Use of encapsulated organic compounds as biostimulants for mycelia and mushrooms

Abstract

The present invention relates to the use of specific organic compounds encapsulated in a specific manner as biostimulants in mycelium production and mushroom culture. More particularly, the invention relates to the use of at least one organic compound encapsulated with an inert solid material and fat, selected from the group consisting of beta-pinene, delta-3-carene, or beta-phellandrene, or one of the mixtures thereof, for stimulating the development and growth of edible mushrooms and mycelia.

Description

[0001] The present invention relates to the use of specific organic compounds as biostimulants in mycelium production and mushroom cultivation. More particularly, the present invention relates to the use of specific organic compounds encapsulated in a specific manner to stimulate the development and growth of mycelium and increase the cultivation yield of edible mushrooms.

[0002] Edible mushrooms have long been used by humans, particularly for their food qualities or medicinal properties. There are many varieties of mushrooms, but the most cultivated worldwide are button mushrooms (Agaricus bisporus), oyster mushrooms (Pleurotus ostreatus, Pleurotus eryngii, etc.), and shiitake mushrooms (Lentinula edodes). Button mushrooms alone account for more than 40% of the world market, oyster mushrooms for 25%, and shiitake for 15%. Globally, the main players in the market are China, the United States, and Europe, each accounting for 50% of production.

[0003] On an industrial scale, the method for producing mushrooms can be divided into two steps: the first step involves obtaining an inoculum of mycelium, and the second step involves cultivating the mycelium obtained during the first step, strictly speaking, mushroom cultivation. These two steps can be carried out independently by different actors (the first by the mycelium producer, the second by the mushroom grower) or by the same actor (the mushroom grower).

[0004] Obtaining the inoculum mycelium begins by inoculating a sterile culture medium with the spores or a portion of the inoculum. The culture medium can be agar, such as potato dextrose agar (PDA, Potato Dextrose Agar); liquid culture medium, such as potato dextrose broth (PDB, Potato Dextrose Broth); or any other nutrient solution with or without a gelling agent. The development and growth of the mycelium on its culture medium is variable, but generally lasts 7 to 28 days, depending on the strain of mushroom. Once the mycelium has completely colonized the culture medium, a portion of the culture medium is taken to inoculate a colonization substrate. Suitable colonization substrates can be synthetic or are generally composed of cereals, typically rye, millet, sorghum, wheat, barley, rice or oats, that have been sterilized in advance and packaged in cans or micro-perforated culture bags.

[0005] The mycelial inoculum thus obtained is then used for the cultivation and production of mushrooms. Thus, the inoculum is used to seed a fructification substrate.

[0006] In the case of button mushrooms, the growing substrate is a compost typically made of straw and animal manure, which is heavily watered to ensure that it matures within two to three weeks. The growing substrate is then pasteurized at a temperature reduced from 60°C to 40°C for several days. After pasteurization, the growing substrate is sown, for example, using a sowing device, by mixing the inoculum contained in its colonization substrate with the growing substrate. The result is an incubation period, during which the inoculated growing substrate is placed in a closed chamber where the temperature, humidity, and oxygen are controlled for two weeks. The temperature is maintained at 22°C to 25°C. The subsequent step is covering with soil, which includes covering the growing substrate with a suitable layer of soil. For example, the covering soil is a mixture of ground and sterilized rough stone and horticultural peat. After the controlled temperature drops, the first mushroom heads emerge from the growing substrate, and harvesting can then begin. Button mushroom harvesting follows several growing cycles, or harvesting periods, alternating with growth dormancy periods, known as rounds. The first two rounds are by far the most productive, with round 1 accounting for 50% of the total harvest and round 2 for 35%. The harvest cycle restarts approximately weekly. Harvesting can continue up to round 3, but yields decline rapidly. Generally, mushroom growers try to produce only two rounds for both productivity and health reasons, as round 3 is often the gateway for disease onset and long-term establishment.

[0007] Consequently, these methods for obtaining inoculum and producing mushrooms can take several weeks, particularly since it takes time for various mycelial strains to colonize their substrate. Furthermore, the development and growth of the mycelium can be disturbed by various factors, such as variations in culture parameters and its ability to colonize. Consequently, the growth rate and quality of the mycelium can become limiting factors, extending the inoculum production time for producers and negatively impacting the mushroom harvest yield for mushroom growers. Consequently, the entire mushroom production chain can be directly affected by the quality of the mycelium used.

