CN112154857A

CN112154857A - Wild-simulated cultivation method for improving amino acid content of winter fungi

Info

Publication number: CN112154857A
Application number: CN202011210574.3A
Authority: CN
Inventors: 毕宁; 侯俊; 周聿; 王彩云; 何忠国; 肖艳; 李斌
Original assignee: Bijie Forestry Research Institute
Current assignee: Bijie Forestry Research Institute
Priority date: 2020-11-03
Filing date: 2020-11-03
Publication date: 2021-01-01
Anticipated expiration: 2040-11-03
Also published as: CN112154857B

Abstract

The invention belongs to the field of edible fungus cultivation, and particularly relates to a wild-simulated cultivation method for improving amino acid content of winter dictyophora; the cultivation material is prepared by adopting the waste gastrodia elata firewood as the main material, so that the connection between the gastrodia elata industry and the winter fungus industry is realized, and the economic benefit is improved; and the wood and bamboo branches in the cultivation material auxiliary materials are respectively treated, so that the dictyophora can fully absorb the nutrition of the cultivation material, the content of the content is improved, the content of the produced amino acid of the dictyophora reaches 16.95 percent, and the content of glutamic acid (Glu) reaches 2.07 percent; meanwhile, management parameters are adjusted according to different growth stages of the winter dictyophora, the cultivation time of the winter dictyophora is shortened, the winter dictyophora is listed in advance, and the economic benefit is maximized.

Description

Wild-simulated cultivation method for improving amino acid content of winter fungi

Technical Field

The invention belongs to the field of edible fungus cultivation, and particularly relates to a wild-simulated cultivation method for improving amino acid content of winter dictyophora.

Background

The winter fungus is also known as subphylum bamboo, bamboo fungus and dictyophora indusiata, and is a fungus of the genus phalloidea in the family of phalloideae. The hyphae grow white, are in goose feather shape, are stout, are large in bamboo egg body shape, are white, are slightly thin at two ends, are in sponge shape and are hollow; the color of the sporocarps is not changed when the sporocarps are exposed to sunlight and stimulated, the sporocarps have a special odor, and the sporocarps mostly grow in forests or on plots under bamboo forests in summer and autumn, grow singly to grow in groups and are mainly distributed in high-altitude areas such as Yunnan, Guizhou, Liaoning and Guangdong and the like. The winter fungus is a rare edible fungus, has delicious taste, crisp mouthfeel and extremely high nutritional value. According to related researches, the dictyophora indusiata fruiting body contains various components such as maltol, uronic acid glycan, formaldehyde, acetaldehyde, phenylacetaldehyde, phenyl crotonaldehyde, methyl mercaptan, hydrogen sulfide, dihydrochalcone, phenylacetic acid, acetic acid and propionic acid. The stipe, the truffle and the fruiting body of the winter fungus are used as medicines, have sweet and light properties and warm property, have the effects of activating blood circulation to stop pain, dispelling wind and removing dampness, and can be used for treating rheumatalgia. The winter fungus can inhibit the growth of putrefying bacteria, and can be used as short-term antiseptic for food. At present, the market mainly depends on wild collection and processing, and the yield cannot meet the market demand. With the increase of market demand, artificial planting of the winter dictyophora becomes an important way for meeting the market demand.

At present, the following problems mainly exist in the cultivation of the winter dictyophora: (1) the strain production is also a method for producing the strain by using the separation of wild edible fungus eggs to prepare mother strains by farmers with mastered technology. (2) The seed sources are disordered, and the winter fungus mainly comprises a honeycomb long-handle type, a smooth and tender type and a short-handle meat type, wherein the honeycomb long-handle type is high in yield, poor in taste and low in market price; the latter two tastes are better, the yield is slightly lower and the price is slightly higher. In production, most strain producers collect and separate bamboo eggs by themselves, and select varieties to directionally culture excellent varieties rarely considered, so that provenance in the market is very disordered. (3) The strain quality standard is lacked, and the strain production quality is difficult to guarantee. Most strain producers have simple and crude production conditions, the strain production by adopting the traditional method has poor production equipment and environment, and the strain of the Chinese fungus (the phallus impudicus) grows slowly, so that the strain pollution rate is high, and the phenomenon of mushroom failure and the like often occurs when the strain is applied to production. (4) The harvesting and processing of the winter fungus also stay in the primary stage, mainly comprising scattered harvesting and respective processing by farmers, and mainly drying with coal fire in the processing process. The baking methods do not form a fixed processing technology, the temperature is not easy to regulate and control, the water-removing time is long, the winter dictyophora can be naturally lengthened, the tissue density is sparse, and the quality is greatly reduced; meanwhile, the sulfur content cannot be guaranteed not to exceed the standard by baking with coal fire, and the yellow and brown baking of the dictyophora phalloidea are easily caused, so that the quality of the dictyophora phalloidea is greatly reduced.

Patent document No. CN201610947621.X discloses a layered three-dimensional cultivation method of Potentilla chinensis, which utilizes agricultural waste fermentation to prepare culture medium of Dictyophora phalloidea, and adopts layered three-dimensional cultivation of Dictyophora phalloidea, so as to shorten cultivation period and improve production benefit. However, during the spawn running management, hyphae grow on the soil surface, and in this case, if the humidity is too high, a large amount of ineffective primordia is formed, which results in waste of nutrients and further affects the yield. The main research of the patent is to adopt a fermentation material bacterial bed to cultivate the winter dictyophora, and the heavy metal content and the content of effective active ingredients of the winter dictyophora are not researched.

