A method for cultivating Poria cocos with bagged materials in a green way

Abstract

The present invention discloses a method for cultivating Poria cocos by using bagged materials, which uses bagged materials with a specific formula and cold-extracted pine wood extract obtained by low-temperature extraction to induce the growth of Poria cocos sclerotia, solving the problems that in the existing cultivation of Poria cocos by using bagged materials, the growth of Poria cocos fungi in the fungus bags filled with conventional bagged materials is slow, the mycelium distribution is uneven, and the success rate of inducing the growth of Poria cocos sclerotia by using fresh pine branches is low; and uses safe and environmentally friendly essential oils and pine needle extracts to repel pests such as termites in the cultivation of Poria cocos, solving the problems of pesticide residues and environmental pollution caused by spraying pesticides to trap pests such as termites in the existing cultivation of Poria cocos.

Description

Technical Field

[0001] The present invention relates to the technical field of fungal cultivation, and specifically relates to a method for cultivating Poria cocos in bags with a green approach. Background Art

[0002] Poria cocos (Schw.) Wolf is a fungus of the family Polyporaceae and belongs to commonly used edible and medicinal fungi, with a relatively large market demand. Due to the sharp decline in wild Poria cocos resources, artificial cultivation of Poria cocos has become the main way to obtain it. Cultivating Poria cocos in an artificially managed manner has the characteristics of a short cultivation cycle and high economic benefits. The existing methods for artificially cultivating Poria cocos mainly include bag cultivation and log cultivation; in bag cultivation, pine sawdust, rice bran, etc. are generally used as cultivation substrates, and after sterilization, Poria cocos strains are inoculated, such as a cultivation method of Poria cocos recorded in Chinese Patent Application CN114467623A; in log cultivation, pine logs, pine stumps, or tree stumps are generally used as cultivation substrates. Due to low resource utilization rate and the need for a large amount of pine forest felling, which seriously damages the environment, its popularization and application have gradually decreased.

[0003] The following problems mostly exist in the conventional artificial cultivation of Poria cocos:

[0004] In the existing artificial cultivation of Poria cocos, the success rate of inducing the growth of Poria cocos sclerotia using Poria cocos mycelial blocks, fresh pine branches, etc. is low. In the artificial cultivation of Poria cocos in the wild environment, such as underforest cultivation of Poria cocos, termite damage is serious. The main damages are boring the fungus material, affecting the growth of hyphae and sclerotia formation; in order to reduce termite damage and trap termites, cultivators often spray pesticides, generally spraying pesticides on the substances that attract termites to play a trapping role. More pesticides are sprayed, such as organochlorine insecticides, organophosphorus insecticides, and / or pyrethroid insecticidal drugs and other organic insecticides, with dichlorvos and phoxim being used the most. Some technical regulations for artificial cultivation of Poria cocos, such as the "Technical Regulations for Cultivating Poria cocos in Bags in Jingzhou", state that dichlorvos and phoxim need to be sprayed to trap termites; there are also uses of insecticides such as arsenic acid, bicholin, chlorpyrifos, and / or carbofuran to trap termites; problems such as pesticide residues and environmental pollution exist. Summary of the Invention

[0005] The present invention provides a method for cultivating Poria cocos in bags with a green approach, which uses a bag material with a specific formula and a cold extract of pine wood obtained by low-temperature extraction to induce the growth of Poria cocos sclerotia, solving the problems that in the existing bag cultivation of Poria cocos, the growth of Poria cocos fungi in the fungus bags filled with conventional bag materials is slow, the distribution of hyphae is uneven, and the success rate of inducing the growth of Poria cocos sclerotia with fresh pine branches is low; it uses safe and environmentally friendly essential oils and pine needle extracts to repel pests such as termites in Poria cocos cultivation, solving the problems of pesticide residues and environmental pollution caused by spraying pesticides to trap pests such as termites in the existing Poria cocos cultivation.

[0006] A method for cultivating Poria cocos in bags with a green approach includes the following steps:

[0007] (1) The bagged material formula includes, by weight parts: 70 - 80 parts of sawdust, 15 - 20 parts of bran, 1 - 2 parts of gypsum, 1 - 2 parts of brown sugar, 6 - 8 parts of corn, 1 - 3 parts of oyster shell, 3 - 5 parts of the first pine branch, and 0.1 - 1 part of pine resin; after mixing evenly, bag it, sterilize, cool, inoculate with Poria cocos fungus, and cultivate until the mycelium fills the fungus bag.

[0008] (2) Make a small opening on one side of the fungus bag filled with mycelium, drop the cold - extracted solution of pine wood into the small opening, then put the fungus bag into a light - proof groove or bury it in soil, fine sand or sandy soil, etc., and cover the top of the fungus bag with one or two of soil and fine sand, and wait quietly for the formation of Poria cocos.

[0009] The preparation method of the cold - extracted solution of pine wood includes:

[0010] By weight percentage, take 40% - 50% of the second pine branch, 10% - 20% of pine trunk, and 30% - 40% of pine needles, dry them at a low temperature of 30℃ - 45℃ until the moisture content is 45% - 55%, crush to obtain pine wood raw materials, add an ethanol - n - hexane solution with an ethanol mass percentage concentration of 70% - 85% in an amount of 3 - 5 times the weight of the pine wood raw materials, extract at a low temperature of 30℃ - 35℃, and vacuum - concentrate and desolventize the extract at a temperature of 35℃ and a pressure of 10KPa to obtain the cold - extracted solution of pine wood.

[0011] To achieve better effects, the following optimizations are carried out:

[0012] In step (1), using the bagged material formula of the present invention, generally, cultivating in an environment with a conventional cultivation temperature of 22℃ - 30℃ for 10 - 14 days can make the mycelium fill the fungus bag, improving the cultivation efficiency, and the mycelium in the fungus bag shows a uniform distribution.

[0013] The oyster shell is processed into long strips with a length of 2mm - 5mm; on the one hand, it increases trace elements, and on the other hand, it helps to form a loose channel in the fungus bag, making it easy for the liquid spawn to penetrate into the fungus bag and be evenly distributed in the bagged material.

[0014] The first pine branch is selected as the pine branch cut into a length of 2cm - 4cm and flattened to a thickness of 0.5mm - 1.5mm. The flattened pine branch, first, forms a loose channel in the fungus bag, making it easy for the liquid spawn to penetrate and be evenly distributed in the bagged material, and second, the loose pine branch after flattening promotes the growth of Poria cocos fungus in the fungus bag. Preferably, the first pine branch is selected as the pine branch with a thickness of 0.5cm - 1.5cm, cut into a length of 2cm - 4cm, and flattened to a thickness of 0.5mm - 1.5mm; selecting a thinner pine branch is a renewable resource with a lower cost.

[0015] The sawdust mainly serves as a culture medium, and there is no particular limitation on its type. It is not required to have pine sawdust, and one or more types of existing sawdust can be selected. For example, one or more types of sawdust from coniferous trees such as masson pine, Korean pine, and Chinese fir, or sawdust from broad-leaved trees can be selected.

[0016] The bran is the main by-product during the processing of the seeds of crops such as rice, wheat, millet, and sorghum, and generally mainly includes the husks or hulls of the seeds of crops such as rice, wheat, millet, and sorghum. There is no strict limitation on the type of crops that produce the bran in the present invention. It can be rice husk bran produced during the process of hulling paddy rice to obtain rice, or wheat bran (such as wheat bran) produced during the processing of wheat. Commercially available products can be selected, or the by-product bran obtained directly from the processing of food crops can also be used.

[0017] The pine resin is a natural resin product collected from the trunks of plants of the genus Pinus in the family Pinaceae such as Korean pine, slash pine, and masson pine; commercially available products can be selected, or it can be directly collected. Preferably, the pine resin is ground to more than 200 meshes and uniformly mixed in the bagged material.

[0018] The Poria cocos fungus for inoculation is preferably: after the Poria cocos fungus strain is cultured in a liquid culture medium to obtain Poria cocos mycelial balls, and then the Poria cocos mycelial balls are dispersed into small mycelial balls (such as small mycelial balls of 2 mm - 3 mm) to obtain a liquid Poria cocos fungus strain. Dispersing the Poria cocos mycelial balls into small mycelial balls can adopt existing technologies such as ultrasonic treatment and mechanical shearing. Among them, the liquid culture medium for the fungus strain uses the existing conventional liquid culture medium for fungus strains. Commercially available products can be selected, or it can be prepared according to the existing preparation method. Using the liquid Poria cocos fungus strain dispersed into small mycelial balls is most conducive to inoculation, can quickly penetrate into the middle of the fungus bag, and promote the rapid spread of the mycelium in the bagged material; the growth rate is faster and the success rate is higher than that of the fungus strain cultured in solid state. The conditions for culturing the Poria cocos fungus strain in the liquid culture medium adopt the existing culture conditions for the Poria cocos fungus strain, such as commonly culturing at 22°C - 30°C for 6 days - 12 days.

