Semi-wild tending method for cordyceps sinensis

Abstract

The invention provides a semi-wild tending method for cordyceps sinensis, which comprises the following steps: hatching and feeding indoor larvae: hatching self-propagating hepialus armoricanus eggs in a controllable indoor environment to obtain larvae; the larvae are sequentially subjected to centralized feeding, infection and inoculation, breeding bag transition feeding and adaptive exercise to be bred to 6-7 years old and wait for putting, and all the steps are carried out in an indoor environment; larva putting: selecting proper putting time, place and weather, adopting a hole digger to dig holes, putting 6-7-age larvae subjected to adaptive exercise into a field environment together with the breeding bags, and backfilling soil and turf dug out by hole digging after putting; management after putting: covering waterproof cloth at the putting position, removing the waterproof cloth after a preset time, covering weeding cloth at the putting position after winter, and uncovering the weeding cloth 1-2 months after the soil is unfrozen; harvesting is conducted before sporocarp is mature but spores are not ejected in quantity, and cordyceps sinensis is obtained. The survival rate of field larvae can be increased, the culture period of field cordyceps sinensis is shortened, and the quality of cordyceps sinensis is improved.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of artificial cultivation of traditional Chinese medicinal materials, in particular to a rearing method for cultivating larvae of the bacteria-susceptible bat moth to old age (6-7 instars) in a controlled indoor environment and then releasing them into a suitable outdoor environment for infection and fruiting body development. BACKGROUND

[0002] Ophiocordyceps sinensis is a rare and precious traditional Chinese medicinal material with extremely high medicinal and economic value. Due to overexploitation and environmental degradation, wild resources are on the verge of exhaustion, and artificial cultivation is urgently needed.

[0003] Bottlenecks in existing cultivation models: Complete indoor simulation: high technical difficulty, high risk, high cost, and the cultivated Ophiocordyceps sinensis has gaps in morphology, active ingredients, and other aspects compared to wild products, with low market acceptance. Traditional semi-wild rearing: usually releasing low-age larvae (1-3 instars) into the wild. The main problems are: low survival rate in the wild: low-age larvae have weak resistance to environmental changes (sudden temperature changes, drought, heavy rain), natural enemies (beetles, spiders, birds), and high mortality. Long rearing cycle: larvae need to complete most of their growth (from low-age to mature larvae) in the wild environment, and the entire growth and infection process takes 3-4 years, resulting in high risk, high cost, and slow capital turnover. Unstable quality: due to the long growth period in the wild and many uncontrollable environmental factors, larvae may develop poorly, and Ophiocordyceps sinensis fruiting bodies may not develop well, resulting in uneven quality of Ophiocordyceps sinensis (such as size, fullness, and active ingredient content). SUMMARY

[0004] The present application provides a semi-wild rearing method for Ophiocordyceps sinensis, which can improve the survival rate of larvae in the wild, shorten the cultivation cycle of Ophiocordyceps sinensis in the wild, and improve the quality of Ophiocordyceps sinensis.

[0005] According to the first aspect of the present application, one or more embodiments of the present application provide a semi-wild rearing method for Ophiocordyceps sinensis, comprising the following steps:

[0006] S1, indoor larva hatching and feeding: hatching larvae from self-reproduced bat moth eggs in a controllable indoor environment; the larvae are sequentially subjected to concentrated feeding, infection inoculation, transition feeding in a cultivation bag, and adaptive exercise to cultivate 6-7 instars for release; the concentrated feeding, infection inoculation, transition feeding in a cultivation bag, and adaptive exercise are all carried out in an indoor environment;

[0007] S2, larva release: selecting appropriate release time, location, and weather, using a hole puncher to punch holes, and then releasing 6-7 instar larvae that have undergone adaptive exercise together with the cultivation bag into the wild environment, and backfilling the soil and turf excavated by the hole punching after release;

[0008] S3. Post-deployment management: Cover the placement site with a waterproof sheet and remove it after a predetermined period of time. Management will be carried out in stages based on the placement time, including:

[0009] After winter, cover the placement area with weed cloth and uncover it 1-2 months after the soil thaws;

[0010] From spring to summer of the following year, shade the area where the seeding is placed, regularly check soil moisture and add water to keep it moisturized;

[0011] S4. Harvesting: Harvesting is done when the fruiting body is mature but before the spores are ejected in large quantities, retaining the intact insect body and fruiting body to obtain Cordyceps sinensis.

