Use of encapsulated organic compounds as biostimulants for mycelia and fungi

Abstract

The present invention relates to the use of specific organic compounds that are encapsulated in a particular way as biostimulants in the production of mycelia and the cultivation of fungi. More particularly, the invention relates to the use of at least one organic compound chosen from among beta-pinene, delta-3-carene or beta-phellandrene, or one of the mixtures thereof, encapsulated with an inert solid material and a fat to stimulate the development and growth of edible fungi and mycelia.

Description

[0001] DESCRIPTION

[0002] USE OF ENCAPSULATED ORGANIC COMPOUNDS AS BIOSTIMULANTS OF MYCELIAS AND MUSHROOMS

[0003] The present invention relates to the use of particular organic compounds as biostimulants in the production of mycelia and the cultivation of mushrooms. More particularly, the invention relates to the use of particular organic compounds encapsulated in a particular manner, in stimulating the development and growth of mycelia and improving the yields of edible mushroom cultures.

[0004] Edible mushrooms have always been used by humans, particularly for their nutritional and medicinal properties. There are many varieties of mushrooms, but the most widely cultivated mushrooms in the world are the button mushroom (Agaricus bisporus), oyster mushrooms (Pleurotus ostreatus, Pleurotus eryngii, etc.), and shiitake mushrooms (Lentinula edodes). The button mushroom alone accounts for more than 40% of the global market, oyster mushrooms 25%, and shiitake 15%. The main players in the global market are China, with 50% of production, the United States, and Europe.

[0005] On an industrial scale, the mushroom production method can be broken down into two stages: the first involves obtaining inoculum, mycelia, and the second involves the actual cultivation of the mushrooms, using the mycelia obtained during the first stage. These two stages can be carried out independently by different parties, the first by a mycelia producer, the second by a mushroom grower, or by a single party, the mushroom grower.

[0006] Obtaining inoculum, mycelia, begins by inoculating a sterilized culture medium from spores or a piece of inoculum. The culture medium can be agar, such as potato dextrose agar (PDA), a liquid medium such as potato dextrose broth (PDB), or any other nutrient solution containing a gelling agent or not. The development and growth of the mycelium on its culture medium is variable but generally extends from 7 to 28 days, depending on the fungal strain. Once the mycelium has completely colonized the culture medium, a piece of the culture medium is then removed for inoculation with a colonization substrate.A suitable colonization substrate can be synthetic, or most often composed of cereals, generally rye, millet, sorghum, wheat, barley, rice or oats, previously sterilized and packaged in jars or micro-perforated culture bags.

[0007] The resulting mycelium inoculum is then used in mushroom cultivation and production. The inoculum is then used to seed a fruiting substrate.

[0008] In the case of the button mushroom, the fruiting substrate is a compost generally composed of straw and animal droppings, watered abundantly to ensure its maturation for two to three weeks. The fruiting substrate is then pasteurized for a few days with decreasing temperatures from 60 to 40°C. The inoculation of the fruiting substrate is done after pasteurization, for example using a larder, by mixing the inoculum contained in its colonization substrate and the fruiting substrate. This is followed by an incubation period during which the inoculated fruiting substrates are placed in an enclosed room whose temperature, humidity and oxygen are controlled for two weeks. The temperature is maintained at 22 to 25°C. The next step is casing, which consists of covering the fruiting substrate with a layer of suitable soil.The casing soil is, for example, a mixture of crushed and disinfected tuffeau and horticultural peat. After a controlled drop in temperature, the first mushroom heads emerge from the fruiting substrate, and harvesting can then begin. Button mushroom picking follows several fruiting cycles called rounds, or alternating periods of harvest and vegetative rest. The first two rounds are by far the most productive, with 50% of the total harvest attributed to the first round and 35% to the second. The harvest cycle is renewed approximately every week. Harvesting can be carried out up to the third round, but yields decline rapidly. Generally speaking, mushroom growers seek to produce in only two rounds for reasons of productivity but also hygiene, because the third round is often a gateway to the development of diseases and their longer-term establishment.These methods of obtaining inoculum and producing mushrooms thus extend over several weeks, in particular due to the time required for different strains of mycelia to colonize their substrates. The development and growth of mycelia can also be disrupted by various factors, such as a variation in the parameters of the culture conditions and its colonization capacity. The growth rates and the quality of mycelia development can thus become a limiting factor, extending inoculum production times for producers and negatively impacting mushroom harvest yields for mushroom growers. Thus, the entire mushroom production chain can be directly impacted by the quality of the mycelium used.

