CN110896782A

CN110896782A - Artificial cultivation method of fossa orchioides

Info

Publication number: CN110896782A
Application number: CN201911264859.2A
Authority: CN
Inventors: 孔德平; 王增池; 阎旭东
Original assignee: Cangzhou Academy Of Agriculture And Forestry Sciences
Current assignee: Cangzhou Academy Of Agriculture And Forestry Sciences
Priority date: 2019-12-11
Filing date: 2019-12-11
Publication date: 2020-03-24

Abstract

The invention provides an artificial cultivation method of fossa stuartii, which comprises the following steps: (1) selecting wild fossa comosum strains, separating and obtaining fossa comosum sporocarp as a fossa comosum stock; (2) preparing a couch grass root-PDA composite culture medium, placing the couch grass root-PDA composite culture medium in an aseptic culture container, inoculating original seeds, and culturing mother seeds; (3) mixing crushed fresh couch grass, cottonseed hull, wheat bran and lime, bagging, sterilizing, cooling, inoculating parent strain of Hierochloes Adoratae, and culturing to obtain culture strain of Hierochloes Adoratae; (4) mixing fresh couch grass, cottonseed hull, wheat bran and lime, adding water, and fermenting to obtain culture medium; spreading the culture medium on a ground culture bed in a greenhouse, sowing the culture seeds of the fossa comosum, covering a film, and controlling the greenhouse condition to ensure the growth of the fossa comosum; (5) after the fossa comosum hyphae grow fully, removing the membrane, covering a strip-shaped fine soil belt, and spraying moisture on the covering soil layer for thorough watering; controlling the greenhouse condition, and harvesting the fruiting body when the fossa alnoides fruiting body is flat to obtain the fossa alnoides product.

Description

Artificial cultivation method of fossa orchioides

Background

Large-area saline-alkali wasteland cover plants in the Bohai Bay mainly comprise cogongrass, accompanying plants comprise reed, bicolor limonium, piemarker, asclepiadaceae, zoysia japonica, Chinese limonium, apocynum venetum, green bristlegrass, herba cephalanoploris, Chinese tamarisk and the like, and the total coverage degree is about 70%; the festuca arundinacea is succession of halophyte meadow vegetation to the last herbaceous vegetation of woody plants. A unique edible fungus, namely nest fungus, grows in the grassland of saline-alkali land, local people strive for picking in rainy days when the nest fungus grows out, or directly eat the nest fungus in fresh form, or dry the nest fungus in the air to form dry products for storage.

The fossa comosum consists of mycelium and sporocarp, wherein the mycelium is a vegetative organ and is white and filamentous; the fossa comosum fruiting body is a reproductive organ, is grey white, and consists of pileus, stipe, fold and the like, and the diameter of the pileus is about 0.5-2 cm. The fossa majoris generally parasitizes at the base of the plant of the thatch, grows singly or in clusters, is easy to grow after continuous raining in summer, is limited by natural resource conditions such as environmental temperature, humidity, climate, thatch land conditions and the like, has small and unstable yield, and is difficult to meet the requirements of people, and particularly dried fossa majoris becomes a treasure for being presented to relatives and friends.

At present, the fossa majus grows in a wild way, and the breeding and cultivation of the fossa majus are carried out by a conventional edible fungus artificial breeding method, so that the effect is not ideal enough; so far, no report on the success of artificially cultivating the fossa orchioides exists.

Disclosure of Invention

In order to solve the technical problem, the invention provides an artificial cultivation method of fossa orchioides, which comprises the following steps:

(1) selecting a wild fossa alnoides strain with typical properties and good growth vigor, obtaining fossa alnoides fruiting bodies by adopting a tissue isolation method, and preserving as fossa alnoides stock seeds or directly using the fossa alnoides stock seeds in the inoculation operation in the step (2);

(2) taking cut or crushed fresh cogongrass rhizome, adding 3-5 times of water by mass, boiling the water, keeping slightly boiling for 20-60min, and filtering to obtain filtrate which is cogongrass rhizome extracting solution; adding required amount of potato slices, sugar and agar into the lalang grass rhizome extractive solution, boiling, keeping boiling for 20-40min, filtering, and cooling the filtrate to obtain lalang grass rhizome-PDA composite culture medium; placing the couch grass root-PDA composite culture medium in an aseptic culture container, inoculating the raw seeds of the couch grass fungi collected or stored in the step (1), and culturing at 22-24 ℃ for 4-6 days until hyphae grow over the culture container to obtain mother seeds of the couch grass fungi;

(3) taking 30 parts by mass of cut or crushed fresh couch grass on a dry basis, adding 60-65 parts of cottonseed hulls, 3-10 parts of wheat bran and 0.5-2 parts of lime, mixing uniformly according to the proportion, adding water until the water content accounts for 65-70% of the total mass, uniformly treating, bagging, sterilizing the bagged material by using an autoclave method, cooling to natural temperature after sterilization to obtain bagged culture materials, inoculating the broomrape mother strain obtained in the step (2) under an aseptic condition, and culturing for 20-25 days at 23-26 ℃ until hypha grows in the bag to obtain broomrape strain culture seeds;

(4) according to the mass portion percentage, the fresh fescue is mixed and evenly mixed according to the proportion of 40-45 percent of dry basis, 45-55 percent of cottonseed hull, 5-10 percent of wheat bran and 0.5-2 percent of lime, water is added until the water content is 60-65 percent, the mixture is treated evenly, the mixture is piled and fermented for 5-8 days to prepare a culture medium, when the temperature of the material pile reaches 65-70 ℃, the pile is turned, and the material pile is perforated from top to bottom for oxygen introduction; spreading the fermented culture medium on a ground culture bed of a greenhouse, firstly spreading the culture medium with the thickness of 5-7cm, then broadcasting, dibbling or drilling the culture seeds of the fossa orchioides obtained in the step (3), then spreading the culture medium with the thickness of 1-2cm, finally covering with a film, and pressing the periphery tightly for moisturizing; before sowing, processing the culture seeds of the fossa pulchra to the size of 0.5-2cm, wherein the sowing amount is 1kg of the culture seeds of the fossa pulchra, and sowing 1-3 square meters; after sowing, controlling the temperature of a greenhouse to be 22-28 ℃ to grow fossa comosus hyphae;

(5) removing the film when the culture medium is full of fossa comosum hyphae, and covering a fine soil band with the width of 8-12cm, the thickness of 0.2-0.3cm and the interval of 4-6cm by adopting a strip covering mode; after covering soil, spraying moisture on the covering soil layer by using a sprayer and completely watering; controlling the temperature of the greenhouse to be 24-28 ℃ and the relative humidity to be more than 85%, and keeping proper ventilation and shading; when the fossa comosum fruiting body is flat, harvesting the fruiting body to obtain the fossa comosum product.

