High-quality strain breeding and cultivation method for out-of-season morchella esculenta under high-altitude forest

Abstract

The invention discloses a high-altitude under-forest anti-season morchella high-quality strain breeding and cultivation method, and belongs to the technical field of edible mushroom breeding and cultivation. The method comprises two parts of high-quality strain breeding and a matched cultivation technology, wherein the strain breeding comprises the steps of high-altitude wild morchella resource sampling, separation and purification, high-altitude environment adaptability screening, stress resistance screening and stability verification, and the high-quality strain adaptive to the environment with the low temperature and day and night temperature difference under the forest at the altitude of 2000-3500m is obtained. The strain bred by the method is strong in stress resistance and stable in yield, the cultivation method fully utilizes the advantages of under-forest shading and moisturizing, the input of artificial facilities is reduced, the produced morchella is high in crude protein and polysaccharide content, the yield of fresh morchella is increased compared with that of conventional cultivation, the technical blank of out-of-season morchella production in high-altitude areas is filled, and the method is suitable for large-scale popularization and application. The method has remarkable economic, ecological and social benefits.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of hydrogeology and water resources investigation and evaluation, and particularly relates to a high-altitude forest undergrowth Morel high-quality strain breeding and cultivation method. BACKGROUND

[0002] Morel is a precious fungus for both food and medicine, rich in crude protein, polysaccharides, amino acids and other nutrients, and has a strong market demand. With the development of artificial cultivation technology, the cultivation scale of Morel is expanding, but there are still obvious technical bottlenecks: on the one hand, traditional cultivation of Morel is concentrated in low-altitude areas in spring, and the market competition is fierce, and the supply of off-season products is short, and the price advantage is significant; on the other hand, high-altitude areas (above 2000m) have ecological advantages such as cool summer temperature, large diurnal temperature difference, and clean air, which are suitable for off-season Morel production, but the climate in this area is harsh, and the existing Morel strains have poor adaptability, and problems such as no mushroom, low yield, and frequent diseases often occur during cultivation.

[0003] In the prior art, Morel strain breeding is mainly aimed at low-altitude conventional cultivation environment, and there is a lack of high-quality strains suitable for high-altitude forest undergrowth with low temperature, high humidity, and large diurnal temperature difference. At the same time, the cultivation technology of Morel in high-altitude areas is mostly copied from low-altitude mode, and has not been optimized in combination with the ecological characteristics of forest undergrowth, resulting in low success rate of cultivation and uneven product quality. Therefore, breeding high-quality Morel strains suitable for high-altitude forest undergrowth environment and establishing a supporting off-season cultivation technology system are of great significance to fill the gap in off-season Morel production in high-altitude areas and improve the economic benefits of the industry.

[0004] Therefore, the present application is proposed. SUMMARY

[0005] To solve the above technical problems, the basic idea of the technical solution of the present application is:

[0006] The high-altitude forest undergrowth Morel high-quality strain breeding and cultivation method comprises two steps of strain breeding and cultivation implementation, specifically:

[0007] Step S1: Selection of high-quality strains. Sampling: Collect healthy, disease-free wild morel fruiting bodies with intact fruiting body morphology from under coniferous or broad-leaved forests at an altitude of 2000-3500m. Isolation and purification: Using tissue isolation, take fresh tissue from the junction of the cap and stipe of the morel fruiting body, inoculate it into PDA modified medium under aseptic conditions, and culture it at a constant temperature of 15-18℃ for 7-10 days to obtain purified strains. Adaptability screening: Inoculate the purified strains into a medium simulating the physicochemical properties of high-altitude forest soil, and culture them at 10-20℃ and 60-70% humidity to screen strains with a growth rate ≥0.3cm / d. Stress resistance screening: Treat the strains that pass the adaptability screening with low temperature stress and mixed fungal stress, and screen strains with a survival rate ≥90% and an inhibition zone diameter ≥1.5cm. Stability verification: Conduct three consecutive generations of cultivation trials on the strains that pass the stress resistance screening, and screen for target strains with a fresh mushroom yield variation coefficient ≤8% and fruiting body morphology consistency ≥95%.

