MIXTURE OF BACTERIA LYSAT AND ISOLATED METABOLITES FROM CURCUMA LONGA OF BHUTAN AND TO INCREASE THE YIELD AND ANTI-INFLAMMATORY PROPERTIES OF AGRICULTURAL PRODUCTION.

Abstract

TITLE: MIXTURE OF BACTERIA, LYSATE, AND ISOLATED METABOLITES FROM BUHTAN'S CURCUMA LONGA TO INCREASE YIELD AND ANTI-INFLAMMATORY PROPERTIES IN AGRICULTURAL PRODUCTION. The present invention relates to a plant treatment composition characterized in that it contains a mixture of live bacteria, bacterial lysate, and metabolites from the rhizomes of Curcuma longa. Spraying or applying said composition to plants increases plant fertility and enhances the anti-inflammatory properties of cereals, fruits, and vegetables.

Description

Title of the invention: MIXTURE OF LYSATE BACTERIA AND ISOLATED METABOLITES FROM CURCUMA LONGA BHUTAN AND TO INCREASE THE YIELD AND ANTI-INFLAMMATORY PROPERTIES OF AGRICULTURAL PRODUCTION. FIELD OF INVENTION

[0001] The present invention relates to a plant treatment composition intended to increase the yield of agricultural products such as cereals, fruits, and vegetables. Advantageously, the treatment according to the invention enhances the beneficial properties of agricultural production on human health. The treatment composition according to the invention may contain a mixture of live bacteria obtained from the microbiota extraction of turmeric rhizomes (Curcuma longa), preferably from Bhutan, a lysate of these bacteria, and / or metabolites resulting from the slow fermentation of these same bacteria. Advantageously, the bacteria according to the invention persist durably in the soil. STATE OF THE ART

[0002] Just like humans, plants possess a microbiome composed of bacteria, yeasts, fungi, and viruses that colonize all accessible plant tissues. These microorganisms can form complex co-associations with plants and play an important role in promoting plant health in the natural environment. The plant microbiota consists of numerous and varied species of microorganisms that perform several functions essential to the plant. For example, the microbiota forms a symbiotic relationship with plant roots, allowing them to both benefit from their environment and enrich it. The plant root system explores the soil to find nutrients and is therefore exposed to the various microorganisms present in the soil.Plants provide sugar to microorganisms, while microorganisms supply nitrogen and phosphorus, essential for healthy plant growth. Microorganisms also produce plant hormones that stimulate plant growth.

[0003] The microbial population can be altered by the environment and by agricultural methods, in particular by intensive agriculture, monoculture, and the massive use of agrochemicals such as herbicides, fungicides, insecticides, nematicides, molluscicides, rodenticides, and chemical fertilizers. The use of Chemicals significantly decrease the diversity and composition of the soil and plant microbiome. This alteration of the diversity and composition of the beneficial microbial community can be detrimental to plant growth and development, either by reducing nutrient availability or by increasing disease incidence (Meena, RS et al. “Impact of Agrochemicals on Soil Microbiota and Management. A Review.” Land 2020, 9, 34). Alterations in the microbial community can also impact plant fertility and the yield of seeds, fruits, and plant matter. Microbes play a key role in ecosystems and influence many important ecosystem processes, including plant nutrient acquisition, nitrogen and carbon cycling, and soil formation and composition.The soil microbiome forms a symbiosis with plants, which is essential for their nutrient absorption and growth.

[0004] Fertilizer-based solutions containing bacteria of the genus Bacillus (WO2017105238A1) or Bacillus, Pseudomonas (WO2015118516A1), intended to fertilize the soil and promote faster plant growth, have been proposed. However, their effects are limited. These products contain single bacteria from a different environment that cannot compete with the native microbiome, which has already formed a stable and balanced ecosystem. The study by ES Jensen and LH Sprensen ("Survival of Rhizobium leguminosarum in soil after addition as inoculant," FEMS Microbiology Ecology, Volume 3(4), August 1987, Pages 221-226) demonstrates that the population of inoculated bacteria in the soil decreases over time and that the half-life of an inoculant is approximately 1.5 to 2.1 years.This demonstrates that plant treatment solutions containing only bacteria are not sustainable and require continuous application, generally every year, to have a significant effect, resulting in a waste of time and money. Similarly, while existing solutions improve plant growth, they do not impact the benefits of these plants for human health.

