CN106305138B - Bionic cultivation method of purple mushroom and application of bionic cultivation method in planting trees and forests - Google Patents
Bionic cultivation method of purple mushroom and application of bionic cultivation method in planting trees and forests Download PDFInfo
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- 241000939957 Lactarius vividus Species 0.000 claims abstract description 4
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- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 6
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Classifications
-
- 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
- A01G17/00—Cultivation of hops, vines, fruit trees, or like trees
- A01G17/005—Cultivation methods
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- Life Sciences & Earth Sciences (AREA)
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Abstract
A bionic cultivation method of purple mushroom comprises the steps of mycorrhizal seedling raising and field bionic cultivation. The purple mushroom (Lactarius vividus) is a fungus of Rugu of Russulaceae of Russula of Russulaceae, mainly distributed in the middle and south of China, and is a rare wild edible fungus. The strain has low natural yield and a descending trend, the price is always high, and the artificial cultivation is an urgent requirement at present. The invention discloses a bionic cultivation technology of the purple-flower fungi from the aspects of mycorrhizal seedling cultivation, field bionic cultivation, tree planting and afforestation in stony desertification areas and the like for the first time, and provides a new technology for stony desertification control, barren mountain afforestation, returning to farming and the like.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a method for cultivating ectomycorrhizal fungi and planting trees in the field, and more particularly relates to the technical problems of strain separation of purple-flowered fungi, mycorrhizal seedling synthesis and field planting trees.
Background
At present, most mycorrhizal fungi adopt a mode of cultivating mycorrhizal seedlings in an indoor sterile state and then cultivating soil, and the chanterelle (Canthraeus cibarius), the Lactarius deliciosus (Lactarius), the Lactarius hatsutake (Lactarius) and the like are successful. The purple mushroom (Lactarius viridus) is widely distributed in the middle and south of China and is eaten by a large number of people, in recent years, because the weather becomes warm, and human activities are added, the purple mushroom is difficult to find in mountain forests around cities, the trend of yield reduction is obvious, and no report about artificial bionic cultivation is provided at present, so that the yield of the purple mushroom can be increased and the vegetation can be recovered by combining bionic cultivation of the purple mushroom with afforestation, the ecological environment is improved, and the income is created for local farmers.
Disclosure of Invention
The invention aims to provide a bionic cultivation method of purple mushroom and application thereof in planting trees and forests. At present, no report about artificial bionic cultivation is available, and the bionic cultivation of the purple mushroom and the afforestation can increase the yield of the purple mushroom, restore vegetation, improve the ecological environment and create income for local farmers.
In order to achieve the above purpose of the present invention, the present invention provides the following technical solutions:
a bionic cultivation method of purple mushroom comprises the steps of mycorrhizal seedling raising and field bionic cultivation.
According to the bionic cultivation method of the purple mushroom, the mycorrhizal seedling raising step comprises the following steps:
1) preparing a host plant seedling: selecting host plant pinus massoniana seeds of purple fungi, cleaning the seeds for three times by using tap water, soaking the seeds for 30 minutes by using 75% alcohol for surface disinfection, flushing the seeds for three times by using sterile water, sowing the seeds in a plastic basket filled with 1:1 vermiculite and perlite, culturing the seeds under the conditions of accurately controlled greenhouse of 20-25 ℃ of air temperature and 60-70% of substrate humidity, irrigating the seeds by using sterile water, and preparing for inoculation after culturing the seeds in the host plant substrate for 3 months; sterilizing the seedling culture substrates vermiculite and perlite for 1h at 121 ℃ and 0.1MPa, standing for 3 days after sterilization, and sterilizing again for 1 time under the same conditions for use;
2) obtaining the strains of the purple fungi: under the aseptic condition, taking the flesh tissue at the joint of the pileus and the stipe of the purple mushroom, inoculating the flesh tissue on an MMN culture medium, and culturing in a dark place at 28 ℃, wherein the MMN culture medium comprises the following formula: MgSO (MgSO)4.7H2O：1.5g/L，KH2PO4: 3g/L, yeast extract: 5g/L, glucose: 20g/L, VB1：0.01g/L，NaCl：0.058g/L，CaCl2: 1.11g/L, ferric citrate: 1.5X 10- 2g/L, agar: 20g/L, and culturing for 20 days to obtain mycelium.