In the steps (1) to (3), the wild litsea cubeba strain, the fresh grass root and the fresh grass can be taken from saline-alkali grassland or saline-alkali-free grassland, but the common saline-alkali land is easy to grow the grass, and the common grass is seldom seen in the non-saline-alkali land. The wild litsea cubeba strain and the fresh grass root are found to have better effect when being taken from the saline-free alkali grass land.

In the above steps (2) - (4), the length of the cut or smashed couchgrass root or couchgrass is preferably 0.5-2.0cm, and the couchgrass is harvested in north China preferably 4 months later so as to have the required maturity and nutritional ingredients; in the steps (3) and (4), the cotton seed hulls are the original size of the cotton seed peels, and are not crushed, the fresh fescue is too short or ground into powder on a dry basis, or the cotton seed hulls are not well crushed, one of the defects is that the void ratio of ingredients is reduced, hypha growth is not facilitated, and the other defect is that the fermentation process in the step (4) is too fast or difficult to control, and the yield of the fossa arundinacea in the step (5) is reduced.

In the step (2), the couch grass root-PDA composite culture medium is prepared by adding 200g of potato slices with the thickness of 1-3mm, 20g of sugar and 15-20g of agar into 1000ml of couch grass root extract.

In the step (4), the film is mainly used for moisturizing and cannot contain herbicide; a light-transmitting film may be used; the black film has better effect and has double functions of moisture preservation and shading.

In the step (5), fossa comosum sporocarp grows out from the position near the boundary of the earthing band of the culture substrate powder, namely the fruiting position is at the edge of the culture substrate outside the earthing band; the earthing tape with the width of 8-12cm and the thickness of 0.2-0.3cm basically has no fossa comosum sporocarp. After the fruiting body, namely the fossa stulata product, is harvested for the first time, the temperature of a greenhouse is continuously controlled to be 24-28 ℃, the relative humidity is more than 85%, proper ventilation and shading are kept, and fossa stulata fruiting bodies can continuously grow out from the edge of the culture medium which is not covered with soil between the fine soil belts with the width of 4-6cm and can be harvested to a mature state; generally, 4 crops can be continuously harvested, and the yield of the fossa stuartii per square meter of the culture bed can reach 1.2-1.6 kg.

In the step (2), if the cogongrass rhizome is not separately boiled and filtered to prepare the cogongrass rhizome extracting solution, and then the required amount of potato slices, sugar and agar are added into the cogongrass rhizome extracting solution, but the cogongrass rhizome, the potato slices, the sugar and the agar are directly soaked in water, boiled in water and filtered to prepare the cogongrass rhizome-PDA composite culture medium, the raw seeds of the fossa aeruginosa collected or stored in the step (1) are inoculated, the hyphae grow slowly and obviously weakly in the culture process at the temperature of 22-24 ℃, only a small amount of hyphae grow out in 10-15 days of culture and cannot be used as mother seeds of the fossa aeruginosa, and even if the raw seeds are used for inoculation of the high-pressure sterilization bagging material in the step (3), the hyphae grow slowly and obviously weakly in the culture process at the temperature of 23-26 ℃, and even if the hyphae grow greatly, the hyphae cannot grow fully in the bags in 25 days. The method proves that the independent preparation of the couch grass root extracting solution is important, and the culture medium prepared by directly adding water into the couch grass root, the potato slices, the sugar and the agar has poor effect.

In the step (2), if the raw seeds of the fossa lanceolata collected or stored in the step (1) are inoculated in the PDA culture medium, hyphae grow slowly and weakly in a 10-day culture process at 22-24 ℃, the growth vigor is weak, the culture container can not grow well after 20 days of culture, if the raw seeds of the fossa lanceolata are used for inoculation of the high-pressure sterilization bagging material in the step (3), the hyphae grow slowly and weakly at a 23-26 ℃ culture process, and the hyphae can not grow well in the bag after 30 days of culture even if the inoculation amount is increased by multiple times. The result shows that the fresh grass roots or the extracting solution thereof play a key role in the growth of the raw seeds of the fossa aeruginosa in the culture medium and the preparation of the mother seeds of the fossa aeruginosa; the effect of PDA culture medium alone is not good. If the extract of the cogongrass rhizome is directly used as a culture medium, hyphae grow slowly and weakly in the process of inoculating the raw seeds of the cogongrass fungus collected or stored in the step (1) at the temperature of 22-24 ℃ and culturing for 10 days, only a small amount of hyphae grow after culturing for 10 days, if the extract is used as the mother seeds of the cogongrass fungus for inoculating the high-pressure sterilization bagging material in the step (3), the hyphae grow slowly and weakly in the culture process at the temperature of 23-26 ℃, and even if the inoculation amount is doubled, the hyphae can not grow in the bag for 25 days of culturing. The culture effect of the parent species of the fossa stulata in the step (2) for preparing the cogongrass rhizome-PDA composite culture medium is obviously higher than the sum of the culture effect of the PDA culture medium alone and the culture effect of the cogongrass rhizome extracting solution serving as the culture medium, and the effect of 1+1>2 is exerted.