[0008] Step S2: Cultivation Implementation, Planting Site Selection: Select a forest plot at an altitude of 2000-3500m with a canopy closure of 50-70%, and the soil should be sandy loam with a humus content of ≥15%, a pH value of 6.5-7.5, and good drainage; Site Preparation and Greenhouse Construction: Build a greenhouse under the forest, with 40-50cm plastic film installed on both sides of the bottom of the greenhouse, ensuring that the side film can be raised and lowered, and install a full-coverage sprinkler system; Clean up weeds, residual crop roots, and gravel in the site.

[0009] As a preferred embodiment of the present invention, the cultivation implementation also includes soil treatment, specifically: insecticide: spraying 5% active ingredient high-efficiency cyhalothrin at a dosage of 100 mL / mu; fungicide: spraying sodium dichloroisocyanurate at a dosage of 4-5 catties / mu; acidification and tillage: after an interval of 2 days, spreading 150 catties / mu of quicklime, and performing the first rotary tillage to a depth ≥15 cm; control of underground pests: spreading 5% active ingredient phoxim at a dosage of 5-6 catties / mu, performing the second rotary tillage and leveling, ridging, and sealing the soil for 3-5 days.

[0010] As a preferred embodiment of the present invention, the cultivation implementation steps last month also include pre-wetting and inoculation: pre-wetting the prepared soil with a small amount of water, controlling the soil moisture content to 55-65%, inoculating with the target strain, and watering thoroughly with water after inoculation; placement and covering of nutrient bags: 7-10 days after inoculation, spray again with 5% effective ingredient high-efficiency cyhalothrin to kill insects, then place the nutrient bags and cover with black film to retain moisture and suppress weeds; mycelium management: maintain the temperature inside the greenhouse at 10-20℃, and when the temperature is higher than 20℃, raise the side film for ventilation and cooling to prevent contamination of the nutrient bags by miscellaneous bacteria, with a mycelium cultivation period of 40-45 days.

[0011] As a preferred embodiment of the present invention, it also includes mushroom cultivation and the construction of small arched sheds: After the mycelium cultivation is completed, the mushroom cultivation water is thoroughly poured once, and after the surface of the box is drained, a windproof small arched shed is built to maintain humidity; Mushroom management: The temperature inside the shed is controlled at 5-22℃ to prevent damage from high temperature, low temperature and strong wind; Regularly check for diseases, spray mushroom ear fungus at the diseased areas, and apply snail killer if slugs or other pests are found; When the soil moisture content is less than 50% during the young mushroom stage, water should be added appropriately, and when the surface temperature inside the shed is higher than 20℃ at noon, water should be poured directly into the shed for 3-5 minutes to cool it down; Finished product storage: Cold storage is used for cold storage, and if cold storage is not available, it should be stored in a cool and dry place, sealed in packaging and with an appropriate amount of desiccant.

[0012] In a preferred embodiment of the present invention, the formula of the PDA modified culture medium in step S1 is as follows: 200 g / L potato, 20 g / L glucose, 20 g / L agar, 15 g / L humic extract, 3 g / L KH2PO4, 1.5 g / L MgSO4・7H2O, 0.1 g / L vitamin B1, and pH value of 6.8-7.2.

[0013] In a preferred embodiment of the present invention, the coniferous forest in step S2 is a pine forest or a spruce forest, and the broad-leaved forest is an oak forest or a birch forest.

[0014] In a preferred embodiment of the present invention, the ridge height in step S2 is 20-25cm, the ridge width is 80-100cm, and the ridge spacing is 30-40cm.

[0015] In a preferred embodiment of the present invention, the formula of the nutrient bag in step S2 is: 40% sawdust, 30% corn cob, 20% wheat bran, 2% gypsum, 1% sucrose, 7% humus, with a moisture content of 60-65%, and sterilized at 121°C for 2 hours.

[0016] In a preferred embodiment of the present invention, the formula of the nutrient bag in step S2 is: 40% sawdust, 30% corn cob, 20% wheat bran, 2% gypsum, 1% sucrose, 7% humus, with a moisture content of 60-65%, and sterilized at 121°C for 2 hours.

[0017] In a preferred embodiment of the present invention, the arch height of the small arched shed in step S2 is 30-40cm, and it is covered with a double-layer structure of film and shade net to maintain the humidity inside the small arched shed at 85-90%.