3) Infection of host plants by the purple-flowered fungi: transferring the non-pollution mycelia of the purple mushrooms obtained by the separation in the step 2) into an MMN liquid culture medium, containing no agar, standing and culturing for 4 weeks at 28 ℃, dividing the mycelia into 1cm × 1cm squares under aseptic operation, taking out 1) culturing 3 half-and-half-month host seedlings, cutting off main root tips, wherein the height of a nutrition pot is 14cm, the bottom is 5cm × 5cm, and the top is 6.5cm × 6.5cm, putting the host seedlings into the pot, adding a substrate to 1/3 parts of the root length, putting 2) cultured mycelia into the pots, cutting the mycelia into 1cm × 1cm squares, putting 4 blocks of the nutrition pots close to the root, then continuously adding the substrate to 2cm of the pot mouth, adding 1:1 sterilized vermiculite and perlite as the substrate, and culturing for 10-11 months to obtain the plants infected by the purple mushrooms.
According to the bionic cultivation method of the purple fungi, in the preparation step of the host plant seedlings, the pinaceae plants are selected as host plants of the purple fungi.
According to the bionic cultivation method for the purple mushroom, the outdoor bionic cultivation refers to transplanting mycorrhizal seedlings, infected by the purple mushroom, which are cultivated for 12-18 months, into natural soil, wherein the soil is red soil, yellow soil and purple soil, and the pH value of the soil is 5-6.5.
According to the bionic cultivation method of the purple mushroom, the soil is slate development soil.
The bionic cultivation method of the purple mushroom is applied to plant afforestation, a plant infected by the purple mushroom cultured for 12-18 months, namely a mycorrhizal sapling, is transplanted into natural soil, purple mushroom fruiting bodies are harvested in 2-3 years, the soil is red soil, yellow soil and purple soil, and the pH value of the soil is 5-6.5; the mycorrhizal sapling is obtained by the following method:
1) preparing host plant seedlings: selecting host plant pinus massoniana seeds of purple fungi, cleaning the seeds for three times by using tap water, soaking the seeds for 30 minutes by using 75% alcohol for surface disinfection, flushing the seeds for three times by using sterile water, sowing the seeds in a plastic basket filled with 1:1 vermiculite and perlite, culturing the seeds under the conditions of accurately controlled greenhouse of 20-25 ℃ of air temperature and 60-70% of substrate humidity, irrigating the seeds by using sterile water, and preparing for inoculation after culturing the seeds in the host plant substrate for 3 months; sterilizing the seedling culture substrates vermiculite and perlite for 1h at 121 ℃ and 0.1MPa, standing for 3 days after sterilization, and sterilizing again for 1 time under the same conditions for use;
2) obtaining the strains of the purple fungi: under the aseptic condition, taking the flesh tissue at the joint of the pileus and the stipe of the purple mushroom, inoculating the flesh tissue on an MMN culture medium, and culturing in a dark place at 28 ℃, wherein the MMN culture medium comprises the following formula: MgSO (MgSO)4.7H2O：1.5g/L，KH2PO4: 3g/L, yeast extract: 5g/L, glucose: 20g/L, VB1：0.01g/L，NaCl：0.058g/L，CaCl2: 1.11g/L, ferric citrate: 1.5X 10- 2g/L, agar: 20g/L, and culturing for 20 days to obtain mycelium.