[0008] Biostimulants are increasingly being used in agricultural production methods. Biostimulants are substances that stimulate the metabolism of plants or mushrooms and their natural processes for nutrient absorption. More precisely, Regulation (EU) 2019 / 1009 of the European Parliament and of the Council of 5 June 2019, which entered into force on July 22, 2022, defines a biostimulant as "a product that stimulates plant nutrient processes independently of the nutrients it contains, with the aim of improving one or more of the following characteristics of the plant or its rhizosphere:

[0009] a) Utilization efficiency of nutrients

[0010] b) Tolerance to abiotic stress

[0011] c)Quality characteristics

[0012] d) limiting the availability of nutrients in the soil or rhizosphere"

[0013] The object of the present invention is therefore to identify and propose biostimulants that are effective in methods for producing mycelial inoculum and for cultivating mushrooms from mycelial inoculum. In particular, the biostimulants according to the invention make it possible to shorten the time required for producing mycelial inoculum and to increase the yield of cultivated mushrooms, or at least to avoid prolonging this time and thus losing this yield. Finally, the biostimulants according to the invention can be used in conventional inoculum production and mushroom production methods.

[0014] For this purpose, the applicant company has conducted studies and discovered that certain organic compounds, chosen from one of β-pinene, δ-3-carene or β-phellandrene, or a mixture thereof, have an unexpected biostimulatory capacity on the development and growth of mycelium and mushrooms, in particular when they are added to their culture medium or their substrate, and can therefore be used in processes and methods for obtaining mushroom inoculum or culture.

[0015] The use of natural biostimulants is a real innovation for mushroom production. The use of products with biostimulant properties has a significant impact on mycelium and mushroom production. The main limitation for inoculum producers is their ability to quickly produce, store, and distribute their mycelium to mushroom producers. The use of the biostimulants according to the present invention makes it possible to reduce the time between the start of production and distribution, thereby increasing producers' productivity. Similarly, during the mushroom production phase, the use of pre-biostimulated mycelium and the addition of biostimulants to the culture matrix increase yields and boost mushroom producer production without negatively impacting the culture. Thus, biostimulants can be added during the inoculation of the substrate bags or during the various stages of compost incubation, more particularly, when supplementing (nutritional) supplements.

[0016] Thus, during the tests carried out on composts, by stimulating the mycelium with the organic compounds selected according to the invention, it was possible to observe an increase in the yield of the culture, an increase in the average mass of each mushroom and, in general, an improvement in the quality of the harvested mushrooms.

[0017] The organic compounds selected in the context of the present invention have the property of having a strong biostimulatory effect within a concentration range that is wide enough for the producer.

[0018] However, when they are applied to mycelium growth bags in their natural form, overstimulation, or local overstimulation phenomena, or temporary inhibition zones at the contact points can be observed. In this form, the active substance is released immediately and completely, and the mycelium develops significantly but often unevenly, which is something mycelium producers would like to avoid and which can cause problems with uniformity in mushroom cultivation.

[0019] To solve this problem, the Applicant Company selected and developed a specific encapsulation technology adapted to the physical and chemical characteristics of the selected organic compounds. This encapsulation technology makes it possible to preserve all the biostimulant properties of the organic compounds while releasing their active substances gradually over time and under controlled conditions suitable for both mycelial production and mushroom cultivation.

[0020] Therefore, the present invention also relates to the use of at least one terpene in encapsulated form, selected from one of β-pinene, δ-3-carene or β-phellandrene, or a mixture thereof, for stimulating the development and growth of mycelium or mushrooms.

[0021] Treatment by encapsulating the selected organic compound allows for increased addition of biostimulant substances without any adverse effects on the mycelium and its development. Compared to mycelium not treated with the encapsulated organic compound according to the invention, the biostimulated mycelium exhibits a higher growth rate, more uniform development, and more promising behavior. Addition in encapsulated form also avoids localized over-stimulation, which can lead to uneven colonization of the colonization matrix bags. A single application is sufficient for effective, uniform, and complete inoculation of the colonization matrix. Thus, the use of encapsulated biostimulants according to the invention allows for a reduction in the time between production start and distribution, thereby increasing the productivity of mycelium producers. The main limitation for inoculum producers is their ability to quickly produce, store, and distribute high-quality mycelium to compost providers and mushroom producers.

[0022] Thus, the addition of encapsulated selected organic compounds makes it possible to increase the yield of mushroom cultures, particularly when compost (a solid substrate) is inoculated directly with biostimulated mycelium. The increase in yield is even more pronounced when supplements (nutrients) are added to the compost. This is because the application of the encapsulated biostimulant allows the mycelium to permanently increase its efficiency in utilizing these nutrients.