[0012] According to the above technical solution of the present invention, the following improvements can also be made:

[0013] Among them, in step S1, the insect eggs are fully mixed with the bacterial soil and plant feed during infection and inoculation, and the weight ratio of the number of insect eggs to the bacterial soil and plant feed is 1:1:3.

[0014] Among them, plant feed includes ginseng fruit, Polygonum villosa, Fagopyrum serrata, and Potentilla scabra.

[0015] The plant feed comprises, by weight percentage, 20-30% of human fruit, 25-30% of Polygonum villosa, 25-30% of golden buckwheat, and 10-30% of Herba Potentillae.

[0016] Wherein, in said step S1, the larvae are stably cultured to 3-4 years old during the concentrated rearing stage, and the larvae are cultured to 6-7 years old during the transitional rearing stage in the breeding bag, and the duration of adaptive training is at least one week.

[0017] During the centralized rearing stage, the larvae are cultivated in a square rearing box with a lid. The weight ratio of plant feed to substrate in the rearing box is 3:1. The substrate is a mixture of soil and coconut coir in a weight ratio of 1:1, with an average density of 2-3 heads per cubic centimeter. When the larvae develop to 3-4 years old, the centralized reared larvae are transferred to the breeding bags for rearing. At this time, there are 1-2 larvae in each breeding bag, and the plant feed in the breeding bag is ginseng fruit and golden buckwheat in a weight ratio of 3:2.

[0018] Among them, adaptive training includes simulating the humidity, light and day and night temperature difference in the environment where the larvae are released. After the larvae adapt to the initial temperature, the day and night temperature difference range is gradually adjusted from 5-10℃ to above 10℃. The daily temperature difference is controlled at 2-3℃ compared with the previous day. When the day and night temperature difference reaches above 10℃, this state is maintained for at least one week.

[0019] Among them, the adaptive training before release lasts for 7-15 days, the daytime temperature gradually drops to 15℃ for 14 hours, the nighttime temperature gradually drops to 5℃ for 10 hours, and the humidity fluctuates between 65% and 85%.

[0020] Among them, the breeding bag is made of degradable, breathable and water-permeable material and can be bitten by larvae; the breeding bag is a cylindrical bag with a diameter of 5 cm and a height of 10 cm, the bottom is sealed, and several annular supports are arranged at intervals at the bottom and middle of the bag, and the annular supports are made of degradable materials; the breeding bag 1 is filled with plant feed and matrix, the plant feed includes ginseng fruit and golden buckwheat in a weight ratio of 3:2, and the matrix is ​​a mixture of soil and coconut bran in a weight ratio of 1:1. The bag mouth of the breeding bag can be sealed by bundling, while the bottom and middle maintain their original shape under the action of the annular supports to provide space for larvae to move.

[0021] The larvae are placed at a density of 30-50 per square meter in step S2.

[0022] The beneficial effects of the present invention are as follows: the present invention provides a semi-wild cultivation method for Cordyceps sinensis, which significantly shortens the field cultivation period compared with the existing technology:

[0023] The most vulnerable, slow-growing young stages (ages 1-6) are completed efficiently and safely indoors, while the field only requires the later growth and fruiting body development stages of age 6-7. This shortens the field cycle to approximately six months (or even less), significantly reducing long-term field risks and management costs, and accelerating capital turnover.