3) Infection of host plants by the purple-flowered fungi: transferring the non-pollution mycelia of the purple mushrooms obtained by the separation in the step 2) into an MMN liquid culture medium, containing no agar, standing and culturing for 4 weeks at 28 ℃, dividing the mycelia into 1cm × 1cm squares under aseptic operation, taking out 1) culturing 3 half-and-half-month host seedlings, cutting off the tips of main roots, wherein the height of a nutrition pot is 14cm, the length of the bottom is 5cm × 5cm, the width of the top is 6.5cm × 6.5cm, putting the host seedlings into the pot, adding a substrate to 1/3 parts of the root length, putting the cultured mycelia into 1cm × 1cm squares, placing 4 blocks in each nutrition pot and closing to the root, then continuously adding the substrate to 2cm of the pot mouth, adding 1:1 sterilized vermiculite and perlite into the substrate, and culturing for 10-11 months to obtain the plants infected by the purple mushrooms.
The invention provides a bionic cultivation technology of the purple mushroom, and the technology is combined with afforestation, so that the yield of the purple mushroom is stable, the vegetation recovery is facilitated, the natural environment is friendly, and the local farmer is created.
Detailed Description
The following examples are provided to further illustrate the essence of the present invention and are not intended to limit the present invention.
Example 1
Bionic cultivation of purple mushroom (Lactarius vividus) and masson pine (Pinus massoniana) and application thereof in planting forests.
Identifying fruiting bodies of the purple mushrooms: the diameter of the pileus is 3-10cm, and when the pileus is rolled inwards, the center is quickly sunken and even turns outwards; the surface has slight annular stripe, water immersion, slight oily, light yellow, orange, gray orange, golden yellow, brown yellow, orange color of the mixture of central ochre and light gray green, orange color of the mixture of orange color and light blue green color of the mixture of orange color; some Basidion fruits are jagged at their edges when they are fully ripe, and occasionally split locally. The thickness of the mushroom meat is 2-8mm, the mushroom meat is light orange or lighter than the mushroom cap, the mushroom meat does not change color after being scratched, and the mushroom meat is light green after being aged. The fungus folds are 2-8mm wide and thick, are arranged crowded when young, are separated gradually along with time, and grow to a short and prolonged life, have bright orange color, golden yellow color and orange color, and do not fade to dark green and green when old or scratched. The stipe is 3-4.5 multiplied by 0.8-2cm, equally thick or gradually thinned downwards; the surface is slightly water-immersed, is slightly sunken, is bright orange to golden yellow, is almost the same as the color of a mushroom fold, has light local color and near white or lighter top, and does not change into green after being scratched; the bottom is provided with short, light orange or light green bristles. Less milk, orange to golden yellow, faded slowly to dark reddish blood after 2-10 minutes or drying. The spores were printed in light ochre.
1) Obtaining the pinus massoniana sterile seedlings: cleaning Pinus massoniana seeds with tap water for three times, soaking in 75% alcohol for 30 minutes for surface disinfection, performing aseptic water washing for three times, sowing in a plastic basket filled with 1:1 vermiculite and perlite, culturing under the conditions of a precisely controlled greenhouse of air temperature of 20-25 ℃ and matrix humidity of 60-70%, and performing aseptic water washing; sterilizing vermiculite and perlite as seedling culture medium at 121 deg.C under 0.1MPa for 1 hr, standing for 3 days, sterilizing again for 1 time under the same conditions, culturing in host plant matrix for 3 months, and preparing for inoculation;
2) obtaining the strains of the purple fungi: under the aseptic condition, taking the flesh tissue at the joint of the pileus and the stipe of the purple mushroom, inoculating the flesh tissue on an MMN culture medium, and culturing in a dark place at 28 ℃, wherein the MMN culture medium comprises the following formula: MgSO (MgSO)4.7H2O：1.5g，KH2PO4: 3g, yeast extract: 5g, glucose: 20g, VB1：0.01g，NaCl：0.058g，CaCl2: 1.11g, ferric citrate: 1.5X 10-2g, agar: 20g (1L); after 20 days mycelium grew.