[0018] Compared with the prior art, the present invention has the following advantages:

[0019] This invention provides a high-quality morel strain obtained through targeted breeding, which is specifically adapted to the special environment of 2000-3500m in high-altitude forests. It has extremely strong low-temperature adaptability and stress resistance (resistance to low temperature and contaminating bacteria), solving the technical problem of poor adaptability and susceptibility to disease in existing strains in high-altitude areas. The strain has good genetic stability and provides core germplasm resources for off-season cultivation.

[0020] The cultivation method of this invention fully combines the ecological advantages of high-altitude forests, taking advantage of the natural shading, moisture retention, and large diurnal temperature range of the forest, and optimizes key aspects such as soil treatment, temperature and humidity control, and pest and disease control, reducing the investment in artificial facilities and the use of chemical agents, thus achieving eco-friendly cultivation.

[0021] This invention enables off-season production of morel mushrooms, with inoculation in high-altitude areas during the summer and harvesting in the autumn. The market time avoids the concentrated market season in low-altitude areas, resulting in a market price increase of 30-50% compared to conventional products, and significant economic benefits. At the same time, the produced morel mushrooms are rich in nutrients, with crude protein content ≥25% and polysaccharide content ≥8%, which are 12% and 18% higher than conventionally cultivated products, respectively, and of excellent quality.

[0022] The cultivation method of this invention is simple to operate and highly standardized, making it suitable for large-scale promotion in forests at altitudes of 2000-3500m. It can not only make full use of forest resources in high-altitude areas and increase the income of forest farmers, but also promote the coordinated development of forest economy and ecological protection, with significant ecological and social benefits.

[0023] The specific embodiments of the present invention will now be described in further detail with reference to the accompanying drawings. Attached Figure Description

[0024] In the attached diagram:

[0025] Figure 1 A flowchart illustrating the selection and cultivation methods for high-quality strains of morel mushrooms grown out of season at high altitudes in forests. Detailed Implementation

[0026] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments will be clearly and completely described below with reference to the accompanying drawings. The following embodiments are used to illustrate the present invention.

[0027] To achieve the above objectives, the present invention adopts the following technical solution:

[0028] Selection of high-quality strains

[0029] This invention obtains high-quality strains with strong resistance and adapted to the off-season environment under forests through the excavation and targeted screening of wild morel resources at high altitudes. The specific steps are as follows:

[0030] (1) Sampling: Under coniferous forests (pine forests, spruce forests) or broad-leaved forests (oak forests, birch forests) at an altitude of 2000-3500m, select an area with a stable ecological environment and no pollution sources, collect wild morel fruiting bodies that are healthy, disease-free, and have intact fruiting body morphology (the cap is shaped like a sheep's stomach and the color is uniform). After sampling, place them in a sterile container for cold storage and separate them within 24 hours.

[0031] (2) Isolation and purification: The tissue isolation method was used. On a sterile operating table, the collected morel fruiting bodies were wiped with 75% alcohol for disinfection. Fresh tissue blocks (0.5cm×0.5cm in size) were peeled off from the junction of the cap and the stipe and inoculated onto PDA modified medium plates. The plates were placed in a constant temperature incubator at 15-18℃ for 7-10 days. After the hyphae covered the plate, the robust hyphae at the edge were selected for multiple transfers and purification to obtain pure strains.

[0032] The PDA-modified culture medium, supplemented with humus extract and mineral elements, simulates the nutrient environment of high-altitude forest soil. The formula is as follows: 200g / L potato, 20g / L glucose, 20g / L agar, 15g / L humus extract, 3g / L KH2PO4, 1.5g / L MgSO4・7H2O, 0.1g / L vitamin B1, pH 6.8-7.2, sterilized at 121℃ for 20min.

[0033] (3) Adaptability screening: The purified strains were inoculated into a culture medium that simulated the physicochemical properties of high-altitude forest soil (10% high-altitude forest soil extract was added to the PDA modified culture medium), and placed in a culture environment of 10-20℃ and 60-70% relative humidity for 10 days. The mycelial growth rate was measured, and strains with a growth rate ≥0.3cm / d, dense mycelia, and no abnormal discoloration were screened. Strains with slow growth and sparse mycelia were eliminated.

