Cultivation method for promoting growth of tricholoma matsutake by using biological activation substrate and application thereof

Abstract

The invention provides a cultivation method for promoting growth of tricholoma matsutake by using a biological activation substrate and application of the cultivation method. The invention provides a tricholoma matsutake cultivation process which takes bamboo shoot shells as a main material and is matched with a growth-promoting strain vv6. The method comprises the following steps: crushing and pre-wetting raw materials, carrying out aerobic high-temperature composting, cooling to a safety window, adding vv6 for pre-activation, inoculating tricholoma matsutake, covering rice husks, and controlling a fruiting environment. A ground isolating membrane structure is overlaid on a field cultivation scene, a soil-borne disease and pest ascending channel is cut off in a physical mode, and experimental results show that the red tricholoma matsutake planted through the method is higher in yield, the nutrient content of sporocarp is richer, and application and popularization are better facilitated.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of edible mushroom cultivation, in particular to a cultivation method for promoting the growth of Stropharia rugosoannulata by using a bio-activated substrate and application thereof. BACKGROUND

[0002] Stropharia rugosoannulata is a typical grass-decomposing edible mushroom, which can efficiently utilize rice husk, corn, wheat and other crop straw, as well as sugarcane residue, rice husk, peanut shell and other types of cellulose biomass as main carbon source and skeleton raw materials. In recent years, the consensus of production and research is to optimize the base material around the available straw resources in different regions: the formula technology system based on rice husk in some areas has been relatively mature, and there are also explorations of using unconventional raw materials such as ramie straw and edible mushroom cultivation by-products to replace or blend, showing potential in improving substrate availability and biological conversion efficiency. Overall, the main material mode of rice husk, sawdust and the like is stable and reliable, while the formula based on other fiber raw materials is still in continuous improvement.

[0003] However, the existing technology still has many limitations. Firstly, the seasonality and regionalism of the main material are strong, and the source and price are easy to fluctuate, affecting stable supply and cost control; secondly, the C / N ratio and buffering capacity of commonly used fiber raw materials are difficult to keep consistent between different batches, and the mycelium growth potential and primordium uniformity are easily affected by fluctuations; thirdly, under the conditions of forest land or ground cultivation, the substrate is in direct contact with the surface soil layer, and soil-borne pathogens and soil insects can form an upward channel of "soil-substrate-fruiting body", leading to increased contamination rate, rotten roots and handles, and affecting the stability of flushes; fourthly, the potential of traditional formula based on rice husk in improving nutritional and flavor quality is limited.

[0004] Bamboo shoot shell is a by-product of bamboo shoot processing, which has stable source, low price and broad utilization prospect. However, it often has problems such as slow mycelium whitening and high contamination rate when directly used as a base material; at the same time, if the conventional method of directly laying the substrate on the ground is still used, the risk of pests and diseases from the ground cannot be effectively inhibited. In the existing public data, there is still a lack of a complete and reproducible process scheme from "de-inhibition fermentation" to "introduction window and dosage of growth-promoting microorganisms" for this specific raw material system of bamboo shoot shell; there is also a lack of systematic description of the parameterization design and application of the ground physical isolation structure in the Stropharia rugosoannulata bed cultivation scenario. Therefore, how to build a stable micro-ecology in the bamboo shoot shell substrate system, determine the addition window and dosage of growth-promoting strains, and cut off the transmission channel of soil-borne pests and diseases through the ground isolation membrane structure, so as to balance the growth potential, uniformity, yield and commodity quality, has become a technical problem to be solved in the field. SUMMARY

[0005] The purpose of the present application is to overcome the shortcomings and deficiencies of the prior art, and to provide a bio-activated substrate.

[0006] Another object of the present application is to provide a cultivation method for promoting the growth of Tricholoma matsutake by using a biological activation substrate.

[0007] The object of the present application is achieved by the following technical solutions:

[0008] A biological activation substrate comprises the following components in parts by mass:

[0009] Bamboo shoot shell 0-81%, rice husk 0-81%, bamboo chips 2-6%, bran 4-10%, corn flour 3-10%, lime 1.0%, gypsum 1.0%; after preparation, the total water content is about 60-68%, and the pH is 7.5-8.4.