In the step (3), if fresh couch grass is not counted on a dry basis but is only counted by cotton seed hulls, wheat bran and lime, the fresh couch grass is replaced by the cotton seed hulls and the wheat bran in a corresponding proportion on the dry basis, the amount or the proportion of the lime is not changed, the materials are mixed and uniformly mixed, water is added until the water content is 65-70 m%, the mixture is uniformly processed and then bagged, the mixture is sterilized by an autoclaving method and cooled to natural temperature, the brood pit mushroom mother strain obtained in the step (2) is inoculated by the bagged compost without the couch grass, the growth of hypha is slow and weak obviously in the culture process at 23-26 ℃, and the hypha can not grow in the bag after 30 days of culture even if the inoculation amount is increased by multiple times. The fresh couch grass plays a key role in the growth of the fossa ampelopsis raw seeds in the bagged culture material; that is, the culture effect of the aporthe species cultured by using the cotton seed hulls, the wheat bran and the lime as the culture materials is not good. If the water content of the fresh fescue is dried to 65-70m percent, the cotton seed hulls and the wheat bran are replaced by the fresh fescue in a corresponding proportion on a dry basis without the cotton seed hulls and the wheat bran, the amount or the proportion of lime is not changed, the materials are mixed and uniformly mixed and then bagged, the bagged material without the cotton seed hulls and the wheat bran is sterilized by an autoclaving method, cooled to the natural temperature and inoculated with the fossa arundinacea mother strain obtained in the step (2), the growth of hyphae is slower in the culture process at 23-26 ℃, the growth vigor is obviously weak, and the hyphae can not grow in the bag when the bag is cultured for 30 days even if the inoculation amount is increased by multiple times. The result shows that the cottonseed hulls and the wheat bran play a great role in the growth of the raw seeds of the fossa ampelopsis in the bagged culture materials and can provide most of nutrient components required by the growth of hypha; the culture effect of the culture species of the fresh couch grass and lime culture materials is not good. And (3) preparing a bagged culture material by using a similar method and only airing the fresh lemongrass until the moisture content reaches 65-70 m% without adding lime, inoculating the mother strain of the fossa orchioides obtained in the step (2), wherein the culture effect of the cultivated strain of the fossa orchioides is inferior or lower than that of the fresh lemongrass and lime culture material. The culture effect of the fresh couch grass, cottonseed hull, wheat bran, lime and water high-pressure sterilization bagging culture material prepared in the step (3) to inoculate the nest fungus mother strain obtained in the step (2) is obviously higher than the sum of the culture effect of the cottonseed hull, wheat bran, lime and water high-pressure sterilization bagging culture material and the culture effect of the couch grass, lime or couch grass high-pressure sterilization bagging culture material, and the effect of 1+1>2 is also exerted.

In the step (4), if fresh couch grass is not used but only the cottonseed hulls, the wheat bran and the lime are used, the fresh couch grass is replaced by the cottonseed hulls and the wheat bran in corresponding proportion on a dry basis, the amount or the proportion of the lime is unchanged, the materials are mixed and uniformly mixed, water is added until the water content is 60-65 m%, the uniform treatment is carried out, the culture medium is prepared by piling and fermenting for 5-10 days, the pile turning is carried out when the temperature of the pile reaches 65-70 ℃, and the hole punching is carried out on the pile from top to bottom to carry out oxygen aeration; the fermented couch grass-free culture medium is also paved on a ground culture bed of a greenhouse, a culture medium with the thickness of 5-7cm is paved, the culture medium obtained in the step (3) is sowed, dibbled and drilled, the culture medium with the thickness of 1-2cm is paved, and finally, the culture medium is covered by a film, and the periphery of the culture medium is pressed tightly to preserve moisture; before sowing, the culture seeds of the fossa orchioides are treated to the same size, the same sowing quantity is adopted, and the same temperature conditions of the greenhouse are controlled after sowing; as a result, hyphae grow slowly in the culture process, the growth vigor is obviously weak, and hyphae cannot grow in the culture medium for the same days when the seeding rate is doubled; and only a small amount of fossa comosum sporocarp can grow in the process of the step (5). The fresh couch grass plays a great role in the growth of the brooch mushroom cultivated species in the cultivation medium; the cultivation effect of the fossa arundinacea by using the fermented cottonseed hull, wheat bran and lime cultivation medium without adding fresh couch grass is poor. If the fresh fescue is only used for properly drying water without adding the cottonseed hulls and the wheat bran, the cottonseed hulls and the wheat bran are replaced by the fresh fescue with corresponding proportion, the amount or the proportion of lime is not changed, the materials are mixed and evenly mixed, the water content is controlled to be 60-65 percent, the materials are uniformly treated, the materials are piled and fermented for 5-10 days to prepare the culture substrate, the pile turning is also carried out when the temperature of the pile reaches 65-70 ℃, and the holes are punched from top to bottom to carry out oxygen ventilation; the fermented culture medium containing the fresh couch grass and the lime is also paved on a ground culture bed of a greenhouse, the culture medium with the thickness of 5-7cm is paved, the culture medium obtained in the step (3) is broadcast sown, dibbled and drill sown, the culture medium with the thickness of 1-2cm is paved, and finally, the culture medium is covered by a film, and the periphery of the culture medium is pressed tightly to preserve moisture; before sowing, the culture seeds of the fossa orchioides are treated to the same size, the same sowing quantity is adopted, and the same temperature conditions of the greenhouse are controlled after sowing; as a result, hyphae grow slowly and weakly in the culture process, and the hyphae cannot grow fully in the culture medium when the seeding rate is doubled; and only a small amount of fossa comosum sporocarp can grow in the process of the step (5). And (3) properly airing the fresh couch grass by an approximate method without adding lime for fermentation to prepare the cultivation material, inoculating the nest fungus culture obtained in the step (3), wherein the growth effect of the nest fungus hypha is inferior or lower than that of the cultivation material prepared by fermentation of the fresh couch grass, the lime and the water mixture. The culture and cultivation effects of the couch grass, cottonseed hull, wheat bran, lime and water fermentation cultivation matrix prepared in the step (4) and inoculated to the nest fungus culture obtained in the step (3) are obviously higher than the sum of the culture and cultivation effects of the cottonseed hull, wheat bran, lime and water fermentation cultivation matrix and the culture and cultivation effects of the couch grass, lime or couch grass fermentation cultivation matrix, and the effect of 1+1>2 is also exerted.

