A mixed cultivation material for producing pilose antler mushroom and a preparation method thereof

Abstract

The application provides a mixed cultivation material for producing pilose antler mushroom and a preparation method thereof, and belongs to the technical field of edible mushroom cultivation. The mixed cultivation material for producing pilose antler mushroom comprises the following components in percentage by mass: pilose antler mushroom residue 20%, chaff 5-10%, and pilose antler mushroom cultivation new material 70-75%. The pilose antler mushroom cultivated by the mixed cultivation material has a good growth cycle and fruiting body character, and the reuse of the pilose antler mushroom residue can save the cost of cultivation material raw materials and solve the problem of residue disposal and conversion.

Description

Technical Field

[0001] This invention relates to the field of edible fungi cultivation technology, specifically to a mixed cultivation substrate for the production of *Agaricus esculentus* and its preparation method. Background Technology

[0002] Deer antler mushroom ( LyopHyllum decastes As one of the main types of edible fungi cultivated in industrialized factories, its industrialized production has been increasing year by year. In order to make efficient use of industrialized cultivation facilities, only one harvest of mushrooms is produced in industrialized cultivation of deer antler mushrooms.

[0003] Mushroom residue, also known as mushroom bran or waste mushroom sticks, refers to the remaining material after cultivating various edible fungi. It contains mycelial remnants of edible fungi, coarse fiber and other nutrient complexes whose structure has changed after enzymatic hydrolysis.

[0004] Along with the increased production of factory-grown deer antler mushrooms, there are also challenges such as the disposal of large quantities of deer antler mushroom residue after production and the cost control of raw materials for deer antler mushroom production. Summary of the Invention

[0005] The purpose of this application includes, for example, providing a mixed culture medium for the production of *Pleurotus ostreatus* and a method for preparing the same, to solve at least some of the aforementioned problems.

[0006] This application provides a mixed cultivation substrate for the production of *Pleurotus ostreatus*, which comprises the following components by weight percentage:

[0007] The mixture consists of 20% deer antler mushroom substrate, 5%-10% rice husks, and 70%-75% new substrate for deer antler mushroom cultivation.

[0008] Preferably, the new substrate for cultivating *Pleurotus ostreatus* in the mixed cultivation medium comprises, by weight percentage, the following components:

[0009] The ingredients are: 60% sawdust, 12% wheat bran, 15% corn cob, 3% soybean meal, 5% corn flour, and 5% soybean hulls.

[0010] Preferably, the moisture content of the new substrate for cultivating *Pleurotus ostreatus* in the mixed cultivation medium is 63%-65%, and the pH is 7.0-7.5.

[0011] Preferably, the preparation method of the new substrate for antler mushroom cultivation in the mixed cultivation material includes the following steps: weighing sawdust, wheat bran, corn cob, soybean meal, corn flour and soybean hulls in proportion and mixing them evenly, adding water and stirring evenly, adjusting the moisture content to 63%-65%, and adjusting the pH to 7.0-7.5 with light calcium carbonate.

[0012] Preferably, the method for preparing the new substrate for *Agaricus esculentus* cultivation in the mixed cultivation substrate further includes the following steps: the cultivation substrate is packaged into mushroom bags and then sterilized; after sterilization, it is cooled to 50-60°C and then transferred to a clean environment to cool to below 22°C.

[0013] Preferably, the method for preparing the new substrate for *Agaricus esculentus* cultivation in the mixed cultivation substrate involves packaging each bag of mushrooms with a weight of 1150g-1250g; sterilizing by high-pressure steam at a pressure of 0.125MPa-0.30MPa for 80min-120min.

[0014] This application provides a method for preparing a mixed cultivation substrate for the production of *Agaricus esculentus*, comprising the following steps:

[0015] S1 mushroom residue fermentation: Remove the mushroom stems from the mushroom residue inside the mushroom bags of the first harvest of deer antler mushrooms, pile the material for fermentation for 5-14 days, and remove the remaining mushroom blocks after fermentation.

