Preparation method of stropharia rugoso-annulata cultivated species

Abstract

The invention discloses a preparation method of stropharia rugoso-annulata cultivated species. The preparation method comprises the following steps: selecting pollution-free stropharia rugoso-annulata slant mother species with white and robust hyphae and a PDA (potato dextrose agar) substrate; preparing a shake flask liquid culture medium, sterilizing and inoculating; carrying out shake-flask culture and hypha scattering; preparing a cultivation bag and sterilizing; and carrying out sterile inoculation and spawn running culture and the like. According to the method, by optimizing a culture medium formula, an inoculation process and culture conditions, the seed production time of the stropharia rugoso-annulata is effectively shortened, efficient preparation is achieved, the quality of the obtained cultivated seeds is good, and the efficient and high-quality requirements of modern agriculture are met.

Description

Technical Field

[0001] This invention relates to the field of edible fungi cultivation technology, specifically a method for preparing a cultivar of *Stropharia macrocarpa* (red pine mushroom). Background Technology

[0002] King Oyster Mushroom, also known as Red Matsutake, is a nutritious edible fungus that is highly sought after in the market due to its unique taste and rich nutritional value. However, conventional methods for cultivating King Oyster Mushroom spawn suffer from problems such as mycelial non-spreading in the middle of the cultivation process, long cultivation cycles, and low efficiency, making it difficult to meet the demands of modern agriculture for high efficiency and high quality. Therefore, developing an efficient and simple method for preparing King Oyster Mushroom spawn is of great significance. Summary of the Invention

[0003] 1. Purpose of the invention: This invention aims to provide a method for preparing *Agaricus bisporus* spawn. By optimizing the culture medium formula, inoculation process, and culture conditions, and by maintaining the mycelial viability of *Agaricus bisporus* through initial liquid culture, the spawn production time of *Agaricus bisporus* is effectively shortened, achieving efficient preparation of *Agaricus bisporus* spawn.

[0004] 2. Technical Solution: The method for preparing the *Agaricus bisporus* cultivar of the present invention includes the following steps: (1) Prepare slant mother culture and PDA substrate: Select slant mother culture of large-flowered mushroom with clean, strong and uncontaminated mycelium, and PDA substrate as initial culture medium.

[0005] (2) Preparation of shake flask liquid culture medium: Prepare MP300 or potato glucose liquid culture medium according to product requirements. Fill each 500mL shake flask with 300mL of liquid and place a 3-4cm long magnetic rotor inside. Cover with four layers of gauze and one layer of kraft paper, and sterilize at 121℃ for 20 minutes. After cooling, inoculate the test tube mother culture into the shake flask under aseptic conditions, with an inoculation volume of 2-3cm inoculum per flask.

[0006] (3) Shake flask culture and mycelial homogenization treatment: Place the inoculated shake flasks in a constant temperature environment of 20-30℃ for 12-24 hours, then transfer them to a shaker and culture them at a constant speed of 120 rpm and a suitable temperature of 25℃ for 6-8 days. During the culture process or after the culture cycle, the shake flasks can be removed and the mycelial pellets inside the flasks can be broken up with a magnetic stirrer to ensure that uniformly dispersed mycelial fragments are obtained, thus forming the desired liquid inoculum A.

[0007] (4) Preparation of cultivation bags: Use 50% sawdust, 40% cottonseed hulls, and 10% wheat bran (or other suitable formula) as the culture medium, and adjust the moisture content to 70%-75%. Fill the culture medium into the mushroom bags, about 1 kg per bag, put on a ring and cover with a non-woven fabric to seal tightly. Then sterilize at 121℃ for 4 hours to obtain mushroom bag B.

[0008] (5) Inoculation and mycelial growth: Under aseptic conditions, inoculate liquid spawn A into spawn bags B. Each bottle of A can inoculate 8-10 bags of B. After inoculation, mixed spawn bags C are obtained. Place C in a dark incubation room at 25°C for mycelial growth. After about 25-35 days, the spawn for *Agaricus bisporus* can be obtained.

[0009] 3. Beneficial effects: (1) Significantly shortened seed production time: Through optimized culture medium formula and culture conditions, the seed production cycle of giant king mushroom is significantly shortened from the long time of traditional methods, thus improving production efficiency.

[0010] (2) Ensure mycelial vitality: Liquid culture and proper operation can effectively maintain the vitality of the mycelium of Pleurotus ostreatus, laying a good foundation for subsequent growth and development.

[0011] (3) Improved cultivation success rate: Strict aseptic operation and reasonable inoculation process reduced the risk of contamination and improved the success rate of preparing giant king mushroom spawn.

[0012] (4) Easy to operate and promote: The preparation method of the present invention has clear steps and is easy to operate. It does not require complex equipment and advanced technology, and is easy to promote and apply in actual production.

[0013] (5) Improve the quality of cultivars: The optimized cultivation process results in high-quality cultivars of giant king mushrooms, which is conducive to improving the yield and quality of giant king mushrooms.