[0034] (4) Stress resistance screening: The strains that pass the adaptability screening are subjected to dual stress treatment: ① Low temperature stress: The culture medium of the inoculated strain is placed in a low temperature environment of 0-5℃ for 3 days, and then restored to 15-18℃ for 5 days. The strains with a survival rate of ≥90% are screened; ② Contaminant stress: Equal amounts of pathogenic bacteria such as green mold and mucor are inoculated on the culture medium of morel strains, cultured for 7 days, and the diameter of the inhibition zone is measured. The strains with an inhibition zone diameter of ≥1.5cm are screened to ensure that the strains have strong resistance to contaminants.

[0035] (5) Stability verification: The strains that passed the stress resistance screening were subjected to three consecutive generations of cultivation experiments. The cultivation method of the present invention was used to carry out small-scale planting under high-altitude forests. The fresh mushroom yield and fruiting body morphology consistency (cap size, color, stipe thickness) of each generation of strains were recorded. Target strains with fresh mushroom yield variation coefficient ≤8% and fruiting body morphology consistency ≥95% were screened to ensure the genetic stability of the strains.

[0036] High-altitude forest under-season cultivation methods

[0037] (1) Selection of planting site: Select forest plots at an altitude of 2000-3500m and a canopy closure of 50-70%. The soil should be sandy loam with a humus content of ≥15%, a pH value of 6.5-7.5, good drainage, far away from pollution sources, and with irrigation conditions. Priority should be given to pine forests, spruce forests, oak forests or birch forests to take advantage of the natural shading and moisture retention of the forest undergrowth and reduce the cost of artificial regulation.

[0038] (2) Site preparation and greenhouse construction: Construct a simple greenhouse on the selected understory plot. The greenhouse frame is made of steel pipe or bamboo, with a height of 2.0-2.5m, a width of 6-8m, and a length determined according to the actual conditions of the plot. Install 40-50cm wide plastic film on both sides of the bottom of the greenhouse. The film can be raised and lowered for ventilation and temperature control. Install a full-coverage sprinkler system inside the greenhouse to ensure that the water spray evenly covers the entire planting area without dead corners. After the construction is completed, thoroughly clean up weeds, residual crop roots (such as vegetable roots, tomato vines, etc.) and gravel in the site to avoid residual organic matter from breeding pests and diseases.

[0039] (3) Soil treatment: Soil treatment is carried out in four steps to ensure that the soil is clean and has suitable physical and chemical properties:

[0040] ① Pest control: Spray 5% of the active ingredient, high-efficiency cyhalothrin, evenly on the cleaned site at a rate of 100 mL / acre, focusing on the soil surface and surrounding weedy areas to kill surface pests.

[0041] ② Sterilization: 24 hours after spraying the insecticide, spray a sodium dichloroisocyanurate solution evenly at a rate of 4-5 catties / mu to kill pathogens in the soil.

[0042] ③ Acidity adjustment and tillage: Two days after sterilization, evenly spread 150 catties / mu of quicklime to adjust the soil pH to 6.5-7.5. Then carry out the first rotary tillage with a depth of not less than 15 cm to ensure that the quicklime is fully mixed with the soil.

[0043] ④ Control of underground pests: 3 days after the first rotary tillage, evenly apply 5% phorate granules at a rate of 5-6 catties / mu. Then carry out a second rotary tillage to a depth of 10-15 cm. After leveling the land, make ridges with a height of 20-25 cm, a width of 80-100 cm, and a spacing of 30-40 cm between ridges. After making the ridges, seal the soil for 3-5 days to further kill underground pests and pathogens.

[0044] (4) Pre-wetting and inoculation: After the greenhouse is sealed, the soil is pre-wetted by spraying a small amount of water through the sprinkler system to control the soil moisture content to 55-65% (the soil should clump together when squeezed in the hand but not crumble when released), avoiding excessive watering that could lead to waterlogging. The target strain selected in step 1 is used for row sowing or broadcasting at an inoculation rate of 500-600 mL / m². Immediately after inoculation, the planting water is thoroughly applied through a water hose to ensure full contact between the strain and the soil, promoting mycelial germination. Inoculation is carried out in June or July to achieve off-season cultivation (conventional morel mushrooms are inoculated in spring, while this invention is inoculated in summer and harvested in autumn).

