IT201900019412A1 - HYDROLYZED TO PROMOTE VEGETABLE GROWTH, BIOSTIMULATION AND BIOCONTROL, AND ITS USE IN AGRICULTURE - Google Patents

Description

Filing of the patent application for industrial invention entitled: "HYDROLYZED TO FAVOR VEGETABLE GROWTH, BIOSTIMULATION AND BIOCONTROL, AND ITS USE IN AGRICULTURE"

FIELD OF THE INVENTION

The present invention relates to a hydrolyzate from natural raw material, which has been shown to be significantly effective in stimulating plant growth, or in protecting the plant against phytopathogenic agents. The hydrolyzate from natural raw material therefore finds an advantageous application in the agricultural sector.

STATE OF THE TECHNIQUE

Plant growth is dictated by both internal and external factors. The internal mechanisms originate in the genetic composition of the plant and influence the extent and timing of its growth. These internal mechanisms are regulated by various types of signals transmitted within the plant cells, or around the plant itself. External factors are directly related to the environment surrounding the plant. These external influences affect plant growth and include factors such as light, temperature, water and nutrients. The external environment can place constraints on the extent to which internal mechanisms can allow the plant to grow and develop, with two of the most important factors related to the availability of water and nutrients in the soil. Cell expansion is directly related to water supply and therefore any deficit results in a smaller plant. Mineral nutrients are necessary for the biochemical processes of the plant. When nutrients are insufficient, growth will be less vigorous or, in extreme cases, stop altogether. The nutrients needed for plant growth include: the primary macronutrients nitrogen (N), phosphorus (P) and potassium (K); the secondary macronutrients calcium (Ca), sulfur (S) and magnesium (Mg); and trace minerals or trace minerals boron (B), chlorine (CI), manganese (Mn), iron (Fe), zinc (Zn), copper (Cu), molybdenum (Mo) and selenium (Se).

To adequately support proper plant growth, especially in agriculture, the use of fertilizers is known. In general, fertilizers can come in the forms of pure liquid, suspensions or solid. They can be introduced into plants by fertilizing the soil or by applying them to the foliage of plants, such as by spraying, irrigation and the like. In recent years, foliar fertilizers have gradually replaced the common use of soil-applied fertilizers in agricultural areas, as they have minor negative impacts on the environment. Research has shown that the traditional fertilization process through soil fertilization has contributed to the contamination of surface water and groundwater. This is mainly due to the leaching of soluble fertilizer nutrients, such as nitrogen, into the aquifers. Such circumstances can lead to insufficient transport of nutrients to plant cells. Therefore, delivery of nutrients to a plant directly through the foliage of the plants has proved to be more desirable. Foliar fertilizers seem to overcome the disadvantages of the soil fertilization process, however, incorrect application of such foliar fertilizers to plants, for example when a high concentration of nutrients is applied directly to the foliage, can result in damage to the foliage in form of necrotic areas or leaf burns, thus causing a reduction in crop yields. It is speculated that foliar fertilizer with a reduced amount of nutrients can prevent damage to the foliage. However, this is not very functional, as labor-intensive activity is required to fertilize the plants with the low-nutrient foliar fertilizer.

It is therefore an object of the present invention to effectively promote plant growth, without causing damage to foliage and to the plant in general, and in particular by preserving human and animal health, crops and the environment.

SUMMARY OF THE INVENTION

Said object has been achieved by means of a hydrolyzate from natural raw material, as reported in claim 1, as well as a process for its preparation.

Under another aspect, the present invention relates to a composition comprising said hydrolyzate from natural raw material, and a microbial inoculant.

Furthermore, the present invention relates to the use of this hydrolyzate from natural raw material, as well as of this composition, as a promoter of plant growth and fruit production in agriculture, or for biocontrol.

In an additional aspect, the present invention relates to an agro-chemical product comprising the hydrolyzate from natural raw material or the composition, and agro-chemical additives.

In a further aspect, the present invention relates to a method for promoting vegetable growth and fruit production, said method comprising the step of applying the hydrolyzate from natural raw material or the composition or the agro-chemical product to a vegetable or to a soil for vegetables.

The term "vegetable" indicates a plant or plants that can be grown and harvested for profit or subsistence purposes, thereby encompassing crops, cereals, vegetables, fruit and flowers, as well as grown and harvested for gardening or personal use. .

The term "soil for vegetables" indicates the soil in which the vegetable is grown or in which the vegetable is sown or in which the vegetable will be sown, thus including soils, lands and means devoid of soil, including hydroculture and hydroponics.

The characteristics and advantages of the present invention will become apparent from the following detailed description and from the embodiments provided by way of illustrative and non-limiting examples.

DETAILED DESCRIPTION OF THE INVENTION

The invention therefore relates to a hydrolyzate from natural raw material comprising 1-25% of free amino acids and up to 55% of dry residue, in which said vegetable raw material is chosen from corn liqueur, lupine, algae, molasses, coriander, cocoa , pomace, and their combinations.

With "%" or "percentage", we mean "g / 100mL".

It has surprisingly been found that the hydrolyzate from natural raw material of the present invention allows not only to stimulate plant growth and productivity in terms of fruits, but also protects plants from phytopathogenic and weed agents.

Corn liquor (or corn steep liquor, or CSL) is a liquid byproduct of the wet milling process of corn used to make corn starch and high fructose corn syrup (HFCS). CSL consists of soluble corn concentrates extracted during a process in which the corn, after being shelled and purified from the air, is immersed in water (wet), and then divided into its main components by a combination of flotation and screening. wet. During maceration, the soluble materials are dissolved, the corn is softened and its structure weakened and broken, thus facilitating the grinding and further separations of its components. The resulting concentrate is a raw corn liqueur, which can be further combined with gluten and fibrous materials to be sold as feed, or it can be used for other purposes, with or without further processing. In addition to being used as a nutrient for ruminants, CSL has also been used in the penicillin industry as a culture medium for the production of penicillin. The corn liqueur (n.CAS 66071-94-1) is commercially available in aqueous solution (about 50% water), while the rest is made up of natural nutrient materials of corn, such as water-soluble proteins, amino acids (eg. alanine, arginine, arginine, aspartic acid, cysteine, glutamic acid, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, threonine, tyrosine, valine), vitamins (eg complex B), carbohydrates, organic acids (eg. lactic acid), minerals (eg. Mg, P, K, Ca, Ca, S), enzymes and other nutrients. CSL is a viscous slurry with a light brown to dark brown color, and has a pH of about 4.0.

Lupine is a legume from the Mediterranean basin, East Africa and the Americas, where it was already cultivated over 4,000 years ago. Today, however, it is grown in Australia, Europe and South America for food purposes, both for human and animal nutrition, and is also used as green manure (a crop that enriches the soil with precious elements, which are synthesized by bacteria nitrogen fixers present on the roots of the lupine plant). In addition to seeds, lupine can also be consumed in the form of flour, obtained by grinding dry seeds.

From a nutritional point of view, lupine stands out for its very high content of soluble and insoluble protein and fiber, and for a low content of carbohydrates and fats. Lupine flour contains at least 40% protein, 31% fiber, 10% sugar, 10% fat and various micronutrients. The fats contained in lupine seeds are mostly "good" fats, selling out 67% of monounsaturated and 19% polyunsaturated fatty acids, and only 14% of saturated fatty acids. Lupine seeds are a good source of thiamine (vitamin B1) and folic acid (vitamin B9), as well as some mineral salts, especially manganese, copper, magnesium, phosphorus, zinc and potassium.

Among the first and best known natural biostimulant raw materials, there are algae and in particular algae extracts. They have been used for hundreds of years in agriculture as soil improvers to improve soil fertility. Just over 50 years ago, the production of liquid extracts began to enhance the biostimulating properties of algae. Today there are numerous biostimulant products based on algae extracts available on the market.

The extracts are obtained starting from green, red or brown algae, especially of the type Ascophyllum nodosum, Ecklonia maxima, Laminaria digitata and Fucus spp.

The algae are harvested manually or mechanically along the ocean coasts and subjected to washing, cutting and extraction. The extraction can take place in different ways and with the use of solvents of various kinds. Techniques for the production of algae extracts have also been proposed which involve microbial fermentation of the starting plant matrix. The type of seaweed used, the harvesting period and the extraction process greatly influence the chemical characteristics of the extract and therefore its biostimulating properties.