If the root or the grass of the fresh cogongrass is replaced by one or more of reed, limonium bicolor, piemarker, asclepias, zoysia japonica, limonium sinense, apocynum venetum, setaria japonica, field thistle and the root or the fresh stem and leaf of tamarisk in the same amount on a dry basis respectively, and the composite culture medium of the step (2), the bagged culture medium of the step (3) and the culture medium of the step (4) are prepared respectively, the problems that the fossa lata does not survive and grows hyphae or the hyphae grows very slowly after the stock collected or stored in the step (1) is inoculated by the prepared composite culture medium, the problems that the hyphae does not grow and grows very slowly after the fossa lattusa mother strain prepared in the step (2) is inoculated by the prepared bagged culture medium, the problems that the hyphae does not grow and grows very slowly after the prepared culture medium is sown in the fossa lattuo mao, the accompanying plants of the couch grass, such as reed, bicolor limonium, piemarker, asclepias, zoysia japonica, Chinese limonium, apocynum venetum, setaria japonica, field thistle, Chinese tamarisk and the like have no utilization value, and the couch grass is a suitable host of the couch grass fungi hypha.

The above shows that the root or the herb of the citronella provides the material factors which are necessary for promoting the breeding and the growth of the mycelium and the fruiting body of the fossa comosum, the culture material or the culture medium prepared by respectively combining the PDA culture medium and the cottonseed hull/wheat bran/lime with the root or the herb of the citronella can provide the material conditions which are necessary for promoting the normal breeding and the growth of the mycelium and the fruiting body of the fossa comosum, and the PDA culture medium, the cottonseed hull/wheat bran/lime culture material or the culture medium can not provide the material conditions which are necessary for the normal breeding and the growth of the mycelium and the fruiting body of the fossa comosum. The characteristic may be the reason that the common nest fungus is not easy to breed and cultivate by the conventional edible fungus artificial breeding method or the effect is not ideal.

Interchanging the formulas of the autoclaved bagged culture material and the fermentation culture medium in the steps (3) and (4), inoculating the parent strain of the Hierochloes Adoratae in the step (2) when preparing the autoclaved bagged culture material according to the mixture ratio in the step (4), and culturing until more white hyphae grow in each bag, wherein the hyphae can grow over the material bag but are not strong integrally, which indicates that the compost in the autoclaved bagged culture material in the step (3) contains too much and is not good; when the fermentation culture medium is prepared according to the mixture ratio in the step (3), seeding [0004] sections of the culture seeds of the fossa comosum in the step (3), performing film-covering culture and performing the operation in the step (5) find that the number of continuously harvested stubbles of the fossa comosum is small, the total yield of the fossa comosum in each square meter of culture bed is slightly low and is less than 0.8kg, and the fact that a little more fresh grass is in the culture medium in the step (4) is helpful for improving the number of continuously harvested stubbles and the total yield of the fossa comosum.

In the step (5), if the culture medium is not subjected to the earthing and water spraying operation, no fossa comosum sporocarp grows out basically; if the operation of covering the fine soil band is not carried out, and only water spraying is carried out at the same position, only a small amount of fossa comatus sporocarp can grow on the culture substrate; if the operation of covering the fine soil band is only carried out without spraying water, a small amount of fossa comamosa sporocarp grows on the culture substrate which is not covered with soil at the outer edge of the fine soil band. The effect of the covering soil and the water spraying on promoting the formation of the fruiting body is large, and the effect is derived from the physical stimulation of the covering soil and the water spraying on the surface hyphae of the fruiting body, and the physical stimulation is beneficial to the kinking and the differentiation of the fossa comosum hyphae. If the complete soil covering is carried out without adopting strip soil covering, namely, fine soil with the thickness of 0.2-0.3cm is covered on the culture substrate full of the fossa comatus hyphae, and then the water spraying operation is carried out on the soil covering, so that the fossa comatus sporocarp basically does not grow out. If the thickness of the strip-shaped covering soil is too large, if fine soil with the thickness of 0.5-1cm is covered on the culture medium full of the fossa comosus hyphae, and the covering soil layer is sprayed by a sprayer to be wet and completely poured, fossa comosus sporophores can grow on the culture medium of the last covering soil between the fine soil zones, but the fruiting quantity is much lower and the yield of the fossa comosus is much lower compared with the thickness of the strip-shaped covering soil with the thickness of 0.2-0.3cm, which indicates that the stimulation is too large.

The invention realizes the artificial cultivation production of wild fossa stuartii, provides a novel edible fungus material, and also increases a novel rare edible fungus variety capable of being artificially cultivated. The cultivated fossa stuartii is compared with wild fossa stuartii, and the cultivated fossa stuartii is almost equivalent to wild fossa stuartii in color, texture, taste and fragrance, has slightly larger size and unit weight of pileus, is generally stronger, has better consistency and ensures the yield.

Detailed Description

In the raw materials used in the following examples and comparative examples, the root and the grass of the festuca arundinacea are respectively taken from a saline grass land or a saline-free grass land in the Bohai Bay at the southeast of the northeast China of Hebei 4 Mey, and are dug out or harvested on the day of use or the day before; removing weed roots from the fresh couch grass roots, washing the fresh couch grass roots with water, and shearing the fresh couch grass roots into 0.5-1.0 cm-long crushed couch grass roots for use, removing weeds from the fresh couch grass roots, and shearing the fresh couch grass roots into 0.5-1.0 cm-long crushed couch grass for use; the wild L.imperata strains of examples 1, 3 were harvested in mid 4 months from the saline grassland and the non-saline grassland, respectively. The cottonseed hull is the original size of the cottonseed hull without crushing, and the lime is 60-mesh fresh lime powder (the content of calcium oxide is higher than 99 m%). The operations of the steps of the embodiments and the comparative examples are not mechanically implemented, but are properly arranged according to the needs, so that the operations of the steps can be reasonably connected; and (4) adopting operation time and conditions with comparison significance as much as possible in comparison in each corresponding step.