[0010] The bamboo shoot shell is crushed to a size of 3-5 cm.

[0011] The bamboo shoot shell and the rice husk are soaked in a cellulase solution before being added to the substrate, and the concentration of the cellulase is 150-250 U·g - 1. The soaking time is 3-6 h, based on the dry basis of the shell.

[0012] The bamboo shoot shell substrate for cultivating Tricholoma matsutake is subjected to a pre-fermentation treatment for 48-72 h after preparation.

[0013] The bamboo shoot shell substrate for cultivating Tricholoma matsutake is further added with a growth-promoting biological agent after pre-fermentation.

[0014] The growth-promoting biological agent is a biological agent prepared from Brevibacillus nitrificans vv6 strain.

[0015] The addition amount of the Brevibacillus nitrificans vv6 strain preparation is 1×10 6 -1×10 7 CFU per g of raw material.

[0016] Application of the biological activation substrate in planting Tricholoma matsutake.

[0017] A cultivation method for promoting the growth of Tricholoma matsutake by using a biological activation substrate comprises the following steps:

[0018] The bamboo shoot shell substrate for cultivating Tricholoma matsutake is subjected to pre-fermentation after preparation, and then Tricholoma matsutake is inoculated and cultured to obtain Tricholoma matsutake.

[0019] The Tricholoma matsutake is Stropharia rugosoannulata of the genus Stropharia in the family Strophariaceae.

[0020] The present application has the following advantages and effects compared with the prior art:

[0021] The present application proposes a set of cultivation process of Pinus densiflora tricholoma with bamboo shoot shell as main material and combined with growth promoting strain vv6. The process is: raw material crushing and pre-wetting-aerobic high temperature stacking-cooling to the safety window-adding vv6 pre-activation-inoculating Pinus densiflora tricholoma-covering straw-mushroom environment control; The ground planting scene superimposes the ground isolation film structure to cut off the upward channel of soil-borne diseases and insects in a physical way, and the experimental results show that the Pinus densiflora tricholoma planted by this method has higher yield, richer nutritional content of fruiting body, and is more conducive to popularization and application. BRIEF DESCRIPTION OF DRAWINGS

[0022] Figure 1 is the experimental record photo of substrate inoculation in example 2.

[0023] Figure 2 is the mycelium photo taken during the mycelium growth in example 2.

[0024] Figure 3 is the fruiting body photo in growth in example 2.

[0025] Figure 4 is the fruiting body photo picked in example 2. DETAILED DESCRIPTION

[0026] The present application will be further described in conjunction with the embodiments and drawings, but the embodiments of the present application are not limited thereto.

[0027] In the following implementation, if the specific test conditions are not specified, the general test conditions or the test conditions recommended by the reagent company are usually used. If no special instructions are given, the materials, reagents, etc. used are reagents and materials obtained from commercial channels.

[0028] The test Pinus densiflora tricholoma strain was provided by the Resource Garden of the Edible Fungus Research Room of the Vegetable Research Institute of the Guangdong Academy of Agricultural Sciences. The mother culture medium was PDA medium produced by Guangdong Huanke Microbial Technology Co., Ltd. (product number: 021052), 39 g of PDA medium dry powder was dissolved in 1 L of deionized water, and then 121℃ high pressure sterilization for 15 min was used. The rice husks used in the experiment were purchased from Guangzhou City, Guangdong Province. The experiment was carried out in Nanxi Town, Jieyang City, Guangdong Province, and the experimental time was from October 2024 to April 2025.

[0029] Example 1 Preparation and treatment of bamboo shoot shell substrate

[0030] 1.1 Pretreatment of raw materials

[0031] The bamboo shoot shell was crushed to 2-5 cm pieces, and half of it was used as the pretreatment group. The bamboo shoot shell was soaked in cellulase solution at room temperature for 4 h, and the concentration of cellulase solution was 150-250 U·g -1. Based on the dryness of the bamboo shoot shells, drain them after the process without rinsing, and adjust the moisture content to 60%–65%; the other half is not pretreated.