[0019] In step (1), after the bagged material is mixed evenly, it is generally sprayed with water until the moisture content of the material is 60% - 70% and then bagged.

[0020] Preferably, in step (1), an ant repellent liquid is sprayed on the outer surface of the fungus bag that has grown mycelium, and then step (2) is carried out.

[0021] The ant repellent liquid is composed of a compound essential oil and a pine needle ethanol reflux extract.

[0022] The preparation method of the compound essential oil includes: by weight percentage, 20% of the whole plant of rue, 10% of the whole plant of chrysanthemum, 10% of the whole plant of litsea cubeba, 30% of camphor tree leaves, and 30% of citrus peel are used as raw materials, and essential oil is extracted by water distillation to obtain the compound essential oil. Preferably, the weight ratio of water to the raw materials is preferably 5 - 10:1. The time for distillation extraction is preferably 2 hours - 3 hours.

[0023] The weight percentage of the compound essential oil in the ant repellent liquid is 0.9%-10%, and the weight percentage of the ethanol reflux extraction liquid of pine needles is 90%-99.1%.

[0024] The ethanol reflux extraction liquid of pine needles is obtained by heating and refluxing pine needles with ethanol for extraction. Preferably, the weight ratio of ethanol to pine needles is preferably 2-3:1. The time for reflux extraction is preferably 2-3 hours.

[0025] The spraying amount of the ant repellent liquid is more than 5 mL on the outer surface of each fungus bag.

[0026] In step (2), 2.0-5.0 mL of the cold extract of pine wood is dropped into each fungus bag.

[0027] The low-temperature extraction at 30°C-35°C is preferably carried out by low-temperature ultrasonic extraction at 30°C-35°C. Preferably, the power of the ultrasonic wave is 2 KW-5 KW.

[0028] Preferably, the pine wood raw material is selected as particles with a particle size of 2 mm-5 mm, which is convenient for extraction and high-efficiency filtration.

[0029] The cultivation method of the present invention is applicable to both the outdoor cultivation and the indoor cultivation of Poria cocos, and is particularly applicable to the outdoor cultivation of Poria cocos.

[0030] A green substrate for cultivating Poria cocos in bags comprises, by weight: 70-80 parts of sawdust, 15-20 parts of bran, 1-2 parts of gypsum, 1-2 parts of brown sugar, 6-8 parts of corn, 1-3 parts of oyster shell, 3-5 parts of the first pine branch and 0.1-1 part of pine resin;

[0031] The oyster shell is processed into long strips with a length of 2 mm-5 mm;

[0032] The first pine branch is selected as the pine branch cut into a length of 2 cm-4 cm and flattened to a thickness of 0.5 mm-1.5 mm. Preferably, the first pine branch is selected as a pine branch with a thickness of 0.5 cm-1.5 cm.

[0033] In the present invention, the sources of pine branches, pine trunks (i.e., pine tree trunks), and pine needles are pine (a general term for plants of the genus Pinus in the family Pinaceae). The pine can be one or more of Pinus massoniana, Pinus koraiensis, Pinus thunbergii, Pinus taiwanensis, Pinus yunnanensis and other plants of the genus Pinus in the family Pinaceae.

[0034] The raw materials, reagents, culture media, etc. used in the present invention can all adopt existing products, such as commercially available products, or can be prepared according to existing preparation methods.

[0035] Ruta graveolens is the whole herb of Ruta graveolens L. of the genus Ruta in the family Rutaceae.

[0036] Chrysanthemum is the whole herb of the perennial root herbaceous plant Chrysanthemum (Dendranthema morifolium) of the Compositae family and Chrysanthemum genus.

[0037] Litsea cubeba is the whole plant of the plant Litsea cubeba (Lour.) Pers. of the Litsea genus in the Lauraceae family, including fruits (Piper cubeba), roots and leaves. The whole plant is harvested when the fruits are ripe in autumn.

[0038] Cinnamomum camphora leaves are the leaves or branches and leaves of the plant Cinnamomum camphora (L.) Presl. of the Cinnamomum genus in the Lauraceae family.

[0039] Tangerine peel is one or more of the peels of various citrus fruits such as Citrus chachiensis Hort. and / or Ougan of the Rutaceae family, and the peels of the plant Citrus reticulata Blanco of the Rutaceae family and / or its cultivated varieties.

[0040] Pinus massoniana Lamb., Pinus koraiensis Sieb. et Zucc., Pinus thunbergii Parl., Pinus taiwanensis Hayata, and Pinus yunnanensis are all arbors of the Pinaceae family and Pinus genus. Pine needles are the needles of pine.

[0041] Oyster shell is the shell of oyster (Ostreidae), commonly known as sea oyster, oyster, etc.

[0042] In the present invention, the tuber formation rate refers to the percentage of the number of fungus bags with Poria cocos tubers formed in the total number of cultivated fungus bags.

[0043] Advantages of the green Poria cocos cultivation method with bagged materials of the present invention:

[0044] The present invention has a high raw material utilization rate; the raw materials of the fungus bag bagged material formula are common and easy to obtain; there are no high requirements for wood chips and it is not necessary for them to be pine wood chips or must contain pine wood chips. Using broad-leaved wood chips can still ensure a high tuber formation rate, reducing the demand for pine wood. The adaptability range of wood chips is wide, and it is easier to achieve large-scale cultivation compared with the existing Poria cocos fungus bag bagged material formula.

[0045] The existing indoor artificial cultivation method of Poria cocos is not applicable to outdoor natural environment cultivation. The green bagged cultivation method of Poria cocos in the present invention is applicable to both outdoor natural environment cultivation (such as under-forest cultivation, etc.) and indoor artificial cultivation of Poria cocos (such as artificial cultivation in greenhouses, etc.). The applicable cultivation sites are extensive, which is convenient for popularization and application. When using the green bagged cultivation method of Poria cocos in the present invention for indoor artificial cultivation of Poria cocos, it is easy to manage, control temperature and humidity, and can be cultivated and harvested throughout the year. When using the green bagged cultivation method of Poria cocos in the present invention for outdoor natural environment cultivation of Poria cocos, it can be cultivated in late spring and early summer and harvested in autumn.

[0046] For the cultivation temperature and humidity of Poria cocos in the present invention, the existing technologies can be directly adopted. The time for Poria cocos to form sclerotia and be harvested in the present invention: Generally, sclerotia will start to form 20 to 25 days after the fungus bags are placed in a light-shielded groove or buried in soil, fine sand or sandy soil, etc. The Poria cocos is mature and can be harvested 110 to 130 days after the fungus bags are buried. The cultivation cycle is short and controllable.

[0047] The specific bagged formula in the present invention can significantly promote the growth of hyphae. The hyphae have filled the fungus bags evenly within 14 days, realizing high-efficiency cultivation during the hypha cultivation period and saving the cultivation cost.

[0048] The cold-extracted solution of pine wood in the present invention can play a very good triggering role in inducing the formation of Poria cocos sclerotia, that is, it acts as an inducing factor for the growth of Poria cocos sclerotia. Combining with the specific bagged formula in the present invention and the cultivation method of placing the fungus bags in a light-shielded groove or burying them in soil, fine sand or sandy soil, etc. and covering the top of the fungus bags with soil, fine sand or both, the success rate is nearly 100%, and the sclerotia formation rate is above 97%, which is significantly higher than the existing success rate using fresh pine branches, Poria cocos fungus blocks, etc. as induction.

[0049] The ant repellent solution sprayed outside the fungus bags in the present invention is composed of a compound essential oil and an ethanol reflux extract of pine needles. The terpene and other volatile components of the compound essential oil are mixed with components such as tar and resin in the ethanol reflux extract of pine needles. After being sprayed on the outside of the fungus bags and the ethanol volatilizes, it presents a slightly sticky state. The smell of the essential oil and pine wood is stronger than that when sprayed. It is preliminarily determined that after the ethanol volatilizes, the high-molecular viscous components such as tar and resin in the ethanol reflux extract of pine needles prevent the rapid volatilization of the compound essential oil, and can play a long-term slow-release ant repellent role to avoid the damage of the fungus bags by termites; moreover, the components of the ant repellent solution are natural, biodegradable, harmless to the environment, environmentally friendly, the raw materials are cheap, and the cost is low.

[0050] In the present invention, the fungus bags are placed in a light-shielded groove or buried in soil, fine sand or sandy soil, etc., and one or both of soil and fine sand are covered on the top of the fungus bags. By using physical compression and / or avoiding light, the cracking of Poria cocos is greatly reduced or even avoided, and the quality of Poria cocos is greatly improved.