[0023] Furthermore, when cultivating mushrooms, the addition of the encapsulated organic compound according to the invention makes it possible to increase the yield during the first round and improve the quality and average quality of the harvested mushrooms. Thus, at each stage, the impact on the mushroom cultivation industry is accompanied by economic and practical benefits for the producer:

[0024] -Energy savings associated with earlier sales and reduced storage time for mycelium producers.

[0025] - Reduced labor time for mushroom growers during harvesting,

[0026] - The application is easily adaptable to all production methods and can be combined with supplements (nutritional supplements) and therapeutic products.

[0027] According to the present invention, the selected organic compound is adsorbed or absorbed on the support, or encapsulated in the support, and coated with fat to achieve the coating effect.

[0028] Therefore, the present invention also relates to microcapsules comprising:

[0029] - a core comprising at least one organic compound and a solid support, wherein the organic compound is selected from β-pinene, δ-3-carene, or β-phellandrene, or a combination thereof, the organic compound being adsorbed or absorbed on or encapsulated in the solid support, the solid support being an inert solid material, a material composed of silica or silicon dioxide, and

[0030] - at least one coating layer formed of vegetable fat, said layer coating said core.

[0031] The selected organic compounds are interchangeable, for example in case of shortage of one of them.

[0032] Selected organic compounds may also be combined to obtain synergistic effects and enhanced resilience in certain cases.

[0033] The organic compounds are also chosen so as to act on the growth of mycelium and mushrooms under conditions that are less favourable than normal (eg a decrease in incubation temperature by 1 or 2° C. or aging effects).

[0034] It is therefore the combination of the organic compound chosen as a precaution and the form in which it is distributed that makes it possible to achieve the objects of the present invention.

[0035] All combinations of encapsulated organic compounds are possible, for example β-pinene with δ-3-carene, or β-pinene with β-phellandrene, or δ-3-carene with β-phellandrene, or β-pinene and δ-3-carene and β-phellandrene.

[0036] However, according to a preferred embodiment, the encapsulated organic compound is β-pinene.

[0037] The organic compound used in the context of the present invention is advantageously and preferably in the form of a liquid extract comprising at least 90%, at least 92%, preferably at least 95% of said organic compound. The organic compound selected according to the present invention is a terpene, typically an extract of an essential oil. Such extracts are commercially available.

[0038] According to one embodiment of the invention, the mycelium or mushroom is chosen from basidiomycetes, in particular from button mushrooms, oyster mushrooms and shiitake mushrooms, more particularly from Agaricus bisporus, Pleurotus ostreatus, Pleurotus eryngii and Shiitake mushrooms.

[0039] When the mycelium or mushroom is Agaricus bisporus, the organic compound used is preferably β-pinene, δ-3-carene or β-phellandrene.

[0040] When the mycelium or mushroom is Pleurotus ostreatus, the organic compound used is preferably β-pinene, δ-3-carene or β-phellandrene.

[0041] When the mycelium or mushroom is Pleurotus eryngii, the organic compound used is preferably β-pinene or β-phellandrene.

[0042] When the mycelium or mushroom is shiitake mushroom, the organic compound used is preferably β-pinene.

[0043] According to one embodiment of the present invention, the solid support is a spherical particle.

[0044] Preferably, the solid support is an inert solid material, a material composed of silica or a material composed of silicon dioxide.

[0045] According to one embodiment of the invention, the vegetable fat consists of fatty acids or lipids, preferably one or more vegetable oils.

[0046] The microcapsules according to the present invention can be prepared according to methods known to those skilled in the art.

[0047] For example, microcapsules are prepared as follows:

[0048] - loading the solid support into the homogenizer,

[0049] - adding the selected organic compound according to the present invention to a solid support at low temperature,

[0050] - Add vegetable fat,

[0051] - sieving at 1 mm to obtain particle size less than 1 mm,

[0052] -Package.

[0053] The invention also relates to a composition comprising at least one microcapsule as described above.

[0054] Preferably, the composition is in powder form. More preferably, the composition comprises only the microcapsules according to the present invention.

[0055] According to one embodiment, the composition comprises at least 10%, 20%, 30%, 40% or 50% by weight of at least one said organic compound relative to the total weight of the composition.

[0056] According to a preferred embodiment, the composition is composed of 50% of at least one organic compound (chosen from one of β-pinene, δ-3-carene or β-phellandrene, or a mixture thereof), 35% of a support (preferably a silica or silicon dioxide support) and 15% of a fat coating.