3) Inoculation: after 6 months of cultivation of the pinus massoniana seedlings, the pinus massoniana seedlings are ready for inoculation. Before inoculation, the root tip of the main root of the seedling needs to be cut off by 1-2cm so as to facilitate the growth and infection of the lateral root; after the purple mushroom mycelium flat plate is full, under the aseptic condition, dividing the mycelium into squares with the size of 1cm multiplied by 1cm for standby application by using an inoculating knife, putting host seedlings into bowls, adding a matrix to the position with the root length being about 1/3, putting cut mycelium blocks, putting 4 blocks in each nutrition bowl and enabling the nutrition bowls to be close to the root, then continuously adding the matrix to the position with the bowl mouth being 2cm, wherein the added matrix is sterilized vermiculite and perlite with the ratio of 1:1, and culturing for 10-11 months to obtain the Chinese red mushroom infected masson pine;
4) field transplanting: transplanting the pinus massoniana seedlings cultured for 12-18 months into a region suitable for the growth of pinus massoniana and purple mushroom, wherein the pH value of soil is 5-6.5, the development soil of slate is optimal, during the mushroom production period, a cloud precious region is 6 months to 8 months of rainy season, a large amount of rainfall and large amount of temperature reduction are generated, the maximum temperature is lower than 20 ℃, the minimum temperature is above 10 ℃, the ground temperature is about 10-18 ℃, the average rainfall is 6-8mm per day, and the relative humidity reaches 80%. And after 2-3 years, the purple-flower mushroom fruiting bodies can be harvested. Five years after transplantation, 569 purple-flowered sporocarp (30kg) were produced from 32 pine trees.
The field transplanting of the purple-flower mycorrhizal masson pine seedlings has the advantages that the survival rate and the growth rate of the pine seedlings are improved compared with those of nonmycorrhizal masson pine seedlings, the edible fungi cultivated in a forest plantation represent a novel agricultural production activity, the increasing demand on delicious food can be met, the natural ecosystem can be maintained, and extra environmental and social economic benefits can be provided.
Example 2
Bionic cultivation of purple mushroom (Lactarius vividus) and Taiwan pine (Pinus taiwanensis) and application thereof in planting trees and forests.
Identifying fruiting bodies of the purple mushrooms: the diameter of the pileus is 3-10cm, and when the pileus is rolled inwards, the center is quickly sunken and even turns outwards; the surface has slight annular stripe, water immersion, slight oily, light yellow, orange, gray orange, golden yellow, brown yellow, orange color of the mixture of central ochre and light gray green, orange color of the mixture of orange color and light blue green color of the mixture of orange color; some Basidion fruits are jagged at their edges when they are fully ripe, and occasionally split locally. The thickness of the mushroom meat is 2-8mm, the mushroom meat is light orange or lighter than the mushroom cap, the mushroom meat does not change color after being scratched, and the mushroom meat is light green after being aged. The fungus folds are 2-8mm wide and thick, are arranged crowded when young, are separated gradually along with time, and grow to a short and prolonged life, have bright orange color, golden yellow color and orange color, and do not fade to dark green and green when old or scratched. The stipe is 3-4.5 multiplied by 0.8-2cm, equally thick or gradually thinned downwards; the surface is slightly water-immersed, is slightly sunken, is bright orange to golden yellow, is almost the same as the color of a mushroom fold, has light local color and near white or lighter top, and does not change into green after being scratched; the bottom is provided with short, light orange or light green bristles. Less milk, orange to golden yellow, faded slowly to dark reddish blood after 2-10 minutes or drying. The spores were printed in light ochre.
