treatment, the lignin can be precipitated in a similar manner as above.

[0048] Preferably, the separation of lignin from biomass involves an enzymatic treatment process. The enzymatic treatment alters the lignin extracted from the biomass. The lignin separated from the pure biomass is substantially sulfur-free (sulfur content less than 3%) and therefore valuable for further processing. Preferably, the wood is pretreated to remove hemicellulose and then the cellulose is hydrolyzed. The resulting insoluble lignin fraction contains up to 30 wt% cellulose.

[0049] Preferably, the separated lignin is subjected to a further depolymerization process to further reduce the weight average molecular weight of the fragments.

[0050] In some embodiments, the separated lignin is subjected to a depolymerization process to further reduce the weight and number average molecular weight of the fragments.

[0051] Suitable depolymerization processes include base-catalyzed depolymerization, acid-catalyzed depolymerization, metal-catalyzed depolymerization, ionic liquid assisted depolymerization, and supercritical fluid assisted lignin depolymerization.

[0052] In a preferred embodiment, said lignin fraction is obtained by base-catalyzed depolymerization.

[0053] Preferably, said lignin fraction is obtained by subjecting the separated lignin to base-catalyzed depolymerization at a temperature below 300° C. and a pressure below 30 MPa.

[0054] The pH is set between 11 and 14 by adding a base such as NaOH, KOH, Ca(OH)2, LiOH, K2CO3, or mixtures thereof.

[0055] The weight average molecular weight (M w) is measured by size exclusion chromatography (or "SEC"). In SEC, the stationary phase is the liquid present within the pores of the beads, and the moving liquid is the mobile phase. The mobile phase can therefore flow between the beads and also enter and exit the pores of the beads. The separation mechanism is based on the size of the polymer molecules in solution.

[0056] Larger molecules elute first. Smaller molecules that can fit into the many pores in the beads take longer to pass through the column and therefore exit the column slowly. To measure the molecular weight of the components of a polymer sample, a calibration with standard polymers of known weight must be performed. The value of the unknown sample is then compared to the calibration graph. Retention times vary depending on the column material used, the eluent, and how similar the standards used are compared to the sample. Preferably, the eluent is 0.1M NaOH.

[0057] Preferably, the lignin fraction contains fragments having a weight average molecular weight of 2,000 to 20,000 daltons.

[0058] More preferably, the lignin fraction contains fragments having a weight average molecular weight of 3,000 to 20,000 daltons.

[0059] More preferably, the lignin fraction contains fragments having a weight average molecular weight of 4,000 to 15,000 daltons.

[0060] In some preferred embodiments, the lignin fraction comprises fragments having a weight average molecular weight of 4,000 to 6,000 daltons.

[0061] In another preferred embodiment, the lignin fraction comprises fragments having a weight average molecular weight of 9,000 to 11,000 daltons.

[0062] Preferably, in these embodiments, the fragments contain, on average, from 11 to 111 phenylpropane units, and more preferably, on average, from 22 to 111 phenylpropane units.

[0063] The molecular weights of the three phenylpropanoid monomer precursors vary between 150 Da for coumaryl alcohol, 180 Da for coniferyl alcohol, and 210 Da for sinapyl alcohol. Therefore, the average weight was 180 Da, and this value was used as the "phenylpropane unit". w The value was divided by 180 Da to give the weight average number of phenylpropane units.

[0064] In a further embodiment, the lignin fraction has a polydispersity index (PDI) of 1.25-12.

[0065] Polydispersity index (PDI) or heterogeneity index, or simply dispersity, is a measure of the distribution of molecular weights in a particular polymer sample. PDI is the weight average molecular weight (M w ) to the number average molecular weight (M n ) which gives the distribution of individual molecular weights within a batch of polymer.

[0066] Preferably, the lignin fraction further comprises up to 30 wt% cellulose, more preferably 10-30 wt% cellulose, based on the weight of the lignin fraction. Trichoderma fungi produce cellulolytic enzymes such as exoglucanase (EXG), endoglucanase (EG) and β-glucosidase (BGL). Cellulase is the most efficient enzyme system for completely hydrolyzing cellulosic substrates into glucose monomers, a fermentable sugar. Since sugars aid in plant cellular respiration and cell growth, the presence of cellulose in the composition of the present invention can be advantageous to further improve the overall efficiency of plant growth promotion.