[0005] It was while studying turmeric bacteria that the Applicant unexpectedly discovered that these problems can be easily solved by using a composition intended to increase the yield of agricultural productions containing a mixture of live bacteria, a bacterial lysate and / or metabolites from the extraction of the microbiota from the rhizomes of turmeric (Curcuma longa), preferably from Curcuma longa grown in Bhutan. SUMMARY

[0006] The invention relates to a plant treatment composition comprising a mixture of live bacteria from the microbiota of turmeric (Curcuma longa) rhizomes, preferably from Bhutan, a lysate of said bacteria and of metabolites produced by the fermentation of these same bacteria. This composition is intended to increase the yield of agricultural production, preferably cereals, fruits and vegetables, and to enhance their beneficial properties for human health.

[0007] The plant treatment composition according to the invention may comprise one or more strains of bacteria selected from the following: Lactobacillus fermentum, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus acidophilus, Bifidobacterium longum, Bifidobacterium bifidum, Bifidobacterium adolescentis, Bifi-dobacterium infantis, Lactobacillus brevis, Lactobacillus Jensenii, Lactobacillus paracasei, Christensenella minuta, Lactiplantibacillus plantarum, Lactiplantibacillus paraplantarum, Mycobacterium vaccae, Pseudomonas japonica, Pseudomonas fluorescens, Archaeospora trappei, Methylobacterium crusticola, Bradyrhizobium elkanii, Bradyrhizobium japonicum, Gluconacetobacter diazotrophicus, Azospirillum brasilense.

[0008] The plant treatment composition may be in the form of liquids, foams, pastes, emulsions, oils, gels, jellies, syrups, solids, powders, sprays, or aerosols.

[0009] The invention also relates to a method for manufacturing the plant treatment composition as described below, comprising the steps of:

[0010] - isolate bacteria from Curcuma longa rhizomes, preferably from a plantation located in Bhutan and not using chemical fertilizers or herbicides; then

[0011] - inoculate at least one substrate with said bacteria and ferment the mixture of bacteria with said at least one substrate.

[0012] According to another aspect, the invention relates to the use of the plant treatment composition according to the invention for:

[0013] - to increase soil fertility by increasing the concentration of nutrients and in reducing the amount of toxic substances.

[0014] - to increase the yield of cereals, fruits and vegetables, by increasing the plant growth, fertility and increasing the number and weight of seeds, fruits and vegetables.

[0015] - to increase the beneficial properties on human health of cereals, fruits and vegetables.

[0016] - to increase the anti-inflammatory and regulatory properties of the blood sugar levels of cereals, fruits and vegetables.

[0017] - to increase the dietary fibre content of cereals, fruits and vegetables.

[0018] According to a final aspect, the invention relates to a method for treating plants intended to increase the yield of at least one plant, in which the composition according to the invention is sprayed or spread on the soil and / or the plant. DETAILED DESCRIPTION

[0019] The object of the present invention therefore relates to a plant treatment composition intended to increase agricultural production yields. The invention also relates to the use of the composition, as described below, for treating at least one plant, preferably a plant intended for agricultural production. In one embodiment, agricultural production refers to cereals, fruits, and / or vegetables. In a specific embodiment, agricultural production refers to cereals, preferably rice. In a specific embodiment, agricultural production refers to fruits, preferably apples.

[0020] According to one embodiment, the treatment composition makes it possible to increase the beneficial properties of agricultural production on human health.

[0021] The composition may include a mixture of live bacteria from the microbiota of fermented turmeric (Curcuma longd) rhizomes, preferably from Bhutan, a bacterial lysate and metabolites from the extraction.

[0022] In a first embodiment, the composition comprises a mixture of live bacteria from the microbiota of turmeric rhizomes (Curcuma longa, preferably from Bhutan). Advantageously, the bacteria applied according to the invention have a lasting persistence in the soil.

[0023] In a second embodiment the composition comprises a lysate of bacteria from the microbiota of turmeric rhizomes (Curcuma longa), preferably from turmeric from Bhutan.

[0024] In a third embodiment, the composition comprises metabolites from the extraction of fermented bacteria from the microbiota of turmeric rhizomes (Curcuma longa, preferably turmeric from Bhutan.

[0025] In a preferred embodiment, the composition comprises or is made up of the constituents of the three aforementioned embodiments.

[0026] The composition can be used to increase soil fertility. The invention also relates to a composition characterized in that it is used to modulate human blood glucose levels by increasing the amount of dietary fiber in cereals, fruits, and vegetables thus cultivated.

[0027] According to one embodiment, the use of the composition of the invention increases plant growth and / or yield. It should be noted that the composition is characterized in that spraying or applying said composition to plants or to the soil surrounding plants increases plant growth and yield. The invention also relates to a composition characterized in that the va The application of this composition to plants advantageously enhances its health benefits for consumers of the cereals, fruits, and vegetables thus cultivated. In one embodiment, the use of the composition of the invention increases plant yields of nutrients beneficial to human health, such as fiber, vitamins, minerals, polyphenols, terpenes, nitrogen compounds, alkaloids, steroids, terpenoids, flavonoids, omega-3, omega-6, and omega-9 fatty acids, linoleic acid, L-carnitine, choline, or sphingomyelin. In another embodiment, the use of the composition of the invention increases plant yields of nutrients beneficial to human health, such as vitamins, minerals, polyphenols, omega-3, omega-6, and omega-9 fatty acids, and / or linoleic acid.According to a specific embodiment, the use of the composition of the invention increases the fiber yield of plants.