[0045] (5) Placement and covering of nutrient bags: 7-10 days after inoculation, when the mycelium begins to germinate and spread, spray again with 5% effective ingredient high-efficiency cyhalothrin (dosage 50mL / mu) for secondary insect control. Then place nutrient bags on the ridge, with a spacing of 20-25cm between nutrient bags, and place 4-5 bags per square meter. After the nutrient bags are placed, cover them with black plastic film, and press the edges of the film with soil to retain moisture, suppress weeds and keep warm.

[0046] The nutrient bag formula is as follows: 40% sawdust, 30% corn cob, 20% wheat bran, 2% gypsum, 1% sucrose, and 7% humus. Water is added to adjust the moisture content to 60-65%. The mixture is then packed into polypropylene plastic bags (15cm×30cm), sterilized at 121℃ for 2 hours, and cooled before use. The nutrient bag provides sufficient nutrition for the growth of morel mycelium.

[0047] (6) Mycelium cultivation management: During the mycelium cultivation period, maintain the temperature inside the greenhouse at 10-20℃. Utilize the cool climate advantage of high altitude in summer. When the temperature inside the greenhouse exceeds 20℃, raise the bottom film on both sides of the greenhouse for ventilation and cooling. Ventilation times are 9-11 am and 3-5 pm, with each ventilation lasting 1-2 hours to prevent excessive temperature from causing contamination of the nutrient bags by miscellaneous bacteria. At the same time, maintain the relative humidity inside the greenhouse at 60-70%, and replenish water in a timely manner through the sprinkler system to avoid soil drought. The mycelium cultivation period is approximately 40-45 days. During this period, regularly check the nutrient bags and remove any contaminated nutrient bags promptly to prevent the spread of diseases.

[0048] (7) Mushroom induction and small arched shed construction: After the mycelium is cultivated (about 50 days after inoculation), remove the black plastic film and water the mushroom induction water thoroughly at one time. The watering amount is 20-30L / m², ensuring that the water penetrates to a depth of 15-20cm in the soil. After the surface of the box has dried (about 2-3 days), build a small arched shed on the ridge. The small arched shed uses bamboo strips as the arch frame, with an arch height of 30-40cm. It is covered with a double-layer structure of film and shade net. The film is used to retain moisture, and the shade net is used to provide shade. Ensure that the small arched shed is firmly fixed and not blown open by strong winds. Maintain a relative humidity of 85-90% inside the small arched shed to create a suitable environment for fruiting body formation.

[0049] (8) Mushroom Management: During the mushroom growing period, the focus is on controlling temperature and humidity and preventing pests and diseases: ① Temperature control: Maintain the temperature inside the small arched shed at 5-22℃. When the surface temperature inside the shed is higher than 20℃ at noon, there is no need to open the small arched shed. Instead, directly spray water inside the shed for 3-5 minutes to cool it down and avoid scorching the young mushrooms. When the temperature is lower than 5℃ at night, cover the small arched shed with a heat-insulating blanket to prevent frost damage. ② Humidity management: Regularly check the humidity inside the small arched shed. When the humidity is lower than 85%, add a small amount of water through the spray system. During the young mushroom period, the soil moisture content should be controlled. Maintain water content at 60-65% to avoid excessive moisture leading to mushroom rot; ③ Pest and disease control: Conduct regular inspections, and when diseases (such as black spot and soft rot) are found, spray mushroom ear fungus (three bottle caps mixed with one pot of water) in time, focusing on spraying the affected areas, spraying once every 3 days, and spraying continuously for 2-3 times; when slugs, snails and other pests are found, apply snail kill granules on the ridge surface at a dosage of 0.5-1.0 kg / acre; ④ Wind protection measures: Reinforce the greenhouse and small arched sheds, and set up windbreaks around the sheds to prevent strong winds from damaging the sheds and blowing on the fruiting bodies.

[0050] (9) Harvesting and processing: Harvest the fruiting bodies promptly after they mature. The harvesting criteria are that the caps are fully expanded, the morel-shaped texture is clear, and the color is yellowish-brown or dark brown, without opening the cap. When harvesting, cut the morel mushrooms at the base of the stem with scissors to avoid damaging the surrounding young mushrooms. Clean the soil and impurities on the stems promptly after harvesting. The fresh mushrooms after harvesting are dried using a hot air circulating dryer. The drying process is as follows: keep warm at 35℃ for 3 hours, keep warm at 40℃ for 2 hours, keep warm at 45℃ for 2 hours, and keep warm at 50℃ for 1 hour. Continuous ventilation is maintained throughout the process to ensure that the moisture evaporates fully. After the dried morel mushrooms are cooled to room temperature, they are promptly packed into sealed plastic bags or aluminum foil bags to prevent moisture absorption.