[0057] The present invention also relates to the use of at least one microcapsule as described above or a composition containing said microcapsule for stimulating the development and growth of mycelium or mushrooms.

[0058] The organic compounds selected and encapsulated in the context of the present invention can be used simply and effectively in conventional inoculum production and mushroom production methods without making these methods more complicated.

[0059] Therefore, the present invention also relates to a method for obtaining a mycelial inoculum, characterized in that said method comprises the following steps:

[0060] Therefore, the present invention also relates to a method for obtaining a mycelial inoculum, characterized in that said method comprises the following steps:

[0061] - Preparation of colonization matrix,

[0062] - adding to the colonization matrix at least one microcapsule as described above or a composition comprising said microcapsules,

[0063] - inoculation of the colonization substrate with mycelium,

[0064] - obtaining mycelial inoculum,

[0065] - Packaging the inoculum.

[0066] Preferably, in the method according to the invention, the encapsulated organic compound is added to the colonization substrate or compost so that the concentration of the organic compound is 20 to 400 μL / L colonization substrate. For example, this corresponds to about 0.5 to 10 g microcapsules per 15 liter growth bag.

[0067] The method according to the invention makes it possible to shorten by several days the time required to obtain an inoculum or mycelium that can be marketed by the producer, compared to conventional methods. Growth can be stimulated and increased by more than 20%, or even more than 40%, compared to growth obtained using conventional methods (i.e., without the use of the selected and encapsulated organic compounds of the present invention). The stimulation of mycelial growth can be observed by its behavior and the uniformity of its development on the surface of the growth bag.

[0068] According to one embodiment of the invention, in the method, the mycelium or mushroom is selected from the class Basidiomycetes, in particular from button mushrooms, oyster mushrooms and shiitake mushrooms, more particularly from Agaricus bisporus, Pleurotus ostreatus, Pleurotus eryngii and Shiitake mushrooms.

[0069] According to a preferred embodiment, in the method, the organic compound is β-pinene, in particular when the mycelium or mushroom is Agaricus bisporus.

[0070] Therefore, the present invention also relates to a method for cultivating mushrooms using a mycelial inoculum, characterized in that the method comprises the following steps:

[0071] - Preparation of a solid matrix,

[0072] - optionally incorporating in a solid matrix at least one microcapsule as described above or a composition comprising said microcapsules,

[0073] - sowing the hardening substrate with the mycelial inoculum obtained according to the invention and the method described above,

[0074] - Optionally incorporating into the fruiting matrix a supplement or a mixture of nutrients specific to mushroom cultivation.

[0075] - incubation, wherein at least one microcapsule according to one of the preceding claims or a composition comprising said microcapsules is incorporated, optionally in a solid matrix,

[0076] -solid,

[0077] -Harvest.

[0078] According to one embodiment of the present invention, the amount of microcapsules added to the compacting substrate is 50 to 500 g per ton of compost or compacting substrate. In this embodiment, the concentration of the organic compound is 28 to 280 mL per 1000 kg of compacting substrate.

[0079] According to one embodiment of the invention, in the method, the mycelium or mushroom is selected from the class Basidiomycetes, in particular from button mushrooms, oyster mushrooms and shiitake mushrooms, more particularly from Agaricus bisporus, Pleurotus ostreatus, Pleurotus eryngii and Shiitake mushrooms.

[0080] According to a preferred embodiment, in the method, the organic compound is β-pinene, in particular when the mycelium or mushroom is Agaricus bisporus.

[0081] The above-mentioned features and other features of the present invention will become more apparent from a reading of the following description of exemplary embodiments.

[0082] Example 1: Evaluation of the Effect of the Composition According to the Present Invention on the Harvest Yield of Agaricus bisporus 1.1 Preparation of the Composition According to the Present Invention

[0083] The composition is formulated using the following ingredients:

[0084] Organic compound: β-pinene (CAS No 127-91-3), which is usually isolated from turpentine by fractional distillation (β-pinene content > 95%)

[0085] Solid support: silica.

[0086] Vegetable fats: vegetable oils.

[0087] [Table 1]

[0088] Element Amount: (% of total weight) β-pinene 50 Silicon dioxide 35 vegetable oil 15

[0089] The composition is obtained as follows:

[0090] - loading silica into the mixer,

[0091] - Add β-pinene by spraying at a temperature of 20°C to 25°C,

[0092] - Mix for 7 minutes

[0093] -Add vegetable oil

[0094] -Mix for 1 minute

[0095] - sieving at 1 mm to obtain particle size less than 1 mm,

[0096] -Package.

[0097] The composition obtained is in the form of a powder composed of microcapsules with a size of less than 1 mm.