[0009] The use of biostimulants in agricultural production methods is growing rapidly. Biostimulants are substances capable of stimulating the metabolism of a plant, or a fungus, and its natural nutrient absorption processes. More specifically, Regulation (EU) 2019 / 1009 of the European Parliament and of the Council of 5 June 2019, which comes into force on 22 July 2022, defines a biostimulant as “a product that stimulates plant nutrition processes independently of the nutrients it contains, with the aim of improving one or more of the following characteristics of plants or their rhizosphere: a) nutrient use efficiency b) tolerance to abiotic stress c) quality characteristics d) availability of nutrients confined in the soil or rhizosphere.”

[0010] The aim of the present invention is thus to identify and propose biostimulants that are effective in methods for producing mycelium inoculum and in methods for cultivating mushrooms from mycelium inoculum. In particular, the biostimulants according to the invention should make it possible to shorten the production times for mycelium inocula and to improve mushroom cultivation yields or, at the very least, to avoid the lengthening of these times and the loss of these yields. Finally, the biostimulants according to the invention should be usable in conventional methods for producing inoculum and for producing mushrooms.

[0011] It is for this purpose that the applicant company carried out its research and discovered that certain organic compounds, chosen from beta-pinene, delta-3-carene or beta-phellandrene, or one of their mixtures, exhibited surprising biostimulation capacities on the development and growth of mycelia and fungi, in particular when added to their culture media or substrates, and could therefore be used in processes and methods for obtaining inoculum or culturing fungi.

[0012] The use of natural biostimulants is a real innovation for mushroom production. The use of products with a biostimulant effect has a major impact on the production of mycelia and mushrooms. The main limitation of inoculum producers lies in their ability to quickly produce, store and distribute their mycelia to mushroom growers. The use of biostimulants according to the invention makes it possible to reduce the time between the start of production and distribution, thus increasing the production capacity of producers. Similarly, during the mushroom production phase, the use of previously biostimulated mycelium and the addition of biostimulants to the growing medium makes it possible to improve yields and increase the production of mushroom growers, without negatively impacting the crop.The addition of biostimulants can thus be carried out when inoculating the substrate bags or during the different phases of compost incubation, more particularly when adding supplements (nutrient supplements).

[0013] Thus, during tests carried out on composts, the mycelia stimulated by the organic compounds selected according to the invention, it was possible to observe and measure an increase in crop yield, an increase in the average mass per mushroom and, generally speaking, an improvement in the quality of the mushrooms harvested.

[0014] The organic compounds selected within the scope of the invention have the particularity of exhibiting a strong biostimulating effect, over a sufficiently wide range of concentrations for producers. However, when they are applied in their natural form in mycelium culture bags, excess stimulation, or localized hyper-stimulation phenomena, or temporary inhibition zones at the points of contact have been observed. In this form, the active substances are released immediately and completely, and the mycelium develops in an impressive but often heterogeneous manner, which is to be avoided by mycelium producers, and can create problems of homogeneity in mushroom cultivation.

[0015] To solve this problem, the applicant company has selected and improved a specific encapsulation technique, adapted to the physicochemical characteristics of the selected organic compounds. This encapsulation technique makes it possible to preserve all the biostimulant properties of the organic compounds, while releasing their active ingredients gradually over time, and under controlled conditions suitable for both the production of mycelium and the cultivation of mushrooms.

[0016] Thus, the present invention relates to the use of at least one organic compound chosen from beta-pinene, delta-3-carene or beta-phellandrene, or one of their mixtures, in an encapsulated form, to stimulate the development and growth of a mycelium or a fungus.

[0017] The treatment by encapsulation of the selected organic compounds makes it possible to increase the supply of biostimulant substances without deleterious effects on the mycelium and its development. Compared to mycelium not treated with the encapsulated organic compound(s) according to the invention, the growth rate of the biostimulated mycelium is higher, its development is more homogeneous, and its behavior is more prospective. The supply in encapsulated form also makes it possible to avoid localized excesses of biostimulation which lead to heterogeneous colonization of the colonization substrate bags. A single application is sufficient for inoculation for an effective, homogeneous and complete action of the colonization substrate. The use of the encapsulated biostimulants according to the invention thus makes it possible to reduce the time between the start of production and distribution, thereby increasing the production capacity of mycelium producers.The main limitation of inoculum producers lies in their ability to quickly produce, store and distribute quality mycelium to composters and mushroom growers. Also, the contribution of selected encapsulated organic compounds makes it possible to improve mushroom crop yields, particularly when the compost (fruiting substrate) is directly inoculated with biostimulated mycelium. This increase in yield is even more marked when supplements (nutrient supplements) are added to the compost. Indeed, the application of encapsulated biostimulants allows the mycelium to sustainably improve its efficiency in using these nutrients.

