Culture medium synthesized by truffle mycorrhiza and application thereof

Abstract

The invention provides a culture medium synthesized by truffle mycorrhiza and application of the culture medium. Belongs to the technical field of plant and microorganism crossing. According to the method, mushroom dregs for planting edible mushrooms are used as raw materials, after aerobic fermentation, the mushroom dregs, vermiculite and perlite are evenly mixed according to a certain proportion, the pH is adjusted to 7.4-7.6, the culture medium synthesized by truffle mycorrhiza is prepared, after fermentation, the edible mushroom dregs have good water retention and air permeability and have nutrient substances needed by growth of aseptic seedlings, and the nutrient substances are rich in nutrient substances needed by growth of the aseptic seedlings. The culture medium is an ideal culture medium for mycorrhizal seedling culture; high-temperature sterilization treatment is carried out on the fungus sticks before spawn running in the early stage, other mycorrhizal fungi competing with truffles do not exist, and symbiotic culture of aseptic seedlings and truffle mycorrhiza can be effectively promoted. The invention further provides application of the culture medium in truffle mycorrhiza production and a specific application method, and technical support is provided for further development of truffle mycorrhiza production.

Description

Technical Field

[0001] The invention relates to the field of plant and microorganism cross technology, and more specifically to a cultivation substrate synthesized by truffle mycorrhiza and application thereof. Background Art

[0002] Truffles are edible fungi, and their hyphae must combine with the host plant's root system to form a symbiotic mycorrhizal relationship in order for the hyphae to survive and eventually grow into truffles. Therefore, in order to grow truffles, European countries first cultivate sterile seedlings, then inoculate truffle spores on the roots of the sterile seedlings, and plant the seedlings inoculated with fungi in sterilized cultivation substrates. After 5-6 months of co-cultivation, mycorrhizal seedlings will be synthesized, and finally the mycorrhizal seedlings will be transplanted into the wild for afforestation. After many years, truffles can be produced under the forest.

[0003] At present, the cultivation substrates for truffle mycorrhizal seedlings are mostly artificially prepared from a variety of substrates such as peat soil and sand. On the one hand, these substrates need to be sterilized at high temperature before use, which consumes a lot of energy and has high costs. On the other hand, peat soil is mostly produced in forests. There are many wild fungi in the forests, and a large number of spores are scattered in the soil. Most of these fungi spores can also form mycorrhizae with the roots of trees. In the mycorrhizal synthesis of truffles, these spores will compete with truffle spores for the ecological niche on the root system, which seriously affects the quality of truffle mycorrhizal synthesis. In production, many non-truffle mycorrhizae formed by contamination are mostly caused by incomplete sterilization of peat soil.

[0004] The edible fungus residue is the waste residue after the edible fungus is grown or the mushroom sticks cannot be used due to mold contamination during the cultivation process, and the waste residue becomes agricultural waste and cannot be effectively utilized. In addition, there is no application of edible fungus residue in the production of truffle mycorrhiza in the prior art.

[0005] Therefore, how to provide an edible mushroom residue for relevant application in truffle mycorrhiza production is a technical problem that those skilled in the art urgently need to solve. Summary of the invention

[0006] In view of this, the present invention provides a cultivation substrate synthesized by truffle mycorrhiza and its application. The residue of edible fungi is used as raw material, and after aerobic fermentation, it is mixed with vermiculite and perlite in a certain proportion, and the pH is adjusted to 7.4-7.6 to prepare a cultivation substrate synthesized by truffle mycorrhiza. After fermentation, the residue of edible fungi has good water retention and air permeability, and has the nutrients required for the growth of sterile seedlings, which is an ideal cultivation substrate for the cultivation of mycorrhizal seedlings; the mushroom sticks are sterilized at high temperature before spawning, and there are no other mycorrhizal fungi that compete with truffles, which can effectively promote the symbiotic culture of sterile seedlings and truffle mycorrhiza.

[0007] In order to achieve the above object, the present invention adopts the following technical solution:

[0008] A cultivation substrate synthesized by truffle mycorrhiza is prepared by mixing fermented edible fungus residue, vermiculite and perlite in a volume ratio of 1-2:1-2:0.5-1, and adjusting the pH to 7.4-7.6.

[0009] Preferably, the edible fungus residue is one or more of golden ear, black fungus, white fungus, shiitake mushroom, mulberry linterus and / or ganoderma lucidum.

[0010] Further preferably, the edible fungus residue is one or more of Fungus fuciformis, Auricularia auricula-judae or Phellinus igniarius.

[0011] Preferably, the edible fungus residue is waste residue produced after the mushroom sticks have been fruited or have been contaminated by mold.