[0098] This composition will hereinafter be referred to as the "encapsulated composition".

[0099] 1.2 Experimental protocol for application in mycelium growth bags

[0100] Application of the composition during the industrial production of mycelium

[0101] - Non-encapsulated β-pinene (CAS No 127-91-3) was obtained.

[0102] - At the time of mycelial inoculation, 300 μL and 600 μL of the non-encapsulated β-pinene were placed per 15 L bag of colonization substrate (ie, 20 and 40 μL / L substrate).

[0103] - Preparation of the encapsulated composition (as described in point 1.1).

[0104] - At the time of mycelial inoculation, 2, 3 and 4 g of the encapsulated composition were placed per 15 liter bag of colonization substrate (ie approximately 80, 120 and 160 μL / L substrate).

[0105] - The mycelia were allowed to colonize the substrate bags for 21 days according to the method commonly used by mycelia producers.

[0106] Preparation and inoculation of parcelles before culture

[0107] - Use 72g of untreated mycelium to compost a 9kg bag (representing 0.1m 2 mushroom culture) for inoculation.

[0108] - 9 kg compost bales were inoculated with 72 g of mycelium treated with non-encapsulated β-pinene.

[0109] - A 9 kg compost bale was inoculated with 72 g of mycelium treated with the encapsulated composition.

[0110] - 72 g of supplement (nutritional supplement) was added to 9 kg compost bags inoculated with untreated mycelium and treated with non-encapsulated β-pinene and encapsulated compositions (see method shown in the "Supplements" column in Table 2).

[0111] Agaricus bisporus production in a cultivation room

[0112] - Incubate the bag for 13 days

[0113] - 3000 mL of cover soil (90% black peat and 10% calcium carbonate) was placed on each bag at a thickness of more than 3 cm.

[0114] - The bags were subjected to post-incubation for 10 days.

[0115] - Start inducing the first round of fruiting.

[0116] - Harvest, weigh and measure the yield over 3 rounds.

[0117] 1.3 Experimental plan for compost inoculation

[0118] Application of the composition during the mushroom production stage

[0119] - Use 72g of untreated mycelium to compost a 9kg bag (representing 0.1m 2 mushroom culture) for inoculation.

[0120] - Place 3 g of the encapsulated composition per 9 kg compost bag (ie 30 g / 90 kg).

[0121] Agaricus bisporus production in a cultivation room

[0122] - The packs were incubated for 13 days.

[0123] - 3000 mL of cover soil (90% black peat and 10% calcium carbonate) was placed on each bag at a thickness of more than 3 cm.

[0124] - The packs were post-incubated for 10 days.

[0125] - Start inducing the first round of fruiting.

[0126] - Harvest, weigh and measure the yield over 3 rounds.

[0127] Each of the above methods was performed on 6 identical experimental packages, thus corresponding to 6 replicates. The mycelial strain used for all experimental tests was a commercial variety of Agaricus bisporus of the Heirloom series produced by the American company Amycel.

[0128] 1.4 Results

[0129] The table below shows the yields measured for the various experimental methods described above, expressed in kilograms of healthy mushrooms per square meter of culture. All yields are also expressed as a percentage relative to the control method (untreated and unsupplemented compost inoculated with untreated mycelium).

[0130] [Table 2]

[0131]

[0132]

[0133] According to the results of the experimental tests of Example 1, a significant increase in yield was observed when β-pinene was applied in its non-encapsulated and encapsulated forms to mycelium growth bags, as well as when applied in an encapsulated form mixed with compost.

[0134] Increasing the dosage applied in both non-encapsulated and encapsulated forms resulted in increased mushroom yield.

[0135] Application in encapsulated form allows for gradual release of the active ingredient and results in a greater increase in yield that is more sustained over time compared to direct application in non-encapsulated form.

[0136] The encapsulated active ingredients enable the early treated mycelium to increase its efficiency in utilizing nutrients. This ability is even more pronounced when supplements (nutrient supplements) are added to the compost at the time of inoculation.

[0137] In this configuration, the yield obtained in only the first two rounds (24.26 kg) was greater than the yield of three rounds of the control process (23.54 kg) in which no supplements were added to the compost and which was carried out with untreated mycelium.

[0138] The latter point is important since mushroom growers prefer to avoid continuing their cultures to a third round in order to shorten the production cycle, to ensure a more uniform production and of high quality and to avoid the emergence of diseases that may be difficult to remove.