[0018] Furthermore, during mushroom cultivation, the addition of the encapsulated organic compounds according to the invention makes it possible to increase the yields of the first flushes and to improve the quality and average mass of the harvested mushrooms. The effects on the mushroom cultivation sector are therefore accompanied by economic and practical benefits for producers, at each stage:

[0019] - Energy savings linked to earlier marketing and reduced storage time for mycelium producers,

[0020] - Reduction of labor time during picking for mushroom growers,

[0021] - Application easily adaptable to all production methods and combinable with supplements (nutritional supplements) and treatment products.

[0022] According to the invention, the selected organic compounds are adsorbed, absorbed onto a support or encapsulated in a support and coated with fatty material which acts as a coating.

[0023] The invention thus also relates to microcapsules comprising:

[0024] - a core comprising at least one organic compound selected from beta-pinene, delta-3-carene or beta-phellandrene, or one of their combinations, a solid support, said organic compound(s) being adsorbed, absorbed onto said solid support or encapsulated therein, the solid support being an inert solid material, a material composed of silica or silicon dioxide, and,

[0025] - at least one coating layer formed of vegetable fat, said layer coating said core. The selected organic compounds are interchangeable, for example in the event of a shortage of one of them.

[0026] The selected organic compounds can also be combined to obtain in certain cases synergistic effects and increased resilience.

[0027] The selected organic compounds also allow their effects on the growth of mycelia and fungi to be produced under less favorable conditions than normal, such as a lowering of the incubation temperature by 1 or 2°C or an aging effect.

[0028] Thus, it is the combination of carefully selected organic compounds and the form in which they are distributed that makes it possible to achieve the objectives of the invention.

[0029] Any combination of encapsulated organic compounds is possible, such as beta-pinene and delta-3-carene or beta-pinene and beta-phellandrene, or delta-3-carene and beta-phellandrene, or beta-pinene and delta-3-carene and beta-phellandrene.

[0030] However, according to a preferred embodiment, the encapsulated organic compound is beta-pinene.

[0031] The organic compounds used in the context of the invention are advantageously and preferably in the form of liquid extracts comprising at least 90%, at least 92%, preferably at least 95% of said organic compound(s). The organic compounds selected according to the invention are terpenes generally extracted from essential oils. Such extracts are commercially available.

[0032] According to one embodiment of the invention, the mycelium or the fungus is chosen from basidiomycetes, in particular the button mushroom, oyster mushrooms and shiitake, more particularly Agaricus bisporus, Pleurotus ostreatus, Pleurotus eryngii and Lentinula edodes.

[0033] When the mycelium or mushroom is Agaricus bisporus, the organic compound used is preferably beta-pinene, delta-3-carene or beta-phellandrene. When the mycelium or mushroom is Pleurotus ostreatus, the organic compound used is preferably beta-pinene, delta-3-carene or beta-phellandrene.

[0034] When the mycelium or mushroom is Pleurotus eryngii, the organic compound used is preferably beta-pinene or beta-phellandrene.

[0035] When the mycelium or fungus is Lentinula edodes, the organic compound used is preferably beta-pinene.

[0036] According to one embodiment of the invention, the solid support is a spheroidal granule.

[0037] Preferably, the solid support is an inert solid material, a material composed of silica or a material composed of silicon dioxide.

[0038] According to one embodiment of the invention, the vegetable fat is composed of fatty acids or lipids, preferably one or more vegetable oils.

[0039] The manufacture of the microcapsules according to the invention can be carried out according to methods known to those skilled in the art.

[0040] For example, the microcapsules are manufactured as follows: loading the solid support into a homogenizer, adding, at low temperature, the organic compound(s) selected according to the invention to the solid support, adding the vegetable fat, sieving to 1 mm, to obtain a particle size of less than 1 mm, packaging.

[0041] The invention also relates to a composition comprising at least one microcapsule as described previously.

[0042] Preferably, the composition is in powder form. More preferably, the composition only comprises the microcapsules according to the invention. According to one embodiment, the composition comprises at least 10, 20, 30, 40 or 50% of at least one of said organic compounds, by weight relative to the total weight of the composition.