[0051] The method of the present invention has few steps, is simple to operate, easy to control, has low requirements for operation, is green and environmentally friendly, does not pollute the environment, and the tuber formation rate is above 97%, reaching or even exceeding the tuber formation rate of the existing Wolfiporia cocos cultivation methods with many steps and complex operations. It is particularly suitable for large-scale popularization and application, providing a practical solution for the green cultivation of Wolfiporia cocos. Detailed implementation mode

[0052] The following further describes the present invention in detail with specific embodiments, so as to facilitate those skilled in the art to better understand the technical solution of the present invention.

[0053] Example 1: Cultivation in the natural environment under the forest

[0054] Preparation of the outer ant repellent liquid:

[0055] By weight percentage, add 5 kg of fresh whole ruta graveolens plants, 2.5 kg of fresh whole chrysanthemum plants, 2.5 kg of fresh whole litsea cubeba plants, 7.5 kg of camphor tree leaves and 7.5 kg of citrus peels to an essential oil extraction kettle, add 200 kg of water, and distill and extract for 2 hours. Separate with an oil-water separator to obtain 380 g of compound essential oil.

[0056] Cut 10 kg of fresh masson pine needles into 1 cm lengths, add 20 kg of ethanol, and heat under reflux for extraction for 2 hours to obtain 21.3 kg of pine needle ethanol reflux extract. Add 215 g of compound essential oil to 21.3 kg of pine needle ethanol reflux extract, stir and mix evenly to obtain the ant repellent liquid.

[0057] Preparation of the cold extract of pine wood:

[0058] Take 4.5 kg of masson pine branches with bark, 1.5 kg of masson pine tree trunks with bark and 4 kg of masson pine needles, dry at a low temperature of 40 °C until the moisture content is 50%, crush the above materials into particles with a particle size of 5 mm, add them to an extraction kettle, and then add 50 kg of ethanol-n-hexane solution with an ethanol mass percentage concentration of 70%. Extract at a temperature of 35 °C with an ultrasonic power of 3 KW. The extract is vacuum concentrated and desolventized at a temperature of 35 °C and a pressure of 10 KPa to obtain 1.5 kg of cold extract of pine wood (the solvent weight content is less than 1%).

[0059] Preparation of the strain: Put Wolfiporia cocos strain (commercial strain Bio08650, sourced from the Institute of Microbiology, Chinese Academy of Sciences) and liquid culture medium (composition: peptone 5 g / L, yeast extract powder 2 g / L, glucose 20 g / L, potassium dihydrogen phosphate 1 g / L, magnesium sulfate 0.5 g / L, the balance is water) into a liquid culture bottle, shake and culture at 28 °C for 6 days to obtain Wolfiporia cocos mycelial balls. Pour them into a 100 L kettle in a sterile room, and ultrasonically disperse the mycelial balls into small mycelial balls of 2 mm - 3 mm at a power of 500 W and a frequency of 33 KHz to obtain liquid Wolfiporia cocos strain.

[0060] Bag material preparation and inoculation: Take 800 kg of Masson pine sawdust, 200 kg of rice bran, 10 kg of gypsum, 10 kg of brown sugar, 80 kg of corn, 20 kg of oyster shells in long strip shape with a length of 2 mm - 5 mm, 30 kg of Masson pine branches with a thickness of 0.5 cm - 1.5 cm and a length of 2 cm - 4 cm are flattened to a thickness of 0.5 mm - 1.5 mm, and 1.5 kg of 200 - mesh pine resin; after mixing all the raw materials evenly, spray water until the moisture content of the materials in the fungus bag is 60%, and then fill them into fungus bags with a bag body diameter of 20 cm and a length of 40 cm. After sterilization and cooling, inoculate with liquid Poria cocos strain, and place them in an environment of 27°C for 14 days. All (100%) of the fungus bags are covered with mycelium, and it is observed that the mycelium in the fungus bags shows a uniform distribution.

[0061] After the mycelium fills the fungus bag, evenly spray 5.0 mL / bag of ant - repellent liquid on the outer surface of the fungus bag, and let it dry naturally until it shows a slightly sticky state. The essential oil and the smell of pine wood are stronger than when spraying. It is initially determined that after the ethanol volatilizes, the high - molecular viscous components such as tar and resin in the pine needle ethanol reflux extract prevent the rapid volatilization of the compound essential oil; make a small hole with a diameter of 0.5 cm on one side of the fungus bag, make one small hole per bag, and drop 2.0 mL of cold - extracted pine wood liquid into each small hole; dig a ditch in the forest in late spring and early summer, bury the treated fungus bags in the ditch, cover the soil 5 cm on the top of the fungus bags, and wait quietly for Poria cocos to form. After the fungus bags are buried in the soil for 130 days, Poria cocos matures and is harvested. To prevent excessive rain in the wild, drainage ditches can be dug. If it is dry, spray water to keep moisture and conduct routine management.

[0062] By the time of harvest, no termite damage to Poria cocos was found. It shows that the ant - repellent liquid composed of compound essential oil and pine needle ethanol reflux extract sprayed outside the fungus bag in the present invention has a good effect on repelling termites, and because it shows a slightly sticky state after the ethanol volatilizes, it prevents the rapid volatilization of the compound essential oil and can play a role in slow - release ant - repelling for a long time.

[0063] Harvest situation: The Poria cocos formation rate is 97.28%; for the fungus bags with Poria cocos formed (1 fungus bag forms 1 Poria cocos), the weight of a single Poria cocos is 0.8 kg - 2.8 kg / bag, and the average weight is 1.2 kg / bag. There is no situation of Poria cocos cracking, and the appearance of Poria cocos is good.

[0064] Example 2: Artificial cultivation in greenhouse

[0065] There are no harmful pests such as termites in the greenhouse, so no ant - repellent treatment is required.

[0066] Bag material preparation and inoculation: Take 800 kg of Huangshan pine sawdust and Chinese fir sawdust (wherein, Huangshan pine sawdust and Chinese fir sawdust are evenly mixed according to a weight ratio of 1:1), 135 kg of wheat bran, 65 kg of rice husk bran, 10 kg of gypsum, 10 kg of brown sugar, 80 kg of corn, 20 kg of long strip-shaped oyster shells with a length of 2 mm - 5 mm, and 30 kg of Huangshan pine branches with a thickness of 0.5 cm - 1.5 cm and a length of 2 cm - 4 cm are flattened to a thickness of 0.5 mm - 1.5 mm, and 2.0 kg of 200-mesh pine resin; after the raw materials are evenly mixed, water is sprayed until the moisture content of the material is 60%, and it is packed into fungus bags with a bag body diameter of 20 cm and a length of 40 cm. After sterilization and cooling, it is inoculated with the liquid Poria cocos strain prepared in Example 1, and placed in an environment of 23°C for culturing for 10 days. The mycelium has grown in 100% of the fungus bags, and it is observed that the mycelium in the fungus bags is evenly distributed.

[0067] After the mycelium has grown in the fungus bags, a small hole with a diameter of 0.5 cm is opened on one side of each fungus bag, and one small hole is opened per bag. 5.0 mL of the cold-extracted liquid of pine wood prepared in Example 1 is dropped into each small hole; in the greenhouse, a soil trough with a length of 30 m, a width of 2 m, and a height of 0.5 m is built, and the treated fungus bags are buried in the soil trough, and 5 cm of soil is covered on the top of the fungus bags. The temperature in the greenhouse is maintained at 28°C ± 5°C and the humidity is 75% ± 10%. Wait quietly for Poria cocos to form. After 120 days when the fungus bags are buried in the soil, the Poria cocos is mature and harvested.

[0068] Harvest situation: The Poria cocos formation rate is 98.5%; for the fungus bags with Poria cocos formed, the weight of a single Poria cocos is 1.0 kg - 2.9 kg / bag, and the average weight is 1.5 kg / bag. There is no situation of Poria cocos cracking, and the appearance of Poria cocos is good.

[0069] Comparative example 1:

[0070] There are no harmful pests such as termites in the greenhouse, and no termite control treatment is required.

[0071] Obtain fungus bags with mycelium grown according to the bag material preparation and inoculation culture steps in Example 2. A small hole with a diameter of 0.5 cm is opened on one side of each fungus bag, and one small hole is opened per bag. 5.0 mL of the cold-extracted liquid of pine wood prepared in Example 1 is dropped into each small hole; the four sides, top, and door of the greenhouse are made of light-proof materials, and a four-layer bracket with a length of 5 m and a height of 2 m is placed inside. The treated fungus bags are arranged on the bracket without covering with soil. The temperature in the greenhouse is maintained at 28°C ± 5°C and the humidity is 75% ± 10%. Wait quietly for Poria cocos to form. After 120 days when the fungus bags are arranged on the bracket, most of the Poria cocos are basically mature and harvested.