Patent document CN201811580375.4 discloses a strain preparation method and application of three excellent strains of dictyophora indusiata, wherein the three excellent strains of the dictyophora indusiata are obtained by obtaining pure strains by a tissue separation method of the dictyophora indusiata, and performing strain purification, indoor evaluation, fruiting characteristic and the like. Wherein the cultivation material contains wood chips or wood blocks, arrow bamboo leaves or broad leaf leaves. However, the technical purpose is to select and prepare pure culture of the winter dictyophora, and the cultivation of the pure culture of the winter dictyophora can be realized by adopting a traditional method or a fermentation material cultivation method, and the content of the prepared pure culture of the winter dictyophora is not researched.

The main planting modes of the winter dictyophora mainly include the following modes: under the forest, wild planting is imitated, the winter edible fungus is interplanted by corn, and the winter edible fungus is interplanted by hemp. The winter dictyophora is mainly interplanted with other crops, and when the winter dictyophora is interplanted with other crops, the conditions such as humidity required by growth of the winter dictyophora are higher, so that the growth of the interplanted crops is influenced to a certain extent, and if the growth conditions of the interplanted crops are emphasized, the growth vigor and the quality of the winter dictyophora are influenced; in the traditional cultivation, the growth speed of the winter dictyophora is low, and the production conditions of most strain producers are simple and crude, so that pollution, heavy metal accumulation, nutrient content of the winter dictyophora and the like are not emphasized, the quality of the produced winter dictyophora is uneven, the market time is late, and the economic benefit is reduced. The specific planting method comprises a wood block planting method, a wood chip planting method and a wood chip and straw mixed planting method. The methods can realize planting of the winter dictyophora but few research reports are reported at present on how to improve the quality of the winter dictyophora and further improve the economic benefit of the winter dictyophora.

Disclosure of Invention

The invention provides a wild-imitating cultivation method for improving the amino acid content of dictyophora phalloidea to solve the problems.

The method is realized by the following technical scheme:

a wild-imitating cultivation method for improving the amino acid content of dictyophora phalloidea comprises the following steps:

1. selecting land: selecting sandy soil below the waist, cool and moist and well ventilated, and avoiding selecting unventilated pit and pit lands and places with frequent termite movement; the land can be selected under bamboo forest, and the relative humidity of air is maintained at 70-85%.

2. Land preparation: the plots were arranged according to a furrow cultivation method or a nest cultivation method. The furrow cultivation method comprises scraping the rotten leaves of dead branches on the surface layer of the land, digging pits with depth of 15-20cm, width of 50cm and unlimited length according to the conditions of the land, leveling the bottom of the pits, loosening the soil with thickness of 3-4cm (water retention effect).

A small-nest type cultivation method: digging small pits with the depth of 15-20cm, the width of 50cm and the length of 60cm according to the conditions of the land mass, leveling the pit bottom, loosening the soil with the pit bottom thickness of 3-4 cm.

3. Cultivation material treatment

Preparing materials: the main material is 20-30 parts by weight of rhizoma gastrodiae waste firewood; the auxiliary materials comprise the following raw materials in parts by weight: 10-15 parts of broad-leaved tree wood, 5-7 parts of carbonized bamboo branches and 10-15 parts of bamboo leaves.

Preparation of main materials: soaking rhizoma Gastrodiae waste firewood in 2% lime water for 10-15 hr, and washing with clear water to obtain the main material.

Preparing auxiliary materials:

a. preparing carbonized bamboo branches: cutting bamboo branches into segments with length of 4-7cm, soaking in hydrochloric acid, washing with clear water, draining, roasting, sieving excessive carbon powder for use, and reserving bamboo segments to obtain carbonized bamboo branches;

further, the roasting is to roast the drained bamboo branch sections for 2 to 3 hours at the temperature of 180 ℃ and 220 ℃;

b. preparing auxiliary materials: cutting broad leaf tree wood into blocks with side length of 5-8cm, spraying a layer of carbon powder screened in the step a on the surface of the broad leaf tree wood after the surface is wetted by a semi-degradation liquid, placing the broad leaf tree wood for 2-3 days under the conditions that the temperature is 30-40 ℃ and the humidity is 60-70%, disinfecting the broad leaf tree wood by ultraviolet rays with the wavelength of 200-275nm for 50-70min, and mixing the broad leaf tree wood with carbonized bamboo branches and bamboo leaves to obtain the auxiliary material.

Further, the semi-degradation liquid is obtained by inoculating white rot fungi into a liquid culture medium and culturing for 10-15 days; wherein the liquid culture medium is prepared by taking monosaccharide as a carbon source and taking an organic matter as a nitrogen source.

Here, a point is to be emphasized: the prepared main material and the auxiliary material need to be stored respectively and can not be mixed for placement.

Further, the cultivation material can also be prepared by mixing rhizoma gastrodiae waste firewood and fresh firewood blocks in a mass ratio of 1: 2.

4. Sowing:

the cultivation material is paved in a cross lamination mode: laying a layer of main material, a layer of strain and a layer of auxiliary material from bottom to top, laying a layer of main material on the surface, and finally covering soil and covering pine needles or ferns. Wherein the usage amount of the main materials at the bottom layer is just enough to cover the soil in the pit, and the main materials are tightly beaten by a hoe back; breaking the strain into small pieces with diameter of about 3cm, and sowing on the bottom main material at a distance of 3-4cm and a sowing amount of 2-2.5kg/m²Seeding is completed before 10 months to 3 months of the next year; coating a thin layer of adjuvants on the strain, wherein the bamboo leaf has effect of drawing mycelium at a concentration of 0.15kg/m²Spraying a layer of white sugar; laying a layer of main material with the thickness of 6-8 cm; finally covering soil, wherein the soil covering is preferably thin in the place difficult to dry and is controlled to be 3-5 percentThe cm is proper, the thickness of the covering soil is preferably controlled between 5cm and 8cm in the places easy to dry, the covering is preferably thin when the humidity is high in the sowing period, the covering soil is preferably thickened slightly when the humidity is high, and the covering soil is arc-shaped (turtle back shape). The covering is preferably made of pine needles and ferns, and is preferably covered with a cover of about 2cm in thickness.