1) Cleaning Taiwan pine seeds for three times by using tap water, soaking the seeds for 30 minutes by using 75% alcohol for surface disinfection, flushing the seeds for three times by using sterile water, sowing the seeds into a plastic basket filled with 1:1 vermiculite and perlite, culturing the seeds under the conditions that the air temperature is 20-25 ℃ and the substrate humidity is 60-70% and flushing the seeds by using the sterile water; sterilizing vermiculite and perlite as seedling culture medium at 121 deg.C under 0.1MPa for 1 hr, standing for 3 days, sterilizing again for 1 time under the same conditions, culturing in host plant matrix for 3 months, and preparing for inoculation;
2) obtaining the strains of the purple fungi: under the aseptic condition, taking the flesh tissue at the joint of the pileus and the stipe of the purple mushroom, inoculating the flesh tissue on an MMN culture medium, and culturing in a dark place at 28 ℃, wherein the MMN culture medium comprises the following formula: MgSO (MgSO)4.7H2O：1.5g，KH2PO4: 3g, yeast extract: 5g, glucose: 20g, VB1：0.01g，NaCl：0.058g，CaCl2: 1.11g, ferric citrate: 1.5X 10-2g, agar: 20g (1L);
3) inoculation: the Taiwan pine is cultivated for 3 and a half months before being inoculated. Before inoculation, the root tip of the main root of the seedling needs to be cut off by 1-2cm so as to facilitate the growth and infection of the lateral root; after the purple mushroom mycelium flat plate is full, under the aseptic condition, dividing the mycelium into squares with the size of 1cm multiplied by 1cm for standby by using an inoculating knife, putting host seedlings into bowls, adding a matrix to the position with the root length of about 1/3, putting cut mycelium blocks, putting 4 blocks in each nutrition bowl and closing to the root, then continuously adding the matrix to the position with the bowl mouth of 2cm, wherein the added matrix is sterilized vermiculite and perlite with the ratio of 1:1, and culturing for 10-11 months to obtain Taiwan pine seedlings infected by the purple mushroom;
4) field transplanting: transplanting the seedling of the Taiwan pine cultivated for 10-11 months into a region suitable for the growth of the Taiwan pine and the purple mushroom, wherein the pH value of soil is 5-6.5, the development soil of slate is optimal, during the mushroom production period, the cloud precious region is 6 months to 8 months of rainy season, a large amount of rainfall and large amount of temperature reduction are generated, the maximum temperature is lower than 20 ℃, the minimum temperature is higher than 10 ℃, the ground temperature is about 10-18 ℃, the average rainfall is 6-8mm per day, and the relative humidity reaches 80%. And after 2-3 years, the purple-flower mushroom fruiting bodies can be harvested. And after 2-3 years, the purple-flower mushroom fruiting bodies can be harvested. After five years of transplantation, 593 purple-flower fungus sporophores (30kg) can be harvested from 40 pine.
The field transplantation of the purple-flower fungus mycorrhizal Taiwan pine not only improves the survival rate of pine seedlings, but also can harvest purple-flower fungus sporocarp, and is economic and ecological.
Claims (4)
1. A bionic cultivation method of purple mushroom Lactarius vividus is characterized in that the method comprises the steps of mycorrhizal seedling raising and field bionic cultivation;
the mycorrhizal seedling raising step comprises the following steps:
1) preparing a host plant seedling: selecting host plant pinus massoniana seeds of purple fungi, cleaning the seeds for three times by using tap water, soaking the seeds for 30 minutes by using 75% alcohol for surface disinfection, flushing the seeds for three times by using sterile water, sowing the seeds in a plastic basket filled with 1:1 vermiculite and perlite, culturing the seeds under the conditions of accurately controlled greenhouse of 20-25 ℃ of air temperature and 60-70% of substrate humidity, irrigating the seeds by using sterile water, and preparing for inoculation after culturing the seeds in a main plant substrate for 3 months; sterilizing the seedling culture substrates vermiculite and perlite for 1h at 121 ℃ and 0.1MPa, standing for 3 days after sterilization, and sterilizing again for 1 time under the same conditions for use;
2) obtaining the strains of the purple fungi: under the aseptic condition, taking the flesh tissue at the joint of the pileus and the stipe of the purple mushroom, inoculating the flesh tissue on an MMN culture medium, and culturing in a dark place at 28 ℃, wherein the MMN culture medium comprises the following formula: MgSO (MgSO)4·7H2O：1.5g/L，KH2PO4: 3g/L, yeast extract: 5g/L, glucose: 20g/L, VB1：0.01g/L，NaCl：0.058g/L，CaCl2: 1.11g/L, ferric citrate: 1.5X 10- 2g/L, agar: 20g/L, culturing for 20 days to obtain mycelium;
3) infection of host plants by the purple-flowered fungi: transferring the non-pollution mycelia of the purple mushrooms obtained by the separation in the step 2) into an MMN liquid culture medium, containing no agar, standing and culturing for 4 weeks at 28 ℃, dividing the mycelia into 1cm × 1cm squares under aseptic operation, taking out 1) culturing 3 half-and-half-month host seedlings, cutting off main root tips, wherein the height of a nutrition pot is 14cm, the bottom is 5cm × 5cm, and the top is 6.5cm × 6.5cm, putting the host seedlings into the pot, adding a substrate to 1/3 parts of the root length, putting 2) cultured mycelia into the pots, cutting the mycelia into 1cm × 1cm squares, putting 4 blocks of the nutrition pots close to the root, then continuously adding the substrate to 2cm of the pot mouth, adding 1:1 sterilized vermiculite and perlite as the substrate, and culturing for 10-11 months to obtain the plants infected by the purple mushrooms.