[0043] According to a preferred embodiment, the composition is composed of 50% of at least one organic compound chosen from beta-pinene, delta-3-carene or beta-phellandrene, or one of their mixtures, of 35% of support, preferably of silica or silicon dioxide support, and of 15% of fatty material coating.

[0044] The invention also relates to the use of at least one microcapsule as described above, or of a composition containing it, for stimulating the development and growth of a mycelium or a fungus.

[0045] The organic compounds selected and encapsulated within the framework of the invention can be used simply and effectively in conventional methods of inoculum production and mushroom production, without it being necessary to complicate these methods.

[0046] Thus, the invention also relates to a method for obtaining an inoculum of mycelium, characterized in that it comprises the following steps:

[0047] Thus, the invention also relates to a method for obtaining an inoculum of mycelium, characterized in that it comprises the following steps: preparation of a colonization substrate, addition to the colonization substrate of at least one microcapsule as described previously, or of a composition comprising it, inoculation of the colonization substrate with a mycelium, obtaining an inoculum of mycelium, conditioning of the inoculum.

[0048] Preferably in the method according to the invention, the encapsulated organic compound(s) are added to the colonization substrate, or compost, so that the concentration of the organic compound(s) is between 20 and 400 pL / L of colonization substrate. For example, this corresponds to approximately 0.5 to 10 g of microcapsules per 15-liter culture bag.

[0049] The method according to the invention makes it possible to shorten by several days the time required to obtain inocula, or mycelia, marketable by producers, compared to a conventional method. Growth can be stimulated and increased by more than 20%, or even more than 40%, compared to the growth obtained in a conventional method, that is to say without the use of the organic compounds selected and encapsulated within the framework of the invention. The stimulation of the growth of the mycelium can be observed by its behavior and the homogeneity of its development on the surface of the culture bags.

[0050] According to one embodiment of the invention, in this method, the mycelium or the fungus is chosen from basidiomycetes, in particular the button mushroom, oyster mushrooms and shiitake, more particularly Agaricus bisporus, Pleurotus ostreatus, Pleurotus eryngii and Lentinula edodes.

[0051] According to a preferred embodiment, in this method, the organic compound is beta-pinene, in particular when the mycelium or mushroom is Agaricus bisporus.

[0052] The invention also relates to a method for cultivating mushrooms from an inoculum, of mycelium, which is characterized in that it comprises the following steps: preparation of a fruiting substrate, optionally, incorporation into the fruiting substrate of at least one microcapsule as described above, or of a composition comprising it, seeding of the fruiting substrate with an inoculum of mycelium obtained by a method according to the invention and described above, optionally, incorporation, into the fruiting substrate, of supplements or mixtures of nutritive materials specific to the mushroom culture, incubation, with optionally incorporation into the fruiting substrate of at least one microcapsule according to one of the preceding claims, or of a composition comprising it, fruiting, harvesting.

[0053] According to one embodiment of the invention, the quantity of microcapsules added to the fruiting substrate is 50 to 500 g per tonne of compost or fruiting substrate. In this embodiment, the concentration of the organic compound(s) is between 28 and 280 mL per 1000 kg of fruiting substrate.

[0054] According to one embodiment of the invention, in this method, the mycelium or the fungus is chosen from basidiomycetes, in particular the button mushroom, oyster mushrooms and shiitake, more particularly Agaricus bisporus, Pleurotus ostreatus, Pleurotus eryngii and Lentinula edodes.

[0055] According to a preferred embodiment, in this method, the organic compound is beta-pinene, in particular when the mycelium or mushroom is Agaricus bisporus.

[0056] The above-mentioned and other features of the invention will become more apparent upon reading the following description of exemplary embodiments.

[0057] Example 1: Evaluation of the impact of a composition according to the invention on the yields of Agaricus bisporus crops

[0058] 1.1 Preparation of the composition according to the invention

[0059] The composition is formulated from the following elements:

[0060] Organic compound: beta-pinene (CAS No. 127-91-3) most often separated from turpentine oil by fractional distillation (beta-pinene content > 95%).

[0061] Solid carrier: silicon dioxide. Vegetable fat: vegetable oil.

[0062] [Tab.l]

[0063] The composition is obtained as follows:

[0064] - loading silicon dioxide into a mixer,

[0065] - addition by spraying, at a temperature of 20-25°C, of ​​beta-pinene,

[0066] - mix for 7 minutes

[0067] - adding vegetable oil

[0068] - mix for 1 minute

[0069] - sieving to 1 mm, to obtain a particle size of less than 1 mm,

[0070] - packaging.