5. Daily management

And (3) hypha growth stage: the humidity requirement is as follows: the planting material, the soil layer and the surface of the soil layer are preferably not whitened and dried; if the material turns white and dry, a small amount of water can be sprayed and the wood leaves are covered, but in rainy days, too much water cannot flow into the material, so as to avoid hyphae from being drowned due to too high humidity. The temperature is kept at 16-24 ℃, and the light irradiation is optimal for growth under the conditions of half yin and half yang.

And (3) an original base forming stage: culturing by temperature difference method under the condition of unchanged humidity and illumination condition after mycelium is formed, specifically, after the mycelium of the dictyophora phalloidea appears, adjusting the temperature to 18-24 ℃ in the daytime, adjusting the temperature to 7-10 ℃ at night, and adjusting the temperature to the formation of the dictyophora phalloidea primordium every day.

And (3) a sub-entity forming stage: after the winter fungus primordium is formed, keeping the temperature at 8-22 ℃ and the humidity at 75-85%, and culturing by adopting an alternative illumination method, specifically, after the winter fungus primordium is formed, starting illumination for 1-2h by 400-fold-power 500 lx-intensity light at 7 points every day, then switching illumination for 1-2h by 100-fold-power 200 lx-intensity light, and then supplementing illumination for 30-60min by 50-75 lx-intensity light at night.

6. Harvesting and processing

When the winter fungus is collected, the thalli is cut off from the bottom of the fungus tray by a knife without pulling by hands so as to avoid damaging the thalli and influencing the re-balling of the winter fungus. The winter edible fungus can be dried by a drying room and a dryer, and can also be made into a simple drying box or a baking cage. The anthracite coal is required to be roasted as fuel, if the yield is low, the anthracite coal can be directly roasted by using charcoal fire, and firewood fire or sulfur-containing coal is avoided; the method is suitable for being carried out under low temperature conditions, and ventilation and dehumidification are required in the baking process. The baking sieve sheet is washed clean and dried in the air. The fungus cover is required to be lightly handled and lightly placed in the baking process to be prevented from being broken, and the restoration and the integrity of the fungus cover are required to be paid attention to by the winter fungi. And packaging the dried winter fungus by using a plastic bag, and sealing to prevent deterioration and yellowing caused by further moisture absorption.

In conclusion, the beneficial effects of the invention are as follows: according to the method, the cultivation material is prepared by taking the waste gastrodia elata firewood as a main material, so that the connection between the gastrodia elata industry and the winter fungus industry is realized, and the economic benefit is improved; and the wood and bamboo branches in the cultivation material auxiliary materials are respectively treated, so that the dictyophora can fully absorb the nutrition of the cultivation material, the content of the content is improved, the content of the produced amino acid of the dictyophora reaches 16.95 percent, and the content of glutamic acid (Glu) reaches 2.07 percent; meanwhile, management parameters are adjusted according to different growth stages of the winter dictyophora, the cultivation time of the winter dictyophora is shortened, the winter dictyophora is listed in advance, and the economic benefit is maximized.

The Guizhou belongs to plateau terrain, the smooth and fertile land is less, the land is in a karst landform, the land is selected on a hillside or under a bamboo forest when being selected, the under-forest weeds and dead branches and fallen leaves can be fully utilized as culture materials, the wild-imitating effect is achieved, forest fire can be reduced, under-forest waste can generate economic benefits, and meanwhile, the intensive utilization of land resources is facilitated.

The method for planting the winter edible fungus by adopting the waste gastrodia elata fungus material is a good circular economy mode, realizes the connection of the gastrodia elata industry and the winter edible fungus industry, and improves the economic benefit. The bamboo branches are cut into segments, acid leaching and roasting are carried out, redundant sugar and impurities in the bamboo branches can be removed, meanwhile, the specific surface area and pores of the roasted bamboo branches are increased, heavy metals in soil are adsorbed while substances required by the winter dictyophora are provided, the heavy metal accumulation of the winter dictyophora is reduced, and the quality of the winter dictyophora is improved. Although wood chips and other raw materials are disclosed to be adopted in the existing dictyophora phalloidea cultivation material, the wood chips are directly used, the utilization rate is not high, in the application, broad-leaf tree wood is treated, the broad-leaf tree wood is cut into blocks, the surfaces of the broad-leaf tree wood are sprayed with semi-degradation liquid to be wet, then a layer of carbon powder screened out in the process of preparing carbonized bamboo branches is wrapped, the cut broad-leaf tree wood is placed for 2 to 3 days at the temperature of 30 to 40 ℃ and the humidity of 60 to 70 percent and then is subjected to ultraviolet disinfection, the main component of the semi-degradation liquid is white rot fungi, white rot fungi liquid is sprayed on the surfaces of the wood blocks and placed, lignin on the surfaces of the wood blocks is degraded by the white rot fungi, meanwhile, the integrity of most fibers is kept, so that the carbon powder can be better adhered to the wood blocks and can absorb heavy metal substances together with the carbonized bamboo branches; meanwhile, the wood blocks are not completely degraded by the white rot fungi, so that the nutrient components can be absorbed by the winter dictyophora more conveniently, the growth time of the winter dictyophora is further shortened, and the content of the winter dictyophora is improved. The wood blocks are subjected to ultraviolet disinfection treatment to kill white rot fungi and other infectious microbes, so that the influence on the growth of the winter dictyophora caused by nutrition competition with the winter dictyophora is avoided. The cultivation material is laid in a cross lamination mode, so that influence on air permeability caused by the fact that dry and wet materials in the cultivation material are mixed together to form a cluster is avoided, and carbon powder in the auxiliary material and the wet material are easily attached to the roots of the winter dictyophora after being mixed, so that the appearance and the growth of the winter dictyophora are influenced to a certain degree.