[0028] For the sake of clarity in the following explanations, we decide to call QKC a composition intended to increase plant yield and enhance their beneficial properties on human health containing a mixture of live bacteria, bacterial lysate and metabolites from the extraction of the microbiota from the rhizomes of turmeric (Curcuma longa, preferably from Bhutan).

[0029] The applicant describes below a composition intended to increase plant yield and enhance their beneficial properties on human health containing a mixture of live bacteria, bacterial lysate and metabolites from the extraction of the microbiota from turmeric rhizomes (Curcuma longa, preferably from Bhutan).

[0030] Microorganisms have long been used as inoculants for biological control or biostimulation. However, these microorganisms have a limited effect. Indeed, products containing these microorganisms most often contain bacteria from a single, different environment. This leads to inconsistent performance that varies according to climate, soil type, and other factors. In fact, these applied bacteria cannot compete with the native microbiome. The functionality and persistence of microorganisms depend on interactions with the environment as well as with other microorganisms within a community. The applicant has therefore developed a plant treatment composition containing bacteria, a bacterial lysate, and fermentation metabolites thereof.The metabolites allow both the modulation of the existing soil microbiota and the perpetuation of bacteria in the soil by modulating communication between the bacteria of the invention and soil bacteria through quorum sensing. Quorum sensing is the communication mechanism of bacteria that allows a bacterial strain to... to inhibit or stimulate growth as well as modulate gene expression of another strain of bacteria.

[0031] The present invention therefore describes a plant treatment composition intended to increase the yield of agricultural production of cereals, fruits and vegetables and to increase their beneficial properties on human health characterized in that it contains a mixture of live bacteria, bacterial lysate and metabolites from the extraction of the microbiota from the rhizomes of turmeric (Curcuma longa), preferably from Bhutan and having increased persistence in the soil.

[0032] The applicant describes below a means of obtaining the plant treatment composition containing a mixture of live bacteria, bacterial lysate and metabolites from the extraction of the microbiota from the rhizomes of turmeric, preferably turmeric (Curcuma longa) from Bhutan.

[0033] According to another aspect the invention relates to a method of preparing the composition according to the invention.

[0034] In one embodiment, the process consists of several steps comprising:

[0035] - a step of extracting bacteria from Curcuma longa rhizomes, preferably sourced from plantations located in Bhutan and not using chemical fertilizers or herbicides,

[0036] -a selection step by PCR followed by 16S rRNA sequencing, and

[0037] - a fermentation step in the presence of substrate.

[0038] According to one embodiment, the process comprises the steps of

[0039] - isolate bacteria from Curcuma longa rhizomes, preferably from a plantation located in Bhutan and not using chemical fertilizers or herbicides;

[0040] - inoculate at least one substrate with said bacteria and ferment the mixture of bacteria with the substrate.

[0041] In one embodiment, the process is a process for preparing the composition comprising a mixture of live bacteria from the microbiota of turmeric (Curcuma longa) rhizomes, preferably from Bhutan, a lysate of said bacteria, and metabolites from the extraction of said fermented bacteria; said process comprising the steps of i) collecting turmeric rhizomes, preferably grown in a field in Bhutan, even more preferably without having undergone chemical treatment; ii) isolating the bacteria from the microbiome of the rhizomes; iii) culturing the isolated bacteria so as to obtain a mother culture of the bacteria; iv) inoculating a sterilized substrate with bacteria from the mother culture and fermenting, preferably in a fermentation tank for at least 120 hours at a temperature between 30°C and 45°C;(v) sterilization and microfiltration of the fermented mixture in step (iv) resulting in a composition comprising the fermentation metabolites and bacterial lysates; (vi) mixing the composition; including fermentation metabolites and bacterial lysates with bacteria from the mother culture of bacteria isolated from turmeric. The invention also relates to a composition that can be obtained, or is directly obtained, by the process according to the invention.

[0042] In one embodiment, the composition according to the invention comprises from 0.5 to 10% of the mother culture.