[0071] The resulting composition is in the form of a powder composed of microcapsules less than 1 mm in size.

[0072] The composition will be called “encapsulated composition” in the following.

[0073] 1.2 Experimental protocol for application in mycelium culture bags

[0074] Application of the composition in the industrial production phase of mycelium - Obtaining non-encapsulated beta-pinene (CAS No. 127-91-3).

[0075] Deposit of 300 pL and 600 pL of said non-encapsulated beta-pinene, per 15 liter bag of colonization substrate (i.e. 20 and 40 pL / L of substrate), at the time of inoculation by the mycelium.

[0076] Preparation of the encapsulated composition (as described in point 1.1). Deposit of 2, 3 and 4 g of the encapsulated composition per 15 litre bag of colonisation substrate (i.e. approximately 80, 120 and 160 pL / L of substrate), at the time of inoculation by the mycelium.

[0077] Colonization of the substrate bags by the mycelium for 21 days, according to the method usually used by mycelium producers. before worship

[0078] Inoculation of plots of 9 kg of compost (representing 0.1 m 2 mushroom culture) with 72 g of untreated mycelium.

[0079] Inoculation of 9 kg plots of compost with 72 g of mycelium treated with non-encapsulated beta-pinene.

[0080] Inoculation of 9 kg plots of compost with 72 g of mycelium treated with the encapsulated composition.

[0081] Addition in plots of 9 kg of compost of 72 g of supplements (nutrient supplements), inoculated with untreated mycelium and treated with unencapsulated beta-pinene and the encapsulated composition (See methods indicated in the “Supp.” column in Tab.2).

[0082] Management of Aqaricus bisporus production in a culture room

[0083] Incubation of plots for 13 days.

[0084] Deposit 3000 mL of casing soil (90% black peat and 10% calcium carbonate) to a thickness of 3 cm on each plot.

[0085] Post-incubation of the plots for 10 days.

[0086] Start of the induction of fruiting of the first flush.

[0087] Harvest, weigh and measure the yields of the 3 rounds.

[0088] 1.3 Experimental protocol for application to compost inoculation of mushrooms

[0089] Inoculation of plots of 9 kg of compost (representing 0.1 m 2 mushroom culture) with 72 g of untreated mycelium.

[0090] Deposit 3 g of the encapsulated composition per 9 kg plot of compost (i.e. 30 g / 90 kg).

[0091] Management of A aricus bisporus production in the culture room

[0092] Incubation of plots for 13 days.

[0093] Deposit 3000 mL of casing soil (90% black peat and 10% calcium carbonate) to a thickness of 3 cm on each plot.

[0094] Post-incubation of the plots for 10 days.

[0095] Start of the induction of fruiting of the first flush.

[0096] Harvest, weigh and measure the yields of the 3 rounds.

[0097] Each method described above was conducted on 6 identical experimental plots, corresponding to 6 repetitions. The mycelium strain used for all experimental tests is the commercial variety of Agaricus bisporus, Heirloom brand, produced by the American company Amycel.

[0098] 1.4 results

[0099] The table below shows the measured yields, in kilograms of healthy mushrooms per square meter of culture, of the different experimental methods described above. All yields are also expressed as a percentage, relative to the control method (untreated and unsupplemented compost, inoculated with untreated mycelium).

[0100] [Ta b.2]

[0101]

[0102] According to the results of the experimental tests in Example 1, a clear improvement in yields can be observed when beta-pinene is applied in its non-encapsulated and encapsulated form in mycelium culture bags, and in encapsulated form mixed with compost.

[0103] Increasing the doses applied, in both non-encapsulated and encapsulated form, results in improved mushroom yields.

[0104] Application in encapsulated form allows for a gradual release of the active ingredient, and results in a greater and more sustained increase in yields over time than direct application in non-encapsulated form.

[0105] The encapsulated active ingredient allows early treated mycelium to improve its nutrient utilization efficiency. This ability is evident when supplements (nutrient supplements) are added to the compost at the time of inoculation.

[0106] In this configuration, the yields obtained only from the first two rounds (24.26 kg) are higher than the yields from the three rounds of the control method (23.54 kg), carried out with untreated mycelium and without supplements added to the compost. This last point is important, because mushroom growers prefer to avoid continuing their crops until the third round to shorten their production cycle, ensure a more qualitative and homogeneous production, and avoid the appearance of diseases that can be difficult to get rid of.