During the daily management period, the growth process of the dictyophora phalloidea is divided into three stages, and in the hypha growth stage, the temperature and humidity and the illumination conditions of semi-yin and semi-yang are kept so as to achieve the optimal conditions for the dictyophora phalloidea to germinate in the wild; after hypha is formed, stimulating the formation of the edible fungus primordium by adopting a temperature difference method, wherein the temperature difference between day and night reaches 17 ℃ at most, so that the edible fungus primordium can be quickly formed, and the cultivation time of the edible fungus is further shortened; after the primordium is formed, the humidity is adjusted to 75-85%, the rapid formation of fruiting bodies of the winter dictyophora can be guaranteed, meanwhile, the illumination is supplemented after the sun falls off, the cultivation time of the winter dictyophora is further shortened, the cultivation time of the winter dictyophora is shortened to about 7 months from about 9 months in the prior art, the winter dictyophora is listed in advance, the content of nutrient substances of the winter dictyophora is high, the selling price is high, and the economic benefit of farmers is further improved.

Detailed Description

The following is a detailed description of the embodiments of the present invention, but the present invention is not limited to these embodiments, and any modifications or substitutions in the basic spirit of the embodiments are included in the scope of the present invention as claimed in the claims.

Example 1

1. selecting land: selecting sandy soil below the waist, cool and moist and well ventilated, and avoiding selecting unventilated pit and pit lands and places with frequent termite movement; the land can be selected under bamboo forest, and the relative humidity of air should be kept at 70%.

2. Land preparation: the method comprises the steps of arranging the plots according to a ditch type cultivation method, scraping dead branches and rotten leaves on the surface layers of the plots aside, digging pits with the depth of 17cm, the width of 50cm and unlimited lengths according to conditions of the plots, leveling the bottoms of the pits, and loosening the soil with the thickness of 3cm at the bottoms of the pits.

3. Cultivation material treatment

Preparing materials: the main material comprises 25 parts by weight of rhizoma gastrodiae waste firewood; the auxiliary materials comprise the following raw materials in parts by weight: 12 parts of broad-leaved tree wood, 6 parts of carbonized bamboo branches and 12 parts of bamboo leaves.

Preparation of main materials: soaking rhizoma Gastrodiae waste firewood in 2% lime water for 13 hr, and washing with clear water to obtain the main material.

Preparing auxiliary materials:

a. preparing carbonized bamboo branches: cutting bamboo branches into sections with the length of 6cm, soaking the bamboo branches in hydrochloric acid, washing the bamboo branches with clear water, draining the bamboo branches, roasting the bamboo branches, screening redundant carbon powder for later use, and reserving bamboo sections to obtain carbonized bamboo branches;

further, the roasting is to roast the drained bamboo branch sections for 3 hours at the temperature of 200 ℃;

b. preparing auxiliary materials: cutting broad leaf tree wood into 5-8cm square, spraying semi-degradation liquid to wet the surface, wrapping with a layer of carbon powder screened in step a, standing at 35 deg.C and 65% humidity for 3 days, sterilizing with ultraviolet ray of 260nm wavelength for 60min, and mixing with carbonized bamboo branch and bamboo leaf to obtain adjuvant.

Further, the semi-degradation liquid is obtained by inoculating white rot fungi into a liquid culture medium and culturing for 13 days; wherein the liquid culture medium is prepared by taking 1.5 percent of glucose, 0.2 percent of peptone and 10.03 percent of vitamin B as raw materials, and the inoculation amount of the white rot fungi is 10 percent.

4. Sowing:

the cultivation material is paved in a cross lamination mode: laying a layer of main material, a layer of strain, a layer of auxiliary material and a surface from bottom to topLaying a layer of main materials, covering with 4cm thick soil, covering with arc soil, and covering with 2cm thick pine needles. Wherein the usage amount of the main materials at the bottom layer is just enough to cover the soil in the pit, and the main materials are tightly beaten by a hoe back; during sowing, breaking strains into small blocks with the diameter of about 3cm, and sowing the small blocks on the bottom main material, wherein the distance between the strains is 3cm, and the sowing amount is 2kg/m²Completing sowing in 11 months in the year; covering the strain with an adjuvant layer of about 1cm thick, wherein the bamboo leaf has hypha drawing effect of 0.15kg/m²Spraying a layer of white sugar; and laying a layer of main material with the thickness of 6 cm.

5. Daily management

And (3) hypha growth stage: the humidity requirement is as follows: the planting material, the soil layer and the surface of the soil layer are preferably not whitened and dried; if the material turns white and dry, a small amount of water can be sprayed and the wood leaves are covered, but in rainy days, too much water cannot flow into the material, so as to avoid hyphae from being drowned due to too high humidity. The temperature is kept at 20 ℃, and the light is optimally grown under the conditions of half yin and half yang.

And (3) an original base forming stage: after hyphae are formed, cultivating by adopting a temperature difference method under the condition that humidity and illumination conditions are unchanged, specifically, after the hyphae of the dictyophora phalloidea appear, adjusting the temperature to 21 ℃ in the daytime, adjusting the temperature to 8 ℃ at night, and adjusting the temperature to the formation of the dictyophora phalloidea primordium every day.

And (3) a sub-entity forming stage: after the winter fungus primordium is formed, keeping the temperature at 17 ℃ and the humidity at 80%, and cultivating by adopting an alternative illumination method, specifically, after the winter fungus primordium is formed, starting to irradiate for 1h with light with the intensity of 450lx at 7 points every day, then irradiating for 1h with light with the intensity of 150lx, and then supplementing the light for 50min with light with the intensity of 75lx at night.