Currently, the most used method involves cold extraction in high pressure water, in order to prevent chemical alterations of the bioactive molecules.

Seaweed extracts act as biostimulants by improving germination speed, growth, fruit set, production, product quality and resistance to environmental stress. In addition, the algae extracts increase the absorption of macro and micronutrients in various crops. The biostimulating effects are mainly due to the presence of phytohormones, polysaccharides, polyphenols and other organic molecules. The phytohormones identified in algae extracts that stimulate plant growth are auxins, cytokinins, abscisic acid, gibberellins, etc.

Of particular interest is the presence in the extracts of polysaccharides such as laminarine, alginates and fucoidans for their effect of improving the tolerance of plants to environmental stress. For example, applications of algae extracts increase tolerance to abiotic stresses such as drought, salinity and extreme temperatures.

The mechanisms underlying the increased tolerance to abiotic stress induced by applications of algae extracts are many and not yet fully known. These include the increase in root development and especially the roots / aerial part ratio, the improvement of the nutritional status of the crop, contributes to maintaining cell turgor, reducing the activity of free radicals, and increasing the activity. defense enzymes from oxidative stress.

From an application point of view, the positive effects of algae extracts are more marked in crops grown in low fertile soils and with repeated applications during the crop cycle. Foliar applications are generally preferred due to the reduced dosages required and the rapidity of action.

Molasses is a term that identifies, strictly speaking, the by-product of the production process of sucrose (commonly known as sugar), obtained by extraction from beet (Beta vulgaris) in the Mediterranean basin, and from sugar cane (Saccharum officinarum) in particularly in Central and South American countries. In common parlance, molasses are frequently identified, in an improper sense, other by-products or residues of extraction processes for obtaining sugars other than sucrose. On a commercial level, the term black-strap molasses identifies, in generic terms, molasses, regardless of origin: more specifically molasses refers to beet molasses, while black-strap refers to cane molasses. Molasses is an example of a raw material, widely used on an industrial scale as a source of carbon, in numerous microbiological processes and in animal feed. This raw material is presented as a viscous liquid, brown in color, obtained by concentrating the mother liquors, which remain from the sucrose extraction phase, starting from beet cut into strips or from cane subjected to pressure. In some cases the molasses undergo chemical-physical pretreatments before being used. Depending on the process technology and in particular on the parameters that affect the composition of the beet / cane (plant variety, seasonal trend, time elapsed between harvesting and extraction, etc.), molasses can present wide variability in composition. Molasses obtained from beet and cane have a similar total sugar content, but differ in particular in the content of sucrose and invert sugar, which is sucrose hydrolyzed into glucose and fructose. With regard to the content of mineral salts and growth factors, cane molasses has a higher content of vitamins, in particular of biotin (1 ÷ 3 μg / g), compared to that from beet (0.04 ÷ 0.15 μg / g). For this reason, beet molasses is frequently added with cane molasses, regardless of the geographical area of production and therefore of transport costs, with the aim of providing microbial crops with higher levels of biotin. Other by-products identified with the generic and improper term of molasses include other residues such as those of the extraction of sucrose from sorghum (50% of sucrose) and corn molasse or hydrol (sugar content 50 ÷ 60%), mother liquors of the process of production of glucose from corn starch. The molasse high-test (15 ÷ 35% of sucrose and 40 ÷ 60% of invert sugar), on the other hand, is not a residue or by-product of the production of sucrose, but the main product of the process, without the crystallization phase of the sucrose, as it is directly obtained by evaporation of the cane juice and partial inversion with invertase. This product can also derive from the processing of citrus fruits. In a specific case, the product may have the following characteristics:

Coriander (Coriandrum sativum, L.1753) or Chinese parsley or with the Spanish name cilantro, is an annual herbaceous plant of the Apiaceae family (or Umbelliferae). It belongs to the same family as cumin, dill, fennel and parsley. Coriandrum is a Latin word cited by Pliny (Naturalis Historia), which has its roots in the Greek word corys or korios (bedbug) followed by the suffix -ander (similar), referring to the supposed similarity of the smell emanating from unripe fruits or by rubbing leaves.

In this case, coriander can be used both as seed meal, but also the agricultural waste deriving from its preparation can constitute an economic and active starting substrate, albeit with more contained properties. Seventeen components have been identified representing 91.84% of the Egyptian residual essential oil of coriander.

Trans anethol was identified as the main compound of coriander waste oil oil in the percentage of 29.29% followed by, linalool (20.06%), butanoic acid, 2-methyl-, 2-methoxy-4 ester - (2-propyl) phenyl (14.17%), estragon (10.25%) longifolene (6.82%), and carvacrol (5.1%)

By comparing the constituents of green herbs, dry seeds and secondary materials of processing, it was found that, the main constituent was linalool in the oils of green herbs and dry seeds, while trans-anethole was found as the main component oil of the fractions. waste, followed by linalool. The natural raw material used can have the following characteristics:

Cocoa refers to the plant and seeds of Theobroma cacao Linn, an arboreal plant grown in tropical and subtropical regions, belonging to the Sterculiacee family. From the processing of the cocoa beans, different products are obtained in terms of quantity and quality, in addition to the paste, the powder and the cocoa butter, various by-products such as the shell, the pulp, the integuments and the germ are derived. These by-products each year involve many quantities of material that could be used for new production chains, this could be of help for developing countries, where the major cocoa producers are in fact present. The international cocoa market is divided into two: the producing countries, that is the South of the world and the processing and consumer countries, that is the North of the world, in particular Holland and North America. The undisputed leader among the producing countries is the Ivory Coast, with about 40% of world production, followed by Ghana and Indonesia. Also Ecuador, Cameroon, Nigeria and Brazil, which in total account for about 37%. The remaining 23% is divided between the producing countries in the tropical belt. In its production cycle, cocoa requires deep and clayey soils to be cultivated and a temperature between 25 ° C and 35 ° C. It needs abundant rainfall. The cocoa tree reaches about 8 m in height, in the most extreme cases even 18 m, with green leaves, reddish flowers gathered in panicles. It produces about 40-50 fruits over a year. A single fruit weighs about 400-500 g, is yellow-red when fresh and brown when dry. Inside there is an acidulous white mucilaginous pulp, where the seeds are found, which are wrapped in an integument consisting of a film that wraps the bean.

The cultivated species belongs to the genus Teobroma Cacao and the most commercially widespread cultivars are the Criollo, Forastero and Trinitario varieties. When they are ripe they are detached from the tree with a hook knife. Immediately after the harvest, the fruits are broken and the cocoa beans are spread on wooden pallets or on the ground on a layer of banana leaves, in order to facilitate the fermentation triggered by the yeasts. At the end of fermentation, the green cocoa is obtained, it is then left to dry at room temperature, until the humidity is reduced. Then there is the roasting of the beans, at a temperature between 130 ° C and 140 ° C. Then the special machines detach the integument from the germ. The by-products of the processing are the shells, the pulp, the integuments and the germ. The shells are the outermost part of the fruit and are the most significant by-product. Research in Japan has shown that they have an anticaryogenic activity, thanks to polyphenolic compounds and unsaturated fatty acids. In addition, the high mineral content makes the shells suitable for the production of soaps (black soap) and detergents. Thanks to potassium they can be good fertilizers. They can also be used for pig and sheep feed, mixed with other plant by-products. The pulp is removed during fermentation. From the pulp it is possible to obtain: pectins for jams, alcohol for industrial and hospital uses, vinegar and soft drinks. The pulp can be frozen and then used for the production of ice cream, to give yogurt flavor, but it is only marketed locally due to the high cost. From the integuments it is possible to extract a pigment used as a food coloring: the polyflavone glucoside. Furthermore, antioxidants such as flavonoids can be extracted through solvents. The germ is removed to improve the finished product. It is used for the extraction of oil for the food and cosmetic industry. In conclusion, by-products are obtained from cocoa processing that can be used in other fields and be useful for economic development. The quantity of by-products that originate is considerable and can be placed on the market and develop new production chains. In the case described here as a waste product, cocoa husks were preferably used, whose example analysis identified the following characteristic components:

The pomace consists of the solid residue (stones, peel films, parts of pulp) from the pressing of the olive paste. It represents 30-50% of the olives processed. Its composition is:

Oil: 5 - 10%

Water: 25 - 30%

Solid fraction of which 30% is core: 60 - 75%

The acidity of the oil extracted from the pomace ranges from 15 - 80%; a high acidity should be avoided because it complicates the rectification which becomes more expensive. The acidity increases with the stop at the oil mill due to hydrolysis due to lipase, active in the presence of H2O, and to auto-oxidation due to contact with air and considerable surface development. Antifermentatives such as NaHSO3 (2 - 3%) are not recommended because they cause drawbacks to the extraction.