[0016] S2 auxiliary material pretreatment: Weigh the rice husks according to the formula weight, soak them in 2.5% lime water for 4-5 days, and drain them naturally;

[0017] S3 Mixing: Mix the fermented mushroom residue, rice husks, and new material evenly to prepare a mixed cultivation substrate.

[0018] The mixed cultivation substrate for the production of *Flammulina velutipes* and its preparation method provided in this application have at least the following beneficial effects:

[0019] Reusing the waste residue from deer antler mushroom cultivation can, on the one hand, avoid environmental pollution caused by the indiscriminate disposal of deer antler mushroom residue, and on the other hand, effectively reduce the raw material cost of deer antler mushroom production. Attached Figure Description

[0020] To more clearly illustrate the technical solutions in the embodiments of this application, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0021] Figure 1 Experimental images of the mycelial growth process in each group in Example 2 of this application.

[0022] Figure 2 Comparison of the spread distance of the top of the mycelium in different groups at different mycelial growth stages in Example 2 of this application.

[0023] Figure 3 Comparison of the lateral spread distance of mycelia in different groups at different mycelial growth stages in Example 2 of this application.

[0024] Figure 4 Experimental images of the mushroom cultivation management process in each group in Example 2 of this application.

[0025] Figure 5Comparison of fruiting body heights at different fruiting stages in Example 2 of this application.

[0026] Figure 6 Comparison of the diameter of the fruiting body stems of each group at different fruiting stages in Example 2 of this application.

[0027] Figure 7 Comparison of cap diameters of fruiting bodies in different fruiting stages in Example 2 of this application. Detailed Implementation

[0028] The technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of this application, and not all of the embodiments. Based on the embodiments of this application, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of this application.

[0029] Example 1

[0030] 1. Raw material preparation

[0031] (1) Preparation of new materials: Weigh each raw material according to the formula (60% sawdust, 12% wheat bran, 15% corn cob, 3% soybean meal, 5% corn flour, 5% soybean hulls). Mix the dry materials evenly, add water and mix evenly, adjust the moisture content to 63%-65%, and adjust the pH to 7.0-7.5 with light calcium carbonate. Pack the mixed materials into polypropylene bags (17-18cm×37-38cm×0.0045cm-0.0050cm cultivation bags), each bag weighing 1150g-1250g. Sterilize by high-pressure steam, steam pressure 0.125MPa-0.30MPa, sterilization time 80min-120min. After sterilization, cool to 50-60℃ and then transfer to a clean environment to cool to below 22℃.

[0032] (2) Treatment of mushroom residue: Remove the mushroom feet from the mushroom residue in the mushroom bags (from the first harvest of mushrooms from Jiangxi Xiangchuan Biotechnology Co., Ltd.), pile the material for fermentation for 10 days, and remove the residual mushroom blocks after fermentation.

[0033] (3) Rice husk treatment: Soak the rice husks in water for 2-3 days, then soak them in 2.5% lime water for 5 days, and let them drain naturally.

[0034] 2. Preparation of cultivation substrate

[0035] (1) Experimental group design: Gradient experiments were designed according to the mass ratio of mushroom residue, rice husk and new material. A total of 10 experimental groups were formed. 12 portions of cultivation material were prepared for each group and 12 repeated experiments were conducted. The total mass of each portion of cultivation material (the sum of the weights of mushroom residue, rice husk and new material after treatment) was 16 kg.

[0036] Table 1 Experimental group design

[0037]

[0038] (2) Mixing: According to the design in Table 1, weigh the mushroom residue, rice husk and new material separately using an electronic balance, mix them evenly, determine their moisture content and pH, and add distilled water and light calcium carbonate to adjust the moisture content to 63%-65%, and use light calcium carbonate to adjust the pH to 7.0-7.5.