6. Harvesting and processing

The cultivated winter dictyophora can be harvested at the bottom of 5 months and at the beginning of 6 months in the next year, and the thalli is cut off from the bottom of the fungus tray by a knife when the winter dictyophora is harvested without pulling by hands so as to avoid damaging the thalli and influencing the re-balling of the winter dictyophora. Drying the winter edible fungus by adopting a drying room, and baking by using anthracite coal as fuel; ventilating and dehumidifying in the baking process. The fungus cover is required to be lightly handled and lightly placed in the baking process to be prevented from being broken, and the restoration and the integrity of the fungus cover are required to be paid attention to by the winter fungi. And packaging the dried winter fungus by using a plastic bag, and sealing to prevent deterioration and yellowing caused by further moisture absorption.

Example 2

1. selecting land: selecting sandy soil below the waist, cool and moist and well ventilated, and avoiding selecting unventilated pit and pit lands and places with frequent termite movement; the land can be selected under bamboo forest, and the relative humidity of air is maintained at 70%.

2. Land preparation: and (3) arranging the land blocks according to a pit type cultivation method, digging small pits with the depth of 15cm, the width of 50cm and the length of 60cm according to the conditions of the land blocks, leveling the pit bottoms, and loosening the soil with the pit bottoms of 3cm in thickness.

3. Cultivation material treatment

Preparing materials: the main material comprises 20 parts by weight of rhizoma gastrodiae waste firewood; the auxiliary materials comprise the following raw materials in parts by weight: 10 parts of broad-leaved tree wood, 5 parts of carbonized bamboo branches and 10 parts of bamboo leaves.

Preparation of main materials: soaking rhizoma Gastrodiae waste firewood in 2% lime water for 10 hr, and washing with clear water to obtain the main material.

Preparing auxiliary materials:

a. preparing carbonized bamboo branches: cutting bamboo branches into sections with the length of 4cm, soaking the bamboo branches in hydrochloric acid, washing the bamboo branches with clear water, draining the bamboo branches, roasting the bamboo branches, screening redundant carbon powder for later use, and reserving bamboo sections to obtain carbonized bamboo branches;

further, the roasting is to roast the drained bamboo branch sections for 2 hours at the temperature of 180 ℃;

b. preparing auxiliary materials: cutting broad leaf tree wood into 5cm square, spraying semi-degradation liquid to wet the surface, wrapping with a layer of carbon powder sieved in step a, standing at 30 deg.C and 60% humidity for 2 days, sterilizing with 200nm ultraviolet ray for 50min, and mixing with carbonized bamboo branch and bamboo leaf to obtain adjuvant.

Further, the semi-degradation liquid is obtained by inoculating white rot fungi into a liquid culture medium and culturing for 10 days; wherein the liquid culture medium is prepared by taking 1.5% of sucrose, 2% of wheat bran leach liquor and 10.03% of vitamin B as raw materials, and the inoculation amount of the white rot fungi is 10%.

4. Sowing:

the cultivation material is paved in a cross lamination mode: laying a layer of main material, a layer of strain and a layer of auxiliary material from bottom to top, laying a layer of main material on the surface, covering a layer of soil with the thickness of 3cm, covering the soil with an arc shape, and covering a layer of ferns with the thickness of 2cm on the soil. Wherein the usage amount of the main materials at the bottom layer is just enough to cover the soil in the pit, and the main materials are tightly beaten by a hoe back; breaking strains into small pieces with diameter of about 3cm during sowing, and sowing on the bottom layer of firewood, wherein the distance between the strains is 3cm, and the sowing amount is 2kg/m²Seeding is completed 12 months before the current year; covering the strain with a layer of 0.6cm thick adjuvant, wherein the bamboo leaf has hypha drawing effect of 0.15kg/m²Spraying a layer of white sugar; and laying a layer of main material with the thickness of 6 cm.

5. Daily management

And (3) hypha growth stage: the humidity requirement is as follows: the planting material, the soil layer and the surface of the soil layer are preferably not whitened and dried; if the material turns white and dry, a small amount of water can be sprayed and the wood leaves are covered, but in rainy days, too much water cannot flow into the material, so as to avoid hyphae from being drowned due to too high humidity. The temperature is kept at 16 ℃, and the light is optimally grown under the conditions of half yin and half yang.

And (3) an original base forming stage: after hyphae are formed, cultivating by adopting a temperature difference method under the condition that the humidity and the illumination condition are not changed, specifically, after the hyphae of the dictyophora phalloidea appear, adjusting the temperature to be 18 ℃ in the daytime, adjusting the temperature to be 7 ℃ at night, and adjusting the temperature to the formation of the dictyophora phalloidea primordium every day.

And (3) a sub-entity forming stage: after the winter fungus primordium is formed, keeping the temperature at 8 ℃ and the humidity at 75%, and cultivating by adopting an alternative illumination method, specifically, after the winter fungus primordium is formed, starting to irradiate with 400 lx-intensity light for 2h at 7 points every day, then irradiating with 100 lx-intensity light for 2h, and then supplementing with 50 lx-intensity light for 60min at night.

6. Harvesting and processing

The cultivated winter dictyophora can be harvested at the bottom of 7 months in the next year, and the thallophora is cut off from the bottom of the fungus tray by a knife without pulling by hands so as to avoid damaging the fungus and influencing the re-balling of the winter dictyophora. And drying the winter edible fungus by using a dryer. During the drying process, ventilation and dehumidification are required. In the drying process, the edible fungus should be lightly handled and lightly placed to prevent the edible fungus from being broken, and the edible fungus should pay attention to the restoration and integrity of the pileus. And packaging the dried winter fungus by using a plastic bag, and sealing to prevent deterioration and yellowing caused by further moisture absorption.