[0043] The applicant studied turmeric rhizomes from different countries and compared their effectiveness on anti-inflammatory gene expression. Turmeric from Bhutan was found to be more effective in inducing an anti-inflammatory effect. By studying the microbiota of this turmeric, the applicant discovered that the microbiota of Bhutanese turmeric produced small molecules, called metabolites, which gave Bhutanese turmeric its exceptional health effects. The applicant therefore developed a process for extracting this microbiota, as well as a fermentation process for producing the metabolites and bacterial lysates. Initially, turmeric rhizomes cultivated in a field in Bhutan without the use of chemicals are collected. The samples are then mixed at 20% with a phosphate saline buffer at pH 7 and a concentration of 1 mol / L. After centrifugation, the supernatant is collected.The supernatant is cultured in the presence of a bacterial growth-stimulating agent. The species are identified by PCR, followed by 16S rRNA sequencing. PCR, or polymerase chain reaction, is a technique for amplifying DNA fragments in vitro. DNA amplification allows for easier sequencing and better species identification. 16S rRNA is an RNA fragment present in all bacterial species, with sequence variations characteristic of each species. The sequences obtained for the turmeric microbiota are compared to a ribosomal sequence database. The selected bacteria are cultured in the presence of a substrate that promotes their multiplication and constitute the mother culture of bacteria of interest.

[0044] The selected bacteria are then used in a fermentation process. This fermentation process takes place in a fermentation tank in several stages for a total duration of at least 120 hours, including prior sterilization of the ingredients intended to undergo fermentation, inoculation of the bacteria and culture of the mixture at a temperature between 30°C and 45°C.

[0045] The ingredients used as a substrate in the fermentation process are all bio-based and edible, i.e., all plant matter such as fruits, vegetables, and algae, as well as food waste from households, workplaces, and catering establishments, along with compost and manure. The fermented substrate ingredients can be, for example This list is not exhaustive and may include the fruits, peels, leaves, or flowers of figs, olives, rosemary, thyme, beans, sage, melon, pomegranate, soybeans, watermelon, cabbage, green or black tea, turmeric, and pine needles. Materials are selected for their pesticide-free cultivation. A mixture of any number of materials, or a single material, may be used as a fermentation substrate. At the end of fermentation, the contents of the fermentation tank are sterilized and filtered using microfiltration to remove plant debris, retaining only the bacterial metabolites and lysates. The metabolites and lysate mixture is then combined with the mother culture of bacteria isolated from turmeric. The composition is adjusted to contain 0.5% to 10% of the mother culture.

[0046] The bacteria isolated from turmeric, such as for example those from the mother culture, and identified by 16s rRNA sequencing can be, without this being limiting: Lactobacillus fermentum, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus aci-dophilus, Bifidobacterium longum, Bifidobacterium bifidum, Bifidobacterium ado-lescentis, Bifidobacterium infantis, Lactobacillus brevis, Lactobacillus Jensenii, Lactobacillus paracasei, Christensenella minuta, Lactiplantibacillus plantarum, Lactiplantibacillus paraplantarum, Mycobacterium vaccae, Pseudomonas japonica, Pseudomonas fluorescens, Archaeospora trappei, Methylobacterium crusticola, Bra-dyrhizobium elkanii, Bradyrhizobium japonicum, Gluconacetobacter diazotrophicus, Azospirillum brasilense.

[0047] Soil fertility is defined as the capacity of a soil to support plant growth by providing essential plant nutrients.

[0048] In particular, nitrification is an important process in soil fertility management. Nitrogen is an important element for plants. Indeed, it stimulates plant growth and regulates nutrient use. A deficiency of this element manifests itself in poor leaf formation and yellowing, as well as a decrease in crop yield. However, heavy rainfall can cause nitrate to leach into the water, reducing the amount of nitrate in the soil. This nitrate leaching leads to a need for farmers to constantly apply fertilizer containing nitrate, which in turn leads to soil acidification.

[0049] The bacteria of composition QKC are nitrogen-fixing bacteria, that is to say, they are capable of fixing atmospheric nitrogen gas into "fixed nitrogen" compounds, such as ammonia, which can be used by plants.

[0050] The metabolites resulting from the fermentation process also increase soil fertility. Indeed, these metabolites contain nutrients essential to plants. The metabolites produced can be of the following types: minerals, non-ribosomal peptides, lipopeptides, polyketides, terpenes, nitrogenous compounds such as indole, pyrazines, volatile compounds containing sulfur. The terpenes can be of the following types: brassinosteroids, gib- Berellins. Bacterial lysates also contain elements that can improve soil fertility and provide elements necessary for plant growth.