Example 1

The method comprises the following steps of carrying out artificial cultivation of the fossa orchioides:

(1) selecting a wild fossa majus strain with typical properties and good growth vigor in a saline-alkali thatch land, obtaining a plurality of hypha blocks near the mycelial fold junction of a fruiting body by adopting a tissue separation method, wherein each hypha block is mung bean grain in size, and immediately using the hypha blocks as a fossa majus stock seed for the inoculation operation in the step (2);

(2) taking 600g of chopped saline-alkali soil cogongrass rhizome, adding 2000g of water, boiling until the water is boiled, keeping slightly boiling for 30min, and filtering to obtain a filtrate which is a cogongrass rhizome extracting solution; adding 200g of potato slices, 20g of glucose and 20g of agar into the couch grass root extracting solution, boiling with big fire until the water is boiled, keeping the boiling with middle fire for 20min, filtering with four layers of gauze to obtain 1000ml of filtrate, and cooling to obtain the couch grass root-PDA composite culture medium; placing a proper amount of couch grass root-PDA composite culture medium in 5 sterile culture test tubes, respectively inoculating each original species of the fossa comosus collected in the step (1), and culturing at 23-24 ℃ for 4 days until hyphae grow over each test tube to obtain a mother strain of the fossa comosus, wherein the hyphae are white and have strong growth vigor;

(3) taking 600g of chopped fresh couch grass (the dry basis rate is 19%) of saline-alkali soil, adding 1300g of cottonseed hulls, 120g of wheat bran and 20g of lime, uniformly mixing, adding water to 6000g (containing 66%) of total mass, uniformly treating, subpackaging into a plurality of polypropylene plastic bags, filling about 1kg of materials into each bag, sterilizing the bagged materials in an autoclave at 126 ℃ for 2h, cooling to room temperature after sterilization, taking 3 bags of materials, respectively inoculating the parent strains of the nest bacteria obtained in the step (2) under an aseptic condition, culturing for 21 days at 23-25 ℃ until white hypha grows in each bag, and culturing the strong hypha to obtain the culture of the nest bacteria;

(4) taking 40kg of chopped fresh couch grass (the dry basis rate is 20%) of saline-alkali soil, 49kg of cottonseed hull, 10kg of wheat bran and 1kg of lime, uniformly mixing, adding water to reach 263kg of total mass (the water content is 62%), uniformly treating, piling and fermenting for 8 days at the ambient temperature of 23-25 ℃ under ventilation conditions to prepare a culture medium, turning the pile when the temperature of the middle part of the pile reaches 70 ℃, turning the pile for four times in total, and perforating the pile from top to bottom every day to introduce oxygen; spreading the fermented culture medium on ground culture bed of greenhouse for 2.5m²Firstly, a cultivation substrate with the thickness of 6-7cm is paved, the culture seeds of the fossa orchioides obtained in the step (3) are sown, then the cultivation substrate with the thickness of 1.5-2cm is paved, and finally, the cultivation substrate is covered by a transparent plastic film without the herbicide, and the periphery of the cultivation substrate is compacted and moisturized; before sowing, processing the fossa orchioides cultivated species to the size of 0.5-1cm, wherein the sowing amount of the fossa orchioides cultivated species is 1.4 kg; controlling the temperature of a greenhouse to be 22-28 ℃ after sowing, and enabling the fossa comatus hyphae to grow;

(5) removing the plastic film when the culture medium is full of fossa comosum hyphae, and covering a fine soil band with the width of 10cm, the thickness of 0.2-0.3cm and the interval of 5cm by adopting a strip covering mode; after covering soil, spraying moisture on the covering soil layer by using a sprayer and completely watering; controlling the temperature of the greenhouse to be 24-28 ℃ and the relative humidity to be 85-90%, and keeping proper ventilation and shading; the fossa comosum sporocarp gradually grows out of the terminal earthing culture substrate at the outer edge of the thin soil belt with the width of 5cm, and no fossa comosum sporocarp grows out of the earthing belt with the width of 10cm and the thickness of 0.2-0.3 cm; and when the fruiting body pileus of the fossa alnoides reaches or approaches to flat state, namely the mushroom grows to a mature state capable of being harvested, harvesting the fruiting body to obtain a first batch of fossa alnoides product, wherein the average yield of the first batch of fossa alnoides per square meter of the culture bed is 0.45 kg.

Example 2

After the first harvest of the fruiting body, namely the fossa citronella product, in the embodiment 1, the temperature of a greenhouse is continuously controlled to be 24-28 ℃, the relative humidity is controlled to be 85-90%, proper ventilation and shading are kept, and fossa citronella fruiting bodies grow continuously from the culture medium which is not covered with soil between the fine soil belts with the width of 4-6cm and develop to a mature state which can be harvested; 4 crops can be continuously harvested; the average total yield of fossa citronella per square meter of cultivation bed, together with the first crop, amounted to 1.35 kg.

The procedure for preparation of the autoclave pouch material of step (3) of example 1 was repeated to prepare 6 pouches of the autoclave pouch material for use as required.

Example 3

The operation of each step is carried out synchronously with the embodiment 1, and the artificially cultivated fossa stuartii is distinguished by comprising the following steps: selecting a wild fossa majus strain with typical properties and good growth vigor from a saline-free Alternaria grassland, and obtaining a plurality of hypha blocks near the mycelial fold junction of fruiting body stipe by adopting a tissue separation method to be used as a fossa majus stock strain; and (2) preparing the couch grass root-PDA composite culture medium by adopting fresh couch grass roots without saline-alkali soil.