The best conservation is obtained with drying in special rotary ovens, generally fed with pomace; with these, however, anhydrous pomace must never be obtained because cell membranes would be denatured which, forming an impermeable layer, would not allow the oil to be extracted. The optimal humidity value is 7% and in industrial practice it is controlled by listening to the noise produced by a handful of pomace in a hand. If the humidity is higher, the lipase can intervene with an exothermic reaction which can also cause fires due to the self-combustion of the product parked before extraction.

Whisk extraction process (now gone).

It is a physical-mechanical process by which the re-milled pomace is thrown into a tub of water. The light parts (bucchiette) rise to the surface, the stones go to the bottom and are discharged. By means of a stirrer (whisk) the separation of the oil is favored, which is released from the pits for washing in successive cascade tanks. Just over half of the oil is recovered.

The embodiments in which said hydrolyzate from natural raw material is hydrolyzed of corn liqueur are particularly preferred.

Preferably, said hydrolyzed from natural raw material is:

- hydrolyzate of corn liqueur, comprising 6-16% of free amino acids and up to 53% of dry residue,

- lupine hydrolyzate, comprising 7-20% free amino acids and up to 30% dry residue,

- hydrolyzate of algae, comprising 6-8% of free amino acids and up to 10% of dry residue,

- hydrolyzate of molasses, comprising 6-8% of free amino acids and up to 55% of dry residue,

- coriander hydrolyzate, comprising 1-5% free amino acids and up to 3% dry residue,

- cocoa hydrolyzate, comprising 1-2% of free amino acids and up to 3% of dry residue,

- pomace hydrolyzate, comprising 6-8% of free amino acids and up to 55% of dry residue,

or a combination thereof.

From another aspect, the present invention relates to a process for preparing this hydrolyzate from natural raw material, comprising the steps of:

i) supply the natural raw material, and optionally add water,

ii) adding an enzyme complex comprising at least one peptide bond hydrolase and at least one fiber and / or carbohydrate hydrolase, and

iii) obtain a liquid mixture, i.e. the hydrolyzate from natural raw material.

Preferably, in step i) water is added to the natural raw material.

Preferably, in step ii) the temperature is 4-70 ° C for a time period of 2-20 hours, more preferably the temperature is 10-50 ° C for a time period of 4-16 hours. Examples of enzymes active on the protein component: for example the components of class EC 3.4, enzymes capable of acting on the peptide bond, such as for example mono, di or tripepdidases, or their mixtures, proteases and proteinases.

Examples of enzymes active on fibers and / or carbohydrates: for example the components of the EC 3.2 Glycosidase class, enzymes capable of acting O- and S-glycosyl compounds or N-glycosyl compounds, such as for example amylase, cellulase, hemicellulase, laccase, and xylanase.

Examples of enzymes active on lipids: for example the components of class EC 3.1, enzymes capable of acting on the ester bond, such as lipases.

Preferably, in said enzymatic complex, the hydrolase is selected from mono, di or tripepdidases, proteases, proteinases, cellulases, hemicellulases, pectinases, xylanases, amylases, laccases, lipases and their mixtures.

The enzymatic complex comprises a mixture of at least two hydrolases, ie at least one hydrolase active on fibers and / or carbohydrates and at least one hydrolase active on the protein component. It has in fact been observed that this mixture acts synergistically, allowing surprising results to be obtained compared to the use of a single enzyme.

Preferably in step ii), and in any case in compliance with the pH activity range of the specific enzyme, a pH corrector is added, more preferably an organic or inorganic base, such as NaOH or KOH, to adjust the pH to about 6. In in some cases an inorganic or organic acid, such as phosphoric acid, can also be used in order to reach the optimal reaction pH of the specific enzymatic mix.

According to a particularly preferred first preparation procedure, the natural raw material is treated with an anti-fermentation agent and a mixture of enzymes EC 3.2 for 6 hours at about 52 ° C and at pH about 4. Subsequently, KOH was added up to pH 6 and then a mixture of EC 3.2 and 3.4 enzymes for another 6 hours. In the case of raw materials with a lipid part, such as pomace, EC 3.1 is also added in this step, enzymes capable of acting on the ester bond, such as lipases.

According to a second particularly preferred preparation procedure, the natural raw material is treated with an anti-fermentation agent and a mixture of enzymes EC 3.2 and 3.4 for 6 hours at approximately 52 ° C and at pH 4. lipidic, such as pomace, in this step EC 3.1 is also added, enzymes capable of acting on the ester bond, such as lipases. Subsequently, KOH was added up to pH 5.5-6.

From another aspect, the present invention relates to a hydrolyzate from natural raw material obtainable from the process described above, in which said vegetable raw material is selected from corn liqueur, lupine, algae, molasses, coriander, cocoa, pomace, and their combinations, in which the percentage of free amino acids in the hydrolyzate is at least 0.5% higher than the percentage in the starting vegetable raw material, and in which the dry residue in the hydrolyzate is at least 2% lower than the percentage in the vegetable raw material of departure.

With "%" or "percentage", we mean "g / 100mL".

Preferably, said hydrolyzate from natural raw material obtainable from the process described above is:

- corn liqueur hydrolyzate, in which the percentage of free amino acids in the hydrolyzate is 0.5-11% higher than the percentage in the starting corn liqueur, and in which the dry residue in the hydrolyzate is 10-15% less than the percentage in the starting corn liqueur,

- lupine hydrolyzate, in which the percentage of free amino acids in the hydrolyzate is 2-15% higher than the percentage in the starting lupine, and in which the dry residue in the hydrolyzate is 10-15% lower than the percentage in the lupine of departure,

- hydrolyzate of algae, in which the percentage of free amino acids in the hydrolyzate is 4-6% higher than the percentage in the starting algae, and in which the dry residue in the hydrolyzate is 2-4% lower than the percentage in the algae of departure,

- hydrolyzate of molasses, in which the percentage of free amino acids in the hydrolyzate is 4-6% higher than the percentage in the starting molasses, and in which the dry residue in the hydrolyzate is 10-20% lower than the percentage in the molasses of departure,

- coriander hydrolyzate, in which the percentage of free amino acids in the hydrolyzate is 0.5-4.5% higher than the percentage in the starting coriander, and in which the dry residue in the hydrolyzate is 10-15% lower than percentage in the starting coriander, - cocoa hydrolyzate, in which the percentage of free amino acids in the hydrolyzate is 0.5-1.5% higher than the percentage in the starting cocoa, and in which the dry residue in the hydrolyzate is 10 -15% lower than the percentage in the starting cocoa,

- pomace hydrolyzate, in which the percentage of free amino acids in the hydrolyzate is 4-6% higher than the percentage in the starting pomace, and in which the dry residue in the hydrolyzate is 5-10% lower than the percentage in the pomace of departure,

or a combination thereof.

From another aspect, the present invention also relates to the use of this hydrolyzate from natural raw material, as a promoter of plant growth and fruit production in agriculture, or for biocontrol.

In particular, this hydrolyzate is preferably to be used in quantities of 0.5-50 liters per hectare of soil if liquid, or in equivalent proportional quantities for other liquid or solid formulations, such as for example solid granules dispersible in water or non-dispersible in water.

From another aspect, the present invention relates to a composition comprising a microbial and hydrolyzed inoculant from natural raw material as described above, wherein said microbial inoculant is selected from bacterial inoculant, fungal inoculant or a combination thereof.