[0014] (6) Meeting market demand: The efficient and high-quality preparation method of giant king mushroom spawn can better meet the growing market demand for giant king mushroom and promote the development of the edible fungi industry. Detailed Implementation

[0015] The following is an example of a specific implementation method based on the above-described invention: Preparation of experimental materials 1. Slant mother culture: Select slant mother cultures of large-cap mushrooms with clean, robust, and uncontaminated mycelium to ensure good growth and suitability as inoculation source.

[0016] 2. PDA matrix: Used for the initial culture of test-tube mother cultures to ensure the purity and nutrition of the culture medium.

[0017] 3. Liquid culture medium: Use MP300 medium (or potato glucose medium), prepare according to product instructions, and fill 300mL of liquid into each 500mL shake flask. Before filling, place a 3-4cm long magnetic rotor inside, cover with four layers of gauze and one layer of kraft paper, and then place in an autoclave to sterilize at 121℃ for 20 minutes. Cool before use.

[0018] 4. Cultivation bag material: Use 50% sawdust, 40% cottonseed hulls, and 10% wheat bran (or other suitable formula) as the culture medium, and adjust the moisture content to 70%-75%.

[0019] Experimental steps 1. Inoculation vial shaking: Under aseptic conditions, inoculate the test tube mother culture into sterilized liquid culture medium shake flasks, with the inoculation volume per flask controlled at 2-3 cm in volume.

[0020] After inoculation, place the shake flask in a constant temperature environment of 25°C for 16 hours to allow the strain to adapt to the new environment and begin to grow.

[0021] 2. Shake flask culture and mycelial homogenization treatment: After settling, transfer the shaker to a shaker, set a constant speed of 120 rpm, and maintain the temperature at 25°C.

[0022] The culture was carried out for 7 days, during which the mycelial growth was checked regularly.

[0023] After the culture period is over, use a magnetic stirrer to break up the mycelial pellets in the bottle to ensure that the mycelial fragments are evenly dispersed and form the required liquid inoculum A.

[0024] 3. Preparation and sterilization of cultivation bags: - Mix the raw materials such as sawdust, cottonseed hulls, and bran evenly according to the formula ratio, and adjust the moisture content to 72%.

[0025] Pack the culture medium into the bags, each weighing about 1 kg, and then put a ring over them and cover them with a non-woven fabric to seal them tightly.

[0026] Place the culture bag filled with culture medium in a pressure cooker and sterilize it at 121℃ for 4 hours to obtain culture bag B.

[0027] Inoculation and mycelial culture: Under aseptic conditions, liquid bacterial culture A is inoculated into bacterial bags B, with each bottle of A capable of inoculating 9 bags of B.

[0028] After inoculation, the bags are placed in a dark incubation room at 25°C for mycelial growth.

[0029] Regularly observe the mycelial growth and maintain a stable culture environment.

[0030] After about 30 days of mycelial growth and cultivation, the mycelium will fill the bag, and the spawn for the giant king mushroom can be obtained.

Claims

1. A method for preparing a cultivation species of Stropharia rugosoannulata, characterized by, The method comprises the following steps: Preparation of white and robust mycelium and PDA medium as initial culture medium.

2. According to the product requirements, configure MP300 or potato glucose liquid medium, 300mL per 500mL flask, and put in a 3-4cm long magnetic rotor, cover four layers of gauze and a layer of cowhide paper, sterilize at 121℃ for 20 minutes, cool, then inoculate the test tube mother strain in the flask under sterile conditions, 2-3cm of mycelium per bottle.

3. Place the inoculated flask in a constant temperature environment of 20-30℃ for 12-24 hours, then transfer it to a shaker, with a constant speed of 120rpm and a suitable temperature of 25℃, for a period of 6-8 days of culture. During the culture process or at the end of the culture period, the flask can be taken out and the mycelium block in the bottle can be dispersed by a magnetic stirrer to ensure that uniform dispersed mycelium fragments are obtained, i.e. the required liquid strain A is formed.

4. Use 50% sawdust, 40% cottonseed hulls, and 10% bran as culture medium, adjust the moisture content to 70%-75%, fill the culture medium into the fungus bag, about 1kg per bag, cover the ring and tighten the non-woven fabric cover, then sterilize at 121℃ for 4 hours to obtain fungus bag B.

5. Under sterile conditions, inoculate liquid strain A into fungus bag B, 8-10 bags B per bottle A, after inoculation, place it in a 25℃ dark culture room for 25-35 days of mycelium culture to obtain the cultivation of Stropharia rugoso-annulata.

6. The preparation method of the Stropharia rugosannulata cultivation seed according to claim 1, characterized in that, The liquid medium is MP300 or potato glucose.

7. The preparation method of the Stropharia rugosannulata cultivation seed according to claim 1, characterized in that, The static temperature of the inoculated flask is 20-30℃, and the static time is 12-24h.

8. The preparation method of the Stropharia rugosannulata cultivation seed according to claim 1, characterized in that, The rotation speed of the flask culture is 120rpm, the temperature is 25℃, and the culture time is 6-8 days.

9. The preparation method of the Stropharia rugosannulata cultivation species according to claim 1, characterized in that, The formula of the fungus bag culture medium is a suitable formula, and the moisture content is adjusted to 70%-75%.