[0139] In summary, the encapsulation of the selected organic compounds according to the invention makes it possible to increase the dosage of the applied active ingredient without the risk of over-stimulation of the biomass and uneven development of the mycelium in the growing bag. This possible increase in the applied dosage leads to an increase in mushroom yield.

[0140] Example 2: Evaluation of the Effect of the Composition According to the Invention on the Harvest Yield of Agaricus bisporus 2.1 Experimental Protocol for Applying the Composition for Compost Inoculation

[0141] Application of the composition during the mushroom production stage

[0142] - Preparation of the encapsulated composition (as described in point 1.1).

[0143] - Use 72g of untreated mycelium to compost a 9kg bag (representing 0.1m 2 mushroom culture) for inoculation.

[0144] - Place 100g to 250g of the encapsulated composition per ton of compost, ie 0.9g to 2.25g per 9kg compost bag.

[0145] - Add 72 g of supplements (nutritional supplements) per 9 kg compost bag (see method shown in the "Supplements" column in Table 3).

[0146] Agaricus bisporus production in a cultivation room

[0147] - The packs were incubated for 13 days.

[0148] - 3000 mL of cover soil (90% black peat and 10% calcium carbonate) was placed on each bag at a thickness of more than 3 cm.

[0149] - The packs were post-incubated for 10 days.

[0150] - Start inducing the first round of fruiting.

[0151] - Harvest, weigh and measure the yield over 3 rounds.

[0152] Each of the above methods was performed on 6 identical experimental packages, thus corresponding to 6 replicates. The mycelial strain used for all experimental tests was a commercial variety of Agaricus bisporus of the Heirloom series produced by the American company Amycel.

[0153] 2.2 Results

[0154] The table below shows the yields measured for the various experimental methods described above, expressed in kilograms of healthy mushrooms per square meter of culture. All yields are also expressed as a percentage relative to the control method (untreated and unsupplemented compost inoculated with untreated mycelium).

[0155] [Table 3]

[0156]

[0157]

[0158] According to the results of the experimental tests of Example 2, when β-pinene is applied in its encapsulated form mixed with compost, including when supplements are present, a significant increase in yield can be observed.

[0159] Increasing the applied dose resulted in a clear increase in mushroom yield, which was better in only two rounds (19.51 to 20.77 kg) than in three rounds of the control method (19.22 kg).

[0160] Example 3: Evaluation of the Effect of the Composition According to the Invention on the Harvest Yield of Agaricus bisporus 3.1 Experimental Protocol for Applying the Composition for Compost Inoculation

[0161] Application of the composition during the mushroom production stage

[0162] - Preparation of the encapsulated composition (as described in point 1.1).

[0163] - Use 72g of untreated mycelium to compost a 9kg bag (representing 0.1m 2 mushroom culture) for inoculation.

[0164] - Place 200g to 350g of the encapsulated composition per ton of compost, ie 1.8g to 3.15g per 9kg compost bag.

[0165] - Add 72 g of supplements (nutritional supplements) per 9 kg compost bag (see method shown in the "Supplements" column in Table 4).

[0166] Agaricus bisporus production in a cultivation room

[0167] - The packs were incubated for 13 days.

[0168] - 3000 mL of cover soil (90% black peat and 10% calcium carbonate) was placed on each bag at a thickness of more than 3 cm.

[0169] - The packs were post-incubated for 10 days.

[0170] - Start inducing the first round of fruiting.

[0171] - Harvest, weigh and measure the yield over 3 rounds.

[0172] Each of the methods described previously was performed on 9 identical experimental packages, thus corresponding to 9 replicates. The mycelial strain used for all experimental tests was a commercial variety of Agaricus bisporus of the Heirloom series produced by the American company Amycel.

[0173] 3.2 Results

[0174] The table below shows the yields measured for the various experimental methods described above, expressed in kilograms of healthy mushrooms per square meter of culture. All yields are also expressed as a percentage relative to the control method (untreated and supplemented compost inoculated with untreated mycelium).

[0175] [Table 4]

[0176]

[0177] According to the results of the experimental tests of Example 3, when β-pinene is applied in its encapsulated form mixed with compost, including when supplements are present, a significant increase in yield is observed.

[0178] Example 4: Evaluation of the Effect of the Composition According to the Invention on the Harvest Yield of Pleurotus ostreatus 4.1 Experimental Protocol for Applying the Composition for Matrix Inoculation

[0179] Application of the composition during the mushroom production stage

[0180] - Preparation of the encapsulated composition (as described in point 1.1).

[0181] - 15 kg of solid substrate blocks were inoculated with 375 g of untreated mycelium, ie a release or inoculation rate of 2.5%.