Example 3

1. selecting land: selecting sandy soil below the waist, cool and moist and well ventilated, and avoiding selecting unventilated pit and pit lands and places with frequent termite movement; the land can be selected under bamboo forest, and the relative humidity of air is kept between 85%.

2. Land preparation: and (3) arranging the land blocks according to a pit type cultivation method, digging small pits with the depth of 20cm, the width of 50cm and the length of 60cm according to the conditions of the land blocks, leveling the pit bottoms, and loosening the soil with the pit bottoms being 4cm thick.

3. Cultivation material treatment

Preparing materials: the main material is 30 parts by weight of rhizoma gastrodiae waste firewood; the auxiliary materials comprise the following raw materials in parts by weight: 15 parts of broad-leaved tree wood, 7 parts of carbonized bamboo branches and 15 parts of bamboo leaves.

Preparation of main materials: soaking rhizoma Gastrodiae waste firewood in 2% lime water for 15h, and washing with clear water to obtain the main material.

Preparing auxiliary materials:

a. preparing carbonized bamboo branches: cutting bamboo branches into segments with the length of 7cm, soaking the segments with hydrochloric acid, washing the segments with clear water, draining, roasting, screening redundant carbon powder for later use, and reserving bamboo segments to obtain carbonized bamboo branches;

further, the roasting is to roast the drained bamboo branch sections for 3 hours at the temperature of 220 ℃;

b. preparing auxiliary materials: cutting broad leaf tree wood into 8cm square, spraying semi-degradation liquid to wet the surface, wrapping with a layer of carbon powder sieved in step a, standing at 40 deg.C and 70% humidity for 3 days, sterilizing with ultraviolet ray of 275nm wavelength for 70min, and mixing with carbonized bamboo branch and bamboo leaf to obtain adjuvant.

Further, the semi-degradation liquid is obtained by inoculating white rot fungi into a liquid culture medium and culturing for 15 days; wherein the liquid culture medium is prepared by taking 1.5 percent of maltose, 1.5 percent of corn grit leaching liquor and 10.004 percent of microorganism B as raw materials, and the inoculation amount of the white rot fungi is 12 percent.

4. Sowing:

the cultivation material is paved in a cross lamination mode: laying a layer of main material, a layer of strain and a layer of auxiliary material from bottom to top, laying a layer of main material on the surface, covering a layer of soil with the thickness of 5cm, covering the soil with an arc shape, and covering a layer of pine needles with the thickness of 2cm on the soil. Wherein the usage amount of the main materials at the bottom layer is just enough to cover the soil in the pit, and the main materials are tightly beaten by a hoe back; breaking the strain into small pieces with diameter of about 3cm, and sowing on the bottom layer of firewood, wherein the distance between the strain and the strain is 4cm, and the sowing amount is 2.5kg/m²Completing seeding 2 months ago; coating a thin layer of adjuvants on the strain, wherein the bamboo leaf has effect of drawing mycelium at a concentration of 0.15kg/m²Spraying a layer of white sugar; and paving a layer of main material with the thickness of 8 cm.

5. Daily management

And (3) hypha growth stage: the humidity requirement is as follows: the planting material, the soil layer and the surface of the soil layer are preferably not whitened and dried; if the material turns white and dry, a small amount of water can be sprayed and the wood leaves are covered, but in rainy days, too much water cannot flow into the material, so as to avoid hyphae from being drowned due to too high humidity. The temperature is kept at 24 ℃, and the light is optimally grown under the conditions of half yin and half yang.

And (3) an original base forming stage: after hyphae are formed, cultivating by adopting a temperature difference method under the condition that the humidity and the illumination condition are not changed, specifically, after the hyphae of the dictyophora phalloidea appear, adjusting the temperature to be 24 ℃ in the daytime, adjusting the temperature to be 10 ℃ at night, and adjusting the temperature to the formation of the dictyophora phalloidea primordium every day.

And (3) a sub-entity forming stage: after the winter fungus primordium is formed, keeping the temperature at 22 ℃ and the humidity at 85%, and cultivating by adopting an alternative illumination method, specifically, after the winter fungus primordium is formed, starting illumination for 1h by 500lx intensity light at 7 points every day, then irradiating for 1h by 200lx intensity light, and then supplementing illumination for 30min by 75lx intensity light at night.

6. Harvesting and processing

The cultivated winter dictyophora can be harvested at the beginning of 9 months, and the thalli is cut off from the bottom of the fungus tray by a knife during harvesting of the winter dictyophora, so that the winter dictyophora is not required to be pulled by hands to avoid damaging the thalli to influence re-balling of the winter dictyophora. The winter fungus is roasted by using smokeless coal as fuel, and ventilation and dehumidification are required in the process. The fungus cover is required to be lightly handled and lightly placed in the baking process to be prevented from being broken, and the restoration and the integrity of the fungus cover are required to be paid attention to by the winter fungi. And packaging the dried winter fungus by using a plastic bag, and sealing to prevent deterioration and yellowing caused by further moisture absorption.

Example 4

2. Land preparation: the land parcel is arranged according to a furrow type cultivation method, dry branches and rotten leaves on the surface layer of the land parcel are scraped aside, then pits with the depth of 18cm, the width of 50cm and the length of no limit are dug according to the conditions of the land parcel, the bottom of each pit is flat, and loose soil with the thickness of 3cm is loosened.

3. Cultivation material treatment

The rhizoma gastrodiae waste firewood and fresh firewood blocks are mixed according to the mass ratio of 1: 2.

4. Sowing:

the cultivation material is paved in a cross lamination mode: spreading a layer of cultivation material, a layer of strain, a layer of bamboo leaf, a layer of cultivation material on the surface, and finally covering a layer of cultivation material with a thickness of 3cmCovering soil with arc shape, and covering a layer of fern with thickness of 2 cm. Wherein the amount of the cultivation material at the bottom layer is just enough to cover the soil in the pit, and the soil is tightly beaten by a hoe back; breaking the strain into small pieces with diameter of about 3cm, and sowing on the bottom cultivation material, wherein the distance between strain and strain is 3cm, and the sowing amount is 2.5kg/m²Seeding is completed 10 months before the current year; adding 0.15kg/m of folium Bambusae²Spraying a layer of white sugar; and laying a layer of main material with the thickness of 6 cm.

