CN113875493A - Breeding and stable-yield cultivation method for black-vein strain morchella strain - Google Patents
Breeding and stable-yield cultivation method for black-vein strain morchella strain Download PDFInfo
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- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/20—Culture media, e.g. compost
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/40—Cultivation of spawn
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Abstract
The invention discloses a breeding and stable-yield cultivation method of black-vein strain morchella strains, which is technically characterized in that a wild morchella sporophore called as the black-vein strain in taxonomy is utilized, and the wild morchella sporophore is utilized through strain breeding, absorption and utilization of nutrient media and innovation of sclerotic mushroom acceleration and conservation management technology, so that the wild morchella sporophore is applied to the facility artificial cultivation of morchella, the stability of the morchella strains and the cultivation technology can be greatly improved, and the effects of stable yield and high quality are achieved. Compared with the traditional morel tissue isolation breeding, simple spore breeding and the existing cultivation management technical method, the technology has the advantages that the production efficiency can be improved by 3-5 times, a series of problems of unstable fruiting, low yield, high cost and the like in morel strain production and depending on the day and seasonal cultivation are solved, and the technology has extremely high industrial commercial value and prospect for stabilizing morel market supply, providing farmer income and the like.
Description
Technical Field
The invention belongs to the technical field of edible fungi, and particularly relates to a black vein strain morchella strain breeding and stable yield cultivation method.
Background
Morchella, also known as "Yangguang fungus" or "Baogu fungus", belongs to the phylogenetic group of Ascomycotina, Panzhi, Morchella, and is a native edible fungus. The common types of the morchella esculenta are cones, thick legs, black veins, morchella esculenta and the like. Morchella esculenta is one of the most precious rare edible fungi in the world and belongs to high-grade fungus foods. Has the effects of tonifying kidney, strengthening yang, nourishing brain and refreshing, can prevent and resist cancer, prevent cold and enhance human immunity, and has important development value in medicine. At present, the artificial cultivation technology of the morchella is not mature, a bionic cultivation method is mainly adopted, most of strains are obtained by tissue separation of local wild morchella, and cultivation time consistent with the growth season of the local wild morchella is adopted. Although the commercial planting is realized, the yield is low and unstable due to the influence of genetic instability of strains and regional climate change, huge loss is caused to cultivators, and the method becomes the biggest problem of hindering the development of the morchella industry in China.
Disclosure of Invention
The first purpose of the invention is to provide a breeding method of black vein strain morchella strain, and the second purpose of the invention is to provide a stable yield cultivation method of black vein strain morchella strain.
The first object of the present invention is achieved by comprising the steps of:
s 1: selecting morchella sporocarp called black-vein strain in taxonomy as an object for collection and breeding, digging out the morchella sporocarp and a matrix with a root square and round size of 5 cm, and quickly filling the morchella sporocarp and the matrix into a collection container for sealing;
s 2: peeling the root soil under aseptic conditions, picking up white villous mass of the root by using an aseptic scalpel, connecting the white villous mass to a PDA culture medium, and transferring the culture medium to a constant temperature box for culture to form mycelium;
s 3: obtaining polysporus mycelia from the selected morchella sporocarp according to a conventional spore separation technology;
s 4: inoculating the mycelium obtained in the steps s2 and s3 to a new PDA culture medium at intervals of 1-2 cm, and then transferring to a thermostat to culture the new mycelium;
s 5: after the two mycelia are fused and crossed, picking the mycelia at the binding part in time, transferring the mycelia to a new PDA culture medium for continuous culture, observing and detecting the mycelia or formed sclerotia by using a microscope after the mycelia grow to be mature, identifying the mycelia with vigorous mycelium production and cells in a multi-core state, and then carrying out mating type detection and confirmation by using an its technology as a standard for detecting high-quality strains to obtain production type safe strains for artificial culture.