Microbial inoculants also known as soil inoculants or bio-inoculants are agricultural corrective agents that use beneficial rhizospheric or endophytic microbes to promote plant health. Many of the microbes involved form symbiotic relationships with target crops that both parties benefit from (mutualism). While microbial inoculants are applied to improve plant nutrition, they can also be used to promote plant growth by stimulating the production of plant hormones. When added to seeds, soils or leaves, microbial inoculants have proven useful in all crops both in the open field and in the greenhouse. Microbial inoculants can also be used to initiate acquired systemic resistance (SAR) of plant species to several common crop diseases.

As mentioned, the microbial inoculant is chosen from bacterial inoculant, fungal inoculant or a combination thereof.

Preferred kinds of bacterial microorganisms are Azospirillum, Rhizobium, Bacillus, Pseudomonas, Streptomyces, Zooglia, Agrobacterium, and combinations thereof.

Preferably, the bacterial inoculant belongs to the species Bacillus subtilis, Bacillus megaterium, Bacillus velezensis, Bacillus amyloliquefaciens, Azotobacter vinelandi, Azospirillum brasilense, Ensifer meliloti, Pseudomonas vancouverensis, Paenibacillus polymyxa, and their combinations.

For non-legume crops, Azospirillum has been shown to be beneficial for nitrogen fixation and plant nutrition. Rhizobium is a genus of soil bacteria that fix nitrogen and form symbiotic associations within the nodules on the roots of legumes. This increases nitrogen nutrition and is important for the cultivation of soy, chickpeas and many other legumes. Bacillus, Pseudomonas and Streptomyces provide some, if not all, of the following benefits: increased plant growth, decomposition of organic matter and pesticide residues, increased nutrient cycle and nitrogen fixation, increased resistance to biotic and abiotic, greater solubility of meso- and microelements, greater production of natural hormones useful for plant growth, better soil structure and better germination and vitality of seeds. Furthermore, to improve the bioavailability of phosphorus, solubilizing phosphate bacteria (PSB) are also used, which act on the inorganic phosphates of the soil in simpler forms that allow their absorption by plants.

In other preferred embodiments, the composition of the invention comprises forms of microorganisms in concentration from 1x10 <5> CFU / ml to 1x10 <10> CFU / ml of said bacterial inoculant.

Preferred genera of fungal microorganisms are Ascomycetes and Basidiomycetes.

Preferably, the fungal inoculant belongs to the species Trichoderma asperellum, Trichoderma longibrachiatum, Metarhizium anisopliae, Pochonia chlamidospora, and their combinations. These fungi provide many plant health benefits of the above bacteria, including increasing the plant's resistance to environmental stresses and producing natural hormones.

In other preferred embodiments, the composition of the invention comprises forms of microorganisms in concentration from 1x10 <5> CFU / ml to 1x10 <10> CFU / ml of said fungal inoculant.

In preferred embodiments, said microbial inoculant comprises up to 5 different microorganisms.

More preferably, said microbial inoculant comprises Bacillus subtilis Bacillus megaterium, Bacillus velezensis, Bacillus amyloliquefaciens, Azotobacter vinelandi, Azospirillum brasilense, Ensifer meliloti, Pseudomonas vancouverensis, or a combination thereof.

In other embodiments, the composition essentially consists of a microbial and hydrolyzed inoculant of natural raw material, as described above. For the purposes of the present invention, the expression "essentially consists of" indicates that said microbial inoculant and said corn liqueur hydrolyzate are the only active ingredients which act as plant growth and fruit production promoters present in the compositions, the possible other components exhibiting different activities which do not interfere with those of microbial and hydrolyzed inoculant, or being simple co-formulants.

In further embodiments, the composition consists of a microbial and hydrolyzed inoculant of natural raw material, as described above.

Preferably, the composition of the invention further comprises liqueur of corn, lupine, algae, molasses, coriander, cocoa, pomace, and their combinations.

Optionally, the composition of the invention further includes additional ingredients glycerol, humates (from humic acid), fulvates (from fulvic acid), acetic acid, propionic acid, citric acid, lactic acid or combinations thereof.

Optionally, the composition of the invention further comprises additional ingredients such as botanical extracts, fermented plant extracts, or combinations thereof.

Optionally, the composition of the invention further comprises additional ingredients such as amino acids. Amino acids can act as an energy source to increase plant metabolism and improve plant nutrient absorption. Such additional amino acids can be tryptophan, asparagine, glutamine, glycine, selenocysteine, serine, ornithine, taurine or combinations thereof.

Optionally, the composition of the invention further includes additional ingredients such as vitamins and minerals. Vitamins can act as catalysts for beneficial enzymes and improve plant metabolism. Folic acid and biotin (two components of the vitamin B complex) also work to improve microbial and plant growth. These vitamins and minerals can be Boron, Copper, Iron, Iron, Manganese, Zinc, Molybdenum, Chlorine, Phosphorus, Potassium, Calcium, Magnesium, Sulfur, Sulfur, or combinations thereof.

Optionally, the composition of the invention further comprises additional ingredients such as carbohydrates. Such carbohydrates can be glucose, galactose, galactose, fructose, arabinose, xylose, sucrose, lactose, maltose, amylase, amylopectin, glycogen, glycogen, glyceraldehyde, ribose or combinations thereof.

Optionally, the composition of the invention further comprises additional ingredients such as enzymes. Such enzymes can be phospholipases, lipases, proteases, amylases, cellulases, catalases, laccases or combinations thereof.

Preferably, the composition can be applied in an amount of 0.5-50 liters / hectare, more preferably with periodic treatments depending on the crops and seasonality. In an additional aspect, the present invention relates to an agro-chemical product comprising the composition and agro-chemical additives.

Suitable additives are pH regulators, acidity regulators, water hardness regulators, mineral oils, vegetable oils, fertilizers, natural leaf fertilizers and their combinations.

It is to be understood that all the possible combinations of the preferred aspects of the components of the composition, as indicated above, are also described, and therefore similarly preferred.

It should also be understood that all the aspects identified as preferred and advantageous for the composition and its components are to be considered similarly preferred and advantageous also for the preparation and uses of the composition itself.

Examples of embodiments of the present invention are given below, provided for illustrative and non-limiting purposes.

EXAMPLES

Example 1. Preparation of the hydrolyzates of the invention

The natural raw material was analyzed for the content in organic nitrogen and starches and fibers, in order to evaluate the best execution of the enzymatic digestion of the components. On the basis of the results, different hydrolases able to act on the components were tested, evaluating the necessary units on the basis of the composition.

Examples of enzymes active on the protein component: for example the components of class EC 3.4 enzymes capable of acting on the peptide bond, such as for example mono, di or tripepdidases, or their mixtures, proteases and proteinases.

Examples of enzymes active on fibers and carbohydrates: for example the components of the EC 3.2 Glycosidase class, enzymes capable of acting O- and S-glycosyl compounds or N-glycosyl compounds, such as for example amylase, cellulase and xylanase.

Each hydrolase was used in the optimum pH and temperature ranges, developing different processes, which can all lead to the hydrolysis of the different components present, regardless of the path.

1A) Hydrolysis of CSL

For the hydrolysis of CSL one of the mixtures of enzymes used and effective consists of: EC 3.2 Blend in equal parts of arabanase, cellulase, beta-glucanase, hemicellulase, and xylanase, eg. from Tricoderma 5-10% weight / volume

EC 3.2 Cellulase, eg. from Tricoderma 2-10% weight volume

EC 3.4 Protease, eg. from Subtilisin 2-10% weight / volume

EC 3.2 alpha amylase, eg. from Bacillus Licheniformis 1-2.5% Weight / volume

The production recipe is as follows:

CO O O Q A '% /

According to a first preparation procedure, the CSL is treated with an antifermentative and a mixture of enzymes EC 3.2 for 6 hours at about 52 ° C and at pH about 4. Subsequently, KOH was added up to pH 6 and then a mixture of EC enzymes 3.2 and 3.4 for another 6 hours.

Instead, following a second preparation procedure, the CSL is treated with an antifermentative and a mixture of enzymes EC 3.2 and 3.4 for 6 hours at about 52 ° C and at pH about 4. Subsequently, KOH was added up to pH 5.5-6 .

The amino acid analysis shows that there is a general increase in free amino acids between 0.5 and 11%, and specifically, these are the amino acids made more bioavailable.

These results are obtained both when the enzymes of class 3.4 acting on the peptide bond are used alone, and in a mixture with enzymes of the other classes.