2. The bionic cultivation method of purple mushroom according to claim 1, characterized in that the field bionic cultivation is that the mycorrhizal seedlings of the purple mushroom infected plants cultivated for 12-18 months are transplanted into natural soil, the soil is red soil, yellow soil or purple soil, and the pH value of the soil is 5-6.5.
3. The biomimetic culture method of purple mushroom according to claim 2, characterized in that the soil is slate development soil.
4. The application of the bionic cultivation method of purple mushroom in the plant forestation of claim 1, which is characterized in that the plant infected by the purple mushroom cultured for 12-18 months, namely mycorrhizal sapling, is transplanted into natural soil, and the purple mushroom fruiting bodies are harvested in 2-3 years, wherein the soil is red soil, yellow soil or purple soil, and the pH value of the soil is 5-6.5; the mycorrhizal sapling is obtained by the following method:
1) preparing host plant seedlings: selecting host plant pinus massoniana seeds of purple fungi, cleaning the seeds for three times by using tap water, soaking the seeds for 30 minutes by using 75% alcohol for surface disinfection, flushing the seeds for three times by using sterile water, sowing the seeds in a plastic basket filled with 1:1 vermiculite and perlite, culturing the seeds under the conditions of accurately controlled greenhouse of 20-25 ℃ of air temperature and 60-70% of substrate humidity, irrigating the seeds by using sterile water, and preparing for inoculation after culturing the seeds in the host plant substrate for 3 months; sterilizing the seedling culture substrates vermiculite and perlite for 1h at 121 ℃ and 0.1MPa, standing for 3 days after sterilization, and sterilizing again for 1 time under the same conditions for use;
2) obtaining the strains of the purple fungi: under the aseptic condition, taking the flesh tissue at the joint of the pileus and the stipe of the purple mushroom, inoculating the flesh tissue on an MMN culture medium, and culturing in a dark place at 28 ℃, wherein the MMN culture medium comprises the following formula: MgSO (MgSO)4·7H2O：1.5g/L，KH2PO4: 3g/L, yeast extract: 5g/L, glucose: 20g/L, VB1：0.01g/L，NaCl：0.058g/L，CaCl2: 1.11g/L, ferric citrate: 1.5X 10- 2g/L, agar: 20g/L, culturing for 20 days to obtain mycelium;
3) infection of host plants by the purple-flowered fungi: transferring the non-pollution mycelia of the purple mushrooms obtained by the separation in the step 2) into an MMN liquid culture medium, containing no agar, standing and culturing for 4 weeks at 28 ℃, dividing the mycelia into 1cm × 1cm squares under aseptic operation, taking out 1) culturing 3 half-and-half-month host seedlings, cutting off the tips of main roots, wherein the height of a nutrition pot is 14cm, the length of the bottom is 5cm × 5cm, the width of the top is 6.5cm × 6.5cm, putting the host seedlings into the pot, adding a substrate to 1/3 parts of the root length, putting the cultured mycelia into 1cm × 1cm squares, placing 4 blocks in each nutrition pot and closing to the root, then continuously adding the substrate to 2cm of the pot mouth, adding 1:1 sterilized vermiculite and perlite into the substrate, and culturing for 10-11 months to obtain the plants infected by the purple mushrooms.
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