[0067] In a preferred embodiment, the composition of the present invention comprises a fungus of the genus Trichoderma and a lignin fraction comprising: - the fungus is selected from Trichoderma harzianum, Trichoderma atroviride, Trichoderma virens, and mixtures thereof; - the lignin fraction comprises fragments having a weight average molecular weight of 3,000 to 20,000 daltons as measured by size exclusion chromatography, the fragments comprising, on average, 16 to 111 phenylpropane units; and Fungus is 1×10 6 ~8×10 6 The concentration is in spores / ml of composition and the lignin fraction is at a concentration of 0.1 wt% to 4 wt% based on the weight of the composition.

[0068] More preferably, the composition of the present invention comprises a fungus of the genus Trichoderma and a lignin fraction comprising: - the fungus is selected from Trichoderma harzianum, Trichoderma atroviride, Trichoderma virens, and mixtures thereof; - the lignin fraction comprises fragments having a weight average molecular weight of 3,000 to 20,000 daltons as measured by size exclusion chromatography, the fragments comprising, on average, 16 to 111 phenylpropane units; and Fungus is 2 x 10 6 ~6×10 6 The concentration is of spores / ml of composition and the lignin fraction is at a concentration of 0.5 wt% to 3 wt% based on the weight of the composition.

[0069] In some preferred embodiments, the compositions of the present invention comprise a fungus of the genus Trichoderma and a lignin fraction: - the fungus is selected from Trichoderma harzianum HK2, Trichoderma atroviride HK4, Trichoderma virens GV41, and mixtures thereof; - the lignin fraction comprises fragments having a weight average molecular weight of 4,000 to 6,000 daltons as measured by size exclusion chromatography, the fragments comprising, on average, 22 to 33 phenylpropane units; and Fungus is 3×10 6 ~5×106 The concentration is of spores / ml of composition and the lignin fraction is at a concentration of 1 wt% to 2 wt% based on the weight of the composition.

[0070] In another preferred embodiment, the composition of the present invention comprises a fungus of the genus Trichoderma and a lignin fraction: - the fungus is selected from Trichoderma harzianum HK2, Trichoderma atroviride HK4, Trichoderma virens GV41, and mixtures thereof; - the lignin fraction comprises fragments having a weight average molecular weight of 9,000 to 11,000 daltons as measured by size exclusion chromatography, the fragments comprising, on average, 50 to 61 phenylpropane units; and Fungus is 3×10 6 ~5×10 6 The concentration is of spores / ml of composition and the lignin fraction is at a concentration of 1 wt% to 2 wt% based on the weight of the composition.

[0071] In a most preferred embodiment, the composition of the present invention comprises a fungus of the genus Trichoderma and a lignin fraction comprising: - the fungus is Trichoderma virens GV41; - the lignin fraction comprises fragments having a weight average molecular weight of 9,000 to 11,000 daltons as measured by size exclusion chromatography, the fragments comprising, on average, 50 to 61 phenylpropane units; and Fungus is 3×10 6 ~5×10 6 The concentration is of spores / ml of composition and the lignin fraction is at a concentration of 1 wt% to 2 wt% based on the weight of the composition.

[0072] In another embodiment, the composition consists essentially of a fungus of the genus Trichoderma and a lignin fraction: - the fungus is selected from Trichoderma aggressiveum, Trichoderma asperellum, Trichoderma atroviride, Trichoderma citrinoviride, Trichoderma cremeum, Trichoderma harzianum, Trichoderma koningii, Trichoderma longibrachiatum, Trichoderma reesei, Trichoderma virens, Trichoderma viride, Trichoderma viridescens, protoplast fusions thereof, and mixtures thereof; - the lignin fraction comprises fragments having a weight average molecular weight of at most 10,000 Daltons as measured by size exclusion chromatography, the fragments comprising a weight average of at most 55 phenylpropane units; and Fungi have a maximum of 1×10 7 The concentration is in spores / ml of composition and the lignin fraction is at a concentration of up to 5 wt% based on the weight of the composition.

[0073] For the purposes of the present invention, the expression "consists essentially of" means that the fungus and the lignin fraction are the only active ingredients present in the composition acting as plant growth promoters and fruit production promoters, possible other ingredients having different activities or simply being co-formulants.