[0107] In conclusion, the encapsulation of the organic compounds selected according to the invention makes it possible to increase the application doses of active ingredients, without risking excessive biostimulation and heterogeneous development of the mycelium in the growing bags. This possible increase in application doses results in an improvement in mushroom yields.

[0108] Example 2: Evaluation of the impact of a composition according to the invention on the yields of Agaricus bisporus crops

[0109] 2.1 Experimental protocol for applying the composition to the inoculation of compost

[0110] Application of the composition in the mushroom production phase

[0111] Preparation of the encapsulated composition (as described in point 1.1). - Inoculation of plots of 9 kg of compost (representing 0.1 m 2 mushroom culture) with 72 g of untreated mycelium.

[0112] Deposit of 100 g to 250 g of the encapsulated composition per tonne of compost, or 0.9 g to 2.25 g per 9 kg plot of compost.

[0113] Addition of 72 g of supplements (nutrient supplements) per 9 kg plot of compost (see details indicated in the “Supp.” column in Tab.3).

[0114] Management of Agaricus bisporus production in the growing room

[0115] Incubation of plots for 13 days.

[0116] Deposit 3000 mL of casing soil (90% black peat and 10% calcium carbonate) to a thickness of 3 cm on each plot.

[0117] Post-incubation of the plots for 10 days.

[0118] Start of fruiting induction of the first round. Harvest, weighing and measurement of the yields of the 3 rounds.

[0119] Each method described above was conducted on 6 identical experimental plots, corresponding to 6 repetitions. The mycelium strain used for all experimental tests is the commercial variety of Agaricus bisporus, Heirloom brand, produced by the American company Amycel.

[0120] 2.2 Results

[0121] The table below shows the measured yields, in kilograms of healthy mushrooms per square meter of culture, of the different experimental methods described above. All yields are also expressed as a percentage, relative to the control method (untreated and unsupplemented compost, inoculated with untreated mycelium). [Ta b.3]

[0122] According to the results of the experimental tests in Example 2, a clear improvement in yields can be observed when beta-pinene is applied in its encapsulated form mixed with compost, even in the presence of supplements. The increase in the applied doses clearly results in an improvement in mushroom yields, which are higher in only two rounds (19.51 to 20.77 kg) than the control modality in three rounds (19.22 kg).

[0123] Example 3: Evaluation of the impact of a composition according to the invention on the yields of Agaricus bisporus crops

[0124] 3.1 Experimental protocol for applying the composition to the inoculation of compost

[0125] Preparation of the encapsulated composition (as described in point 1.1).

[0126] Inoculation of plots of 9 kg of compost (representing 0.1 m 2 mushroom culture) with 72 g of untreated mycelium.

[0127] Deposit of 200 g to 350 g of the encapsulated composition per tonne of compost, or 1.8 g to 3.15 g per 9 kg plot of compost.

[0128] Addition of 72 g of supplements (nutrient supplements) per 9 kg plot of compost (see details indicated in the “Supp.” column in Tab.4).

[0129] Production management of in the growing room

[0130] Incubation of plots for 13 days.

[0131] Deposit 3000 mL of casing soil (90% black peat and 10% calcium carbonate) to a thickness of 3 cm on each plot.

[0132] Post-incubation of the plots for 10 days.

[0133] Start of the induction of fruiting of the first flush.

[0134] Harvest, weigh and measure the yields of the 3 rounds.

[0135] Each method described above was conducted on 9 identical experimental plots, corresponding to 9 repetitions. The mycelium strain used for all experimental tests is the commercial variety of Agaricus bisporus of the Heirloom brand, produced by the American company Amycel.

[0136] 3.2 Results

[0137] The table below shows the measured yields, in kilograms of healthy mushrooms per square meter of culture, of the different experimental methods described above. All yields are also expressed as a percentage, relative to the control method (untreated and supplemented compost, inoculated with untreated mycelium). [Ta b.4]

[0138] According to the results of the experimental tests in Example 3, a clear improvement in yields can be observed when beta-pinene is applied in its encapsulated form mixed with compost, including in the presence of supplements.

[0139] Example 4: Evaluation of the impact of a composition according to the invention on the yields of Pleurotus ostreatus crops

[0140] 4.1 Experimental protocol for applying the composition to the substrate inoculation Application of the composition in the mushroom production phase

[0141] Preparation of the encapsulated composition (as described in point 1.1). Inoculation of 15 kg blocks of fruiting substrate with 375 g of untreated mycelium, i.e. a release or inoculation rate of 2.5%.