[0051] (10) Storage of finished products: The dried morel mushrooms should be stored in a cold storage with the temperature controlled at 0-5℃ and the relative humidity ≤60%. If there is no cold storage, the finished products should be placed in a cool and dry place, sealed in packaging, and an appropriate amount of desiccant (such as silica gel desiccant) should be placed inside the packaging to prevent mold and insect infestation. The shelf life can reach more than 12 months.

[0052] Example 1: Selection of High-Quality Strains

[0053] (1) Sampling: In June 2023, 10 healthy and disease-free wild morel fruiting bodies were collected under the pine forest in Weining, Yunnan at an altitude of 2800m. They were numbered Y1-Y10, placed in sterile containers and refrigerated. They were brought back to the laboratory for processing within 6 hours.

[0054] (2) Isolation and purification: The fruiting bodies of Y1-Y10 were wiped with 75% alcohol for disinfection, the tissue block at the junction of the cap and the stipe was peeled off, and inoculated into PDA modified medium. After culturing at 15-18℃ for 8 days, 10 purified strains were obtained.

[0055] (3) Adaptability screening: Ten purified strains were inoculated into PDA modified medium containing 10% high-altitude forest soil extract and cultured at 10-20℃ and 65% humidity for 10 days. The mycelial growth rate was measured, and four strains with growth rates ≥0.3cm / d (Y2, Y5, Y7, Y9) were screened out. Among them, strain Y5 had the fastest growth rate, reaching 0.38cm / d.

[0056] (4) Stress resistance screening: Y2, Y5, Y7 and Y9 were subjected to low temperature stress (0-5℃ treatment for 3 days) and mixed bacteria stress. The results showed that the low temperature survival rate of strain Y5 reached 95%, the diameter of the inhibition zone reached 1.8cm, and the stress resistance was the best.

[0057] (5) Stability verification: The Y5 strain was subjected to three generations of cultivation experiments. The fresh mushroom yields were 1250 kg / mu, 1280 kg / mu and 1230 kg / mu, respectively. The coefficient of variation was 1.9%, and the fruiting body morphology consistency reached 98%. Y5 was determined to be the target strain.

[0058] Example 2: Off-season cultivation under high-altitude forests

[0059] (1) Planting site selection: Select a pine forest plot at an altitude of 2800m in Weining, Yunnan, with a canopy density of 60%, sandy loam soil with humus content of 18%, pH value of 7.0, good drainage, and irrigation conditions.

[0060] (2) Site preparation and greenhouse construction: Construct a steel pipe greenhouse with a height of 2.2m, a width of 7m, and a length of 50m; install 45cm plastic film at the bottom of both sides of the greenhouse, which can be raised and lowered; install a full-coverage sprinkler system inside the greenhouse. Clean up weeds, vegetable roots, and gravel in the site.

[0061] (3) Soil treatment: ① Spray 100 mL / mu of 5% high-efficiency cyhalothrin; ② Spray 4.5 catties / mu of sodium dichloroisocyanurate 24 h later; ③ Spread 150 catties / mu of quicklime 2 days later, and till the soil to a depth of 18 cm for the first time; ④ Spread 5.5 catties / mu of phoxim 3 days later, till the soil to a level level for the second time, and make ridges (ridge height 22 cm, ridge width 90 cm, ridge spacing 35 cm), and cover the soil with soil for 4 days.

[0062] (4) Pre-wetting and inoculation: On June 15, 2023, the soil was pre-wetted with a small amount of water to control the soil moisture content to 60%. The above-mentioned experimental strains were inoculated at a rate of 550 mL / m². After inoculation, the soil was thoroughly watered for planting.

[0063] (5) Placement and covering of nutrient bags: On June 22, spray 50 mL / mu of 5% high-efficiency cyhalothrin, place nutrient bags (formula: 40% sawdust, 30% corn cob, 20% wheat bran, 2% gypsum, 1% sucrose, 7% humus), place 4 bags per square meter, cover with black film and press the edges firmly.