The second object of the present invention is achieved by a: preparation of culture medium and nutrient material
a 1: preparing a culture medium by selecting broad-leaved tree barks, wheat and/or fruit residues of different varieties, and requiring fresh drying without mildew; the method comprises the steps of naturally drying fresh broad-leaved tree barks, wheat and/or fruit residues, then crushing and weighing, adding water according to the proportion of 1:1.2, mixing and stirring uniformly, adjusting the pH to 8-9 by using lime powder 2%, stirring and fermenting in a solid fermentation stirrer, starting stirring, ventilating and radiating heat when the fermentation temperature exceeds 65 ℃, stopping stirring and continuing fermenting when the fermentation temperature is reduced to 50 ℃, repeating the fermentation for 3 times in the way, wherein the time is 7-10 days, and when the color of a substrate raw material is observed to be brownish red and has obvious acid fragrance, the substrate can be determined to be mature in fermentation and is used as a culture substrate suitable for growth and development of morchella;
a 2: after the matrix in the step a1 of preparing the nutrient material is fermented to be mature, putting the wheat into a water pool for soaking 12 hours in advance to ensure that the wheat fully absorbs enough water, then fishing out and mixing the wheat with the fermented matrix, and simultaneously adding 0.5 percent of lime powder, 2 percent of thiophanate bactericide and 20 percent of soil to be uniformly stirred together to obtain the nutrient material suitable for growth and development of morchella;
b: mycelium culture
b 1: building a culture pond, paving a layer of plastic film or a shading net at the bottom of the pond, uniformly paving the culture medium obtained in the step a2 on the plastic film or the shading net in the pond, after the culture medium is paved, injecting water into the pond, wherein the water injection amount is less than the medium, sowing is started when the water content of the medium to be measured is 60-65%, sowing is carried out by adopting a hole sowing method, nutrient materials with the thickness of 3-5 cm are covered on strains after sowing, a shed door is closed after the sowing is finished, and a mycelium cultivation stage is carried out, wherein the temperature in the pond is kept at 14-16 ℃ and the humidity is kept within 55-60%;
b 2: indoor cultivation: selecting a plastic basket as a cultivation device, filling a cultivation substrate into the basket according to the thickness of 14-16 cm, then sowing by adopting a hole sowing method, covering nutrient materials with the thickness of 8-12 cm on strains after sowing, moving the strains into an indoor cultivation base frame, and controlling by an air conditioner, a humidifier and a fan during the sowing process to keep the indoor temperature at 14-16 ℃, the humidity at 55-60% and the carbon dioxide concentration within 0.2%;
c: cultivation of fruiting bodies
c 1: when a large amount of fungus frost begins to be converted into sclerotium and mycelia turns brown from white, removing the nutrient materials laid on the cultivation matrix layer in time;
c 2: when the temperature in the culture medium is not higher than 8-10 ℃ and not lower than 4 ℃ in outdoor cultivation, after covering a shading net on the culture medium, beginning to inject a large amount of water into the culture medium, and stopping injecting water when the water consumption exceeds 2cm of the surface of the culture medium;
c 3: the indoor cultivation is that a cultivation substrate which is mature in fermentation is placed in water to be soaked for 12-16 hours, then excessive moisture is drained, the cultivation substrate is moved back to a cultivation base frame, the indoor temperature is adjusted to be 8-10 ℃, the humidity is kept to be about 85-95%, the carbon dioxide concentration is within 0.2%, an illuminating lamp is turned on to accelerate mushroom management, white point-shaped kinks are formed on the surface of the cultivation substrate after 72 hours under the condition that the conditions of temperature, humidity, air, light and the like are met, namely morchella larvas are young mushrooms, and then a mushroom fruiting management stage is carried out, wherein the outdoor cultivation needs moisture preservation, the indoor cultivation needs ventilation until sporocarp is mature.
The invention has the beneficial effects that: the invention utilizes a wild morchella sporocarp called black vein strain in taxonomy, and realizes artificial yearly and stable cultivation of morchella by a manual separation breeding technology, a fungus growing and fruiting sectional cultivation management method and modern agricultural facilities. Thoroughly solves the problems of unstable heredity of strains, low yield and limitation to seasonal cultivation in the artificial morchella cultivation at present. Compared with the traditional morel tissue isolation breeding, simple spore breeding and the existing cultivation management technical method, the technology has the advantages that the production efficiency can be improved by 3-5 times, a series of problems of unstable fruiting, low yield, high cost and the like in morel strain production and depending on the day and seasonal cultivation are solved, and the technology has extremely high industrial commercial value and prospect for stabilizing morel market supply, providing farmer income and the like.
Detailed Description
The invention is further illustrated by the following examples, which are not intended to be limiting in any way, and any variations or modifications which are based on the teachings of the present invention are intended to be within the scope of the present invention.