[0039] (3) Dispensing and sterilization: The culture medium of each experimental group was separately packed into polypropylene bags (each bag weighs 1200g, the material level inside the bag is about 15cm high and about 10cm in diameter, and there are 12 bags in each group; when filling the bags, pinch the two sides of the bag opening and shake the bag up and down to use gravity to press the lower half of the culture medium tightly, and the top surface can be pressed tightly). After filling the bags, punch holes about 2cm away from the bottom of the bag, and then sterilize them by high pressure steam, with a steam pressure of 0.125MPa-0.30MPa and a sterilization time of 80min-120min. After cooling to 60℃ in the sterilizer, the bags were transferred to a clean room to cool to below 22℃.

[0040] Example 2

[0041] 1. Microbial strains

[0042] Liquid bacterial strains independently propagated by Jiangxi Xiangchuan Biotechnology Co., Ltd.

[0043] 2. Vaccination

[0044] In a sterile inoculation workshop, an automatic inoculation machine was used to spray equal amounts of inoculated material into the culture medium of each experimental group.

[0045] 3. Cultivation and Management

[0046] All groups of mushroom bags were transferred to the same incubation room for cultivation. During the cultivation process, the temperature, light intensity, humidity, and CO2 concentration conditions in the edible mushroom production workshop were uniformly controlled (management methods refer to DB36 / T 1741—2023 "Technical Specification for Factory Production of Deer Antler Mushrooms") for mycelial growth, bud induction, and fruiting management until the fruiting bodies were harvested. From the time of inoculation, the growth of deer antler mushroom mycelium, buds, and fruiting bodies was observed and recorded.

[0047] 4. Data observation

[0048] (1) Mycelial growth

[0049] After inoculating *Pleurotus ostreatus*, the mycelial growth was observed and recorded every 2-7 days. The mycelial top spread distance from the inoculation point to 3 cm from the top edge of the bag was measured, as well as the lateral spread distance from 3 cm from the top edge to the point of full colony coverage. The average mycelial spread distance for each group was calculated, and the time required for mycelial colonization from inoculation to full colony coverage (referred to as mycelial growth time) was compared among the groups. The recorded experimental data were collected and organized, and one-way ANOVA was performed using SPSS (the same applies below). The results are shown in Tables 2 and 3. Based on Tables 2 and 3, comparative charts of mycelial top spread distances for each group at different mycelial growth stages were plotted. Figure 2 , 3 As shown.

[0050] Table 2: Summary of indicators for mycelial spread distance at the top

[0051]

[0052] Note: Data are x̄ ± s (n=12); data in the same column without identical letters in the superscript indicate significant differences (P < 0.05), while data with identical letters indicate no significant differences (P > 0.05).

[0053] Table 3: Summary of mycelial lateral spread distance and mycelial full-scale growth time

[0054]

[0055] Note: Data are x̄ ± s (n=12); data in the same column without identical letters in the superscript indicate significant differences (P < 0.05), while data with identical letters indicate no significant differences (P > 0.05).

[0056] Depend on Figure 1-3 As shown in Tables 1-3, mycelial growth consists of three stages: slow growth germination stage, rapid growth and spread stage, and mycelial maturation stage. During the germination stage, there were no significant changes in mycelial growth among the groups. After 5 days, the slope of the curves showed significant differences, and the time required for the mycelium to eventually cover the entire culture medium varied among the groups: 10 groups ≈ 2 groups (42 days) > 1 group ≈ 3 groups (43 days) > 5 groups (44 days) > 4 groups ≈ 6 groups (45 days) > 7 groups ≈ 8 groups ≈= 9 groups (47 days). This indicates that different formulations of culture medium have different effects on the mycelial growth process of *Pleurotus ostreatus*. Compared with pure fresh culture medium, culture medium with 20% mycelial residue has little effect on the mycelial growth of *Pleurotus ostreatus*, which is almost negligible. However, experimental groups with other contents of mycelial residue have a significant effect on the mycelial growth of *Pleurotus ostreatus*, inhibiting the mycelial growth rate and prolonging the mycelial growth period.