[0182] - 100 g of the encapsulated composition is placed per ton of substrate, ie 1.5 g / 15 kg of substrate mass.

[0183] Production of Pleurotus ostreatus in a culture chamber

[0184] - The blocks were incubated for 2 weeks.

[0185] - Post-incubation in a hardiness chamber for 2 weeks.

[0186] - Start inducing the first round of fruiting.

[0187] - Harvest, weigh and measure yield until the end of harvest.

[0188] Each of the methods described previously was performed on 6 identical experimental blocks, thus corresponding to 6 replications. The mycelial strain used for all experimental tests was a commercial variety of Pleurotus ostreatus of the 3015 series produced by the American company Sylvan.

[0189] 4.2 Results

[0190] The table below shows the yields measured for the various experimental methods described above, expressed in kilograms of healthy mushrooms per ton of fruiting substrate. All yields are also expressed as a percentage relative to the control method (untreated substrate inoculated with untreated mycelium).

[0191] [Table 5]

[0192]

[0193] According to the results of the experimental tests of Example 4, a significant increase in yield was observed when β-pinene was applied in its encapsulated form mixed with a fruiting matrix. The composition according to the invention has a positive effect on yield from the first week of harvest, which is particularly beneficial for producers.

[0194] Example 5: Evaluation of the Effect of the Composition According to the Invention on the Harvest Yield of Pleurotus eryngii 5.1 Experimental Protocol for Applying the Composition for Matrix Inoculation

[0195] Application of the composition during the mushroom production stage

[0196] - Preparation of the encapsulated composition (as described in point 1.1).

[0197] - 15 kg of solid substrate blocks were inoculated with 900 g of untreated mycelium, ie a release or inoculation rate of 6%.

[0198] - 100 g to 300 g of the encapsulated composition are placed per ton of substrate, ie 1.5 g to 4.5 g per 15 kg of substrate mass.

[0199] Production of Pleurotus eryngii in a culture chamber

[0200] - The blocks were incubated for 3 weeks.

[0201] - Post-incubation in a firming chamber for 1 week.

[0202] - Start inducing the first round of fruiting.

[0203] - Harvest, weigh and measure yield until the end of harvest.

[0204] Each of the methods described previously was performed on 6 identical experimental blocks, thus corresponding to 6 replications. The mycelial line used for all experimental tests was a commercial variety of Pleurotus eryngii of the 3066 series produced by the American company Sylvan.

[0205] 5.2 Results

[0206] The table below shows the yields measured for the various experimental methods described above, expressed in kilograms of healthy mushrooms per ton of fruiting substrate. All yields are also expressed as a percentage relative to the control method (untreated substrate inoculated with untreated mycelium).

[0207] [Table 6]

[0208]

[0209] According to the results of the experimental tests of Example 5, a significant increase in yield was observed when β-pinene was applied in its encapsulated form mixed with a fruiting matrix. The composition according to the invention has a positive effect on yield from the first week of harvest, which is particularly beneficial for producers.

[0210] Example 6: Evaluation of the Effect of the Composition According to the Invention on the Harvest Yield of Shiitake Mushrooms 6.1 Experimental Protocol for Applying the Composition for Substrate Inoculation

[0211] Application of the composition during the mushroom production stage

[0212] - Preparation of the encapsulated composition (as described in point 1.1).

[0213] - 15 kg of solid substrate blocks were inoculated with 900 g of untreated mycelium, ie a release or inoculation rate of 6%.

[0214] - 200 g to 300 g of the encapsulated composition are placed per ton of substrate, ie 3 g to 4.5 g per 15 kg of substrate mass.

[0215] Production of Shiitake Mushrooms in a Cultivation Room

[0216] - The blocks were incubated for 5 weeks.

[0217] - Post-incubation in a hardiness chamber for 2 weeks.

[0218] - Start inducing the first round of fruiting.

[0219] - Harvest, weigh and measure yield until the end of harvest.

[0220] Each of the methods described previously was performed on 9 identical experimental blocks, thus corresponding to 9 replicates. The mycelial strain used for all experimental tests was a commercial variety of Shiitake mushrooms of the 4325 series produced by the American company Sylvan.

[0221] 6.2 Results

[0222] The table below shows the yields measured for the various experimental methods described above, expressed in kilograms of healthy mushrooms per ton of fruiting substrate. All yields are also expressed as a percentage relative to the control method (untreated substrate inoculated with untreated mycelium).