[0074] In a further embodiment, the composition consists of a fungus of the genus Trichoderma and a lignin fraction, as described above.

[0075] In another aspect, the present invention relates to the use of said compositions as plant growth promoters and fruit production promoters in agriculture.

[0076] Preferably, the composition may be applied in an amount of 100-300 ml per plant every 5-15 days.

[0077] In a further aspect, the present invention relates to an agricultural chemical product comprising the composition and an agricultural chemical additive.

[0078] Suitable additives are pH adjusters, acidity adjusters, water hardness adjusters, mineral oils, vegetable oils, fertilizers, foliar fertilizers, and combinations thereof.

[0079] Exemplary additives include 2-ethylhexanol EO-PO, alkoxylated alcohols, alkoxylated fatty amines, alkoxylated triglycerides, alkyl polyglycosides, sodium alkyl ether sulfates, alkylphenol ethylene oxide condensates, alkylphenyl hydroxypolyoxyethylenes, allyl polyethylene glycol methyl ethers, dipropionic acid type amphoteric surfactants, di-1-p-menthene, dimethylpolysiloxanes, esterified vegetable oils, ethylene oxide condensates, fatty acid esters, fatty alcohol ethylene oxide condensates, fatty alcohol polyalkoxylates, Examples of suitable surfactants include glycerin, lecithin (soybean), methylated rapeseed oil, n-dodecylpyrrolidone, n-methylpyrrolidone, n-octylpyrrolidone, nonionic surfactants, nonylphenol ethylene oxide condensates, paraffin oil, poly(vinylpyrrolidone / 1-hexadecene), polyacrylamide, polyalkylene glycols, polyalkylene oxides, polyether-modified trisiloxanes, polyethylene polypropylene glycols, polyoxyethylene monolaurate, propionic acid, styrene-butadiene copolymers, synthetic latex, tallow amine ethoxylates, vegetable oils, and mixtures thereof.

[0080] Considering the fact that the composition is effective even at greatly reduced concentrations of fungi and lignin fractions, advantageously, the agrochemical product preferably comprises said composition at a concentration of 1-500 g per kg of agrochemical product.

[0081] The pesticide product may be a solid or a liquid.

[0082] When the agrochemical product is a solid, said solid may be a tablet, minitablet, microtablet, granule, granule, pellet, multiparticulate, micronized particle, or powder.

[0083] When the agrochemical product is a liquid, the liquid may be a solution, suspension, emulsion, dispersion, drip or sprayable liquid, and may be water-based or oil-based. The liquid may include a solvent. Suitable solvents are water, glycols, alcohols, polyhydric alcohols, organic acids, and combinations thereof.

[0084] Preferred solvents are water, methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol, allyl alcohol, 1,2-propylene glycol, 1,3-propylene glycol, 1,2-ethylene glycol, polyethylene glycol (PEG), glycerol, lactic acid, polylactic acid, and mixtures thereof. More preferred solvents are water, 1,2-propylene glycol, 1,3-propylene glycol, 1,2-ethylene glycol, polyethylene glycol (PEG), and mixtures thereof.

[0085] Preferably, when the agrochemical product is a liquid, said liquid has a pH of 5-9, more preferably 6-8.

[0086] When the agrochemical product is a liquid, the liquid contains 1-50 wt% of the composition, which means that the agrochemical product is a concentrate and can be appropriately diluted before use as necessary, or directly mixed with other chemical products.

[0087] The agrochemical products include granular bait, aerosol can, liquid (undiluted), bulk bait, matrix, concentrated bait, oil miscible liquid concentrate, encapsulated granule, suspension capsule, dispersible concentrate, powder, powder for dry tanning of seeds, emulsifiable concentrate, electrostatically charged liquid, water in oil emulsion, emulsion for tanning of seeds, water in oil emulsion, smoky jar, jar), granules, smoky candles, smoky cartridges, smoky slats, concentrated suspensions for blackening, smoky tablets, smoking agents (fumigants), smoky granules (or pellets), gas (under pressure), granular bait, gasification products, microgranules, sliding powder, granular, oil paste, hot smoke concentrate, solid / liquid combination packaging, liquid / liquid combination packaging, cold smoke concentrate, solid / solid combination packaging, lacquer, solution for blackening seeds, microemulsion, microgranules, dispersible oil, oil miscible concentrated suspension, oil miscible liquid, oil suspension, paste, flat The composition may be a bait, a concentrated paste or gel, a pour-on, a plant stick, treated or coated seeds, a ready to use bait, a spot-on, a fragmented bait, a concentrated suspension, a suspension-emulsion, a water soluble granular, a soluble concentrate, a film forming oil, a water soluble powder, a soluble powder for blackening seeds, a suspension, a tablet, an industrial material, an industrial concentrate, a tracing powder, an ultra low volume liquid, a water dispersible micro granular, a water dispersible granular, a wettable powder, a wettable powder for blackening seeds, a self-adhesive patch, and combinations thereof.