A black vein strain morchella strain breeding method comprises the following steps:
s 1: selecting morchella sporocarp called black-vein strain in taxonomy as an object for collection and breeding, digging out the morchella sporocarp and a matrix with a root square and round size of 5 cm, and quickly filling the morchella sporocarp and the matrix into a collection container for sealing;
s 2: peeling the root soil under aseptic conditions, picking up white villous mass of the root by using an aseptic scalpel, connecting the white villous mass to a PDA culture medium, and transferring the culture medium to a constant temperature box for culture to form mycelium;
s 3: obtaining polysporus mycelia from the selected morchella sporocarp according to a conventional spore separation technology;
s 4: inoculating the mycelium obtained in the steps s2 and s3 to a new PDA culture medium at intervals of 1-2 cm, and then transferring to a thermostat to culture the new mycelium;
s 5: after the two mycelia are fused and crossed, picking the mycelia at the binding part in time, transferring the mycelia to a new PDA culture medium for continuous culture, observing and detecting the mycelia or formed sclerotia by using a microscope after the mycelia grow to be mature, identifying the mycelia with vigorous mycelium production and cells in a multi-core state, and then carrying out mating type detection and confirmation by using an its technology as a standard for detecting high-quality strains to obtain production type safe strains for artificial culture.
A stable yield cultivation method for black vein strain morchella comprises the following steps:
a: preparation of culture medium and nutrient material
a 1: preparing a culture medium by selecting broad-leaved tree barks, wheat and/or fruit residues of different varieties, and requiring fresh drying without mildew; the method comprises the steps of naturally drying fresh broad-leaved tree barks, wheat and/or fruit residues, then crushing and weighing, adding water according to the proportion of 1:1.2, mixing and stirring uniformly, adjusting the pH to 8-9 by using lime powder 2%, stirring and fermenting in a solid fermentation stirrer, starting stirring, ventilating and radiating heat when the fermentation temperature exceeds 65 ℃, stopping stirring and continuing fermenting when the fermentation temperature is reduced to 50 ℃, repeating the fermentation for 3 times in the way, wherein the time is 7-10 days, and when the color of a substrate raw material is observed to be brownish red and has obvious acid fragrance, the substrate can be determined to be mature in fermentation and is used as a culture substrate suitable for growth and development of morchella;
a 2: after the matrix in the step a1 of preparing the nutrient material is fermented to be mature, putting the wheat into a water pool for soaking 12 hours in advance to ensure that the wheat fully absorbs enough water, then fishing out and mixing the wheat with the fermented matrix, and simultaneously adding 0.5 percent of lime powder, 2 percent of thiophanate bactericide and 20 percent of soil to be uniformly stirred together to obtain the nutrient material suitable for growth and development of morchella;
b: mycelium culture
b 1: building a culture pond, paving a layer of plastic film at the bottom of the pond, uniformly paving the culture medium obtained in the step a2 on the plastic film or shading net in the pond, injecting water into the pond after the culture medium is paved, wherein the water injection amount is less than the medium, measuring the water content of the medium to be 60%, starting seeding, sowing by adopting a hole sowing method, covering nutrient materials with the thickness of 3-5 cm on strains, closing a shed door after the seeding is finished, and transferring to a mycelium cultivation stage, wherein the temperature in the pond is kept at 14-16 ℃ and the humidity is kept within 55-60%;
b 2: indoor cultivation: selecting a plastic basket as a cultivation device, filling a cultivation substrate into the basket according to the thickness of 14-16 cm, then sowing by adopting a hole sowing method, covering nutrient materials with the thickness of 8-12 cm on strains after sowing, moving the strains into an indoor cultivation base frame, and controlling by an air conditioner, a humidifier and a fan during the sowing process to keep the indoor temperature at 14-16 ℃, the humidity at 55-60% and the carbon dioxide concentration within 0.2%;
c: cultivation of fruiting bodies
c 1: when a large amount of fungus frost begins to be converted into sclerotium and mycelia turns brown from white, removing the nutrient materials laid on the cultivation matrix layer in time;
c 2: when the temperature in the culture medium is not higher than 8-10 ℃ and not lower than 4 ℃ in outdoor cultivation, after covering a shading net on the culture medium, beginning to inject a large amount of water into the culture medium, and stopping injecting water when the water consumption exceeds 2cm of the surface of the culture medium;
c 3: the indoor cultivation is that the cultivation substrate which is mature in fermentation is placed in water to be soaked for 12-16 hours, then the excessive moisture is drained, the cultivation substrate is moved back to a cultivation base frame, the indoor temperature is adjusted to be 8-10 ℃, the humidity is kept to be 85-95%, the carbon dioxide concentration is within 0.2%, an illuminating lamp is turned on to urge mushroom management, under the condition that the conditions of temperature, humidity, air, light and the like are met, white point-shaped kinks are formed on the surface of the cultivation substrate after 72 hours, namely morchella larch mushrooms are obtained, and then the fruiting management stage is carried out until the fruit bodies are mature.