[0223] [Table 7]

[0224]

[0225] According to the results of the experimental tests of Example 6, a significant increase in yield was observed when β-pinene was applied in its encapsulated form mixed with a fruiting matrix. The composition according to the invention has a positive effect on yield from the first week of harvest, which is particularly beneficial for producers.

Claims

1. Use of at least one organic compound in encapsulated form, selected from one of β-pinene, δ-3-carene, or β-phellandrene, or a mixture thereof, for stimulating the development and growth of mycelium or mushrooms.

2. Use according to claim 1, wherein the organic compound is in the form of a liquid extract comprising at least 90%, at least 92%, preferably at least 95% by weight of said organic compound relative to the weight of said liquid extract.

3. The use according to claim 1 or 2, wherein the mycelium or mushroom is selected from the class Basidiomycetes, in particular from button mushrooms, oyster mushrooms and shiitake mushrooms, more particularly from Agaricus bisporus, Pleurotus ostreatus, Pleurotus eryngii and Lentinula edodes.

4. A microcapsule comprising: a core comprising at least one organic compound and a solid support, the organic compound being selected from one of β-pinene, δ-3-carene, or β-phellandrene, or a combination thereof, the organic compound being adsorbed or absorbed on or encapsulated in the solid support, the solid support being an inert solid material, a material consisting of silica or silicon dioxide, and - at least one coating layer formed of vegetable fat, said layer coating said core.

5. The microcapsule according to claim 4, characterized in that The organic compound is in the form of a liquid extract comprising at least 90%, at least 92%, preferably at least 95% by weight of the organic compound relative to the weight of the liquid extract.

6. The microcapsule according to claim 4 or 5, characterized in that The organic compound is β-pinene.

7. Microcapsules according to one of claims 4 to 6, characterized in that The solid support is a spherical particle.

8. Microcapsules according to one of claims 4 to 7, characterized in that The solid support is an inert solid material, such as silica or silicon dioxide.

9. Microcapsules according to one of claims 4 to 8, characterized in that The vegetable fat consists 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. The composition according to claim 10, characterized in that The composition comprises at least 10%, 20%, 30%, 40% or 50% of at least one of the organic compounds.

12. Use of at least one microcapsule according to one of claims 4 to 9 or a composition comprising said microcapsule for stimulating the development and growth of mycelium or mushrooms.

13. The use according to claim 12, wherein the mycelium or mushroom is selected from the class of Basidiomycetes, in particular from button mushrooms, oyster mushrooms and shiitake mushrooms, more particularly from Agaricus bisporus, Pleurotus ostreatus, Pleurotus eryngii and Shiitake mushrooms.

14. Use according to one of claims 12 or 13, characterized in that The organic compound is beta-pinene, particularly when the mycelium or mushroom is Agaricus bisporus.

15. A method for obtaining a mycelial inoculum, characterized in that The method comprises the following steps: - Preparation of colonization matrix, - adding to the colonization matrix at least one microcapsule according to one of claims 4 to 9 or a composition comprising said microcapsules, - inoculating the colonization matrix with mycelium, - obtaining mycelial inoculum, - Packaging the inoculum.

16. The method according to claim 15, characterized in that The microcapsules or a composition comprising the microcapsules are added to the colonization matrix such that the concentration of the organic compound is 20 to 400 μL / L of colonization matrix.

17. A method for cultivating mushrooms using mycelial inoculum, characterized in that The method comprises the following steps: - Preparation of a solid matrix, - optionally, incorporating into said solid matrix at least one microcapsule according to one of claims 4 to 9 or a composition comprising said microcapsules, - sowing the fruiting substrate with a mycelial inoculum obtained by the method according to claim 15 or 16, - optionally, incorporating into said firming matrix a supplement or a mixture of nutrients specific to mushroom cultivation, - incubation, wherein optionally in said solid matrix at least one microcapsule according to one of claims 4 to 9 or a composition comprising said microcapsules is incorporated, -solid, -Harvest.

18. The method according to claim 17, characterized in that The amount of microcapsules added to the consolidated matrix is ​​between 50 and 500 g per ton of matrix, so that the concentration of the organic compound is between 28 and 280 mL per 1000 kg of consolidated matrix.

19. Method according to one of claims 15 to 18, characterized in that The mycelium or mushroom is selected from the class Basidiomycetes, in particular from button mushrooms, oyster mushrooms and shiitake mushrooms, more particularly from Agaricus bisporus, Pleurotus ostreatus, Pleurotus eryngii and Shiitake mushrooms.

20. Method according to one of claims 15 to 19, characterized in that The organic compound is beta-pinene, particularly when the mycelium or mushroom is Agaricus bisporus.

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