[0088] Advantageously, the agrochemical product may further comprise a fertilizer comprising a nitrogen, phosphorus, potassium compound, or a mixture thereof.

[0089] In a further aspect, the present invention relates to a method for promoting plant growth and fruit production, said method comprising the step of applying the composition or the agrochemical product to the plant or to the soil of the plant.

[0090] The pesticide product may be applied by one or more of the following procedures: - Mixing the pesticide product with the seeds in the sower hopper; - Applying pesticide products adjacent to the seeding furrow; - Spraying the entire field with agrochemical products before or after the last soil tillage.

[0091] If the pesticide product is a liquid, it may also be applied by one or more of the following procedures: - Spraying tubers, bulbs and seeds with pesticide products; - spraying the above-ground parts, leaves and stems of plants with pesticide products; - Immersing the roots of plants in an aqueous solution containing the pesticide product.

[0092] The agrochemical product may be applied in an amount ranging from 1,000g to 10,000kg of composition per hectare (ha), preferably from 1,000g to 1,000kg of composition, more preferably from 1,000g to 10,000g of composition.

[0093] It should also be understood that all combinations of the preferred embodiments of the compositions of the present invention, and their preparation methods, and their uses, reported above, are considered to be disclosed herein.

[0094] All combinations of the preferred aspects of the compositions, preparation methods and uses of the invention disclosed above are to be understood as being described herein.

[0095] Below are examples of the present invention that are provided for illustrative purposes. EXAMPLES

[0096] In these examples, M w and M n was determined by size exclusion chromatography according to the following procedure.

[0097] "wt%" means weight % based on the weight of the organic-inorganic composite material, unless otherwise specified.

[0098] Reagents and materials - Eluent: 0.1M NaOH, flow rate 0.5ml / min - RI detector calibration: pullulan standard sample, M p (M p is the maximum peak molecular weight): 100,000 to 1,080 (six types of standard samples) - Calibration of UV detector (280 nm): PSS standard sample, polystyrene sulfonate sodium salt, M p : 65,400 to 891 (6 types of standard samples). The standard samples are dissolved in ultrapure water to a concentration of approximately 5 mg / ml. The injection volume is 20 μl. - Quality control sample: M w Lignin with a known distribution is used.

[0099] Facilities and equipment - Dionex Ultimate 3000 Autosampler, Column Compartment, and Pump - Dionex Ultimate 3000 Diode Array Detector - Refractive index detector: Shodex RI-101 - Columns: PSS MCX column: pre-column and two analytical columns: 1000 Å and 100000 Å, the material of the columns is a sulfonated divinylbenzene copolymer matrix. - Syringe filter 0.45μm and glass sample bottle for STD samples. Sample filtration: Mini-Uniprep Syringeless Filter Device PTFE or Nylon, 0.45μm. Optionally, 5μm syringe filter for pre-filtration. - Measuring bottle

[0100] procedure - Preparation of eluent Ideally, the water used to prepare the eluent should be high-quality deionized water with low resistivity (greater than 18 MΩ cm) and as little dissolved carbon dioxide as possible. The water should be free of biological contaminants (e.g., bacteria and mold) and particulate matter. - Needle wash with 10% MeOH-water

[0101] - Liquid samples The strongly alkaline concentrated solution sample is diluted 1:100 and filtered through a PTFE syringe filter (0.45 μm) into a vial. The solid lignin sample is diluted and dissolved in 0.1 M NaOH and filtered through a PTFE (0.45 μm) syringe filter. The prepared sample is placed in the autosampler. The injection volume is 20 μl. 1 M NaOH is injected after the sample as a sample to wash the column.