[0072] Harvest situation: The Poria cocos formation rate is 95%; for the fungus bags with Poria cocos formed, the weight of a single Poria cocos is 0.9 kg - 1.8 kg / bag, and the average weight is 1.4 kg / bag; but 15% of the total weight of the harvested Poria cocos is cracked and has a poor appearance.

[0073] Example 3: Artificial cultivation in the greenhouse

[0074] There are no harmful pests such as termites in the greenhouse, and no ant control treatment is required.

[0075] Obtain fungus bags full of mycelium according to the steps of bag material preparation and inoculation culture in Example 2. Make a small hole with a diameter of 0.5 cm on one side of each fungus bag, one small hole per bag, and add 5.0 mL of the cold extract of masson pine prepared in Example 1 dropwise to each small hole. Place a 5-meter-long, 3-meter-high, 5-layer groove bracket in the greenhouse, arrange the treated fungus bags in the grooves of each layer of the bracket, cover the top of the fungus bags with clean fine sand, covering the fungus bags by 3 cm. Keep the temperature in the greenhouse at 28°C ± 5°C and the humidity at 75% ± 10%, and wait quietly for tuber formation. After 120 days of covering the fungus bags with clean fine sand, the poria cocos is mature and harvested.

[0076] Harvest situation: The tuber formation rate is 97.03%; for the fungus bags with tuber formation, the weight of a single poria cocos is 0.9 kg - 2.6 kg per bag, and the average weight is 1.3 kg per bag; there is no situation of poria cocos cracking, and the appearance of poria cocos is good.

[0077] Examples 2 - 3 and Comparative Example 1 show that in the present invention, the operation of placing the fungus bags in light-shieldable grooves or burying them in soil, fine sand, or sandy soil, etc., and covering the top of the fungus bags with one or two of soil and fine sand can greatly reduce or even avoid the cracking of poria cocos by using physical pressure and / or avoiding light, and greatly improve the quality of poria cocos.

[0078] Example 4: Broadleaf wood sawdust

[0079] Bag material preparation and inoculation: Take 800 kg of oak sawdust, 200 kg of rice husk bran, 10 kg of gypsum, 10 kg of brown sugar, 80 kg of corn, 20 kg of long strip-shaped oyster shells with a length of 2 mm - 5 mm, and 30 kg of masson pine branches with a thickness of 0.5 cm - 1.5 cm and a length of 2 cm - 4 cm, flattened to a thickness of 0.5 mm - 1.5 mm, and 2.0 kg of 200-mesh rosin. After mixing all the raw materials evenly, spray water until the moisture content of the material is 60%, and sub-pack it into fungus bags with a bag body diameter of 20 cm and a length of 40 cm. After sterilization and cooling, inoculate with the liquid poria cocos strain prepared in Example 1, and culture it in an environment of 27°C for 14 days. The mycelium has grown in 100% of the fungus bags, and it is observed that the mycelium in the fungus bags is evenly distributed.

[0080] After the mycelium filled the fungus bags, 5 mL / bag of the natural ant repellent liquid prepared according to Example 1 was evenly sprayed on the outer surface of the fungus bags, and then naturally dried until it showed a slightly sticky state. The essential oil and the odor of pinewood were stronger than when they were sprayed. A small opening with a diameter of 0.5 cm was made on one side of each fungus bag, and 2.0 mL of the cold-extracted pinewood liquid prepared according to Example 1 was dropped into each small opening. In late spring and early summer, a ditch was dug in the forest, and the treated fungus bags were buried in the ditch. 5 cm of soil was covered on the top of the fungus bags, and then wait quietly for tuber formation. After the fungus bags were buried in the soil for 130 days, the Poria cocos was mature and harvested. To prevent excessive rain in the wild, a drainage ditch can be dug. If it is dry, spray water to keep the humidity and conduct routine management.

[0081] Until the harvest, no termite damage to the Poria cocos was found. It shows that the ant repellent liquid composed of the compound essential oil and the ethanol reflux extract of pine needles sprayed outside the fungus bags of the present invention has a good effect on repelling termites. And because it shows a slightly sticky state after the ethanol volatilizes, it prevents the rapid volatilization of the compound essential oil and can play a slow-release ant repellent role for a long time.

[0082] Harvest situation: The tuber formation rate was 97.8%; for the fungus bags with tuber formation, the weight of a single Poria cocos was 0.8 kg - 4.2 kg / bag, and the average weight was 1.5 kg / bag. There was no situation of Poria cocos cracking, and the appearance of Poria cocos was good.

[0083] It can be seen that the bag material formula of the fungus bags of the present invention has low requirements for wood chips and does not require them to be pine wood chips or must contain pine wood chips. Using broad-leaved wood chips can still ensure efficient mycelium cultivation and a high tuber formation rate, reduce the demand for pine wood, have a wide adaptability range of wood chips, and are easier to achieve large-scale cultivation.

[0084] Example 5:

[0085] Adjust the weight percentage of the compound essential oil in the ant repellent liquid to be 0.9% - 10% (specifically: 0.9%, 3%, 5%, 7%, 10%), and the weight percentage of the ethanol reflux extract of pine needles to be 90% - 99.1% (specifically: 99.1%, 97%, 95%, 93%, 90%). The rest of the operations are the same as in Example 4. Until the harvest, no termite damage to the Poria cocos was found after spraying the ant repellent liquid with different proportions of the compound essential oil and the ethanol reflux extract of pine needles. It shows that the ant repellent liquid composed of the compound essential oil and the ethanol reflux extract of pine needles sprayed outside the fungus bags of the present invention has a good effect on repelling termites. And because it shows a slightly sticky state after the ethanol volatilizes, it prevents the rapid volatilization of the compound essential oil and can play a slow-release ant repellent role for a long time.

[0086] Example 6:

[0087] Except for using fungus bags with a bag body diameter of 30 cm and a length of 40 cm, the rest of the operations are the same as in Example 1. After the fungus bags were buried in the soil for 130 days, the Poria cocos was mature and harvested.

[0088] Until the harvest time, no termite damage to Poria cocos was found. It shows that the ant repellent liquid composed of compound essential oil and ethanol reflux extract of pine needles sprayed outside the fungus bag in the present invention has a good effect on repelling termites. And because it presents a slightly sticky state after the ethanol volatilizes, it prevents the rapid volatilization of the compound essential oil and can play a role in slow-release ant repellent for a long time.

[0089] Harvest situation: The fruiting rate is 98%; for the fungus bags with fruiting, the weight of a single Poria cocos is 1.0 kg - 5.1 kg per bag, and the average weight is 1.9 kg per bag. No cracking of Poria cocos occurs, and the appearance of Poria cocos is good. It shows that the specification of the fungus bag does not affect the growth rate and fruiting rate of Poria cocos, and may have a moderate impact on the size of Poria cocos. Larger-diameter bags may be more likely to grow larger Poria cocos.

[0090] Comparative Example 2:

[0091] Preparation of pine wood extract by high-temperature extraction at 80°C:

[0092] Take 4.5 kg of pine branches with bark, 1.5 kg of pine trunks with bark and 4 kg of pine needles of Masson pine, dry them at a low temperature of 40°C until the moisture content is 50%, crush the above materials into particles with a particle size of 5 mm, add them to the extraction kettle, and then add 50 kg of ethanol-n-hexane solution with an ethanol mass percentage concentration of 70%. Extract at a temperature of 80°C with an ultrasonic power of 3 KW. The extract is vacuum concentrated and de-alcoholized at a temperature of 80°C and a pressure of 10 KPa to obtain 1.3 kg of pine wood extract by high-temperature extraction at 80°C (the solvent weight content is less than 0.2%).

[0093] According to the bag material preparation and inoculation and cultivation steps in Example 1, several groups of fungus bags full of mycelium with the same number as the fungus bags in Example 1 were obtained, denoted as fungus bag group A, fungus bag group B, fungus bag group C and fungus bag group D; select the same forest environment as in Example 1 for cultivation by digging trenches in the forest. To prevent excessive rain in the wild, trenches can be dug for drainage. If it is dry, spray water to keep moist and do routine management.