[0057] (2) Fruiting body growth and fruiting

[0058] The growth and development process of the fruiting bodies of *Pleurotus ostreatus* was observed, and the time from the appearance of the buds to the final harvest (referred to as the fruiting time) and the morphology of the fruiting bodies (the height of the stipe, the diameter of the cap and the stipe) were recorded. The results are shown in Table 4.

[0059] Table 4: Summary of Fruiting Body Growth Indicators

[0060]

[0061]

[0062] Note: Data are x̄ ± s (n=12); data in the same column without identical letters in the superscript indicate significant differences (P < 0.05), while data with identical letters indicate no significant differences (P > 0.05).

[0063] Depend on Figure 4-7 As shown in Table 4, compared with the cultivation substrate made from pure new substrate, the cultivation substrate with 20% mycelium residue did not have a significant effect on the fruiting time of each group. At the same time, the fruiting body morphology index was close to that of the control group, showing good performance. However, the fruiting body growth of each group of cultivation substrate with high mycelium content was poor, and the fruiting time was significantly prolonged.

[0064] Therefore, when the content of *Mushroom arborescens* residue needs to be controlled between 20% and 40%, the optimal cultivation substrate formula is 20% residue, 7.5% rice husks, and 72.5% new substrate, which has almost no impact on *Mushroom arborescens* production. At the same time, by reusing *Mushroom arborescens* residue, the cost of cultivation substrate raw materials can be saved, and the problem of residue disposal and conversion can be solved.

[0065] Finally, it should be clearly stated that the above description is merely a preferred embodiment of this application and is not intended to limit this application. Various modifications and variations can be made to this application by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the protection scope of this application.

Claims

1. A mixed cultivation substrate for the production of *Flammulina velutipes*, characterized in that, It comprises the following components by mass percentage: The substrate consists of 20% mushroom residue, 5%-10% rice husks, and 70%-75% new substrate for mushroom cultivation. The new substrate for mushroom cultivation is composed of the following components by weight percentage: 60% sawdust, 12% wheat bran, 15% corn cob, 3% soybean meal, 5% corn flour, and 5% soybean hulls.

2. The mixed cultivation substrate as described in claim 1, characterized in that, The moisture content of the new substrate for cultivating *Agaricus esculentus* is 63%-65%, and the pH is 7.0-7.

5.

3. The mixed cultivation substrate as described in claim 1, characterized in that, The preparation method of the new substrate for antler mushroom cultivation includes the following steps: weigh sawdust, wheat bran, corn cob, soybean meal, corn flour and soybean hulls in proportion and mix them evenly, add water and stir evenly, adjust the moisture content to 63%-65%, and adjust the pH to 7.0-7.5 with light calcium carbonate.

4. The mixed cultivation substrate as described in claim 3, characterized in that, The preparation method of the new substrate for antler mushroom cultivation also includes the following steps: the substrate is divided into mushroom bags and then sterilized. After sterilization, it is cooled to 50-60℃ and then transferred to a clean environment to cool to below 22℃.

5. The mixed cultivation substrate as described in claim 4, characterized in that, Each bag of mushrooms weighs between 1150g and 1250g.

6. The mixed cultivation substrate as described in claim 4, characterized in that, Sterilization is performed using high-pressure steam, with a steam pressure of 0.125MPa-0.30MPa and a sterilization time of 80min-120min.

7. The method for preparing the mixed cultivation substrate as described in any one of claims 1 or 6, characterized in that, Includes the following steps: S1 mushroom residue fermentation: Inoculate the mushroom residue from the first harvest of deer antler mushroom bags with compost and ferment for 5-14 days. After fermentation, remove any remaining mushroom blocks and mushroom stems. S2 auxiliary material pretreatment: Weigh the rice husks according to the formula weight, soak them in 2.5% lime water for 4-5 days, and drain them naturally; S3 Mixing: Mix the fermented mushroom residue, rice husks, and new material evenly to prepare a mixed cultivation substrate.

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