[0051] Furthermore, the bacteria comprising QKC ​​make minerals more available for absorption by plants. Plants need minerals to grow, such as phosphorus, potassium, magnesium, sulfur, and calcium. These minerals already exist in nature. However, their bioavailability depends on their solubility in the environment. If the minerals are in a precipitated form in the soil, they cannot be absorbed by plants. The bacteria of the present invention are capable of solubilizing minerals by secreting organic acids that prevent the binding of minerals in an insoluble form. The metabolites in the composition of QKC increase the solubilization capacity of the bacteria by stimulating genes related to this activity. In particular, the metabolites increase the activity of the enzyme carbonic anhydrase, which catalyzes the dissolution of calcium carbonate.

[0052] Soils contain compounds that are toxic to both plants and human health. For example, they contain polychlorinated biphenyls (PCBs), pesticides, industrial solvents, petroleum products, dioxins and furans, heavy metals, explosives, and brominated flame retardants. These compounds can be absorbed by plants and pose a risk to human health. Traditional technologies commonly used for the remediation of contaminated environmental soils include excavation, transport to specialized landfills, incineration, stabilization, and vitrification. These technologies are costly and can damage soils and the landscape. Some soil bacteria have the ability to degrade toxic compounds. For example, PCBs can be degraded by dehalogenation, that is, by removing a halogen from the PCB compound, thereby reducing its toxicity.However, not all bacteria have the genetic machinery necessary to carry out this degradation.

[0053] The bacteria of the present invention are capable of degrading toxic compounds. The combination of bacteria and metabolites triggers the biodegradation of the toxic compounds present by stimulating the genes of enzymes involved in the degradation of toxic chemical compounds.

[0054] The present invention is therefore described as a composition for use in increasing soil fertility by increasing nutrient concentration and decreasing the amount of toxic substances.

[0055] Agricultural production yield is defined as the quantity of product harvested from a given cultivated area. It can be influenced by soil fertility, access to water, as well as diseases and pests.

[0056] The QKC composition increases soil fertility. The QKC composition also prevents diseases by inhibiting pathogenic bacteria. As in humans and Animals and plants can be infected by pathogenic bacteria and fungi. These bacteria can include: Xanthomonas campestris, Clavibacter michiganensis, Pseudomonas spp., Ralstonia solanacearum, and Pseudomonas syringae. Phytopathogenic fungi can cause diseases such as downy mildew caused by the fungus Phytophthora infestons, fruit rot caused by Aspergillus, and fruit rot caused by Altemaria and Botrytis. These infections can lead to fruit rot, resulting in reduced harvests; they can also cause plant death. Thanks to the lysozymes contained in the bacterial lysate, the QKC composition inhibits the establishment of pathogenic bacteria. Lysozymes are enzymes that destroy the cell membranes of pathogenic bacteria.

[0057] The metabolites of the QKC composition also stimulate the native soil and plant microbiota, producing an additional barrier against the establishment of pathogenic bacteria.

[0058] Different strains of bacteria are capable of communicating with each other through chemical signals and modulating the growth and activity of other strains by quorum sensing. Bacterial strains are also capable of modulating fungal growth. Bacteria inhibit the proliferation of bacteria and fungi harmful to plants through competition. The metabolites of the QKC composition stimulate the indigenous microbiome and inhibit pathogenic bacteria by modulating quorum sensing.

[0059] The composition also directly increases plant fertility and production, particularly in rice plants, by inducing a Darwinian effect, namely the activation of the autophagy gene, OsATG8a. The OsATG8a gene selects only functional cells that will produce a fertile stem. Overexpression of OsATG8a significantly increases the level of autophagy and the number of fertile thalli in rice. The bacteria in the QKC composition also increase the OsNPF7.2 and OsSta2 genes, which are involved in the number of thalli (stems bearing rice grains) and rice grain yield.

[0060] The present invention is therefore described as a plant treatment composition for use in increasing the yield of cereals, fruits and vegetables, by increasing plant growth, their fertility and increasing the number and weight of seeds, fruits and vegetables.

[0061] Plants are consumed not only for their energy content but also for their health effects. Indeed, plants are composed of vitamins, minerals, and other molecules, such as polyphenols, terpenes, nitrogenous compounds, alkaloids, steroids, terpenoids, flavonoids, omega-3, 6, and 9, linoleic acid, L-camitine, choline, and sphingomyelin. These molecules are im- These molecules are involved in plant growth and resistance to biotic and abiotic stress, combating diseases and pests. They also have effects on human health. Indeed, some molecules have antioxidant properties and can directly modulate gene expression in human cells. In particular, molecules such as curcumin, colchicine, resveratrol, capsaicin, epigallocatechin-3-gallate (EGCG), and quercetin. Many plants produce only small amounts of these secondary metabolites, but beneficial microbes associated with their host plant can increase their production.

[0062] The present invention contains bacteria that increase the production of such molecules.Indeed, the applicant noted that bacteria isolated from turmeric generate signal transduction networks in plants, inducing modulation of plant gene expression and leading to the accumulation of certain bioactive compounds in plant tissues.