There is a specific example of total CSL analysis before and after hydrolysis, in which specifically the free amino acids using process 1 reach 7%

Following the treatment with enzymes belonging to classes 3.2 that act on fibers and complex carbohydrates, the dry residue, indicating the presence of insoluble material, decreases, and the quantity of polysaccharides or simple sugars present in the mixture increases:

1B) Lupine hydrolysis

For the hydrolysis of CSL one of the mixtures of enzymes used and effective consists of:

EC 3.2 Blend in equal parts of arabanase, cellulase, beta-glucanase, hemicellulase, and xylanase, eg. from Tricoderma 5-10% weight / volume

EC 3.2 Cellulase, eg. from Tricoderma 2-10% weight volume

EC 3.4 Protease, eg. from Subtilisin 2-10% weight / volume

EC 3.2 alpha amylase, eg. from Bacillus Licheniformis 1-2.5% Weight / volume

The production recipe is as follows:

According to a first preparation procedure, the lupine is treated with an anti-fermentative and a mixture of enzymes EC 3.2 for 6 hours at about 52 ° C and at a pH of about 4. Subsequently, KOH was added up to pH 6 and then a mixture of EC enzymes 3.2 and 3.4 for another 6 hours.

Instead, following a second preparation procedure, the lupine is treated with an antifermentative and a mixture of enzymes EC 3.2 and 3.4 for 6 hours at about 52 ° C and at pH about 4. Subsequently, KOH was added up to pH 5.5-6 .

Following the treatment with enzymes belonging to classes 3.2 which act on fibers and complex carbohydrates, the dry residue, indicating the presence of insoluble material, decreases, and the quantity of polysaccharides or simple sugars present in the mixture increases. Viscosity decreases, and probably thanks to this aspect, the protein part becomes more available for the attack of proteolytic enzymes.

The amino acid analysis shows that there is a general increase in free amino acids between 2 and 15%.

These results are obtained both when the enzymes of class 3.4 that act on the peptide bond are used alone, and in a mixture with enzymes of the other classes.

1C) Hydrolysis of Ascophyllum algae

For the hydrolysis of algae, one of the mixtures of enzymes used and effective consists of: EC 3.2 Blend in equal parts of arabanase, cellulase, beta-glucanase, hemicellulase, and xylanase, eg. from Tricoderma 5-10% weight / volume

EC 3.2 Cellulase, eg. from Tricoderma 2-10% weight volume

EC 3.4 Protease, eg. from Subtilisin 2-10% weight / volume

EC 3.2 alpha amylase, eg. from Bacillus Licheniformis 1-2.5% Weight / volume

The production recipe is as follows:

According to a first preparation procedure, the algae are treated with an anti-fermentative and a mixture of enzymes EC 3.2 for 6 hours at about 52 ° C and at a pH of about 4. Subsequently, KOH was added up to pH 6 and then a mixture of EC enzymes 3.2 and 3.4 for another 6 hours.

Instead, following a second preparation procedure, the algae are treated with an anti-fermentation agent and a mixture of enzymes EC 3.2 and 3.4 for 6 hours at about 52 ° C and at pH about 4. Subsequently, KOH was added up to pH 5.5-6 .

Following the treatment with enzymes belonging to classes 3.2 which act on fibers and complex carbohydrates, the dry residue, indicating the presence of insoluble material, decreases, and the quantity of polysaccharides or simple sugars present in the mixture increases. Viscosity decreases, and probably thanks to this aspect, the protein part becomes more available for the attack of proteolytic enzymes

The amino acid analysis shows that there is a general increase in free amino acids between 4 and 6%.

These results are obtained both when the enzymes of class 3.4 that act on the peptide bond are used alone, and in a mixture with enzymes of the other classes.

1D) Hydrolysis of molasses

For the hydrolysis of CSL one of the mixtures of enzymes used and effective consists of:

EC 3.2 Blend in equal parts of arabanase, cellulase, beta-glucanase, hemicellulase, and xylanase, eg. from Tricoderma 5-10% weight / volume

EC 3.2 Cellulase, eg. from Tricoderma 2-10% weight volume

EC 3.4 Protease, eg. from Subtilisin 2-10% weight / volume

EC 3.2 alpha amylase, eg. from Bacillus Licheniformis 1-2.5% Weight / volume

The production recipe is as follows:

According to a first preparation procedure, the molasses is treated with an anti-fermentative and a mixture of enzymes EC 3.2 for 6 hours at about 52 ° C and at a pH of about 4. Subsequently, KOH was added up to pH 6 and then a mixture of EC enzymes 3.2 and 3.4 for another 6 hours.

Instead, following a second preparation procedure, the CSL is treated with an antifermentative and a mixture of enzymes EC 3.2 and 3.4 for 6 hours at about 52 ° C and at pH about 4. Subsequently, KOH was added up to pH 5.5-6 .

Following the treatment with enzymes belonging to classes 3.2 which act on fibers and complex carbohydrates, the dry residue, indicating the presence of insoluble material, decreases, and the quantity of polysaccharides or simple sugars present in the mixture increases. Viscosity decreases, and probably thanks to this aspect, the protein part becomes more available for the attack of proteolytic enzymes

The amino acid analysis shows that there is a general increase in free amino acids between 4 and 6%.

These results are obtained both when the enzymes of class 3.4 that act on the peptide bond are used alone, and in a mixture with enzymes of the other classes.

1E) Coriander hydrolysis

For the hydrolysis of cocoa, one of the mixtures of enzymes used and effective consists of: EC 3.2 Blend in equal parts of arabanase, cellulase, beta-glucanase, hemicellulase, and xylanase, eg. from Tricoderma 5-10% weight / volume

EC 3.2 Cellulase, eg. from Tricoderma 2-10% weight volume

EC 3.4 Protease, eg. from Subtilisin 2-10% weight / volume

EC 3.2 alpha amylase, eg. from Bacillus Licheniformis 1-2.5% Weight / volume

The production recipe is as follows:

According to a first preparation procedure, the cocoa is treated with an anti-fermentative and a mixture of enzymes EC 3.2 for 6 hours at about 52 ° C and at a pH of about 4. Subsequently, KOH was added up to pH 6 and then a mixture of EC enzymes 3.2 and 3.4 for another 6 hours.

Instead, following a second preparation procedure, the cocoa is treated with an anti-fermentative and a mixture of enzymes EC 3.2 and 3.4 for 6 hours at about 52 ° C and at pH about 4. Subsequently, KOH was added up to pH 5.5-6 .

Following the treatment with enzymes belonging to classes 3.2 which act on fibers and complex carbohydrates, the dry residue, indicating the presence of insoluble material, decreases, and the quantity of polysaccharides or simple sugars present in the mixture increases. Viscosity decreases, and probably thanks to this aspect, the protein part becomes more available for the attack of proteolytic enzymes

The amino acid analysis shows that there is a general increase in free amino acids between 0.5 and 4.5%.

1F) Hydrolysis of cocoa

For the hydrolysis of cocoa, one of the mixtures of enzymes used and effective consists of:

EC 3.2 Blend in equal parts of arabanase, cellulase, beta-glucanase, hemicellulase, and xylanase, eg. from Tricoderma 5-10% weight / volume

EC 3.2 Cellulase, eg. from Tricoderma 2-10% weight volume

EC 3.4 Protease, eg. from Subtilisin 2-10% weight / volume

EC 3.2 alpha amylase, eg. from Bacillus Licheniformis 1-2.5% Weight / volume

The production recipe is as follows:

According to a first preparation procedure, the cocoa is treated with an anti-fermentative and a mixture of enzymes EC 3.2 for 6 hours at about 52 ° C and at a pH of about 4. Subsequently, KOH was added up to pH 6 and then a mixture of EC enzymes 3.2 and 3.4 for another 6 hours.

Instead, following a second preparation procedure, the cocoa is treated with an anti-fermentative and a mixture of enzymes EC 3.2 and 3.4 for 6 hours at about 52 ° C and at pH about 4. Subsequently, KOH was added up to pH 5.5-6 .

Following the treatment with enzymes belonging to classes 3.2 which act on fibers and complex carbohydrates, the dry residue, indicating the presence of insoluble material, decreases, and the quantity of polysaccharides or simple sugars present in the mixture increases. Viscosity decreases, and probably thanks to this aspect, the protein part becomes more available for the attack of proteolytic enzymes.