[0102] Equipment parameters: - Flow rate 0.5ml / min - Eluent 0.1M NaOH - Column oven temperature 30°C - Isocratic elution - Run time 48 minutes

[0103] - Solid samples Dry solid samples (lignin) overnight in a 60 °C oven if necessary. Weigh approximately 10 mg into a 10 ml weighing bottle. Dissolve and dilute the sample in 0.1 M NaOH solution and fill to the mark. Filter the sample through a PTFE 0.45 μm filter. If the sample does not dissolve properly, it can be placed in an ultrasonic water bath or filtered through a 5 μm syringe filter.

[0104] - Calibration standards Approximately 50 mg of each standard is weighed into a 10 ml weighing bottle and filled to the mark with ultrapure water. The standards are filtered through a PTFE 0.45 μm syringe filter. After running the calibration samples, the calibration results are integrated into the processing method for processing and storage. The calibration is a linear first order calibration.

[0105] - Quality control samples The lignin samples were wLignin with known distribution is used as a quality control sample. The lignin is dissolved in 0.1 M NaOH and has a concentration of approximately 1 mg / ml.

[0106] Example 1 Organosolv lignin from beech tree (Fagus sylvatica) was subjected to enzymatic pretreatment to remove hemicellulose and then hydrolyze cellulose. The lignin fraction thus isolated from the pure biomass has the following characteristics: >95% of total solids Single species: Beech M w : 9,000-11,000 Da (50-61 phenylpropane units) Essentially sulfur free (less than 3% sulfur content) Contains 23-29 wt% cellulose.

[0107] This solid lignin fraction is called "EOS" for short.

[0108] Example 2 The following lignin fractions were extracted from kraft black liquor, said lignin fractions having the following characteristics: >95% of total solids Single species: Southern pine M w : 4400-5000 Da (24-28 phenylpropane units) M n : 1200-1300 Da (6-7 phenylpropane units) Structure of OH group: Aliphatic 2.1mmol / g Carboxyl group 0.5mmol / g Condensed and syringyl 1.7mmol / g Guaiacil 2.0mmol / g Catechol and p-OH-phenyl 4.0mmol / g

[0109] This solid lignin fraction is called "OXO" for short.

[0110] Example 3 Materials and Methods Seedlings sprouted 10 days after sowing in polystyrene boxes were transplanted into soil under a plastic tunnel and sprayed with 50 ml of the composition shown in Table 1 below (i.e., treatment). A second spray was performed 10 days later using 50 ml of the same composition. Subsequent sprays were performed every 10 days for 2 months, with each plant receiving 150-200 ml of the aqueous composition (Table 1).

[0111] If the treatment included a fungal consortium, the strains were mixed 1:1 or 1:1:1 to a final concentration of 4 × 10 6 Spores ml -1 It was decided.

[0112] A reference (i.e., negative control) was obtained by spraying the soil with water only.

[0113] Each treatment set in Table 1 consisted of 21 plants (three biological replicates of seven plants in randomly selected soil areas).

[0114] [Table 1]

[0115] Fully ripened fruits were harvested three times over a two-month period (August and September) starting three months after transplanting.

[0116] Tomato productivity was evaluated by measuring the yield in grams of tomato fruit per plant (considering the first two fruiting branches from the bottom up) and the number of fruits per plant in each treatment.

[0117] result In this example, the compositions were applied to tomato seedlings and the effect on plant growth and yield was evaluated.

[0118] The results obtained from field experiments and shown in Figures 1-4 showed that the composition is better in terms of promoting growth and yield, resulting in increased production in terms of number and kg of tomato fruits per plant compared to single component compositions.

Claims

[Claim 1] 1. A composition comprising a fungus of the genus Trichoderma and a lignin fraction: - said fungus is selected from Trichoderma species, their protoplast fusions, and mixtures thereof; - the lignin fraction comprises fragments having a weight average molecular weight of up to 20,000 Daltons as measured by size exclusion chromatography, the fragments comprising a weight average of up to 111 phenylpropane units; and The maximum fungal concentration is 1 x 10 ７ spores / ml of composition and a concentration of the lignin fraction is up to 5 wt% based on the weight of the composition.

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