5. Daily management

And (3) hypha growth stage: the humidity requirement is as follows: the planting material, the soil layer and the surface of the soil layer are preferably not whitened and dried; if the material turns white and dry, a small amount of water can be sprayed and the wood leaves are covered, but in rainy days, too much water cannot flow into the material, so as to avoid hyphae from being drowned due to too high humidity. The temperature is kept at 18 ℃, and the light is optimally grown under the conditions of half yin and half yang.

And (3) an original base forming stage: after hyphae are formed, cultivating by adopting a temperature difference method under the condition that humidity and illumination conditions are unchanged, specifically, after the hyphae of the dictyophora phalloidea appear, adjusting the temperature to be 20 ℃ in the daytime, adjusting the temperature to be 7 ℃ at night, and adjusting the temperature to the formation of the dictyophora phalloidea primordium every day.

And (3) a sub-entity forming stage: after the winter fungus primordium is formed, keeping the temperature at 16 ℃ and the humidity at 85%, and cultivating by adopting an alternative illumination method, specifically, after the winter fungus primordium is formed, starting to irradiate with light with the intensity of 400lx for 1h at 7 points every day, then irradiating with light with the intensity of 100lx for 1h, and then supplementing the light with the intensity of 75lx for 40min at night.

6. Harvesting and processing

The cultivated winter fungi can be harvested 7 months after sowing, and the fungi cables are cut off from the bottom of the fungi tray by a knife without pulling the fungi cables by hands so as to avoid damaging the fungi cables and influencing the re-balling of the winter fungi. The winter fungus is dried by baking with charcoal fire, and ventilation and dehumidification are carried out during the baking. The fungus cover is required to be lightly handled and lightly placed in the baking process to be prevented from being broken, and the restoration and the integrity of the fungus cover are required to be paid attention to by the winter fungi. And packaging the dried winter fungus by using a plastic bag, and sealing to prevent deterioration and yellowing caused by further moisture absorption.

First, screening comparative experiment

1.1 Experimental materials

Experiment 1: in the experiment, under the same conditions as those in the example 1, the main material in the cultivation material is not soaked in lime water, the carbonized bamboo branches in the auxiliary material are replaced by common bamboo branches, and broad-leaf tree wood is directly used after being cut into pieces;

experiment 2: in the experiment, under the same conditions as those in example 1, the cultivation material prepared in the patent document with the application number of CN201610947621.X is adopted for cultivation of the dictyophora phalloidea;

experiment 3: in the experiment, under the same conditions as those in example 1, the temperature and humidity conditions in the patent document with the application number of CN201610947621.X are adopted for daily management;

experiment 4: in the experiment, under the same conditions as those in example 1, temperature, humidity and illumination conditions in a hypha growth stage are adopted for management until the winter dictyophora is harvested;

experiment 5: in this experiment, incubation was not carried out by the temperature difference method at the primordial formation stage under the same conditions as in example 1;

experiment 6: in this experiment, the seed bodies were grown without alternate light irradiation at the seed body formation stage under the same conditions as in example 1.

1.2 Experimental methods

The amino acid content, polysaccharide content and total flavone content of the winter dictyophora cultivated in experiments 1-6 were measured and compared with those of examples 1-4, and the results are shown in table 1. It should be noted that only a few amino acids with higher contents are listed in table 1.

The method for measuring the amino acid content refers to GB 5009.124-2016 determination of amino acids in food.

The method for measuring the polysaccharide content comprises the following steps: pulverizing dried Dictyophora Indusiata, extracting with ethanol under reflux, filtering, volatilizing ethanol, and extracting the residue with water extraction and ethanol precipitation to obtain polysaccharide; preparing a sample solution by using water as a solvent, measuring the absorbance at 490nm, and calculating the polysaccharide content according to a standard curve, wherein the standard curve is obtained by drawing the absorbance and the concentration of a glucose solution.

The method for measuring the content of the total flavonoids comprises the following steps: pulverizing dried Dictyophora Indusiata, extracting with ethanol under reflux, filtering, collecting filtrate, evaporating to dryness under reduced pressure, adding 50% ethanol to constant volume to 25ml volumetric flask to obtain sample solution, measuring absorbance at 510nm wavelength, and calculating according to standard curve to obtain total flavone content, wherein the standard curve is drawn by absorbance and concentration of rutin control solution.

1.3 results of the experiment

TABLE 1

From the experimental results, it can be seen that the growth time of the dictyophora phalloidea is longer than that of the example 1 because the temperature in management is lower in the example 2, but from the detection result, the polysaccharide content of the dictyophora phalloidea cultivated in the example 2 is higher than that of the example 1, which is probably caused by the accumulation of polysaccharide substances due to low temperature, large temperature difference and long growth time. Example 4 the cultivation material was changed to a mixture of waste gastrodia elata firewood and fresh firewood, the growth time was long, and the content of the content was lower compared with that of example 1 in combination though the content of individual amino acid was higher. The contents of various contents of the winter edible fungus cultivated in the experiment 1 are low, and the cultivation material is not processed in the experiment 1, so that the winter edible fungus can be cultivated in a traditional mode of directly mixing and cultivating wood chips and bamboo chips, but the contents of amino acid, polysaccharide and the like of the winter edible fungus are low. Experiment 2 was superior to experiment 1, but still inferior to example 1 in view of data. Experiment 3 had a lower amino acid content, but the total flavone and polysaccharide content was higher than in example 1. Experiment 4 has an amino acid content comparable to that of example 1, but a lower polysaccharide content. The total flavone content of experiment 5 and experiment 6 is comparable to that of example 1, but the amino acid content and polysaccharide content are lower. Thus, overall, example 1 is most preferred.