As a result, the preparation effects of the parent strains and the cultivated strains of the fossa ampelopsis Grossdentata in the steps (2) and (3), and the growth and output effects of the fossa ampelopsis Grossdentata in the steps (4) and (5) are slightly superior to those of the embodiment 1; the average yield of the first nest of fossa pulchra in each square meter of cultivation bed is 0.51kg, the temperature of a greenhouse is continuously controlled to be 24-28 ℃ and the relative humidity is controlled to be 85-90% after the first nest of fossa pulchra is harvested, proper ventilation and shading are kept, and fossa pulchra sporocarp grows continuously from the culture medium with soil covering at the end of the fine soil zone with the width of 4-6cm and grows to a harvestable mature state; the total harvest of 4 crops is carried out, and the average total yield of the fossa stuartii per square meter of the cultivation bed is 1.47 kg.

Example 4

The operations of steps (4) to (5) of example 1 were substantially repeated while the operations of the two steps were performed, except that a black plastic film containing no herbicide was used in step (4). The average yield of the first batch of the fossa stuartii products per square meter of the cultivation bed is 0.52 kg; after the first crop is harvested, the temperature of the greenhouse is controlled to be 24-28 ℃ and the relative humidity is controlled to be 85-90% according to the method in the embodiment 2, proper ventilation and shading are kept, and 4 crops of the Erythrochloe odorata products can be continuously harvested; the average total yield of the fossa orchioides per square meter of the cultivation bed is 1.56 kg.

Comparison of the L.pulmonarius harvested in examples 1-4 with wild L.pulmonarius revealed almost equivalent color, texture, mouthfeel, aroma, slightly larger pileus size and weight per unit, generally stronger, and more consistent.

Comparative example 1

While the operations of the steps (2) to (3) of example 1 are carried out, the operations of the two steps are basically repeated, except that in the step (2), chopped fresh grass roots are directly soaked in 200g of potato slices, 20g of sugar and 20g of agar with 2000g of water for 30min without being boiled in water, the mixture is boiled with big fire until the water is boiled and kept boiling for 26min with middle fire, four layers of gauze are filtered to obtain 1000ml of filtrate, and the filtrate is cooled to room temperature to prepare the couch grass root-PDA culture medium; placing a proper amount of couch grass root-PDA culture medium in an aseptic culture test tube, inoculating one seed of the newly collected fossa comosus in the step (1) in the example 1, culturing at 23-24 ℃, finding that hyphae grow slowly and grow obviously weakly, only growing a small amount of hyphae after culturing for 10 days, trying to serve as the mother seed of the fossa comosus to be inoculated in two sterilization bag materials in the step (3) in the example 1, wherein the quantity of inoculated hyphae in one bag material is basically the same as that in the step (3) in the example 1, the hyphae grow slowly and grow obviously weakly in the culture process at 23-26 ℃, the quantity of inoculated hyphae in the other bag material is doubled, and the bag can not grow full of hyphae after culturing for 25 days.

Comparative example 2

While the operations of steps (2) to (3) of example 1 were carried out, the operation of the two steps was substantially repeated except that in step (2), the extract of couch grass root was not prepared, but only potato slices 200g, sugar 20g, agar 20g, water 1400g, boiled with strong fire until the water was boiled and kept boiling with medium fire for 23min, four layers of gauze were filtered to obtain 1000ml of filtrate, and cooled to room temperature to prepare a PDA medium; placing a proper amount of PDA culture medium in an aseptic culture test tube, inoculating one of the fossa maerosa stock seeds newly collected in the step (1) in the example 1, culturing for 10 days at 23-24 ℃, finding that hyphae grow slowly and weakly, and culturing for 20 days to ensure that a culture container can not be overgrown, using all the hyphae grown in the test tube as the fossa maerosa mother strain to be inoculated in a sterilization bag material in the step (3) in the example 1, wherein the hyphae grow slowly and obviously weakly in the culture process at 23-26 ℃, and the bag can not be overgrown with hyphae in 30 days of culturing.

Comparative example 3

The procedure of the steps (2) to (3) of example 1 was carried out while substantially repeating the procedure except that the extract solution of the couchgrass roots prepared in the step (2) of example 1 was used as a medium directly, the culture medium was placed in a sterile culture tube, a new seed of the couchgrass fungus newly collected in the step (1) of example 1 was inoculated, and the growth of the hyphae was found to be slow and weak when the culture was carried out at 23 to 24 ℃, only a small amount of hyphae grew after 10 days of culture, all the hyphae grown in the tube were used as a mother strain of the couchgrass fungus to be inoculated in a sterile bag material of the step (3) of example 1, and the hyphae also grew slowly and weakly during 23 to 26 ℃ of culture, and only a small amount of hyphae grew in the bag when the culture was carried out for 25 days.

Comparative example 4

While the operation of the step (3) in the example 1 is carried out, the operation of the step is basically repeated, the difference is that the high-pressure sterilization bag material is prepared by using only the cotton seed hulls, the wheat bran and the lime instead of the fresh couch grass on a dry basis, the cotton seed hulls and the wheat bran are replaced by the fresh couch grass in the corresponding proportion, and the amount of the lime is unchanged, and specifically: uniformly mixing 1850g of cottonseed hull, 170g of wheat bran and 20g of lime, adding water to 6000g of total mass (containing 66% of water), uniformly processing, subpackaging into a plurality of polypropylene plastic bags, filling about 1kg of the materials into each bag, sterilizing the bagged materials at 126 ℃ X2h in an autoclave, cooling to room temperature after sterilization, and respectively inoculating the parent strains of the fossa orchis obtained in the step (2) in the example 1 into two sterilized bag materials under aseptic conditions, wherein the inoculation bacterial amount of one bag material is basically the same as that of the step (3) in the example 1, the growth of hyphae is slow and weak in the culture process at 23-26 ℃, the inoculation bacterial amount of the other bag material is doubled, only a small amount of hyphae grows in the bag when the bag is cultured for 25 days, and the bag is far from growing hyphae.