[0094] For fungus bag group A, evenly spray 5 mL / bag of the natural ant repellent liquid prepared according to Example 1 on the outer surface of the fungus bag, and let it dry naturally until it presents a slightly sticky state; then make a U-shaped opening at one end of the fungus bag full of mycelium and insert a 30-cm-long fresh pine branch as the induction part. Bury the treated fungus bag in the soil trench, cover 5 cm of soil on the top of the fungus bag, and wait for fruiting quietly. Most of the Poria cocos are basically mature for harvest 130 days after the fungus bag is buried in the soil. Harvest situation: The fruiting rate is 72.05%, the weight of Poria cocos is 0.8 kg - 1.5 kg per bag, and the average weight is 1.1 kg per bag. No cracking of Poria cocos occurs.

[0095] For the mushroom bag group B, evenly spray 5 mL / bag of the natural ant-repellent liquid prepared according to Example 1 on the outer surface of the mushroom bags, and let it dry naturally until it shows a slightly sticky state; then make a small hole with a diameter of 0.5 cm on one side of each mushroom bag, one small hole per bag; bury the treated mushroom bags in the soil ditch under the forest, cover the top of the mushroom bags with 5 cm of soil, and wait quietly for the formation of poria cocos. After 130 days of burying the mushroom bags in the soil, most of the poria cocos are basically mature and harvested. Harvest situation: The fruiting rate is only 27.82%, the weight of poria cocos is 0.3 kg - 1.5 kg / bag, and the average weight is 0.9 kg / bag. There is no situation of poria cocos cracking.

[0096] For the mushroom bag group C, evenly spray 5 mL / bag of the natural ant-repellent liquid prepared according to Example 1 on the outer surface of the mushroom bags, and let it dry naturally until it shows a slightly sticky state; then make a small hole with a diameter of 0.5 cm on one side of each mushroom bag, one small hole per bag, and drop 2.0 mL of the pine wood extract obtained by high-temperature extraction at 80 °C at each small hole; dig a ditch in the forest, bury the treated mushroom bags in the soil ditch, cover the top of the mushroom bags with 5 cm of soil, and wait quietly for the formation of poria cocos. After 130 days of burying the mushroom bags in the soil, most of the poria cocos are basically mature and harvested. Harvest situation: The fruiting rate is only 61.14%, the weight of poria cocos is 0.7 kg - 1.6 kg / bag, and the average weight is 1.3 kg / bag. There is no situation of poria cocos cracking.

[0097] For the mushroom bag group D, evenly spray 5 mL / bag of the natural ant-repellent liquid prepared according to Example 1 on the outer surface of the mushroom bags, and let it dry naturally until it shows a slightly sticky state; then make a small hole with a diameter of 0.5 cm on one side of each mushroom bag, one small hole per bag, and stick the poria cocos fungus block cut into 0.5 cm × 0.5 cm at the small hole and tie it firmly; dig a ditch in the forest, bury the treated mushroom bags in the soil ditch, cover the top of the mushroom bags with 5 cm of soil, and wait quietly for the formation of poria cocos. After 130 days of burying the mushroom bags in the soil, most of the poria cocos are basically mature and harvested. Harvest situation: The fruiting rate is 88.43%, the weight of poria cocos is 0.9 kg - 2.1 kg / bag, and the average weight is 1.3 kg / bag. There is no situation of poria cocos cracking.

[0098] By the time of harvest, since all four groups used the ant-repellent liquid of the present invention, no phenomenon of termite damage to poria cocos was found. It shows that spraying the ant-repellent liquid composed of the compound essential oil and the pine needle ethanol reflux extract on the outer surface of the mushroom bags in the present invention has a good effect of repelling termites, and because it shows a slightly sticky state after the ethanol volatilizes, it prevents the rapid volatilization of the compound essential oil and can play a role of slow-release ant-repellent for a long time.

[0099] Observation on the growth of Wolfiporia cocos sclerotia induced by the above several different materials found that: The cold extract of pinewood at low temperature (35°C) extracted in the present invention has the best effect on inducing the growth of Wolfiporia cocos sclerotia and the highest sclerotia formation rate, which is significantly better than the induction by fresh pine branches commonly used in the prior art (P < 0.01) and the induction by Wolfiporia cocos sclerotia blocks (P < 0.05); When using the pinewood extract extracted at a high temperature of 80°C, the sclerotia formation rate is significantly reduced, which is inferior to the cold extract of pinewood at low temperature (35°C) (P < 0.01), fresh pine branches (P < 0.05) and Wolfiporia cocos sclerotia blocks (P < 0.01). See Table 1 for details.

[0100] Table 1

[0101]

[0102] It shows that the cold extract of pinewood in the present invention can play a very good triggering role in inducing the sclerotia formation of Wolfiporia cocos, that is, it acts as an inducing factor for the growth of Wolfiporia cocos sclerotia. Combined with the specific bag material formula of the present invention and the cultivation method of putting the fungus bags into light-shielding grooves or burying them in soil, fine sand or sandy soil, etc. and covering the top of the fungus bags with one or two of soil and fine sand, the success rate is almost 100%, and the sclerotia formation rate is above 97%, which is significantly higher than the success rate of using fresh pine branches, Wolfiporia cocos sclerotia blocks, and high-temperature pinewood extract as induction.

[0103] Comparative Example 3:

[0104] Preparation of solid culture strains: The Wolfiporia cocos strains (commercial strain Bio08650, sourced from the Institute of Microbiology, Chinese Academy of Sciences) were inoculated into the solid medium in a wide-mouth bottle (composition: peptone 5 g / L, yeast extract powder 2 g / L, glucose 20 g / L, potassium dihydrogen phosphate 1 g / L, magnesium sulfate 0.5 g / L, agar 14 g / L, the balance being water), and cultured at 28°C for 8 days to obtain solid Wolfiporia cocos strains.

[0105] Preparation of liquid culture strains: Wolfiporia cocos strains (commercial strain Bio08650, sourced from the Institute of Microbiology, Chinese Academy of Sciences) and liquid medium (composition: peptone 5 g / L, yeast extract powder 2 g / L, glucose 20 g / L, potassium dihydrogen phosphate 1 g / L, magnesium sulfate 0.5 g / L, the balance being water) were placed in a liquid culture bottle and cultured with shaking at 28°C for 6 days to obtain undispersed liquid Wolfiporia cocos strains (the mycelial balls therein are relatively large, with sizes basically at 1 cm).

[0106] The mycelia of Wolfiporia cocos in the bag materials were cultured respectively according to the following several methods:

[0107] Method 1: Bag material preparation and inoculation: Take 800 kg of masson pine sawdust, 200 kg of rice husk bran, 10 kg of gypsum, 10 kg of brown sugar, 80 kg of corn, and 1.5 kg of 200-mesh pine resin; after mixing all the raw materials evenly, spray water until the moisture content of the materials in the fungus bag is 60%, and divide them into fungus bags with a bag body diameter of 20 cm and a length of 40 cm. After sterilization and cooling, divide them into two groups. One group is inoculated with the liquid Poria cocos strain prepared according to Example 1 and cultured in an environment of 27°C for 18 days. The mycelium has basically grown in all the fungus bags, but the distribution of the mycelium in the fungus bags is not as uniform as that in Example 1. The other group is inoculated with the solid Poria cocos strain and cultured in an environment of 27°C for 25 days. The mycelium has basically grown in all the fungus bags, but the distribution of the mycelium in the fungus bags is significantly uneven and is relatively more concentrated at the inoculation site.

[0108] Method 2: Bag material preparation and inoculation: Take 800 kg of masson pine sawdust, 200 kg of rice husk bran, 10 kg of gypsum, 10 kg of brown sugar, 80 kg of corn, 1.5 kg of 200-mesh pine resin, and 20 kg of long strip-shaped oyster shells with a length of 2 mm - 5 mm; after mixing all the raw materials evenly, spray water until the moisture content of the materials in the fungus bag is 60%, and divide them into fungus bags with a bag body diameter of 20 cm and a length of 40 cm. After sterilization and cooling, divide them into two groups. One group is inoculated with the liquid Poria cocos strain prepared according to Example 1 and cultured in an environment of 27°C for 17 days. The mycelium has basically grown in all the fungus bags, but the distribution of the mycelium in the fungus bags is not as uniform as that in Example 1. The other group is inoculated with the solid Poria cocos strain and cultured in an environment of 27°C for 23 days. The mycelium has basically grown in all the fungus bags, but the distribution of the mycelium in the fungus bags is significantly uneven and is relatively more concentrated at the inoculation site.