[0024] Significantly improve the survival rate of larvae in the wild: the larvae released are older (6-7 years old), which have thicker body walls, stronger activity, and significantly better stress resistance (cold resistance, drought resistance, disease resistance, and mechanical damage resistance) than younger larvae. They can effectively resist adverse outdoor environments (temperature and humidity fluctuations, short-term droughts / rainfalls) and attacks by some natural enemies. The most critical feature is the use of breeding bags to further improve the larvae's adaptability and survival ability in the wild, and the survival rate can be increased by more than 30% compared to releasing younger larvae.

[0025] Significantly improves Cordyceps harvesting efficiency: In the wild, Cordyceps sinensis often grows in underbrush, making it difficult to spot and harvest. However, by covering the area with weed cloth, the herbaceous plant grows slowly. However, Cordyceps sinensis fruiting bodies develop normally in darkness, while strong light inhibits their growth. Therefore, by removing the weed cloth after the fruiting bodies emerge, the Cordyceps sinensis can be easily found and harvested.

[0026] Guarantee and improve the quality of Cordyceps sinensis: ①Strong and well-nourished insect body: Controllable indoor environment ensures the healthy and rapid growth of larvae to old age, large and full insect body, and rich nutrient reserves (outdoor release of small age larvae, slow growth, at least 3 years to sclerotize, and small and thin insect body), which provides an optimal nutritional basis for the later infection of insect body by Cordyceps sinensis and the development of fruiting body. ②Significantly reduce the content of heavy metal arsenic: Due to the shortening of the wild rearing period, heavy metals accumulate on the larvae for a short time, and the content of heavy metal arsenic in Cordyceps sinensis formed is also low. ③High infection success rate: Indoor centralized infection can greatly improve the infection rate of larvae. ④Good development of fruiting body: The combination of high-quality insect body and natural outdoor environmental pressure (such as low temperature, UV, etc.) is conducive to inducing the development of fruiting body to be closer to the wild form (such as thick and natural color of stipe) and accumulating specific secondary metabolites (effective ingredients), and the quality is better and more stable.

[0027] Reduce overall risk and cost: Although there is a certain cost in indoor stage, the cycle is controllable, efficient and loss is small. The shortening of outdoor stage and high survival rate greatly reduces the overall failure risk caused by uncontrollable factors (extreme weather, serious pests and diseases, human damage) in long-term outdoor environment, and the comprehensive cost-effectiveness is higher. BRIEF DESCRIPTION OF DRAWINGS

[0028] Figure 1 The figure is a schematic diagram of the rearing process of the embodiment of the present application;

[0029] Figure 2 The figure is a schematic diagram of the cross-sectional structure of the breeding bag of the embodiment of the present application;

[0030] Figure 3 The figure is a schematic diagram of the step flow of the method of the embodiment of the present application.

[0031] Explanation of reference signs: breeding bag 1, annular support 2. DETAILED DESCRIPTION

[0032] In order to make the purpose, technical scheme and advantages of the present disclosure clearer, the present disclosure is further described in detail below in combination with specific embodiments and with reference to the drawings.

[0033] It should be noted that the technical terms or scientific terms used in one or more embodiments of the present application should be understood as the general meaning understood by those skilled in the art to which the present disclosure belongs, unless otherwise defined. The terms "first", "second", and similar words used in one or more embodiments of the present application do not represent any order, quantity or importance, but are only used to distinguish different components. "Include" or "contain" and similar words mean that the elements or objects before the word cover the elements or objects listed after the word and their equivalents, and do not exclude other elements or objects. "Connected" or "connected" and similar words are not limited to physical or mechanical connections, but can include electrical connections, whether direct or indirect. "Up", "down", "left", "right" and the like are only used to represent the relative positional relationship, and when the absolute position of the described object changes, the relative positional relationship may also change accordingly.