[0142] Deposit of 100 g of the encapsulated composition per ton of substrate, or 1.5 g per block of 15 kg of substrate.

[0143] Management of Pleurotus ostreatus production in the culture room

[0144] Incubation of the blocks for 2 weeks.

[0145] Post-incubation in the fruiting room for 2 weeks.

[0146] Start of the induction of fruiting of the first flush.

[0147] Harvesting, weighing and measuring yields until the end of harvest.

[0148] Each modality described above was conducted on 6 identical experimental blocks, therefore corresponding to 6 repetitions. The mycelium strain used for all experimental tests is the commercial variety of Pleurotus ostreatus brand 3015, produced by the American company Sylvan.

[0149] 4.2 Results

[0150] The table below shows the measured yields, in kilograms of healthy mushrooms produced per tonne of fruiting substrate, of the different experimental methods described above. All yields are also expressed as a percentage, relative to the control method (untreated substrate, inoculated with untreated mycelium).

[0151] [Ta b.5] According to the results of the experimental tests in Example 4, a clear improvement in yields can be observed when beta-pinene is applied in its encapsulated form mixed with the fruiting substrate. The composition according to the invention positively impacts yields from the first weeks of harvest, which is particularly beneficial to producers.

[0152] Example 5: Evaluation of the impact of a composition according to the invention on the yields of Pleurotus eryngii crops

[0153] 5.1 Experimental protocol for applying the composition to the inoculation of the substrate

[0154] Preparation of the encapsulated composition (as described in point 1.1).

[0155] Inoculation of 15 kg blocks of fruiting substrate with 900 g of untreated mycelium, i.e. a release or inoculation rate of 6%.

[0156] Deposit of 100 g to 300 g of the encapsulated composition per ton of substrate, or 1.5 g to 4.5 g per block of 15 kg of substrate.

[0157] Management of Pleurotus eryngii production in the culture room

[0158] Incubation of the blocks for 3 weeks.

[0159] Post-incubation in the fruiting room for 1 week.

[0160] Start of the induction of fruiting of the first flush.

[0161] Harvesting, weighing and measuring yields until the end of harvest.

[0162] Each modality described above was conducted on 6 identical experimental blocks, therefore corresponding to 6 repetitions. The mycelium strain used for all experimental tests is the commercial variety of Pleurotus eryngii brand 3066, produced by the American company Sylvan. 5.2 Results

[0163] The table below shows the measured yields, in kilograms of healthy mushrooms produced per tonne of fruiting substrate, of the different experimental methods described above. All yields are also expressed as a percentage, relative to the control method (untreated substrate, inoculated with untreated mycelium).

[0164] [Ta b.6]

[0165] According to the results of the experimental tests in Example 5, a clear improvement in yields can be observed when beta-pinene is applied in its encapsulated form mixed with the fruiting substrate. The composition according to the invention positively impacts yields from the first weeks of harvest, which is particularly beneficial to producers. Example 6: Evaluation of the impact of a composition according to the invention on the yields of Lentinula edodes crops

[0166] 6.1 Experimental protocol for applying the composition to the inoculation of the substrate

[0167] Application of the composition in the mushroom production phase

[0168] Preparation of the encapsulated composition (as described in point 1.1).

[0169] Inoculation of 15 kg blocks of fruiting substrate with 900 g of untreated mycelium, i.e. a release or inoculation rate of 6%. Deposit of 200 g to 300 g of the encapsulated composition per tonne of substrate, i.e. 3 g to 4.5 g per 15 kg blocks of substrate.

[0170] Management of Lentinula edodes production in a culture room

[0171] Incubation of the blocks for 5 weeks.

[0172] Post-incubation in the fruiting room for 2 weeks.

[0173] Start of the induction of fruiting of the first flush.

[0174] Harvesting, weighing and measuring yields until the end of harvest.

[0175] Each modality described above was conducted on 9 identical experimental blocks, therefore corresponding to 9 repetitions. The mycelium strain used for all experimental tests is the commercial variety of Lentinula edodes brand 4325, produced by the American company Sylvan.

[0176] 6.2 Results

[0177] The table below shows the measured yields, in kilograms of healthy mushrooms produced per tonne of fruiting substrate, of the different experimental methods described above. All yields are also expressed as a percentage, relative to the control method (untreated substrate, inoculated with untreated mycelium).

[0178] [Tab.7] According to the results of the experimental tests in Example 6, a clear improvement in yields can be observed when beta-pinene is applied in its encapsulated form mixed with the fruiting substrate. The composition according to the invention positively impacts yields from the first weeks of harvest, which is particularly beneficial to producers.