Detection of heavy metal content of Dictyophora indusiata

2.1 materials of the experiment

Example 1, experiment 1-2;

experiment 7: in the experiment, the winter edible fungus is collected and then baked and dried by firewood under the same condition as that of the experiment 1.

2.2 Experimental methods

Carrying out mixed acid digestion on the winter dictyophora, and then determining the heavy metal content of the winter dictyophora by adopting an atomic absorption spectrophotometry and a graphite furnace.

2.3 results of the experiment

TABLE 2(mg/kg)

Item	Pb	Cd	Cr	Hg	As	Cu
							Example 1	0.11	0.42	0.96	0.07	0.10	4.33
Experiment 1	0.34	0.60	1.28	0.14	0.37	6.19
							Experiment 2	0.28	0.57	1.03	0.08	0.21	5.22
Experiment 7	0.12	0.46	1.01	0.10	0.18	4.67

According to the experimental results, the heavy metal content of the winter dictyophora cultivated in the experiments 1 and 2 is high, and the composition and preparation of the cultivation material have great influence on the heavy metal content of the winter dictyophora. The heavy metal content of the winter dictyophora cultivated in the experiment 7 is not much different from that of the embodiment 1, but the sulfur dioxide content of the winter dictyophora cultivated in the experiment 7 is additionally detected (the detection method is carried out according to GB 5009.34-2016 (determination of sulfur dioxide in national food safety standards)), and the result shows that the sulfur dioxide content of the winter dictyophora cultivated in the experiment 7 is about 1.7 times of the sulfur dioxide content of the winter dictyophora cultivated in the embodiment 1 due to drying of the winter dictyophora by using firewood. Therefore, the embodiment of example 1 is most preferable in view of the above.

Claims

1. A wild-imitating cultivation method for improving the amino acid content of dictyophora phalloidea comprises the following steps: selecting land, preparing land, treating cultivation materials, sowing, performing daily management, harvesting and processing; the method is characterized in that the cultivation material is prepared by taking waste gastrodia elata firewood as a main material; arranging according to a ditch type cultivation method or a nest type cultivation method during soil preparation; during sowing, the cultivation material is paved in a cross lamination mode and the strains are interpenetrated for sowing; during the daily management period, the hyphae are cultured by adopting a temperature difference method and an alternative illumination method after appearing.

2. The wild-imitating cultivation method for improving the amino acid content of the winter fungi as claimed in claim 1, wherein the cultivation material is composed of main materials and auxiliary materials, and the prepared main materials and auxiliary materials are stored separately and can not be mixed; wherein the main material is 20-30 parts by weight of rhizoma gastrodiae waste firewood; the auxiliary materials comprise the following raw materials in parts by weight: 10-15 parts of broad-leaved tree wood, 5-7 parts of carbonized bamboo branches and 10-15 parts of bamboo leaves.

3. The wild-imitating cultivation method for improving the amino acid content of dictyophora phalloidea as claimed in claim 2, wherein the preparation method of the main material comprises the following steps: soaking rhizoma Gastrodiae waste firewood in lime water for 10-15h, and washing with clear water to obtain main material;

the preparation method of the auxiliary materials comprises the following steps:

b. preparing auxiliary materials: cutting broad-leaf tree wood into 5-8cm square blocks, spraying a layer of carbon powder on the surface of the wood with semi-degradation liquid, wrapping the wet carbon powder with the carbon powder screened in the step a, placing the wood for 2-3 days at the temperature of 30-40 ℃ and the humidity of 60-70%, sterilizing the wood with ultraviolet rays, and mixing the wood with carbonized bamboo branches and leaves to obtain the auxiliary material.

4. The wild-like cultivation method for improving the amino acid content of winter dictyophora as claimed in claim 3, wherein the roasting is carried out by roasting the drained bamboo branch segments at the temperature of 180-220 ℃ for 2-3 h.

5. The wild-imitating cultivation method for improving the amino acid content of winter fungi as claimed in claim 3, wherein the semi-degradation liquid is obtained by inoculating white rot fungi into a liquid culture medium and culturing for 10-15 days.

6. The wild-like cultivation method for improving the amino acid content of the dictyophora phalloidea as claimed in claim 1, wherein the furrow cultivation method is to dig a pit with the depth of 15-20cm and the width of 50cm according to the conditions of the plot, the pit bottom is flat, and loose soil with the thickness of 3-4cm is loosened;

the pit type cultivation method is characterized in that small pits with the depth of 15-20cm, the width of 50cm and the length of 60cm are dug according to the conditions of a land parcel, the pit bottom is flat, and loose soil with the thickness of 3-4cm is loosened.

7. The wild-imitating cultivation method for improving the amino acid content of the winter fungi as claimed in claim 1, wherein the cross lamination is that a layer of main material, a layer of strain, a layer of auxiliary material and a layer of main material are paved from bottom to top during sowing, and finally soil is covered and pine needles or ferns are covered.

8. The wild-like cultivation method for improving the amino acid content of the dictyophora phalloidea as claimed in claim 1, wherein the temperature difference method is characterized in that after the occurrence of the dictyophora phalloidea hyphae, the temperature is adjusted to 18-24 ℃ in the daytime, the temperature is adjusted to 7-10 ℃ in the evening, and the temperature is adjusted to the formation of the dictyophora phalloidea primordium every day.

9. The wild-simulated cultivation method for increasing the amino acid content of winter dictyophora as claimed in claim 1, wherein the alternate illumination method is to start illumination with 500lx light with intensity of 400-.