[0032] 1.2 Preparation of the substrate

[0033] Prepare culture media according to the different formulas in Table 1. After mixing, the target moisture content should be 60%–68%, and the initial pH should be 7.5–8.4. It is recommended to crush the bamboo shoot husks to 2–5 cm to facilitate mycelial penetration. Pre-moisten for 1–2 days and turn the medium intermittently to ensure even moisture penetration. The appropriate moisture content can be visually judged by whether the medium can be formed into a ball by hand, without dripping water between the fingers, and crumbling when dropped.

[0034] Table 1. Bamboo shoot husk-rice husk main ingredient formula (dry basis mass percentage, Total = 100%)

[0035]

[0036] 1.3 Pre-fermentation of the substrate

[0037] For substrate preparation, the substrate pile should ideally be a long trapezoid, with a base width of 2.0–2.5 m, a top width of approximately 1.0 m, and a height of 1.2–1.5 m (or a height of 1.6–1.8 m and a width of 2.0–2.5 m). Ventilation holes should be made from top to bottom to prevent oxygen deficiency. Under normal temperature conditions, the core temperature should rise to 58–68℃ within 48–72 hours and be maintained at ≥62℃ for at least two consecutive days. During this period, the pile should be turned 2–3 times and water added as needed. When the air temperature is low, a transparent or black-and-white film can be used for short-term heating and insulation; after completion, timely cooling should be carried out to reduce the core temperature to ≤40℃.

[0038] 1.4 Add biological agents

[0039] The introduction of growth-promoting strain VV6 is a key step. In the substrate after pre-fermentation (step 1.3), half is added to the biological agent, while the other half is left unadded. After the compost cools, VV6 is evenly sprayed and mixed in, resulting in a dry-base viable cell count of 1×10⁻⁶. 7 CFU / g, humidified with water to 65% and treated for 12 hours. vv6 refers to the Brevibacillus nitrificans vv6 strain preparation, which has been disclosed in Chinese invention patent CN112280705A.

[0040] Example 2 Inoculation and Culture

[0041] 2.1 Material preparation and inoculation

[0042] Inoculation and cultivation follow the principles of "sufficient inoculation, appropriate aeration, and uniform soil covering," with a typical layer of material at 20 kg / m². 3The substrate for cultivation was 15–22 cm thick. During the cultivation period, the temperature was 22–25℃, the relative humidity was 65%–75%, and the environment was characterized by slight wind and low light. Rice husks were then used to cover the substrate for moisture and warmth. After the substrate was laid, *Matsutake* strains were inoculated, and mycelial growth was observed and recorded daily. The results are shown in Table 2.

[0043] Table 2. Mycelial growth in the substrate after inoculation.

[0044]

[0045] Overall, the formula with "vv6 + pretreatment" performed best among all formulas, with the most significant improvement in T2 to T4 (high proportion of bamboo shoot husks); both "pretreatment only" and "vv6 only" showed improvement compared to the control, but the improvement was less than the sum of the two.

[0046] 2.2 Management during the training period

[0047] To reduce the risk of soil-borne diseases and pests, a ground-level isolation film structure is set up for ground planting: After leveling and clearing obstacles, a PE film is laid, with the edges folded down and pressed into the soil 5-10cm to form a continuous seal. Figure 1 A layer of fermented, qualified culture medium and a soil cover layer are then laid on top, which can significantly reduce the probability of re-contamination and damage from terrestrial insects; irrigation can be carried out by laying micro-sprinkler tape along the rows to ensure that the water supply is uniform, moist, and does not accumulate.

[0048] During the fruiting period, maintain a temperature of 15–22℃, relative humidity of 85%–95%, and diffused light of 200–500 lx. Harvest according to the density and uniformity of primordia, from the appearance of needles to the opening of the cap. To maintain surface moisture and marketability, cover the soil with about 2 cm of clean rice husks as a moisture-retaining layer, and lightly spray water as needed according to the weather to avoid the formation of standing water. For overly dense needles, thin out the mushrooms appropriately, maintaining a spacing of 2–4 cm between individual mushrooms to reduce competition and improve uniformity and individual quality.