[0063] The interaction of the microbiome with plants also increases the absorption of nutrients from the soil by plants, promoting their accumulation in plant tissues.

[0064] The present invention is therefore described as a plant treatment composition for use in increasing the beneficial properties on human health of cereals, fruits and vegetables.

[0065] Some molecules have low bioavailability and efficacy if ingested as is and must undergo digestion by the gut microbiota to be activated. However, not everyone possesses the necessary bacteria to activate polyphenols. For example, S-equol is formed by the digestion of daidzein by gut bacteria. S-equol has superior anti-inflammatory efficacy compared to other isoflavones. Only 25 to 30% of the population possesses the bacteria necessary for this digestion. The bacteria composing the present invention have the genes and enzymes necessary to transform daidzein into S-equol. By spraying the QKC composition onto plants, the bacteria present in the composition digest the daidzein into S-equol and thus increase the anti-inflammatory efficacy of the plants on which the composition is used.The present invention is therefore described as a plant treatment composition for use in increasing the beneficial properties on human health of cereals, fruits and vegetables, characterized in that it increases the anti-inflammatory properties of cereals, fruits and vegetables.

[0066] Hyperglycemia (high blood sugar) means that there is too much sugar in the blood and is caused by consuming a diet high in sugar and low in fiber. Associated with diabetes, hyperglycemia can cause vomiting, excessive hunger and thirst, a rapid heart rate, vision problems, and other Symptoms. Untreated hyperglycemia can lead to serious health problems and premature death.

[0067] The consumption of soluble dietary fiber reduces the rate of glucose absorption after the consumption of a high carbohydrate load due to its beneficial viscosity properties. By modulating nutrient absorption from the soil and modifying gene expression in plants, the present invention alters the nutritional qualities of the plants to which it is applied and, in particular, increases the proportion of fiber and resistant starch and decreases the amount of sugars.

[0068] The present invention is therefore described as a plant treatment composition for use in increasing the blood glucose modulation properties by increasing the amount of dietary fiber and resistant starch in cereals, fruits and vegetables.

[0069] The present invention can be used in the form of liquids, foams, pastes, emulsions, oils, gels, jellies, syrups, solids, powders, sprays, aerosols.

[0070] The composition described in the present invention is sprayed or spread on the soil and / or the plant according to the natural plant growth cycle. It can be used either in winter on the soil to improve soil quality and prepare it for sowing, or during plant growth to stimulate plant growth. EXAMPLES

[0071] The present invention will be better understood by reading the following examples which illustrate the invention in a non-limiting way. Example 1: List of QKC metabolites

[0072] By way of example, the following is a non-exhaustive list of metabolites present in a QKC type composition described by the present invention: jasmonic acid, coumarin, benzoazinoids, brassinosteroids, gibberellins, indole, pyrazine, catechin, octanoic acid, sitosterol, citric acid, phloretic acid, erucic acid, glucosamine, methyl jasmonate, lobeline, pyridoxine, trigonelline, syringic acid, procyanidins, geraniin, pro-delphinidin, castalagin, punicalagin, tannic acid, resveratrol, apigenin-7-glucoside, luteorin, rutin, eriodictyol, kaempferol, phloretic acid, homovanillic acid, pelargonic acid, butyric acid, baicalin, saponarin, pyruvic acid, fumaric acid, hypoxanthine, spermine, ornithine, homovanillic acid.

[0073] Example 2: Increased productivity, plant growth and nutritional value

[0074] A study was conducted to evaluate the effect of the QKC composition on the growth, productivity, and nutritional value of rice plants. For this study, QKC and three other types of fertilizers were used on rice plants.

[0075] The fertilizers tested are:

[0076] GROUP 1: control - no fertilizer

[0077] GROUP 2: QKC

[0078] GROUP 3: Commercial composition of bacteria and fungi (yeasts)

[0079] GROUP 4: BIOCHAR (Rice bran charcoal obtained by pyrolysis of biomass)

[0080] GROUP 5: Biofertilizer composed of bacteria of the genus Azobacter

[0081] To facilitate understanding of the study, the definitions are provided below:

[0082] Grain yield: quantity by weight of grains produced in a given area

[0083] Biological yield: quantity by weight of biological matter produced on a given surface

[0084] Spikelet: inflorescence containing the rice grain

[0085] Panicle: a cluster of spikelets

[0086] The results on growth and productivity are presented in Table 1 below:

[0087] [Tables 1] TREATMENT Number of spikelets per panicle Hollow spikelets Grain yield (kg / hectare) Biological yield (kg / hectare) GROUP 1 119 6.37 3190.09 11975.34 GROUP 2: QKC 170 5.17 4284.74 13931.29 GROUP 3 135 5.96 3161.67 9568.33 GROUP 4 128 5.83 3667.06 11690.39 GROUP 5 136 6.62 3477.42 13204.09

[0088] It is observed that the number of spikelets per panicle, grain yield, and biological yield are highest with the treatment using the QKC composition. It is also observed that the number of hollow spikelets decreases with QKC. The QKC treatment increases the number of spikelets per panicle by 43%, grain yield by 34%, and biological yield by 16% compared to the control.