[0012] Preferably, the fermentation process of the waste edible fungus residue is as follows:

[0013] (1) taking edible fungus residue, spraying lime water until the water content is 65-70%, and preparing a pre-treated base material;

[0014] (2) The base material of step (1) is used to build a pile. When the temperature of the pile reaches 55-65° C., the timing is started. After fermentation for 24 hours, the pile is turned once. After turning the pile, the fermentation is continued until the material temperature reaches 55-65° C., the timing is started. After fermentation for 24 hours, the pile is turned a second time. After turning the pile, the fermentation is continued until the material temperature reaches 55-65° C., the timing is started. After fermentation for 24 hours, the fermentation is terminated. When the material temperature drops to below 30° C., a fermented edible fungus residue is obtained.

[0015] The beneficial effects of the above technical solution are as follows: the present invention finds a substitute for peat soil, which, compared with the previous truffle mycorrhizal seedling cultivation technology, gets rid of the dependence on peat soil, reduces the high-temperature sterilization link, reduces energy consumption, and uses edible fungus residues to obtain the culture matrix through natural fermentation, which is more conducive to the promotion and application of production, reduces the interference of competitive bacteria, and improves the quality of mycorrhizal seedlings. It is conducive to the recycling of edible fungus waste residues, reduces the plunder of peat soil natural resources, and is green, low-carbon and environmentally friendly.

[0016] Preferably, the concentration of the lime water in step (1) is 1-2%.

[0017] Preferably, the pile body in step (2) is a trapezoidal pile body with a width of 2.0-2.5m, a height of 1.2-1.5m and an unlimited length; the turning process is as follows: turning the pile body 10-20cm on the surface of the pile body to the middle of the pile body, turning the middle pile body to the bottom and surface of the pile body, and the shape of the pile body after turning the pile body is the same as the initial pile body.

[0018] Another object of the present invention is to provide the use of the above-mentioned cultivation substrate in the production of truffle mycorrhiza.

[0019] Another object of the present invention is to provide a method for producing truffle mycorrhiza, comprising mixing truffle spores with the above-mentioned cultivation substrate, planting the cultured sterile seedlings in the cultivation substrate, and co-culturing with the truffle spores for 2-3 months to obtain mycorrhizal seedlings, which can be transplanted in the field for afforestation to produce truffles.

[0020] It can be seen from the above technical solution that compared with the prior art, the present invention has the following beneficial effects:

[0021] (1) Shorten the time for mycorrhizal formation. The time from inoculation to mycorrhizal formation is shortened from 5-6 months to 2-3 months.

[0022] (2) The mycorrhizal infection rate was increased. The infection rate of mycorrhizal seedlings increased from 80% to over 95%.

[0023] (3) The dosage of the microbial agent was reduced. The dosage of the microbial agent used for mycorrhizal synthesis was reduced from 1.2×10 7 The number of spores / strain was reduced to 1.2×10 6 Spores / strain. DETAILED DESCRIPTION

[0024] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0025] Example 1

[0026] A cultivation medium synthesized by truffle mycorrhiza:

[0027] (1) Selection of edible fungus residue

[0028] The method comprises the following steps: selecting mushroom sticks required for golden ear cultivation, wherein the mushroom sticks are prepared by mixing 60% sawdust, 20% wheat bran, 18% corn cobs, 1% lime and 1% gypsum, and then bagging and sterilizing under high pressure for 2-3 hours; inoculating the mushroom sticks with hairy leather fungus after the hairy leather fungus grows all over the mushroom sticks, and then inoculating golden ear fungus for growth; after the golden ear fungus is harvested after fruiting, the mushroom sticks become waste mushroom residues, which are rich in organic nutrients and can be used for seedling planting and mycorrhizal synthesis;

[0029] (2) Aerobic fermentation of edible fungus residue

[0030] Pretreatment of base material: Use the waste residue of Tremella fuciformis as the fermentation base material, spray 1% lime water, take a small handful of the residue and squeeze it tightly, if water drops seep out of the cultivation material, and the water drops are disconnected, then the moisture content of the base material reaches 70%;

[0031] Aerobic fermentation: pile the base material into a trapezoidal pile with a bottom width of 2.0-2.5m, a height of 1.2-1.5m and an unlimited length, and then use a 5cm diameter wooden stick to punch holes for oxygen permeability every 50cm. The holes are preferably in a herringbone shape. After punching the holes, cover the surface of the pile with straw mats; if covered with plastic film, only cover the surroundings of the material, not the surface, to facilitate ventilation; start timing when the temperature of the pile rises above 55℃, and turn the pile for the first time after 24 hours, and turn the pile upside down, inside and outside, that is, turn the material 10-20cm above the surface of the pile to the middle of the pile, and turn the middle material to the bottom and surface of the pile. After turning the pile, the shape of the pile is the same as the initial pile; when the temperature of the material rises to above 55℃ again for 24 hours, turn the pile for the second time. When the temperature of the material rises to above 55℃ again for 24 hours, the fermentation ends. When the temperature of the material drops to below 30℃, the fermented edible fungus residue is obtained;

[0032] (3) The edible fungus residue fermented in step (2) is mixed with vermiculite (2-4 mm) and perlite (3-6 mm) in a ratio of 2:2:1, and the pH value is adjusted to 7.4-7.6 with lime powder to prepare a cultivation medium synthesized by truffle mycorrhiza.