[0049] During the cultivation period, the mycelium growth period and harvest period were recorded for different substrates. Based on the final yield, the yield per unit area and bioconversion rate were calculated. Photos of the mycelium growth period are shown below. Figures 2-3 As shown, the fruiting bodies obtained after harvesting are as follows: Figure 4 As shown in Table 3, the statistical results are as follows.

[0050] Table 3. Statistical results of growth in different substrates after harvesting.

[0051]

[0052] Among all formulations, the one with "pretreatment + VV6" performed best; the improvement was more significant in T2-T4 formulations with a high proportion of bamboo shoot husks (shortening the mycelium growth period by 4-10 days and increasing yield by 0.3-0.7 kg·m³). -2. The bioconversion rate increased by 2-4 percentage points, which is consistent with and corroborates the results of early growth potential in Table 2.

[0053] Example 3 Physicochemical Testing

[0054] 3.1 Detection Method

[0055] The harvested samples underwent physicochemical and nutritional testing on a dry basis: protein was tested using GB5009.5-2016 / Method 1, crude polysaccharides using NY / T 1676-2008 / 7, crude fiber using GB / T 5009.10-2003, and fat using GB 5009.6-2016 / Method 1. Using "without VV6" and "non-bamboo shoot shell formula" as controls, the time to full mycelial coverage, primordia uniformity, single flush and total yield, incidence of contaminants and pests, commercial grade rate, and the variation range (mean ± standard deviation, n≥3) of indicators such as protein, polysaccharides, and free amino acids were statistically compared to quantify the technical effects of this invention in stabilizing yield, improving quality, and reducing costs.

[0056] 3.2 Test Results

[0057] Under the T1 formulation, the addition of vv6 (A2) significantly increased the protein content of the fruiting bodies, with levels significantly higher than A1 (T1 without vv6) and B1 (T3 pretreatment without vv6), but still lower than the highest level, B3. There were no significant differences in fat, crude fiber, and crude polysaccharides among the treatments, indicating that the main benefit of vv6 was in increasing protein content without altering other major nutrients.

[0058] Table 4. Results of Physicochemical and Nutritional Analysis of Fruiting Bodies

[0059]

[0060] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A bio-activated matrix, characterized in that... Includes the following components in parts by mass: Bamboo shoot husks 0-81%, rice husks 0-81%, bamboo shavings 2-6%, wheat bran 4-10%, corn flour 3-10%, lime 1.0%, gypsum 1.0%; after preparation, the total water content is approximately 60-68%, and the pH is 7.5-8.

4.

2. The bio-activating matrix according to claim 1, characterized in that: The bamboo shoot husks are crushed into pieces of 3-5 cm in size; The bamboo shoot shells are soaked in a cellulase solution at a concentration of 150–250 U·g before being added to the substrate. -1 Based on the dry weight of the bamboo shoots, the soaking time is 3 to 6 hours.

3. The bio-activating matrix according to claim 1, characterized in that: The bio-activated substrate, after being prepared, undergoes a composting and pre-fermentation treatment for 48–72 hours.

4. The bio-activating matrix according to claim 3, characterized in that: The bio-activated matrix, after pre-fermentation, also contains growth-promoting biological agents; The growth-promoting biological agent is a biological agent made from Brevibacillus nitrificans vv6 strain; The dosage of the Brevibacillus nitrificans vv6 strain preparation is 1×10⁶ per gram of raw material. 6 ~1×10 7 CFU.

5. The application of the bio-activated substrate according to any one of claims 1 to 4 in the cultivation of red pine mushrooms.

6. A cultivation method for promoting the growth of *Matsutake* using a bio-activated substrate, characterized in that... Includes the following steps: After preparing the bamboo shoot shell substrate for the cultivation of red pine mushrooms, it is pre-fermented, spread as substrate, inoculated with red pine mushrooms, and then cultivated to obtain red pine mushrooms.

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