The amino acid analysis shows that there is a general increase in free amino acids between 0.5 and 1.5%.

1G) Hydrolysis of pomace

For the hydrolysis of pomace, one of the mixtures of enzymes used and effective consists of:

EC 3.2 Blend in equal parts of arabanase, cellulase, beta-glucanase, hemicellulase, and xylanase, eg. from Tricoderma 5-10% weight / volume

EC 3.2 Cellulase, eg. from Tricoderma 2-10% weight volume

EC 3.4 Protease, eg. from Subtilisin 2-10% weight / volume

EC 3.2 alpha amylase, eg. from Bacillus Licheniformis 1-2.5% Weight / volume

The production recipe is as follows:

According to a first preparation procedure, the pomace is treated with an anti-fermentative and a mixture of enzymes EC 3.2 for 6 hours at about 52 ° C and at a pH of about 4. Subsequently, KOH was added up to pH 6 and then a mixture of EC enzymes 3.2 and 3.4 and 3.1 for another 6 hours.

Instead, following a second preparation procedure, the pomace is treated with an antifermentative and a mixture of enzymes EC 3.2 and 3.4 and 3.1 for 6 hours at about 52 ° C and at pH 4. Subsequently, KOH was added up to pH 5.5. -6.

Following the treatment with enzymes belonging to classes 3.2 which act on fibers and complex carbohydrates, the dry residue, indicating the presence of insoluble material, decreases, and the quantity of polysaccharides or simple sugars present in the mixture increases. Viscosity decreases, and probably thanks to this aspect, the protein part becomes more available for the attack of proteolytic enzymes.

The amino acid analysis shows that there is a general increase in free amino acids between 4 and 6%.

These results are obtained both when the enzymes of class 3.4 that act on the peptide bond are used alone, and in a mixture with enzymes of the other classes.

Example 2.

CSL hydrolysates and efficacy tests

A greenhouse test was carried out on cyclamen; 18 plants were analyzed for each thesis. Each thesis was treated in fertigation, with approximately 25 ml per plant, corresponding to the following doses / hectare. The treatment was carried out three times over three weeks, one treatment per week. The following fertigation protocol was applied 45PPM N - NPK (1: 0.7: 1.5) - 400ML / PLANT x DAY (3 IRR.) - PH 1. CSL 2.5 l / ha

2. CSL hydrolyzed with only enzymes active on carbohydrates 2.5 l / ha

3. CSL hydrolyzed with active enzymes only on a protein fraction 2.5 l / ha

4. CSL hydrolyzed with both enzymes 2.5 l / ha

The methods of preparation of the hydrolysates were those of Example 1.

Example 3.

Lupine hydrolysates and efficacy tests

A greenhouse test was carried out on cyclamen; 18 plants were analyzed for each thesis. Each thesis was treated in fertigation, with approximately 25 ml per plant, corresponding to the following doses / hectare. The treatment was carried out three times over three weeks, one treatment per week. The following fertigation protocol was applied 45PPM N - NPK (1: 0.7: 1.5) - 400ML / PLANT x DAY (3 IRR.) - PH 7

1. lupine 2.5 l / ha

2. Lupine hydrolyzed with only enzymes active on carbohydrates 2.5 l / ha

3. Lupine hydrolyzed with only active enzymes on a protein fraction 2.5 l / ha

4. lupine hydrolyzed with both enzymes 2.5 l / ha

The methods of preparation of the hydrolysates were those of Example 1.

Example 4.

Ascophyllum algae hydrolysates and efficacy tests

A greenhouse test was carried out on cyclamen; 18 plants were analyzed for each thesis. Each thesis was treated in fertigation, with approximately 25 ml per plant, corresponding to the following doses / hectare. The treatment was carried out three times over three weeks, one treatment per week. The following fertigation protocol was applied 45PPM N - NPK (1: 0.7: 1.5) - 400ML / PLANT x DAY (3 IRR.) - PH 7

1. algae extract 2.5 l / ha

2. hydrolyzed algae extract with only enzymes active on carbohydrates 2.5 l / ha

3. hydrolyzed algae extract with active enzymes only on a protein fraction 2.5 l / ha

4. hydrolyzed algae extract with both enzymes 2.5 l / ha

Example 5.

Synergistic blend of CSL hydrolyzate, molasses and algae

A greenhouse test was carried out on cyclamen; 18 plants were analyzed for each thesis. Each thesis was treated in fertigation, with approximately 25 ml per plant, corresponding to the following doses / hectare. The treatment was carried out three times over three weeks, one treatment per week. The following fertigation protocol was applied. 45PPM N - NPK (1: 0,7: 1,5) - 400ML / PLANT x DAY (3 IRR.) - PH 7

1. CSL hydrolyzed with both enzymes 2.5 l / ha

2. Hydrolyzed algae extract with both enzymatic categories 2.5 l / ha

3. molasses 2.5 l / ha

4. algae extract 2.5 l / ha

5.CSL hydrolyzed with both enzymes reduced dose to 33%

6. Hydrolyzed algae extract with both enzymatic categories reduced dose to 33% 7. Molasses reduced dose to 33%

8. Hydrolyzed CSL 33% Hydrolyzed algae 33% molasses 33%

Example 6.

Synergistic blend of lupine hydrolyzate, molasses and algae

A greenhouse test was carried out on cyclamen; 18 plants were analyzed for each thesis. Each thesis was treated in fertigation, with approximately 25 ml per plant, corresponding to the following doses / hectare. The treatment was carried out three times over three weeks, one treatment per week. The following fertigation protocol was applied: 45PPM N - NPK (1: 0.7: 1.5) - 400ML / PLANT x DAY (3 IRR.) - PH 7

1. Lupine hydrolyzed with both enzymes 2.5 l / ha

2. Hydrolyzed algae extract with both enzymatic categories 2.5 l / ha

3. molasses 2.5 l / ha

4. algae extract 2.5 l / ha

5.Lupine hydrolyzed with both enzymes reduced dose to 33%

6. Hydrolyzed algae extract with both enzymatic categories reduced dose to 33% 7. Molasses reduced dose to 33%

8. Hydrolyzed lupine 33% Hydrolyzed algae 33% molasses 33%

Example 7.

Synergistic blend of hydrolyzed lupine and coriander

The test was carried out on tomatoes in greenhouses subjected to SALINE STRESS CULTURE: TOMATO CUORE DI BUE "CUORBENGA" LOCATION: SERRA (AUTUMN)

TREATMENTS, DOSE: The compounds and the hydrolysates were diluted from 1: 100 to 1: 2000, 16 PLANTS / THESIS were used, and the saline stress was provided by means of a 300mM WATER NACL solution IN SUBVASE, to which in the treatise they are biostimulant blends have been added.

TREATMENTS, FREQUENCY: IRRIGATIONS WITH 2,5L / SUB-VESSEL

TREATMENTS, NUMBER: 2 (ONE PER WEEK) TREATMENTS, APPLICATION: RADICAL (UNDER VASE)

The PLANTS purchased in the NURSERY have been REPOTED IN PEAT PERLITE POTS

IRRIGATIONS were carried out WITH 2,5L NACL 300 Mm (TAP WATER) NPK (N 50ppm, 1: 0,5: 1,7) - The growth temperature was the following: 15-25 ° C measured in GREENHOUSE

At the end of the test the wet weight and the dry weight of each plant and of the overall of each thesis were evaluated. From the results listed below it is clear a greater vigor of the plants in which the two compounds are mixed than the single ones, a symptom of a greater ability to overcome the strong saline stress caused.

Example 8.

Hydrolyzed efficacy of algae, coriander, lupine and molasses on tomato

The test was carried out on stressed tomatoes in a climatic growth chamber, 5-week-old seedlings transplanted into a soil made up of 40% sand,

CULTURE: TOMATO HEART OF OX "CUORBENGA"

PLACE: Climatic cell

TREATMENTS, DOSE: 10 liters / hectare

TREATMENTS, FREQUENCY: once a week

TREATMENTS, NUMBER: 4 weeks

TREATMENTS, APPLICATION: RADICAL (SUBVASE)

At the end of the test the seedlings were phenotyped for height of the aerial part and root development.