[0034] Embodiments

[0035] As Figure 1-Figure 3 shown, the method for semi-wild cultivation of Cordyceps sinensis according to one or more embodiments of the present application comprises the following specific steps:

[0036] S1, take the eggs laid by the indoor self-reproducing bat moth and place them in a sterile culture dish in a 12℃ (development period 37-42d), 80% humidity artificial climate box;

[0037] Take out the eggs at 30d of development period (before hatching) and mix them with fungus soil and plant feed, with 500 eggs per box, 1.5kg of plant feed, 500g of matrix soil (40%), and square feeding boxes (30*20*15cm) with covers are used as feeding tools, which are placed in a feeding room with a temperature of 12-15℃ and a humidity of 75%;

[0038] Feed the plant feed (formula: human fruit 25%, pearl bud 30%, gold buckwheat 30%, and white grass 15%);

[0039] The feeding density is controlled at an average of 2cm per larva 3 (calculated according to the hatching rate) ;

[0040] After about 2-3 months of feeding, the larvae develop to 3-4 instars, and the concentrated feeding larvae are transferred to breeding bags 1 for feeding, with 1 larva per bag, and the breeding bags contain a mixture of coconut husk and plant feed (plant feed uses human fruit 6g+white grass 4g), and continue to be fed in the feeding room for about 1 month, with the feeding environment unchanged, and the development is stable to 6-7 instars (body length can reach 4-5cm) ;

[0041] Adaptive exercise: gradually adjust the temperature and humidity of the feeding room, simulate the natural growth environment of the bat moth larvae in the alpine meadow, adjust the temperature to a high temperature difference environment of 5-10℃, finally adjust the temperature to 15℃ for 14h in the daytime and 5℃ for 10h at night, and the humidity fluctuation range is 65%-85%. After 1 week of feeding at the final temperature, it is used for field release.

[0042] S2, field release:

[0043] It can be understood that the release time is selected: the release is carried out 1-2 months before the frozen soil, and the release time is selected in July-October at an altitude of 2000-4000 meters;

[0044] The release time is selected in August, and the altitude of the Sichuan Kangding production area is 3000 meters. At this time, the temperature is suitable (5-20℃), the vegetation is selected as the alpine meadow semi-sunny slope, the soil is loose, the drainage is good, and the area is rich in humus, and the soil layer thickness reaches more than 25 cm.

[0045] The release site is selected: the altitude, vegetation, soil type, temperature and humidity, etc. meet the growth habits of the bat moth larvae and their symbiotic bacteria in the original or close to the original environment. In the altitude of 2000-4000 meters, the meadow shrub vegetation covered by the grass family plants with dwarf shrubs, the soil layer is deep, the soil humidity is moderate 60%-70%, and the semi-sunny slope is not easy to accumulate water;

[0046] The release weather and time are selected: the weather is selected to be sunny in the morning 8-11 o'clock.

[0047] The release operation is selected: a hole puncher with a diameter slightly larger than the breeding bag 1 is selected, the hole depth is 10 cm, the hole diameter is 6 cm, the breeding bag 1 with larvae is put into the hole, and then the grass and soil dug out by the hole puncher are backfilled. In this way, it is ensured that the larvae have sufficient feed in the initial stage of entering the soil, and if there is no feed in the bag, the larvae can also bite the breeding bag 1 to enter the soil layer to find food, thereby improving the survival rate of the larvae. The breeding bag 1 is made of degradable, breathable and water-permeable material, and can be bitten by the larvae; the breeding bag 1 is a cylindrical bag with a diameter of 5 cm and a height of 10 cm, the bottom is sealed, and the bag has a plurality of annular supports 2, and the annular supports are made of degradable material.

[0048] In the selected plot, a hole is punched and dug with a hole puncher, the breeding bag 1 is put into the hole, the breeding bag 1 used in the embodiment is made of non-woven fabric; the part of the soil and grass dug out is backfilled. The release density is about 30 heads per square meter.