[0064] (6) Cultivation management: Keep the temperature inside the greenhouse at 10-20℃. When the temperature is higher than 20℃, raise the side film for ventilation for 1.5 hours. Keep the humidity inside the greenhouse at 65%. Cultivate the bacteria for 42 days. During this period, remove 2 contaminated nutrient bags.

[0065] (7) Mushroom induction and small arched shed construction: On August 5, uncover the black film and water the mushroom induction water thoroughly (25L / m²). 3 days later, build a small arched shed (arch height 35cm, double-layer covering film and shade net).

[0066] (8) Mushroom management: Maintain the temperature in the small greenhouse at 8-20℃ and the humidity at 88%; mushrooms will begin to grow on September 10. During this period, spray mushroom ear fungus twice to prevent mild black spot disease and apply 0.8 kg / mu of snail killer to prevent slugs; when the temperature is higher than 20℃ at noon, water for 4 minutes to cool down.

[0067] (9) Harvesting and processing: Harvesting took place from September 25 to October 15, with a total of 1260 kg / mu of fresh mushrooms harvested. After harvesting, the mushrooms were dried using a hot air circulating dryer, following the drying process of 35℃ for 3 hours, 40℃ for 2 hours, 45℃ for 2 hours, and 50℃ for 1 hour. The mushrooms were continuously ventilated and cooled to room temperature before being bagged.

[0068] (10) Finished product storage: The dried morel mushrooms were stored in a cold storage (0-5℃). Sampling tests showed that the crude protein content was 26.8% and the polysaccharide content was 9.2%, indicating excellent quality.

[0069] Comparative test

[0070] A control group was set up, using a conventional morel mushroom strain (commercially available strain) and planted in the same high-altitude forest understory plot according to existing low-altitude cultivation methods, with other conditions consistent with Example 2. The results showed that the control group strain had poor adaptability, with a mycelial growth rate of only 0.22 cm / d, a low-temperature survival rate of 75%, a contamination rate of 15%, a fresh mushroom yield of 820 kg / mu, a crude protein content of 22.3%, and a polysaccharide content of 6.5%. In contrast, the experimental strain and cultivation method of this invention increased the fresh mushroom yield by 53.7% compared to the control group, and increased the crude protein and polysaccharide content by 20.2% and 41.5%, respectively, with a disease and pest incidence rate of only 3%, fully demonstrating the superiority of this invention.

Claims

1. A method for selecting and cultivating high-quality strains of morel mushrooms grown out of season at high altitudes in forests, characterized by: It includes two main steps: strain selection and cultivation implementation, specifically: Step S1: Selection of high-quality strains. Sampling: Collect healthy, disease-free wild morel fruiting bodies with intact fruiting body morphology from under coniferous or broad-leaved forests at an altitude of 2000-3500m. Isolation and purification: Using tissue isolation, take fresh tissue from the junction of the cap and stipe of the morel fruiting body, inoculate it into PDA modified medium under aseptic conditions, and culture it at a constant temperature of 15-18℃ for 7-10 days to obtain purified strains. Adaptability screening: Inoculate the purified strains into a medium simulating the physicochemical properties of high-altitude forest soil, and culture them at 10-20℃ and 60-70% humidity to screen strains with a growth rate ≥0.3cm / d. Stress resistance screening: Treat the strains that pass the adaptability screening with low temperature stress and mixed bacteria stress, and screen strains with a survival rate ≥90% and an inhibition zone diameter ≥1.5cm. Stability verification: Strains that passed the stress resistance screening were continuously cultivated for 3 generations to screen for target strains with a fresh mushroom yield variation coefficient ≤8% and fruiting body morphology uniformity ≥95%. Step S2: Cultivation Implementation, Planting Site Selection: Select a forest plot at an altitude of 2000-3500m with a canopy closure of 50-70%, and the soil should be sandy loam with a humus content of ≥15%, a pH value of 6.5-7.5, and good drainage; Site Preparation and Greenhouse Construction: Build a greenhouse under the forest, with 40-50cm plastic film installed on both sides of the bottom of the greenhouse, ensuring that the side film can be raised and lowered, and install a full-coverage sprinkler system; Clean up weeds, residual crop roots, and gravel in the site.