The results are summarized in the following table:

The results show how the hydrolysates according to recipes such as Ascophyllum algae, hydrolyzed coriander, hydrolyzed lupine, hydrolyzed molasses and hydrolyzed CSL, give significant results in phenotyping, confirming the goodness of the process developed and the effectiveness of the products subject to the claims.

Example 9.

Hydrolyzed efficacy of Ascophyllum algae, hydrolyzed cocoa and cocoa, hydrolyzed molasses and hydrolyzed pomace

The test was carried out on stressed tomatoes in a climatic growth chamber, 2-month-old seedlings transplanted into a soil made up of 40% sand, CULTURE: TOMATO HEART OF OX "CUORBENGA" LOCATION: Climate cell

TREATMENTS, DOSE: 15 liters / hectare

TREATMENTS, FREQUENCY: 1 time for racking, 1 time after 1 week

TREATMENTS, NUMBER: 2 weeks

TREATMENTS, APPLICATION: RADICAL (SUBVASE)

At the end of the test the seedlings were phenotyped for height of the aerial part and root development. The aerial part showed no significant variations, while the root part, significant in sandy soils, reported the following results.

The positive results of radical phenotyping are summarized in the following table:

The results show how the hydrolysates according to recipes such as Ascophyllum algae, hydrolyzed cocoa and cocoa, hydrolyzed molasses and hydrolyzed pomace, give significant results in phenotyping, confirming the goodness of the process developed and the effectiveness of the products covered by the claims.

Example 10.

Association of hydrolyzed CSL and Microorganisms

The results of a study on leaf physiology are reported, relating to plots cultivated with maize, the seeds of which have been treated with different experimental tanning processes.

To evaluate the effect of the composition of the invention, the corn field was divided as follows:

- Area A, cultivated with seeds tanned with hydrolyzed CSL (Ex.1A)

Zone a1: dose 4 g / kg seed

Zone a2: dose 16 g / kg seed

Zone a2: dose 20 g / kg seed

- Area B, cultivated with seeds tanned with Trichoderma asperellum

Zone b1: dose 4g / kg seed

Zone b2: dose 16g / kg seed

Zone b3: dose 20g / kg seed

- Area C, cultivated with seeds tanned with hydrolyzed Trichoderma asperellum CSL (Ex.

1A)

Zone c1: dose 4g / kg CSL 16g / kg Trichoderma asperellum

Zone c2: dose 16g / kg Trichoderma asperellum 4g / kg CSL

- Area D, Azospirillum brasilense

Zone d1: dose 4g / kg seed

Zone d2: dose 16g / kg seed

Zone d3: dose 20g / kg seed

- Area E, cultivated with seeds tanned with hydrolyzed Azospirillum brasilense CSL (Ex.

1A)

Zone e1: dose 4g / kg CSL 16g / kg Azospirillum brasilense

Zone e2: dose 16g / kg Azospirillum brasilense 4g / kg CSL

- Area F cultivated with seeds tanned with Bacillus megaterium,

Zone f1: dose 4g / kg seed

Zone f2: dose 16g / kg seed

Zone f3: dose 20g / kg seed

- Area G, cultivated with seeds tanned with hydrolyzed Bacillus megaterium CSL (Ex.1A) Zone g1: dose 4g / kg CSL 16g / kg Bacillus megaterium

Zone g2: dose 16g / kg Bacillus megaterium, 4g / kg CSL

- Area H, cultivated with seeds tanned with Glomus intraradices

Zone h1: dose 4g / kg seed

Zone h2: dose 16g / kg seed

Zone h3: dose 20g / kg seed

- Area I, cultivated with seeds tanned with Glomus intraradices CSL hydrolyzed (Ex.1A) Zone i1: dose 4g / kg CSL 16g / kg Glomus intraradices

Zone i2: dose 16g / kg Glomus intraradices 4g / kg CSL

- Area L, cultivated with seeds tanned with the association of Trichoderma asperellum, Azospirillum brasilense, Bacillus megaterium, Glomus intraradices CSL hydrolyzed (Ex.1A)

Zone l1: dose 4 g / kg seed Trichoderma asperellum, 4 g / kg seed Azospirillum brasilense, 4 g / kg seed Bacillus megaterium 4 g / kg seed Glomus intraradices

Zone l2: dose 4 g / kg seed Trichoderma asperellum, 4 g / kg seed Azospirillum brasilense, 4 g / kg seed Bacillus megaterium 4 g / kg seed Glomus intraradices 4g / kg CSL

- NT area, cultivated with untreated seeds.

The microbial cultures used were brought to a concentration of 1x10 ^ 8 CFU / ml, apart from Glomus intraradices which was in a concentration of 500 spores / g.

Over the study areas 150 kg / ha of N in the form of urea were distributed in coverage. No irrigation was required during the cultivation cycle.

The data were collected through three seasonal surveys: raising, flowering and waxy ripening of the cobs.

Leaf physiology

Plant physiology was measured directly in the field by randomly choosing 10 treated plants per area. The measurements were made by taking into consideration a leaf exposed to the sun and located on the stalk above the highest panicle.

Two portable instruments were used:

1- Fluorimeter (model OS5p) instrument composed of a LED lamp that emits light radiation at different frequencies and with different intensities in order to verify 2 parameters:

- Fv / Fm (known as maximum photosynthetic efficiency) whose values fluctuate in an interval between 0 to 1; it is measured on dark-adapted leaves and indicates how readily a leaf responds to light radiation (Baker N. et al., 2004).

- ETR (Electron Transport Rate) is measured on leaves exposed to light and indicates the efficiency of photosystem II (Genty B. et al., 1989).

2- Portable gas exchange analyzer (ADCpro-SD model) instrument that measures the change in concentration of carbon dioxide caused by synthetic photo activity through the "Assimilation rate" parameter (A).

With regard to leaf physiology, starting from the parameters measured in the field, the photosynthetic efficiency (given by the average of several leaves) was calculated, obtained from the ratio between ETR (Electron Transport Rate) and A (Assimilation rate). This ratio expresses the number of µmoles of electrons needed by PII (Photosystem II) to organize one µmole of CO2.

Results

Leaf ecophysiology measured in the field shows appreciable differences in values in plants from different areas.

The stomatal conductance (gs), resulting to be correlated with the activity of photosynthesis, confirms that the plants are not in conditions of water stress.

Example 11.

Biocontrol of Fusarium oxysporum cubense

Test execution method

The tests were performed on PCA medium (0.5% peptone and 0.25% yeast extract) for bacteria, bacteria with hydrolyzed citrus molasses and only hydrolyzed citrus molasses and PDA for fungi, fungi with hydrolyzed citrus molasses and only Hydrolyzed citrus molasses. The dual tests were performed, as regards the fungal cultures, the fungal cultures with the addition of hydrolyzed citrus molasses and only hydrolyzed citrus molasses, placing the F. oxysporum strain on one side of the petri dish on the surface of the soil while on the opposite side the strain or product whose biocontrol activity was to be evaluated was placed. At 10 days of growth at 27 ° C the surface of the plate occupied by F. oxysporum was measured. The inhibition percentage was calculated by making the ratio between the surface occupied by F. oxysporum in the presence of the test strains and the surface occupied by F. oxysporum grown in the absence of a biocontrol agent. Tests with only bacterial cultures, with bacterial cultures with additives with hydrolyzed citrus molasses and with hydrolyzed citrus molasses only were performed by placing the phytopathogen in the center of the petri dish on the surface of the soil while the phytopathogen was swiped on the two opposite sides of the plate. the strain or product for which the biocontrol activity was to be evaluated. At 10 days of growth at 27 ° C, the surface of the plate occupied by F. oxysporum was measured and the percentage of inhibition calculated as described above.

Concentration of use of bacterial cultures: 1x10 ^ 9 CFU / ml

Concentration of use of fungal cultures 5x10 ^ 7 CFU / ml

Hydrolyzed citrus molasses / microbial culture mixture ratio: 50% - 50%

Product dose per plate: 0.1g / plate

Results

The table below shows the results for the strains that have biocontrol activity against Fusarium oxysporum cubense (FOC)

Example 12.