The mass ratio of the raw materials in the nutrient material in the step a1 is as follows: 33% of broad-leaved tree bark, 65% of wheat, 2% of lime powder, 20% of thiophanate bactericide which accounts for 0.5% of the total amount of the added broad-leaved tree bark, 13% of pomace, 65% of wheat, 2% of lime powder and 0.5% of thiophanate bactericide which accounts for 0.5% of the total amount of the added broad-leaved tree bark, wherein the pH value is 8-9% and the water content is 60%.
The mass ratio of the raw materials in a2 is as follows: 20% of fermented bark, 13% of pomace, 65% of wheat, 2% of lime powder, 20% of thiophanate bactericide and soil which account for 0.5% of the total amount, 8% of pH value and 60% of water content.
1. The morchella sporophore stem cell propagation hypha technology and the sporophore spore separation technology are combined with the biological backcross fusion culture technology, so that the cell genetic genes of the morchella are subjected to defect repair, and the genetic stability of the obtained strains can reach 100%. Thoroughly solves the problem that the reliability of the prior artificial cultivation of the morchella is only 40 percent and the reliability of the spore separation does not exceed 50 percent. The strain breeding technology plays a positive promoting role in the development of artificial cultivation of the morchella esculenta, and can reduce the loss of the artificial cultivation of the morchella esculenta caused by strain problems by more than one hundred million yuan per year.
2. The culture medium and the nutrient material are subjected to a segmented culture technology, hypha grows strongly and is aging-resistant in the whole spawn running process, and the hypha has higher activity than the existing cellulose enrichment culture medium. Firstly, the mushroom residues of the needle mushrooms or the pleurotus eryngii after mushroom fruiting are used for raw material selection, so that the waste resources are utilized, the mushroom residues are simple and easy to obtain, the requirements of growth and development of morchella are met, the cost is low, and the yield is improved. And secondly, a culture medium and nutrient material segmented culture method is adopted in the fruiting technology, and the comparison with the conventional method of directly placing nutrient bags through sowing once at present proves that the method has the advantages of quick hypha spawn running, thorough nutrient absorption, regular fruiting and the like. And thirdly, an isolation film is paved on the outdoor ground and an indoor frame type isolation cultivation mode is adopted, and the matrix is treated for 2 times through the outside in the front period and the outside in the rear period and the inside in the later period, so that the damage of plant diseases and insect pests to the morchella is avoided, and the problem of continuous cropping obstacle in cultivation is solved.
3. The strain direct seeding technology is adopted, wherein the dilution refers to breaking the original solid strain, adding purified water to dilute the solid strain into liquid, and then directly scattering the liquid on the soil. Compared with the traditional solid strain cultivation technology, the method has the advantages of fast hypha growth, short spawn running period and the like. The culture medium is only required to be 7 days from inoculation to hypha overgrowth, the culture medium is faster than a pure solid strain by more than 2 times, the growth cycle of the morchella is shortened by more than 20 days, and the method is an ideal mode for industrial morchella cultivation.
Claims (5)
1. A black vein strain morchella strain breeding method is characterized by comprising the following steps:
s 1: selecting morchella sporocarp called black-vein strain in taxonomy as an object for collection and breeding, digging out the morchella sporocarp and a matrix with a root square and round size of 5 cm, and quickly filling the morchella sporocarp and the matrix into a collection container for sealing;
s 2: peeling the root soil under aseptic conditions, picking up white villous mass of the root by using an aseptic scalpel, connecting the white villous mass to a PDA culture medium, and transferring the culture medium to a constant temperature box for culture to form mycelium;
s 3: obtaining polysporus mycelia from the selected morchella sporocarp according to a conventional spore separation technology;
s 4: inoculating the mycelium obtained in the steps s2 and s3 to a new PDA culture medium at intervals of 1-2 cm, and then transferring to a thermostat to culture the new mycelium;
s 5: after the two mycelia are fused and crossed, picking the mycelia at the binding part in time, transferring the mycelia to a new PDA culture medium for continuous culture, observing and detecting the mycelia or formed sclerotia by using a microscope after the mycelia grow to be mature, identifying the mycelia with vigorous mycelium production and cells in a multi-core state, and then carrying out mating type detection and confirmation by using an its technology as a standard for detecting high-quality strains to obtain production type safe strains for artificial culture.