[0049] S3, field management:

[0050] After the release is completed, the waterproof cloth is covered, and the waterproof cloth can be removed after about one week;

[0051] Regular inspections, with a focus on preventing wild hogs and badgers (if repeated, fences can be added) and ant infestations (if ant nests are found, physical removal or the use of environmentally friendly repellents is carried out promptly);

[0052] In November, the herbaceous plants begin to wither and fall over, so cover them with weed cloth. Then, in April of the following year, about a month after the soil thaws, remove the weed cloth.

[0053] S4. Harvesting

[0054] From late April to mid-May of the following year, observe the fruiting bodies (grasses) emerging from the soil. These are approximately 3-6 cm long. Carefully dig out the fruiting bodies, along with the grass and insects, using a specialized tool (such as a small shovel). Remove any debris and dry in the shade or at low temperatures.

[0055] Results: In this example, the larvae were reared indoors for 4-5 months and then raised outdoors for approximately 6-7 months. The survival rate of released larvae in the wild (until overwintering) reached 75%, and the proportion of commercially available Cordyceps sinensis (full larvae and intact stroma) successfully reached 35% of the released quantity. The harvested Cordyceps sinensis had similar appearance (larvae size, stroma color, and plumpness) and content of major active ingredients (such as adenosine and cordycepin) to high-quality wild Cordyceps sinensis from the same region. The average weight of semi-wild-raised Cordyceps sinensis was higher than that of wild Cordyceps sinensis from the same region (average weight of 0.3g / root for semi-wild-raised Cordyceps sinensis compared to 0.25g / root for wild Cordyceps sinensis from the same region). The heavy metal content, particularly arsenic content (less than 1mg / kg, meeting national safety standards), was lower than that of wild Cordyceps sinensis.

[0056] Comparative experiment (traditional low-age delivery):

[0057] Artificially hatched 2nd-3rd instar larvae (approximately 1-1.5 cm in length) were released in the same field area and managed as in Example 1. Results: The field rearing period required more than three years. Larval mortality exceeded 60% within six months of release (primarily due to environmental fluctuations and natural enemies). Only 5% of the Cordyceps sinensis ultimately reached commercial size, and the individual fungi were generally small, with significant quality fluctuations.

[0058] Table 1 Survival rate, grass maturity rate and growth cycle of grass released at different ages

[0059]

[0060] Table 2 Comparison of the quality of Cordyceps sinensis at different ages

[0061]

[0062]

[0063] Conclusion: By comparing the experimental data in Table 1 and Table 2, the strategy of the present application, i.e. precisely cultivating robust older (6-7 years old) larvae indoors and releasing them into suitable wild environments at the best time, effectively solves the core problems of long cycle, low survival rate and unstable quality in traditional semi-wild cultivation, and provides an efficient and reliable technical solution for the sustainable utilization of Cordyceps sinensis resources and the healthy development of the industry.

[0064] While the preferred embodiments of the application have been described, one skilled in the art will appreciate that, given the benefit of the present disclosure, additional modifications and variations to the preferred embodiments can be made without departing from the spirit and scope of the application. Accordingly, it is intended that the appended claims encompass all such modifications and variations as fall within the scope of the present application.

[0065] Obviously, various modifications and changes can be made to the present application by those skilled in the art without departing from the spirit and scope of the present application. Thus, it is intended that the present application encompass all such modifications and changes as fall within the scope of the claims and their equivalents.

Claims

1. A method for semi-wild cultivation of Cordyceps sinensis, characterized in that: The following steps are involved: S1. Indoor larval hatching and rearing: self-propagated bat moth eggs are hatched in a controlled indoor environment to obtain larvae; the larvae are sequentially reared, infected and inoculated, reared in breeding bags, and adapted to be raised to 6-7 years old and ready for release, wherein the centralized rearing, infected and inoculated, reared in breeding bags, and adapted to be trained indoors; S2. Larvae release: Choose the appropriate time, location, and weather for release. Use a hole puncher to dig holes, then release the 6-7th-instar larvae that have undergone adaptive training into the wild environment along with the breeding bags. After release, appropriately backfill the holes with soil and turf. S3. Post-deployment management: Cover the placement site with a waterproof sheet and remove it after the scheduled time; Based on the delivery time, management is carried out in stages, including: After winter, cover the placement area with weed cloth and uncover it 1-2 months after the soil thaws; From spring to summer of the following year, shade the area where the seeding is placed, regularly check soil moisture and add water to keep it moisturized; S4. Harvesting: Harvesting is done when the fruiting body is mature but before the spores are ejected in large quantities, retaining the intact insect body and fruiting body to obtain Cordyceps sinensis.