Botrytis cinerea biocontrol

Test execution method

The tests were performed on PCA medium (0.5% peptone and 0.25% yeast extract) for bacteria, bacteria with hydrolyzed algae extract and hydrolyzed algae extract only and PDA for fungi, fungi with hydrolyzed algae extract only extract hydrolyzed algae. The dual tests were performed, as regards fungal cultures, fungal cultures with additives of hydrolyzed algae extract and hydrolyzed algae extract only, placing the Botrytis cinerea strain on one side of the petri dish on the surface of the soil while on the opposite side it is the strain or product whose biocontrol activity was to be assessed was placed. At 10 days of growth at 27 ° C the plate surface occupied by Botrytis cinerea was measured. The inhibition percentage was calculated by making the ratio between the area occupied by Botrytis cinerea in the presence of the test strains and the area occupied by Botrytis cinerea grown in the absence of a biocontrol agent. Tests with bacterial cultures only, with bacterial cultures with additives of hydrolyzed algae extract and hydrolyzed algae extract only were performed by placing the phytopathogen in the center of the petri dish on the surface of the medium while the two opposite sides of the plate were streaked with the strain or product for which the biocontrol activity was to be evaluated. In addition, the commercial Serenade® and Amylo-x® products were also tested for comparison. At 10 days of growth at 27 ° C, the surface of the plate occupied by Botrytis cinerea was measured and the inhibition percentage calculated as described above.

Concentration of use of bacterial cultures: 1x10 ^ 9 CFU / ml

Concentration of use of fungal cultures 5x10 ^ 7 CFU / ml

Hydrolyzed algae extract / microbial culture mixture ratio: 50% - 50%

Product dose per plate: 0.1g / plate

Results

The table below shows the results for the strains that have biocontrol activity against Botrytis cinerea

Example 13.

Fusarium fujikuroi biocontrol

The tests were performed on PCA medium (0.5% peptone and 0.25% yeast extract) for bacteria, bacteria with polyphenols and only polyphenols and PDAs for fungi, fungi with polyphenols and polyphenols only. The dual tests were performed, as regards the fungal cultures, the fungal cultures with additives with polyphenols and polyphenols only, placing the strain F. fujikuroi on one side of the petri dish on the surface of the soil while on the opposite side the strain o product of which we wanted to evaluate the biocontrol activity. At 10 days of growth at 27 ° C the surface of the plate occupied by F. fujikuroi was measured. The inhibition percentage was calculated by making the ratio between the surface occupied by F. fujikuroi in the presence of the test strains and the surface occupied by F. fujikuroi grown in the absence of a biocontrol agent. Tests with only bacterial cultures, with bacterial cultures with additives with polyphenols and with polyphenols only were performed by placing the phytopathogen in the center of the petri dish on the surface of the medium while the strain or product of which was streaked on the two opposite sides of the plate. we wanted to evaluate the biocontrol activity. The polyphenols used derive from various plant sources, such as pomace hydrolyzate, extracts of vitis vinifera or derivatives of lignocellulose. At 10 days of growth at 27 ° C, the surface of the plate occupied by F. fujikuroi was measured and the inhibition percentage calculated as described above.

Concentration of use of bacterial cultures: 1x10 ^ 9 CFU / ml

Concentration of use of fungal cultures 5x10 ^ 7 CFU / ml

Polyphenol / microbial culture mixture ratio: 50% - 50%

Product dose per plate: 0.1g / plate

Results

Table 1 below reports the results for the strains that have

biocontrol against Fusarium oxysporum cubense (FOC)

Compound / strain Inhibition (%)

Polyphenols 20

Paenibacillus polymyxa 54

Paenibacillus polymyxa polyphenols 81 Bacillus velezensis 47 Bacillus velezensis polyphenols 82 Trichoderma asperellum 70 Trichoderma asperellum polyphenols 94

Claims (10)

CLAIMS 1. Hydrolyzed from natural raw material comprising 1-25% of free amino acids and up to 55% of dry residue, in which said vegetable raw material is chosen from wheat liqueur, lupine, algae, molasses, coriander, cocoa, pomace, and their combinations. 2. The hydrolyzate of claim 1, wherein said hydrolyzate from natural raw material is: - hydrolyzate of corn liqueur, comprising 6-16% of free amino acids and up to 53% of dry residue, - lupine hydrolyzate, comprising 7-20% free amino acids and up to 30% dry residue, - hydrolyzate of algae, comprising 6-8% of free amino acids and up to 10% of dry residue, - hydrolyzate of molasses, comprising 6-8% of free amino acids and up to 55% of dry residue, - coriander hydrolyzate, comprising 1-5% free amino acids and up to 3% dry residue, - cocoa hydrolyzate, comprising 1-2% of free amino acids and up to 3% of dry residue, - pomace hydrolyzate, comprising 6-8% of free amino acids and up to 55% of dry residue, or a combination thereof. 3. Preparation process of the hydrolyzate of claim 1 or 2, comprising the steps of: i) supply the natural raw material, and optionally add water, ii) adding an enzyme complex comprising at least one peptide bond hydrolase and at least one fiber and carbohydrate hydrolase, and iii) obtain a liquid mixture, i.e. the hydrolyzate from natural raw material. 4. The process of claim 3, wherein said at least one hydrolase of the peptide bond is selected from monopepdidase, dipepdidase, tripepdidase, protease, proteinase, and combinations thereof, and said at least one hydrolase of fibers and carbohydrates is selected from amylase, cellulase, hemicellulases, laccases, xylanases and their combinations. The process of claim 3 or 4, wherein the enzyme complex further comprises at least one ester bond hydrolase, preferably selected from lipase. The process of any one of claims 3-5, wherein in step ii) the temperature is 4-70 ° C for a time period of 2-20 hours, preferably the temperature is 10-50 ° C for a time period of 4-16 hours. 7. Hydrolyzed from natural raw material obtainable from the process of any one of claims 3-6, wherein said vegetable raw material is selected from corn liqueur, lupine, algae, molasses, coriander, cocoa, pomace, and combinations thereof, in which the percentage of free amino acids in the hydrolyzate is at least 0.5% higher than the percentage in the starting vegetable raw material, and in which the dry residue in the hydrolyzate is at least 2% lower than the percentage in the starting vegetable raw material . 8. The hydrolyzate from natural raw material of claim 7, said hydrolyzate being: - hydrolyzate of corn liqueur, in which the percentage of free amino acids in the hydrolyzate is 0.5-11% higher than the percentage in the corn liqueur of starting point, and in which the dry residue in the hydrolyzate is 10-15% lower than the percentage in the starting corn liqueur, - lupine hydrolyzate, in which the percentage of free amino acids in the hydrolyzate is 2-15% higher than the percentage in the starting lupine, and in which the dry residue in the hydrolyzate is 10-15% lower than the percentage in the lupine of departure, - hydrolyzate of algae, in which the percentage of free amino acids in the hydrolyzate is 4-6% higher than the percentage in the starting algae, and in which the dry residue in the hydrolyzate is 2-4% lower than the percentage in the algae of departure, - hydrolyzed molasses, in which the percentage of free amino acids in the hydrolyzate is 4-6% higher than the percentage in the starting molasses, and in which the dry residue in the hydrolyzate is 10-20% lower than the percentage in the molasses of departure, - coriander hydrolyzate, in which the percentage of free amino acids in the hydrolyzate is 0.5-4.5% higher than the percentage in the starting coriander, and in which the dry residue in the hydrolyzate is 10-15% lower than percentage in the starting coriander, - cocoa hydrolyzate, in which the percentage of free amino acids in the hydrolyzate is 0.5-1.5% higher than the percentage in the starting cocoa, and in which the dry residue in the hydrolyzate is 10 -15% lower than the percentage in the starting cocoa, - pomace hydrolyzate, in which the percentage of free amino acids in the hydrolyzate is 4-6% higher than the percentage in the starting pomace, and in which the dry residue in the hydrolyzate is 5-10% lower than the percentage in the pomace of departure, or a combination thereof. 9. Agro-chemical product comprising at least one hydrolyzate of claim 1, 2, 7, or 8, and agro-chemical additives, and optionally also a natural raw material selected from wheat liqueur, lupine, algae, molasses, coriander, cocoa, pomace, and their combinations. 10. Use of the hydrolyzate of claim 1, 2, 7, or 8, or of the agro-chemical product of claim 9, as a promoter of plant growth and fruit production in agriculture.