[0109] Method 3: Bag material preparation and inoculation: Take 800 kg of masson pine sawdust, 200 kg of rice husk bran, 10 kg of gypsum, 10 kg of brown sugar, 80 kg of corn, 1.5 kg of 200-mesh pine resin, and 30 kg of masson pine branches with a thickness of 0.5 cm - 1.5 cm and a length of 2 cm - 4 cm, which are flattened to a thickness of 0.5 mm - 1.5 mm; after mixing all the raw materials evenly, spray water until the moisture content of the materials in the fungus bag is 60%, and divide them into fungus bags with a bag body diameter of 20 cm and a length of 40 cm. After sterilization and cooling, divide them into two groups. One group is inoculated with the liquid Poria cocos strain prepared according to Example 1 and cultured in an environment of 27°C for 16 days. The mycelium has basically grown in all the fungus bags, but the distribution of the mycelium in the fungus bags is not as uniform as that in Example 1. The other group is inoculated with the solid Poria cocos strain and cultured in an environment of 27°C for 23 days. The mycelium has basically grown in all the fungus bags, but the distribution of the mycelium in the fungus bags is significantly uneven and is relatively more concentrated at the inoculation site.

[0110] Method 4: Bag material preparation and inoculation: Take 800 kg of masson pine sawdust, 200 kg of rice bran, 10 kg of gypsum, 10 kg of brown sugar, 80 kg of corn, 1.5 kg of 200-mesh pine resin, and 20 kg of oyster powder with a particle size of 0.5 mm; after mixing all the raw materials evenly, spray water until the moisture content of the materials in the fungus bag is 60%, then divide and pack them into fungus bags with a bag body diameter of 20 cm and a length of 40 cm. After sterilization and cooling, divide them into two groups. One group is inoculated with the liquid Poria cocos strain prepared according to Example 1 and cultured in an environment of 27°C for 18 days. The mycelium has basically grown in all the fungus bags, but the distribution of the mycelium in the fungus bags is not as uniform as that in Example 1. The other group is inoculated with the solid Poria cocos strain and cultured in an environment of 27°C for 24 days. The mycelium has basically grown in all the fungus bags, but the distribution of the mycelium in the fungus bags is significantly uneven and is relatively more concentrated at the inoculation site.

[0111] Method 5: Bag material preparation and inoculation: Take 800 kg of masson pine sawdust, 200 kg of rice bran, 10 kg of gypsum, 10 kg of brown sugar, 80 kg of corn, 1.5 kg of 200-mesh pine resin, and 30 kg of pine branch powder made from crushed masson pine branches with a thickness of 0.5 cm - 1.5 cm; after mixing all the raw materials evenly, spray water until the moisture content of the materials in the fungus bag is 60%, then divide and pack them into fungus bags with a bag body diameter of 20 cm and a length of 40 cm. After sterilization and cooling, divide them into two groups. One group is inoculated with the liquid Poria cocos strain prepared according to Example 1 and cultured in an environment of 27°C for 18 days. The mycelium has basically grown in all the fungus bags, but the distribution of the mycelium in the fungus bags is not as uniform as that in Example 1. The other group is inoculated with the solid Poria cocos strain and cultured in an environment of 27°C for 25 days. The mycelium has basically grown in all the fungus bags, but the distribution of the mycelium in the fungus bags is significantly uneven and is relatively more concentrated at the inoculation site.

[0112] Method 6: Bag material preparation and inoculation: Take 800 kg of masson pine sawdust, 200 kg of rice bran, 10 kg of gypsum, 10 kg of brown sugar, 80 kg of corn, 1.5 kg of 200-mesh pine resin, and 30 kg of masson pine branches with a thickness of 0.5 cm - 1.5 cm and a length of 2 cm - 4 cm (not flattened); after mixing all the raw materials evenly, spray water until the moisture content of the materials in the fungus bag is 60%, then divide and pack them into fungus bags with a bag body diameter of 20 cm and a length of 40 cm. After sterilization and cooling, divide them into two groups. One group is inoculated with the liquid Poria cocos strain prepared according to Example 1 and cultured in an environment of 27°C for 17 days. The mycelium has basically grown in all the fungus bags, but the distribution of the mycelium in the fungus bags is not as uniform as that in Example 1. The other group is inoculated with the solid Poria cocos strain and cultured in an environment of 27°C for 24 days. The mycelium has basically grown in all the fungus bags, but the distribution of the mycelium in the fungus bags is significantly uneven and is relatively more concentrated at the inoculation site.

[0113] Method 7. Preparation and inoculation of bagged materials: Take 800 kg of masson pine sawdust, 200 kg of rice husk bran, 10 kg of gypsum, 10 kg of brown sugar, 80 kg of corn, 1.5 kg of 200-mesh pine resin, 20 kg of oyster powder with a particle size of 0.5 mm, and 30 kg of masson pine branches with a thickness of 0.5 cm - 1.5 cm and a length of 2 cm - 4 cm (not flattened); after mixing all the raw materials evenly, spray water until the moisture content of the materials in the mushroom bag is 60%, and then pack them into mushroom bags with a bag body diameter of 20 cm and a length of 40 cm. After sterilization and cooling, divide them into two groups. One group is inoculated with the liquid Poria cocos strain prepared in Example 1 and cultured in an environment of 27°C for 17 days. The mycelium has basically grown in all the mushroom bags, but the distribution of the mycelium in the mushroom bags is not as uniform as that in Example 1. The other group is inoculated with the solid Poria cocos strain and cultured in an environment of 27°C for 23 days. The mycelium has basically grown in all the mushroom bags, but the distribution of the mycelium in the mushroom bags is significantly uneven, and it is relatively more concentrated at the inoculation site.

[0114] Method 8. Preparation and inoculation of bagged materials: Take 800 kg of masson pine sawdust, 200 kg of rice husk bran, 10 kg of gypsum, 10 kg of brown sugar, 80 kg of corn, 1.5 kg of 200-mesh pine resin, 20 kg of oyster powder with a particle size of 0.5 mm, and 30 kg of pine branch powder made from crushed masson pine branches with a thickness of 0.5 cm - 1.5 cm; after mixing all the raw materials evenly, spray water until the moisture content of the materials in the mushroom bag is 60%, and then pack them into mushroom bags with a bag body diameter of 20 cm and a length of 40 cm. After sterilization and cooling, divide them into two groups. One group is inoculated with the liquid Poria cocos strain prepared in Example 1 and cultured in an environment of 27°C for 18 days. The mycelium has basically grown in all the mushroom bags, but the distribution of the mycelium in the mushroom bags is not as uniform as that in Example 1. The other group is inoculated with the solid Poria cocos strain and cultured in an environment of 27°C for 22 days. The mycelium has basically grown in all the mushroom bags, but the distribution of the mycelium in the mushroom bags is significantly uneven, and it is relatively more concentrated at the inoculation site.

[0115] Method 9. Preparation and inoculation of bagged materials: Take 800 kg of masson pine sawdust, 200 kg of rice husk bran, 10 kg of gypsum, 10 kg of brown sugar, 80 kg of corn, 1.5 kg of 200-mesh pine resin, 20 kg of long strip-shaped oyster shells with a length of 2 mm - 5 mm, and 30 kg of masson pine branches with a thickness of 0.5 cm - 1.5 cm and a length of 2 cm - 4 cm flattened to a thickness of 0.5 mm - 1.5 mm; after mixing all the raw materials evenly, spray water until the moisture content of the materials in the mushroom bag is 60%, and then pack them into mushroom bags with a bag body diameter of 20 cm and a length of 40 cm. After sterilization and cooling, inoculate with the solid Poria cocos strain and culture in an environment of 27°C for 22 days. The mycelium has basically grown in all the mushroom bags, but the distribution of the mycelium in the mushroom bags is significantly uneven, and it is relatively more concentrated at the inoculation site.

[0116] Method 10: Preparation of bagged materials and inoculation: Take 800 kg of masson pine sawdust, 200 kg of rice hull bran, 10 kg of gypsum, 10 kg of brown sugar, 80 kg of corn, 1.5 kg of 200-mesh pine resin, 20 kg of oyster shells in the shape of long strips with a length of 2 mm - 5 mm, and 30 kg of masson pine branches with a thickness of 0.5 cm - 1.5 cm and a length of 2 cm - 4 cm, and flatten them to a thickness of 0.5 mm - 1.5 mm; after mixing all the raw materials evenly, spray water until the moisture content of the materials in the fungus bag is 60%, and pack them into fungus bags with a bag body diameter of 20 cm and a length of 40 cm. After sterilization and cooling, inoculate with unbroken liquid Poria cocos strains, and place them in an environment of 27°C for culturing for 16 days. The mycelia have basically grown in all the fungus bags, but the distribution of mycelia in the fungus bags is slightly less uniform than that in Example 1.