[0089] In conclusion, the QKC composition is the most effective for increasing plant growth, fertility, and for increasing grain and biological matter yield.

[0090] The results of the effect of using QKC ​​on the composition of rice are shown in Table 2 below:

[0091] [Tables2] Processing Nutritional Values ​​(per 100g of rice grain) Protein Carbohydrates Fiber Iron Calcium Magnesium GROUP 1 5.7 g 76.1 g 3.4 g 0.82 mg 8.0 mg 122 mg GROUP 2: QKC 6.5 g 74.5 g 3.9 g 0.92 mg 8.5 mg 127 mg GROUP 3 5.5 g 75.5 g 3.3 g 0.85 mg 8.1 mg 119 mg GROUP 4 6.1 g 76.1 g 3.5 g 0.84 mg 8.0 mg 118 mg GROUP 5 5.3 g 74.4 g 3.2 g 0.87 mg 8.1 mg 121 mg

[0092] It is observed that with QKC treatment, the values ​​of protein, fiber, iron, calcium, and magnesium are higher compared to other treatments. Since magnesium has anti-inflammatory properties, it can be concluded that the anti-inflammatory properties of rice were increased with QKC treatment.

[0093] It is also observed that although the amount of fiber is higher, the amount of carbohydrates is lower, implying that the amount of sugars and starch has decreased.

[0094] In conclusion, the QKC treatment improved the nutritional values ​​of the rice and increased the amount of fiber in the rice grains.

[0095] Example 3: Increased persistence of bacteria in the soil.

[0096] This study was carried out to evaluate the effect of the QKC composition on the per Bacterial persistence in soil. For this study, lactic acid bacteria (LAB), either isolated (LAB group) or in a QKC composition (LAB in QKC), were inoculated into soil and their quantity was measured for 28 days.

[0097] Six seed trays were each filled with 100 grams of soil collected from a sugarcane field. Three trays were inoculated with isolated lactic acid bacteria or with a QKC composition at a final soil concentration of 1010 CFU / g. The trays were placed in a plant growth chamber for 28 days. A 25 g sample was taken using a sterile spoon to determine the survival capacity of the lactic acid bacteria at 0, 1, 3, 7, 14, and 28 days post-inoculation.

[0098] The soil sample is serially diluted, then spread onto an agar plate and incubated at The temperature was maintained at 37°C for 48 hours, and the number of colonies was counted. The quantity of bacteria was estimated by averaging the results obtained from the three seed trays.

[0099] The results of the effect of QKC on bacterial persistence in soil are presented in Table 3 below. The results are expressed as log CFU (colony-forming unit) per gram of soil.

[0100] [Tables3] 0 days 1 day 3 days 7 days 14 days 28 days BAL 11.5+0.5 10.2+0.6 8.1+0.7 7.7+0.6 6.9+0.5 6.2+0.7 BAL in QKC 11.4+0.6 10.9+0.4 10.5+0.5 11.8+0.7 12.1+0.6 12.5+0.6

[0101] The results show that without QKC, the number of bacteria decreases rapidly from the first day, probably due to competition with other bacteria present in the soil. Conversely, with the QKC composition, the number of lactic acid bacteria increases by 1.1 log by the end of the study. This study clearly demonstrates that the QKC composition allows the bacteria it contains to persist in the soil for an extended period.

[0102] Example 4: increasing the anti-inflammatory properties of plants.

[0103] This study was carried out to evaluate the effect of the QKC composition on the anti-inflammatory properties of plants.

[0104] To this end, two orchards of the same apple variety, located 1 km apart, were divided. The QKC compound was sprayed on the apple trees of one of the orchards during flowering, while the other orchard received no treatment.

[0105] Apples were selected during harvest and their anti-inflammatory properties were analyzed. To this end, an apple extract from two orchards was prepared and tested on 40 overweight but otherwise healthy volunteers of all sexes, aged between 35 and 55 years, with no known medical conditions. The 30 participants were randomly divided into two groups of 15. The first group ingested 500 mg daily for 30 days of the apple extract from the field sprayed with QKC, and the second group ingested the apple extract from the placebo field. At the beginning and end of the study, participants underwent blood tests for inflammatory markers (IL-6 and TNF-α).