Claims

CLAIMS 1) Use of at least one organic compound chosen from beta-pinene, delta-3-carene or beta-phellandrene, or one of their mixtures, in an encapsulated form, to stimulate the development and growth of a mycelium or a fungus. 2) Use according to claim 1 in which the organic compound(s) are in the form of liquid extracts comprising at least 90%, at least 92%, preferably at least 95% of said organic compound(s), by weight relative to the weight of the liquid extract. 3) Use according to claim 1 or 2 in which the mycelium or the fungus is chosen from basidiomycetes, in particular the button mushroom, oyster mushrooms and shiitake, more particularly Agaricus bisporus, Pleurotus ostreatus, Pleurotus eryngii and Lentinula edodes. 4) Microcapsule comprising: - a core comprising at least one organic compound selected from beta-pinene, delta-3-carene or beta-phellandrene, or one of their combinations, a solid support, said organic compound(s) being adsorbed, absorbed onto or encapsulated in said solid support, the solid support being an inert solid material, a material composed of silica or silicon dioxide, and, - at least one coating layer formed from vegetable fat, said layer coating said core. 5) Microcapsule according to claim 4, characterized in that the organic compound(s) are in the form of liquid extracts comprising at least 90%, at least 92%, preferably at least 95% of said organic compound(s), by weight relative to the weight of the liquid extract. 6) Microcapsule according to claim 4 or 5, characterized in that the organic compound is beta-pinene. 7) Microcapsule according to one of claims 4 to 6, characterized in that the solid support is a spheroidal granule. 8) Microcapsule according to one of claims 4 to 7, characterized in that the solid support is an inert solid material, a material composed of silica or silicon dioxide. 9) Microcapsule according to one of claims 4 to 8, characterized in that the vegetable fat is composed of fatty acids or lipids, preferably one or more vegetable oils. 10) Composition comprising at least one microcapsule as defined in one of claims 4 to 9. 11) Composition according to claim 10, characterized in that it comprises at least 10, 20, 30, 40, or 50% of at least one of said organic compounds. 12) Use of at least one microcapsule according to one of claims 4 to 9, or of a composition containing it, for stimulating the development and growth of a mycelium or a fungus. 13) Use according to claim 12, in which the mycelium or the fungus is chosen from basidiomycetes, in particular the button mushroom, oyster mushrooms and shiitake, more particularly Agaricus bisporus, Pleurotus ostreatus, Pleurotus eryngii and Lentinula edodes. 14) Use according to one of claims 12 or 13, characterized in that said organic compound is beta-pinene, in particular when the mycelium or mushroom is Agaricus bisporus. Tl) Method for obtaining an inoculum of mycelium, characterized in that it comprises the following steps: preparation of a colonization substrate, addition to the colonization substrate of at least one microcapsule according to one of claims 4 to 9, or of a composition comprising it, inoculation of the colonization substrate with a mycelium, obtaining an inoculum of mycelium, conditioning of the inoculum.) Method according to claim 15, characterized in that the microcapsule(s), or the composition comprising it, are added to the colonization substrate so that the concentration of the organic compound(s) is between 20 and 400 pL / L of colonization substrate.) Method for cultivating mushrooms from a mycelium inoculum, characterized in that it comprises the following steps: preparation of a fruiting substrate, optionally, incorporation into the fruiting substrate of at least one microcapsule according to one of claims 4 to 9, or of a composition comprising it, seeding of the fruiting substrate with a mycelium inoculum obtained by a method according to claim 15 or 16, optionally, incorporation, into the fruiting substrate, of supplements or mixtures of nutrients specific to the mushroom culture, incubation, with optional incorporation into the fruiting substrate of at least one microcapsule according to one of claims 4 to 9, or of a composition comprising it, fruiting, harvesting. ) Method according to claim 17, characterized in that the quantity of microcapsules added to the fruiting substrate is 50 to 500 g per tonne of substrate, so that the concentration of the organic compound(s) is between 28 and 280 mL per 1000 kg of fruiting substrate. ) Method according to one of claims 15 to 18, characterized in that the mycelium or fungus is chosen from basidiomycetes, in particular the button mushroom, oyster mushrooms and shiitake, more particularly Agaricus bisporus, Pleurotus ostreatus, Pleurotus eryngii and Lentinula edodes. ) Method according to one of claims 15 to 19, characterized in that said organic compound is beta-pinene, in particular when the mycelium or mushroom is Agaricus bisporus.

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