2. The stable yield cultivation method of the black-vein strain morchella esculenta as claimed in claim 1, which is characterized by comprising the following steps:
a: preparation of culture medium and nutrient material
a 1: preparing a culture medium by selecting broad-leaved tree barks, wheat and/or fruit residues of different varieties, and requiring fresh drying without mildew; the method comprises the steps of naturally drying fresh broad-leaved tree barks, wheat and/or fruit residues, then crushing and weighing, adding water according to the proportion of 1:1.2, mixing and stirring uniformly, adjusting the pH to 8-9 by using lime powder 2%, stirring and fermenting in a solid fermentation stirrer, starting stirring, ventilating and radiating heat when the fermentation temperature exceeds 65 ℃, stopping stirring and continuing fermenting when the fermentation temperature is reduced to 50 ℃, repeating the fermentation for 3 times in the way, wherein the time is 7-10 days, and when the color of a substrate raw material is observed to be brownish red and has obvious acid fragrance, the substrate can be determined to be mature in fermentation and is used as a culture substrate suitable for growth and development of morchella;
a 2: after the matrix in the step a1 of preparing the nutrient material is fermented to be mature, putting the wheat into a water pool for soaking 12 hours in advance to ensure that the wheat fully absorbs enough water, then fishing out and mixing the wheat with the fermented matrix, and simultaneously adding 0.5 percent of lime powder, 2 percent of thiophanate bactericide and 20 percent of soil to be uniformly stirred together to obtain the nutrient material suitable for growth and development of morchella;
b: mycelium culture
b 1: building a culture pond, paving a layer of plastic film or a shading net at the bottom of the pond, uniformly paving the culture medium obtained in the step a2 on the plastic film or the shading net in the pond, after the culture medium is paved, injecting water into the pond, wherein the water injection amount is less than the medium, sowing is started when the water content of the medium to be measured is 60-65%, sowing is carried out by adopting a hole sowing method, nutrient materials with the thickness of 3-5 cm are covered on strains after sowing, a shed door is closed after the sowing is finished, and a mycelium cultivation stage is carried out, wherein the temperature in the pond is kept at 14-16 ℃ and the humidity is kept within 55-60%;
b 2: indoor cultivation: selecting a plastic basket as a cultivation device, filling a cultivation substrate into the basket according to the thickness of 14-16 cm, then sowing by adopting a hole sowing method, covering nutrient materials with the thickness of 8-12 cm on strains after sowing, moving the strains into an indoor cultivation base frame, and controlling by an air conditioner, a humidifier and a fan during the sowing process to keep the indoor temperature at 14-16 ℃, the humidity at 55-60% and the carbon dioxide concentration within 0.2%;
c: cultivation of fruiting bodies
c 1: when a large amount of fungus frost begins to be converted into sclerotium and mycelia turns brown from white, removing the nutrient materials laid on the cultivation matrix layer in time;
c 2: when the temperature in the culture medium is not higher than 8-10 ℃ and not lower than 4 ℃ in outdoor cultivation, after covering a shading net on the culture medium, beginning to inject a large amount of water into the culture medium, and stopping injecting water when the water consumption exceeds 2cm of the surface of the culture medium;
c 3: the indoor cultivation is that a cultivation substrate which is mature in fermentation is placed in water to be soaked for 12-16 hours, then excessive moisture is drained, the cultivation substrate is moved back to a cultivation base frame, the indoor temperature is adjusted to be 8-10 ℃, the humidity is kept to be about 85-95%, the carbon dioxide concentration is within 0.2%, an illuminating lamp is turned on to accelerate mushroom management, white point-shaped kinks are formed on the surface of the cultivation substrate after 72 hours under the condition that the conditions of temperature, humidity, air, light and the like are met, namely morchella larvas are young mushrooms, and then a mushroom fruiting management stage is carried out, wherein the outdoor cultivation needs moisture preservation, the indoor cultivation needs ventilation until sporocarp is mature.
3. The stable yield cultivation method of the black-vein strain morchella esculenta according to claim 2, which is characterized in that: the mass ratio of the raw materials in the nutrient material in the step a1 is as follows: 33% of broad-leaved tree bark, 65% of wheat, 2% of lime powder, 20% of thiophanate bactericide which accounts for 0.5% of the total amount of the added broad-leaved tree bark, 13% of pomace, 65% of wheat, 2% of lime powder and 0.5% of thiophanate bactericide which accounts for 0.5% of the total amount of the added broad-leaved tree bark, wherein the pH value is 8-9% and the water content is 60%.
4. The yield-stabilizing cultivation method of the black-vein strain morchella esculenta according to claim 2, which is characterized in that the raw materials in the a2 are in the following mass ratio: 20% of fermented bark, 13% of pomace, 65% of wheat, 2% of lime powder, 20% of thiophanate bactericide and soil which account for 0.5% of the total amount, 8% of pH value and 60% of water content.
5. The stable yield cultivation method of black-vein strain morchella esculenta according to claim 2, characterized in that solid strains are broken off before sowing, and then purified water is added to dilute the solid strains into liquid state, and then the liquid is directly sprinkled on soil.
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