Comparative example 5

The operation of the step (3) of example 1 was substantially repeated, except that the cut-up cord grass was first dried to remove a part of water to a water content of 67%, and then lime was added thereto and mixed without cottonseed hull and wheat bran to prepare an autoclave bag material, specifically: 2000g of chopped fresh couch grass with a part of water being dried to 67 percent is counted by dry basis, 20g of lime is added to be evenly processed (containing 66.3 percent of water), the materials are subpackaged into a plurality of polypropylene plastic bags, each bag is filled with about 1kg of lime, the materials in the bags are sterilized at a temperature of 126 ℃ and X2h in an autoclave, after the bags are sterilized and cooled to room temperature, two materials in the sterilization bags are respectively inoculated with the fossa orchis virgata mother seeds obtained in the step (2) in the example 1 under an aseptic condition, the inoculation bacterial quantity of one material in the bag is basically the same as that in the step (3) in the example 1, the growth of hyphae is very slow and weak in the culture process at a temperature of 23-26 ℃, the inoculation bacterial quantity in the other material is doubled, only a small amount of hypha grows out in the bag when the bag is cultured for 25.

Comparative example 6

While the operations of the steps (4) to (5) of example 1 were carried out, the operations of the two steps were substantially repeated, except that in the step (4), fresh couch grass was not used but only cotton seed hulls, wheat bran and lime were used, the fresh couch grass was replaced with the cotton seed hulls and wheat bran in the corresponding ratio on a dry basis, and the amount of lime was not changed, and the compounding and the preparation of the culture substrate were carried out, specifically: taking 82.2kg of cotton seed hulls, 16.8kg of wheat bran and 1kg of lime, mixing uniformly, adding water to reach 263kg of total mass (the water content is 62%), treating uniformly, completely absorbing water, piling and fermenting for 8 days under the conditions of ambient temperature of 23-25 ℃ and ventilation to prepare a culture medium, turning the piles when the temperature of the middle part of the piles reaches 70 ℃, carrying out turning for four times in total, and perforating the piles from top to bottom every day to carry out oxygen introduction; spreading the fermented culture medium on ground culture bed of greenhouse for 2.5m²Firstly, a cultivation substrate with the thickness of 6-7cm is paved, the culture seeds of the fossa orchioides obtained in the step (3) in the embodiment 1 are sown, then the cultivation substrate with the thickness of 1.5-2cm is paved, finally, a transparent plastic film without the herbicide is used for covering, and the periphery is pressed tightly for moisturizing; before sowing, processing the fossa orchioides cultivated species to the size of 0.5-1cm, wherein the sowing amount of the fossa orchioides cultivated species is 2.5 kg; controlling the temperature of the greenhouse to be 22-28 ℃ after sowing; namely, before sowing, the culture seeds of the fossa orchioides are treated to the same size as that in the step (4) in the embodiment 1, the sowing quantity is doubled, and the same temperature conditions of a greenhouse are controlled after sowing; the hyphae grow slowly and weakly in the process of culturing, and only a small amount of hyphae grow in the culture medium when the culture medium is cultured for the same days, so that the hyphae cannot grow fully; only a small amount of fossa comosus fruiting bodies can grow in the process of the step (5), and the yield of the fossa comatus per square meter of cultivation bed is less than 0.08 kg.

Comparative example 7

The operations of steps (4) to (5) of example 1 were substantially repeated while carrying out the operations of the two steps, except that in step (4), only fresh couch grass and lime which had been dried off a part of water were used, and cotton seed hulls and wheat bran were replaced with the same amount of fresh couch grass on a dry basis, and the amount of lime was not changed, and the compounding and preparation of the cultivation substrate were carried out, specifically: firstly, airing part of water of cut fresh grass until the water content is 63 percent, adding 1kg of lime into the cut fresh grass according to 99kg of dry basis, uniformly mixing the materials (the water content is 62.4 percent), piling the materials under the conditions of ambient temperature of 23-25 ℃ and ventilation for 8 days to prepare a culture medium, turning the piles when the temperature of the middle part of the pile reaches 70 ℃, carrying out pile turning for four times in total, and perforating the piles from top to bottom every day to carry out oxygen ventilation; spreading the fermented culture medium on ground culture bed of greenhouse for 2.5m²Firstly, a cultivation substrate with the thickness of 6-7cm is paved, the culture seeds of the fossa orchioides obtained in the step (3) in the embodiment 1 are sown, then the cultivation substrate with the thickness of 1.5-2cm is paved, finally, a transparent plastic film without the herbicide is used for covering, and the periphery is pressed tightly for moisturizing; before sowing, processing the fossa orchioides cultivated species to the size of 0.5-1cm, wherein the sowing amount of the fossa orchioides cultivated species is 2.5 kg; controlling the temperature of the greenhouse to be 22-28 ℃ after sowing; namely, before sowing, the culture seeds of the fossa orchioides are treated to the same size as that in the step (4) in the embodiment 1, the sowing quantity is doubled, and the same temperature conditions of a greenhouse are controlled after sowing; in the process of result culture, hyphae grow slowly and weakly, and only a small amount of hyphae grow in the culture medium when the culture medium is cultured for the same days; only a small amount of fossa comosum fruiting bodies can grow in the process of the step (5), and the yield of the first-batch fossa comosum in each square meter of cultivation bed is less than 0.10 kg.

Comparative example 8

While the operations of steps (4) to (5) of example 1 were carried out, the operation of the two steps was substantially repeated, the operation of step (4) was repeated, and a culture substrate was prepared and used for a greenhouse cultivation bed of 3.0m²And seeding and mulching, wherein the thickness is 0.5m²Covering the area with transparent plastic film containing herbicide in another 2.5m²Covering the area with a transparent plastic film without containing herbicide, and controlling the same temperature condition of a greenhouse to 22-28 ℃ to enable the growth of the fossa orchioides; it is found that the coating of the transparent plastic containing the herbicide0.5m of film²The hypha in the culture medium has slow growth, and the quantity and the strength of the hypha are far lower than that of the hypha covered with the transparent plastic film without the herbicide by 2.5m²Area culture of fossa comosum hyphae in the matrix.