[0106] The blood level of IL-6 in the participants at the beginning and end of the study is shown in Table 4 below.

[0107] [Tables4] IL-6 (pg / ml) Day 0 Day 30 QKC 5.9 3.5 Placebo 6.0 4.8

[0108] Blood levels of IL-6 were observed to decrease significantly in both the QKC and placebo groups. However, the anti-inflammatory effect was more pronounced in the QKC group. Blood levels of IL-6 decreased by 40.7% in the QKC group compared to a decrease of 20% in the placebo group.

[0109] The blood level of TNF-a in the participants at the beginning and end of the study is shown in Table 5 below.

[0110] [Tableaux5] TNF-a (pg / ml) Day 0 Day 30 QKC 2.3 1.40 Placebo 2.1 1.6

[0111] Blood levels of TNF-α were observed to decrease significantly in both groups; however, the decrease was greater in the QKC group. Blood levels of TNF-α decreased by 39.1% in the Mito-boost group compared to a decrease of 23.8% in the placebo group.

[0112] This double-blind, placebo-controlled clinical study shows that QKC increases the anti-inflammatory properties of apples.

Claims

Demands

1. A method for manufacturing a plant treatment composition intended to increase the yield of agricultural production, preferably cereals, fruits and vegetables, and to enhance their beneficial properties on human health, characterized in that it comprises a mixture of live bacteria from the microbiota of turmeric (Curcuma longa) rhizomes, preferably turmeric from Bhutan, a lysate of said bacteria and metabolites from the fermentation of said bacteria, comprising the steps of: i) collect turmeric rhizomes, preferably grown in a field in Bhutan, even more preferably without having undergone chemical treatment; ii) isolate bacteria from the microbiome of the rhizomes; iii) culture the isolated bacteria to obtain a mother culture of the bacteria; iv) inoculate a sterilized substrate with bacteria from the mother culture and ferment, preferably in a fermentation tank for at least 120 hours at a temperature between 30°C and 45°C; v) sterilize and microfilter the mixture fermented in step iv), resulting in a composition comprising the fermentation metabolites and bacterial lysates; (vi) mix the composition comprising the fermentation metabolites and bacterial lysates with bacteria from the mother culture of bacteria isolated from turmeric; said substrate being bio-based and edible selected from all plants such as fruits, vegetables and algae; food waste from households, workplaces and catering establishments; compost and manure.

2. Process according to claim 1, in which the bacteria are taken from the following list: Lactobacillus fermentum, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus acidophilus, Bifidobacterium longum, Bifidobacterium bifidum, Bifidobacterium adolescentis, Bifidobacterium infantis, Lactobacillus brevis, Lactobacillus Jensenii, Lactobacillus paracasei, Christensenella minuta, Lactiplantibacillus plantarum, Lactiplantibacillus paraplantarum, Mycobacterium vaccae, Pseudomonas japonica, Pseudomonas fluorescens, Archaeospora trappei, Methylobacterium crusticola, Bradyrhizobium elkanii, Bradyrhizobium japonicum, Gluconacetobacter diazotrophicus, Azospirillum Brazilian.

3. A method according to claim 1 or claim 2, wherein the substrate is selected from the fruits, barks, leaves or flowers of figs, olives, rosemary, thyme, beans, sage, melon, pomegranate, soybeans, watermelons, cabbages, green or black tea, turmeric, pine needles.

4. Use of the plant treatment composition obtained by the process according to any one of claims 1 to 3, to increase soil fertility by increasing the concentration of nutrients and decreasing the amount of toxic substances.

5. Use of the plant treatment composition obtained by the process according to any one of claims 1 to 3, to increase the yield of cereals, fruits and vegetables, by increasing plant growth, their fertility and by increasing the number and weight of seeds, fruits and vegetables.

6. Use of the plant treatment composition obtained by the process according to any one of claims 1 to 3, to increase the yield of nutrients such as fiber, vitamins, minerals, polyphenols, terpenes, nitrogen compounds, alkaloids, steroids, terpenoids, flavonoids, omega-3, omega-6 and omega-9, linoleic acid, L-carnitine, choline or sphingomyelin.

7. Use of the plant treatment composition obtained by the process of any one of claims 1 to 3, to transform daidezin into S-equol in cereals, fruits and vegetables and to increase their dietary fiber and resistant starch content.

8. Use of the plant treatment composition obtained by the process according to any one of claims 1 to 3, to increase the dietary fiber content of cereals, fruits and vegetables.

9. A method for increasing the yield of at least one plant, wherein the composition obtained by the method according to any one of claims 1 to 3 is sprayed or spread on the soil and / or the at least one plant.