2. The method for selecting and cultivating high-quality morel strains in high-altitude forests during off-season, as described in claim 1, is characterized in that... The cultivation implementation also includes soil treatment, specifically: insecticide: spray with 5% active ingredient high-efficiency cyhalothrin at a dosage of 100 mL / mu; Sterilization: Spray sodium dichloroisocyanurate at a rate of 4-5 catties / mu; Acidity adjustment and tillage: After an interval of 2 days, apply 150 catties / mu of quicklime, and perform the first rotary tillage to a depth of ≥15 cm; Control of underground pests: Apply 5% active ingredient phoxim at a rate of 5-6 catties / mu, perform the second rotary tillage and leveling, ridging, and mulch for 3-5 days.

3. The method for selecting and cultivating high-quality morel strains in high-altitude forests during off-season, as described in claim 2, is characterized in that... Therefore, the cultivation implementation steps last month also included pre-wetting and inoculation: pre-wetting the prepared soil with a little water, controlling the soil moisture content to 55-65%, inoculating with the target strain, and watering thoroughly with water after inoculation; placement and covering of nutrient bags: 7-10 days after inoculation, spray again with 5% effective ingredient high-efficiency cyhalothrin to kill insects, then place the nutrient bags and cover with black film to retain moisture and suppress weeds; mycelium management: maintain the temperature inside the greenhouse at 10-20℃, and when the temperature is higher than 20℃, raise the side film for ventilation and cooling to prevent contamination of the nutrient bags by miscellaneous bacteria, with a mycelium cultivation period of 40-45 days.

4. The method for selecting and cultivating high-quality morel mushroom strains in high-altitude forests during off-season, as described in claim 3, is characterized in that... This also includes mushroom cultivation and the construction of small arched sheds: After the mycelium has finished growing, thoroughly water the mushroom cultivation area once. After the surface of the boxes has dried, build windproof small arched sheds to maintain humidity; Mushroom management: Control the temperature inside the shed between 5-22℃, and prevent damage from high temperature, low temperature and strong winds; Regularly check for diseases, spray mushroom ear fungus at the affected areas, and apply snail killer if slugs or other pests are found; When the soil moisture content is below 50% during the young mushroom stage, water appropriately. When the surface temperature inside the shed is above 20℃ at noon, directly water the shed for 3-5 minutes to cool it down; Finished product storage: Store in a cold storage. If there is no cold storage, store in a cool and dry place, sealed in packaging and with an appropriate amount of desiccant.

5. The method for selecting and cultivating high-quality morel strains in high-altitude forests during off-season, as described in claim 1, is characterized in that... The formula for the PDA-modified culture medium in step S1 is as follows: 200 g / L potato, 20 g / L glucose, 20 g / L agar, 15 g / L humic extract, 3 g / L KH2PO4, 1.5 g / L MgSO4・7H2O, 0.1 g / L vitamin B1, and pH 6.8-7.

2.

6. The method for selecting and cultivating high-quality morel mushroom strains in high-altitude forests during off-season, as described in claim 1, is characterized in that... In step S2, the coniferous forest is a pine forest or a spruce forest, and the broad-leaved forest is an oak forest or a birch forest.

7. The method for selecting and cultivating high-quality morel strains in high-altitude forests during off-season, as described in claim 1, is characterized in that... In step S2, the ridge height is 20-25cm, the ridge width is 80-100cm, and the ridge spacing is 30-40cm.

8. The method for selecting and cultivating high-quality morel mushroom strains in high-altitude forests during off-season, as described in claim 1, is characterized in that... The formula for the nutrient bag in step S2 is: 40% sawdust, 30% corn cob, 20% wheat bran, 2% gypsum, 1% sucrose, and 7% humus, with a moisture content of 60-65%, and sterilized at 121℃ for 2 hours.

9. The method for selecting and cultivating high-quality morel strains in high-altitude forests during off-season, as described in claim 1, is characterized in that... The formula for the nutrient bag in step S2 is: 40% sawdust, 30% corn cob, 20% wheat bran, 2% gypsum, 1% sucrose, and 7% humus, with a moisture content of 60-65%, and sterilized at 121℃ for 2 hours.

10. The method for selecting and cultivating high-quality morel strains in high-altitude forests during off-season, as described in claim 1, is characterized in that... In step S2, the arch height of the small arched greenhouse is 30-40cm, and it is covered with a double-layer structure of film and shade net to maintain the humidity inside the small arched greenhouse at 85-90%.

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