2.5m of the transparent plastic film coated with the herbicide-free coating²The procedure of step (5) was carried out for the area, but the difference was that after the culture medium was overgrown with the mycelia of fossa comosum, the following five procedures were carried out simultaneously while controlling the greenhouse temperature at 24-28 ℃ and the relative humidity at 85-90%, and the conditions of keeping appropriate ventilation and shading were substantially the same as those in step (5) of example 1.

At 0.5m²Area without the described earthing and water spraying operations on the culture substrate, it was found that substantially no dimorpholinum sporophore was grown.

At 0.5m²The operation of covering the fine soil zone is not carried out in the area, only water spraying is carried out, a small amount of fossa comosus sporophores grow on the culture substrate, and the yield of the fossa comucopiae is less than 0.07kg per square meter of culture bed.

At 0.5m²Only the operation of covering the fine soil belt is carried out, the fine soil with the thickness of 0.2-0.3cm is covered on the culture substrate full of the fossa comatus hyphae, water is not sprayed, a small amount of fossa comatus sporocarp grows on the edge of the culture substrate covered with soil at the tail end of the fine soil belt, the yield of the fossa comatus per square meter of the culture bed is reduced to be less than 0.12kg, and no fossa comatus sporocarp grows in the soil covering belt.

At 0.5m²And (3) fully covering soil without strip-shaped covering soil, namely covering fine soil with the thickness of 0.2-0.3cm on the culture medium full of the fossa comatus hyphae, and then spraying water on the covering soil, wherein no fossa comatus sporocarp grows out basically.

At 0.5m²Covering the soil in a strip shape in the area, wherein the thickness of the covered soil is large, and thin soil belts with the width of 10cm, the thickness of 0.5-1.0cm and the interval of 5cm are covered; after covering soil, spraying moisture on the covering soil layer by using a sprayer and completely watering; the fossa comosum fruiting bodies are gradually grown from the edges of the culture medium with the thin soil strips with the width of 5cm and the soil not covered between the thin soil strips, but the fruiting quantity is reduced by about 35 percent, and the growth vigor is not high enough.

Comparative example 9

While the operations of the steps (3) to (5) in the example 1 are carried out, the three steps are basically repeated, but the formulas of the autoclaved compost and the fermentation culture medium in the steps (3) and (4) are exchanged, so that after the fossa citronella mother strains obtained in the step (2) are inoculated by the sterilized and cooled bag material in the step (3) under the aseptic condition, the fossa citronella mother strains are cultured for 23 days at the temperature of 23-25 ℃ until more white hypha grows out in each bag, and the bags can grow full of the white hypha, but are not strong enough.

Step (4) spreading the culture substrate prepared by the formula change to a greenhouse ground culture bed with the thickness of 2.5m²Spreading a culture medium with a thickness of 6-7cm, and sowing [0004]]Step (3), flatly paving a culture substrate with the thickness of 1.5-2cm, finally covering the culture substrate with a transparent plastic film without a herbicide, compacting and moisturizing the periphery, and carrying out the operation of the step (5), so that the cavum graveolens can be continuously harvested for only 3 times, the total output of the cavum graveolens per square meter of a culture bed is 0.80kg, which is lower than the total output of 1.2kg of the embodiment 1-2 and the total output of 1.27kg of the embodiment 3; before sowing, processing the fossa orchioides cultivated species to the size of 0.5-1cm, wherein the sowing amount of the fossa orchioides cultivated species is 1.3 kg; after sowing, the temperature of the greenhouse is controlled to be 22-28 ℃.

Comparative example 10

This procedure was substantially repeated while the procedure of step (2) of example 1 was carried out, except that the fresh Japanese lawngrass was replaced with the same amount of Japanese lawngrass on a dry basis and that, as a result, the composite medium prepared by inoculating the stock of the newly collected Japanese lawngrass in step (1) with the stock had problems that the stock could not survive and hyphae did not grow.

Comparative example 11

This procedure was substantially repeated while the procedure of step (3) of example 1 was carried out, except that the fresh couch grass was replaced with zoysia japonica having the same length on a dry basis, and as a result, the sterilized bagged compost prepared had a problem that hyphae did not grow after the inoculation of the pit grass mother species prepared in step (2) of paragraph [0004 ].

Comparative example 12

This procedure was substantially repeated while the procedure of step (4) of example 1 was carried out, except that fresh couch grass was replaced with zoysia japonica of the same length on a dry basis, and as a result, the prepared culture substrate had a problem of extremely slow hypha growth after seeding of the dimorpholinum cultivar prepared in step (3) of [0004 ].

Claims

1. An artificial cultivation method of fossa orchioides comprises the following steps:

2. The artificial cultivation method of the fossa orchioides according to claim 1, wherein the wild fossa orchioides strain, the root of the fresh grass, and the fresh grass are obtained from saline-alkali thatch land in the steps (1) to (3).

3. The artificial cultivation method of the fossa stuartii according to claim 1, wherein the wild fossa stuartii strain, the fresh grass root, is obtained from the grassland of the saline-free Alternaria stuartii in the steps (1) to (2).

4. The artificial cultivation method of fossa orchioides according to claim 1, wherein the cut or crushed roots or pieces of the fresh grass have a length of 0.5 to 2.0cm in the steps (2) to (4).

5. The artificial cultivation method of Erythrophloe odorata as claimed in claim 1, wherein in the step (2), the material ratio of the compound culture medium of Imperata root-PDA is 200g of potato slices with thickness of 1-3mm, 20g of sugar and 15-20g of agar per 1000ml of the extract solution of Imperata.

6. The artificial cultivation method of fossa orchioides according to claim 1, wherein in the step (4), the film used does not contain a herbicide.

7. The artificial cultivation method of fossa orchioides according to claim 1, wherein in the step (4), the film is black.

8. The artificial cultivation method of the fossa stulata as claimed in claim 1, wherein in the step (5), after the first crop of the fossa stulata product is harvested, the greenhouse temperature is continuously controlled to 24-28 ℃ and the relative humidity is more than 85%, proper ventilation and shading are kept, the fossa stulata is continuously grown, and a plurality of crops of the fossa stulata are harvested.