2. The method for semi-wild cultivation of Cordyceps sinensis according to claim 1, characterized in that: In step S1, the insect eggs are fully mixed with the bacterial soil and plant feed during infection and inoculation, and the weight ratio of the number of insect eggs to the bacterial soil and plant feed is 1:1:

3.

3. The method for semi-wild cultivation of Cordyceps sinensis according to claim 2, characterized in that: Plant feed includes ginseng fruit, Polygonum villosa, golden buckwheat, and Herba Potentillae.

4. The method for semi-wild cultivation of Cordyceps sinensis according to claim 3, characterized in that: The plant feed comprises, by weight percentage, 20-30% of human fruit, 25-30% of Polygonum villosa, 25-30% of golden buckwheat, and 10-30% of Potentilla discolor.

5. The method for semi-wild cultivation of Cordyceps sinensis according to claim 1, characterized in that: In step S1, the larvae are stably cultured to 3-4 years old during the concentrated rearing stage, and the larvae are cultured to 6-7 years old during the transitional rearing stage in the breeding bag. The duration of adaptive training is at least one week.

6. The method for semi-wild cultivation of Cordyceps sinensis according to claim 5, characterized in that: During the centralized rearing stage, the larvae are cultivated in square rearing boxes with lids. The weight ratio of plant feed to substrate in the rearing boxes is 3:

1. The substrate is a mixture of soil and coconut coir in a weight ratio of 1:1, with an average density of 2-3 larvae per cubic centimeter. When the larvae develop to 3-4 years old, the centralized reared larvae are transferred to breeding bags for rearing. At this time, there are 1-2 larvae in each breeding bag, and the plant feed in the breeding bags is ginseng fruit and golden buckwheat in a weight ratio of 3:

2.

7. The method for semi-wild cultivation of Cordyceps sinensis according to claim 1, characterized in that: Adaptive training includes simulating the humidity, light and day and night temperature difference in the environment where the larvae are released. After the larvae adapt to the initial temperature, the day and night temperature difference range is gradually adjusted from 5-10℃ to above 10℃. The daily temperature difference is controlled at 2-3℃ compared with the previous day. When the day and night temperature difference reaches above 10℃, this state is maintained for at least one week.

8. The method for semi-wild cultivation of Cordyceps sinensis according to claim 1, characterized in that: The adaptation training before release lasts for 7-15 days. The daytime temperature gradually drops to 12℃ for 14 hours, the nighttime temperature gradually drops to 5℃ for 10 hours, and the humidity fluctuates between 65% and 85%.

9. The method for semi-wild cultivation of Cordyceps sinensis according to claim 1, characterized in that: The breeding bag is made of degradable, breathable and water-permeable material and can be bitten by larvae; the breeding bag is a cylindrical bag with a diameter of 5 cm and a height of 10 cm, with a sealed bottom. Several annular supports are provided at intervals at the bottom and middle of the bag, and the annular supports are made of degradable materials; the breeding bag is filled with plant feed and matrix, the plant feed includes ginseng fruit and golden buckwheat in a weight ratio of 3:2, and the matrix is ​​a mixture of soil and coconut bran in a weight ratio of 1:

1.

10. The method for semi-wild cultivation of Cordyceps sinensis according to claim 1, characterized in that: In step S2, the larvae are placed at a density of 30-50 per square meter.

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