[0117] Method 11: Preparation of bagged materials and inoculation: Take 800 kg of masson pine sawdust, 200 kg of rice hull bran, 10 kg of gypsum, 10 kg of brown sugar, 80 kg of corn (conventional bagged material formula). After mixing all the raw materials evenly, spray water until the moisture content of the materials in the fungus bag is 60%, and pack them into fungus bags with a bag body diameter of 20 cm and a length of 40 cm. After sterilization and cooling, divide them into two groups. One group is inoculated with the liquid Poria cocos strain prepared according to Example 1 and placed in an environment of 27°C for culturing for 22 days. The mycelia have basically grown in all the fungus bags, but the distribution of mycelia in the fungus bags is less uniform than that in Example 1. The other group is inoculated with solid Poria cocos strains and placed in an environment of 27°C for culturing for 26 days. The mycelia have basically grown in all the fungus bags, but the distribution of mycelia in the fungus bags is significantly uneven, and it is relatively more concentrated at the inoculation site.

[0118] Observing the growth of Poria cocos mycelia in the above-mentioned multiple groups of different bagged material formulas and different states of Poria cocos strains, namely solid Poria cocos strains, Poria cocos large mycelial balls (unbroken liquid Poria cocos strains), and broken liquid Poria cocos strains, and the growth of Poria cocos mycelia in Example 1, it is found that:

[0119] When the Poria cocos strain used for inoculation is small mycelial balls cultured in a liquid medium and broken by ultrasound, the inoculation effect is the best, the inoculation success rate is the highest, and it is the easiest to inoculate. The small mycelial balls can quickly penetrate into the middle of the fungus bag, promoting the rapid spread of mycelia in the bagged materials; in the same bagged material formula, the growth rate of the broken liquid Poria cocos strain (small mycelial balls) is significantly faster than that of the solid Poria cocos strain cultured solidly and the Poria cocos large mycelial balls (unbroken liquid Poria cocos strains) (P < 0.05); therefore, the Poria cocos strain used for inoculation in the present invention is preferably a liquid Poria cocos strain. Further preferably: The Poria cocos strain is cultured in a liquid medium to obtain Poria cocos mycelial balls, and the Poria cocos mycelial balls are broken into small mycelial balls by ultrasound and then used as the Poria cocos strain for inoculation.

[0120] Under the same inoculated strains and culture temperature, when the bagged material formula adopts the ordinary bagged material formula + rosin + oyster shells processed into 2 mm - 5 mm long strips + pine branches cut into 2 cm - 4 cm lengths and flattened to 0.5 mm - 1.5 mm thick (the bagged material formula of the present invention), the growth rate of the mycelium in the fungus bag is the fastest (Example 1), significantly better than the formula of the ordinary bagged material formula + rosin (Method 1, P < 0.05), the bagged material formula of the ordinary bagged material formula + rosin + oyster shells processed into 2 mm - 5 mm long strips (Method 2, P < 0.05), the bagged material formula of the ordinary bagged material formula + rosin + pine branches cut into 2 cm - 4 cm lengths and flattened to 0.5 mm - 1.5 mm thick (Method 3, P < 0.05), the bagged material formula of the ordinary bagged material formula + rosin + oyster shell powder (Method 4, P < 0.05), the bagged material formula of the ordinary bagged material formula + rosin + pine branch powder (Method 5, P < 0.05), the bagged material formula of the ordinary bagged material formula + rosin + pine branches cut into 2 cm - 4 cm lengths (not flattened) (Method 6, P < 0.05), the bagged material formula of the ordinary bagged material formula + rosin + oyster shell powder + pine branches cut into 2 cm - 4 cm lengths (not flattened) (Method 7, P < 0.05), the bagged material formula of the ordinary bagged material formula + rosin + oyster shell powder + pine branch powder (Method 8, P < 0.05), and the bagged material formula of the ordinary bagged material formula (Method 11, P < 0.01); this should be because after the combination of rosin, 2 mm - 5 mm long strip oyster shells, and 2 cm - 4 cm length pine branches flattened to 0.5 mm - 1.5 mm thick with ordinary powdery bagged materials, on the one hand, it increases trace elements, and on the other hand, it helps to form a loose channel in the fungus bag, making it easy for the strains, especially liquid strains, to penetrate into the fungus bag and be evenly distributed in the bagged materials, and rosin and the loose pine branches after flattening have a promoting effect on the growth of Poria cocos in the fungus bag. Therefore, the bagged material formula of the present invention preferably adopts the formula of the ordinary bagged material formula + rosin + oyster shells processed into 2 mm - 5 mm long strips + pine branches cut into 2 cm - 4 cm lengths and flattened to 0.5 mm - 1.5 mm thick.

[0121] For the fungus bags with well - grown mycelium in each group of the above - mentioned Methods 1 to 11, operate according to Example 1: evenly spray 5.0 mL / bag of ant - repellent liquid on the outer surface of the fungus bag, and naturally dry it until it shows a slightly sticky state. Make a small hole with a diameter of 0.5 cm on one side of the fungus bag, make one small hole per bag, and drop 2.0 mL of cold - extracted pinewood liquid into each small hole; dig a ditch in the forest in late spring and early summer, bury the processed fungus bags in the ditch, cover the soil 5 cm on top of the fungus bags, and wait quietly for the formation of Poria cocos. After 130 days of burying the fungus bags in the soil, harvest. To prevent excessive rain in the wild, drainage ditches can be dug. If it is dry, spray water to keep moisture and conduct routine management.

[0122] Until the harvest time, no termite damage to Poria cocos was found. Harvest situation: The tuber formation rate is shown in Table 2. There is no situation of Poria cocos cracking, and the appearance of Poria cocos is good.

[0123] Table 2

[0124]

[0125] Comparative Example 4:

[0126] Bag material preparation and inoculation: Take 800 kg of masson pine sawdust, 200 kg of rice hull bran, 10 kg of gypsum, 10 kg of brown sugar, 80 kg of corn, 20 kg of long strip-shaped oyster shells with a length of 2 mm - 5 mm, and 30 kg of masson pine branches with a thickness of 0.5 cm - 1.5 cm and a length of 2 cm - 4 cm are flattened to a thickness of 0.5 mm - 1.5 mm (without pine resin); after mixing all the raw materials evenly, spray water until the moisture content of the materials in the plastic bag is 60%, and then fill them into plastic bags with a diameter of 20 cm and a length of 40 cm. After sterilization and cooling, inoculate with the liquid Poria cocos strain prepared in Example 1, and place it in an environment of 27°C for culturing for 16 days. The mycelia have grown in all the plastic bags, and it is observed that the mycelia in the plastic bags are evenly distributed.

[0127] After the mycelia have filled the plastic bags, spray 5.0 mL / bag of ant repellent liquid evenly on the outer surface of the plastic bags, and let it dry naturally until it shows a slightly sticky state. The essential oil and the smell of pine wood are stronger than when sprayed. It is initially determined that after the ethanol volatilizes, the high-molecular sticky components such as tar and resin in the pine needle ethanol reflux extract prevent the rapid volatilization of the compound essential oil; make a small hole with a diameter of 0.5 cm on one side of the plastic bag, make one small hole per bag, and drop 2.0 mL of cold extract of pine wood into each small hole; dig a ditch in the forest in late spring and early summer, bury the treated plastic bags in the soil ditch, cover the soil 5 cm on the top of the plastic bags, and wait quietly for tuber formation. After 130 days when the plastic bags are buried in the soil, most of the Poria cocos are basically mature and ready for harvest. In case of heavy rain in the wild, drainage ditches can be dug, and if it is dry, water can be sprayed to keep the moisture and conduct routine management.

[0128] Until the harvest time, no termite damage to Poria cocos was found. It shows that in the present invention, spraying the ant repellent liquid composed of compound essential oil and pine needle ethanol reflux extract on the outer surface of the plastic bags presents a slightly sticky state after the ethanol volatilizes, which prevents the rapid volatilization of the compound essential oil and can play a role in slow-release ant repellent for a long time.

[0129] Harvest situation: The tuber formation rate is 90.31%, which is significantly lower than that in Example 1 (P < 0.05); the weight of Poria cocos is equivalent to that in Example 1, with no obvious change. There is no situation of Poria cocos cracking, and the appearance of Poria cocos is good.

[0130] The results of Example 1 and Comparative Examples 3 and 4 show that the type combination and morphological combination of each material in the bagged material formula have a greater impact on the growth rate of mycelium. The growth rate of Wolfiporia extensa mycelium in the specific bagged material formula of the present invention is the fastest, significantly superior to Methods 1-8 and Method 11 in Comparative Example 3 and the bagged material formula in Comparative Example 4; the specific bagged material formula of the present invention combined with the pine wood cold extraction liquid of the present invention is significantly superior to fresh pine branches, Wolfiporia extensa fungus blocks, and the good triggering effect of high-temperature pine wood extraction liquid in inducing Wolfiporia extensa to form sclerotia, realizing high-efficiency cultivation during the mycelium cultivation period and a sclerotia formation rate of more than 97% during the cultivation period.

[0131] Comparative Example 5:

[0132] Several groups of mycelium-covered fungus bags with the same number of fungus bags as in Example 1 were obtained respectively according to the bagged material preparation and inoculation cultivation steps in Example 1, denoted as Fungus Bag Group E, Fungus Bag Group F, and Fungus Bag Group G; the same underforest environment as in Example 1 was selected for cultivation.

[0133] For Fungus Bag Group E, no ant repellent liquid was sprayed. A small hole with a diameter of 0.5 cm was opened on one side of the fungus bag, one small hole was opened per bag, and 3 mL of the pine wood cold extraction liquid prepared according to Example 1 was dropped into each small hole; a ditch was dug in the underforest, the treated fungus bags were buried in the ditch, 5 cm of soil was covered on the top of the fungus bags, and waited for sclerotia formation quietly. The fungus bags were harvested 130 days after being buried in the soil.

[0134] For Fungus Bag Group F, 5 mL / bag of the compound essential oil prepared according to Example 1 was evenly sprayed on the outer surface of the fungus bag and left to dry naturally. The smell of the essential oil was significantly weakened compared with that when it was sprayed, and it was initially determined that most of the essential oil had volatilized; then a small hole with a diameter of 0.5 cm was opened on one side of the fungus bag, one small hole was opened per bag, and 3 mL of the pine wood cold extraction liquid prepared according to Example 1 was dropped into each small hole; a ditch was dug in the underforest, the treated fungus bags were buried in the ditch, 5 cm of soil was covered on the top of the fungus bags, and waited for sclerotia formation quietly. The fungus bags were harvested 130 days after being buried in the soil.

[0135] For Fungus Bag Group G, 5 mL / bag of the pine needle ethanol reflux extract prepared according to Example 1 was evenly sprayed on the outer surface of the fungus bag, and then a small hole with a diameter of 0.5 cm was opened on one side of the fungus bag, one small hole was opened per bag, and 3 mL of the pine wood cold extraction liquid prepared according to Example 1 was dropped into each small hole; a ditch was dug in the underforest, the treated fungus bags were buried in the ditch, 5 cm of soil was covered on the top of the fungus bags, and waited for sclerotia formation quietly. The fungus bags were harvested 130 days after being buried in the soil.

[0136] After the fungus bags were well cultivated in the underforest, the situation of termites and other insects approaching and gnawing on the fungus bags was continuously observed.

[0137] Termites continuously invaded the cultivation area 3 days after the fungus bags in Fungus Bag Group E were well cultivated in the underforest, and it continued until the harvest. By the time of harvest, 35% of the fungus bags in this group failed to form sclerotia due to termite damage. When dug out and checked, serious termite gnawing was found.

[0138] For the fungus bag group F, continuous termite invasion started 5 days after the fungus bags were well-cultivated under the forest and continued until harvest. By the time of harvest, 34% of the fungus bags in this group failed to form poria cocos due to termite damage. When dug out and inspected, serious termite gnawing was found.

[0139] For the fungus bag group G, continuous termite invasion started 3 days after the fungus bags were well-cultivated under the forest and continued until harvest. By the time of harvest, 30% of the fungus bags in this group failed to form poria cocos due to termite damage. When dug out and inspected, serious termite gnawing was found.

[0140] Observing the growth of poria cocos under different termite repellent liquids in the above groups, it was found that: only using the water-extracted compound essential oil might have a relatively good effect in the early stage, but it was easily volatilized and could not achieve long-term slow release, nor could it prevent poria cocos from being damaged; only using the ethanol reflux extract of pine needles had a repellent effect, but the repellent effect was not strong; using the water-extracted compound essential oil + the ethanol reflux extract of pine needles was both ecologically friendly and could play a role in slowly releasing and repelling termites. The termite repellent liquid of the present invention, which is composed of the water-extracted compound essential oil including the whole plant of ruta graveolens, the whole plant of chrysanthemum, the whole plant of litsea cubeba, camphor tree leaves and citrus peel and the ethanol reflux extract of pine needles, has the best termite repelling effect.

[0141] The combination of any parameters within the scope defined by the present invention can achieve the effects of the present invention. They are not listed one by one here. Those skilled in the art can clearly know the technical solution of the present invention and the adjustment rules of each parameter according to the content recorded in the present invention.

Claims

1. A method for cultivating Poria cocos with bagged materials in a green way, characterized in that, Including the steps: (1) The bagged material formula includes, by weight: 70 - 80 parts of sawdust, 15 - 20 parts of bran, 1 - 2 parts of gypsum, 1 - 2 parts of brown sugar, 6 - 8 parts of corn, 1 - 3 parts of oyster shell, 3 - 5 parts of the first pine branch, and 0.1 - 1 part of pine resin; after mixing evenly, bagging, sterilizing, cooling, inoculating with Poria cocos fungus, and culturing until the mycelium fills the fungus bag; the oyster shell is processed into long strips with a length of 2 - 5 mm; the first pine branch is selected as the pine branch cut into a length of 2 - 4 cm and flattened to a thickness of 0.5 - 1.5 mm; (2) Make a small opening on one side of the fungus bag filled with mycelium, drop the cold - extracted pinewood liquid into the small opening, then put the fungus bag into a light - proof groove or bury it in soil, fine sand, or sandy soil, and cover the top of the fungus bag with one or two of soil and fine sand, and wait quietly for the formation of Poria cocos; The preparation method of the cold - extracted pinewood liquid includes: By weight percentage, 40% - 50% of the second pine branch, 10% - 20% of pine trunk, and 30% - 40% of pine needles are dried at a low temperature of 30℃ - 45℃ until the moisture content is 45% - 55%, then crushed to obtain pinewood raw materials. Add an ethanol - n - hexane mixed solution with an ethanol mass concentration of 70% - 85% in an amount of 3 - 5 times the weight of the pinewood raw materials, and extract at a low temperature of 30℃ - 35℃. The extract is vacuum - concentrated to remove the solvent at a temperature of 35℃ and a pressure of 10 KPa to obtain the cold - extracted pinewood liquid.

2. The green poria cocos bag cultivation method according to claim 1, characterized in that, In step (1), the Poria cocos fungus used for inoculation is: the Poria cocos strain is cultured in a liquid medium to obtain Poria cocos mycelial balls, and then the Poria cocos mycelial balls are dispersed into small mycelial balls to obtain liquid Poria cocos seeds.

3. The green tuckahoe bag cultivation method according to claim 1, characterized in that, In step (2), 2.0 - 5.0 mL of the cold - extracted pinewood liquid is dropped into each fungus bag.

4. The green poria cultivation method using bagged materials according to claim 1, characterized in that, Spray the ant - repellent liquid on the outer surface of the fungus bag filled with mycelium, and then carry out step (2); The ant - repellent liquid is composed of compound essential oil and pine - needle ethanol reflux extract; The preparation method of the compound essential oil includes: by weight percentage, 20% of the whole plant of rue, 10% of the whole plant of chrysanthemum, 10% of the whole plant of litsea cubeba, 30% of camphor tree leaves, and 30% of citrus peel raw materials are distilled with water to extract essential oil to obtain compound essential oil.

5. The green Poria cocos bag cultivation method according to claim 4, characterized in that, The weight percentage of the compound essential oil in the ant - repellent liquid is 0.9% - 10%, and the weight percentage of the pine - needle ethanol reflux extract is 90% - 99.1%.

6. The green poria cocos bag cultivation method according to claim 4, characterized in that, The pine - needle ethanol reflux extract is obtained by heating and reflux - extracting pine needles with ethanol.

7. The green poria cocos bag cultivation method according to claim 4, characterized in that, The spraying amount of the ant - repellent liquid is more than 5 mL on the outer surface of each fungus bag.

8. The green tuckahoe bag cultivation method according to claim 1, characterized in that, In step (2), the low - temperature extraction at 30℃ - 35℃ is preferably low - temperature ultrasonic extraction at 30℃ - 35℃.

9. The green Poria cocos bag cultivation method according to any one of claims 1-8, characterized in that, The cultivation method is applicable to the wild - environment cultivation of Poria cocos.

10. A substrate for cultivating Poria cocos by using bagged materials in a green way, characterized in that, Including, by weight: 70 - 80 parts of sawdust, 15 - 20 parts of bran, 1 - 2 parts of gypsum, 1 - 2 parts of brown sugar, 6 - 8 parts of corn, 1 - 3 parts of oyster shell, 3 - 5 parts of the first pine branch, and 0.1 - 1 part of pine resin; The oyster shell is processed into long strips with a length of 2 - 5 mm; The first pine branch is selected as the pine branch cut into a length of 2 - 4 cm and flattened to a thickness of 0.5 - 1.5 mm.

Images