[0033] Example 2

[0034] A cultivation medium synthesized by truffle mycorrhiza:

[0035] (1) Selection of edible fungus residue

[0036] The mushroom sticks required for black fungus cultivation are selected. The mushroom sticks are prepared by mixing 58% of sawdust, 18% of wheat bran, 20% of corn cobs, 2% of soybean meal, 1% of lime and 1% of gypsum evenly, bagging and sterilizing under high pressure for 2-3 hours. After sterilization, the mushroom sticks are inoculated with black fungus strains. After the black fungus is harvested, the mushroom sticks become waste mushroom residues. The mushroom residues are rich in organic nutrients and can be used for seedling planting and mycorrhizal synthesis.

[0037] (2) Aerobic fermentation of edible fungus residue:

[0038] Pretreatment of base material: Use the waste residue of Tremella fuciformis as the fermentation base material, spray 2% lime water, take a small handful of the residue and squeeze it tightly, if water drops seep out of the cultivation material, and the water drops are disconnected, then the moisture content of the base material reaches 65%;

[0039] Aerobic fermentation: pile the base material into a trapezoidal pile with a bottom width of 2.0-2.5m, a height of 1.2-1.5m and an unlimited length, and then use a 5cm diameter wooden stick to punch holes for oxygen permeability every 50cm. The holes are preferably in a herringbone shape. After punching the holes, cover the surface of the pile with straw mats; if covered with plastic film, only cover the surroundings of the material, not the surface, to facilitate ventilation; start timing when the temperature of the pile rises above 60℃, and turn the pile for the first time after 24 hours, and turn the pile upside down, inside and outside, that is, turn the material 10-20cm above the surface of the pile to the middle of the pile, and turn the middle material to the bottom and surface of the pile. After turning the pile, the shape of the pile is the same as the initial pile; when the temperature of the material rises to above 60℃ again for 24 hours, turn the pile for the second time. When the temperature of the material rises to above 60℃ again for 24 hours, the fermentation ends. When the temperature of the material drops to below 30℃, the fermented edible fungus residue is obtained;

[0040] (3) The edible fungus residue fermented in step (2) is mixed with vermiculite (2-4 mm) and perlite (3-6 mm) in a ratio of 2:2:1, and the pH value is adjusted to 7.4-7.6 with lime powder to prepare a cultivation medium synthesized by truffle mycorrhiza.

[0041] Example 3

[0042] A cultivation medium synthesized by truffle mycorrhiza:

[0043] (1) Selection of edible fungus residue

[0044] The mushroom sticks required for the cultivation of Phellinus igniarius are selected. The mushroom sticks are prepared by mixing 40% mulberry branch chips, 20% sawdust, 20% bran, 18% corn cobs, 1% gypsum powder, and 1% brown sugar, and then bagging and sterilizing them under high pressure for 2-3 hours. After sterilization, the mushroom sticks are inoculated with Phellinus igniarius. After the Phellinus igniarius is harvested, the mushroom sticks become waste residues, which are rich in organic nutrients and can be used for seedling planting and mycorrhizal synthesis;

[0045] (2) Aerobic fermentation of edible fungus residue:

[0046] Pretreatment of base material: Use the waste residue of Tremella fuciformis as the fermentation base material, spray 1% lime water, take a small handful of the residue and squeeze it tightly, if water drops seep out of the cultivation material, and the water drops are disconnected, then the moisture content of the base material reaches 70%;

[0047] Aerobic fermentation: pile the base material into a trapezoidal pile with a bottom width of 2.0-2.5m, a height of 1.2-1.5m and an unlimited length, and then use a 5cm diameter wooden stick to punch holes for oxygen permeability every 50cm. The holes are preferably in a herringbone shape. After punching the holes, cover the surface of the pile with straw mats; if covered with plastic film, only cover the surroundings of the material, not the surface, to facilitate ventilation; start timing when the temperature of the pile rises above 65℃, and turn the pile for the first time after 24 hours, and turn the pile upside down, inside and outside, that is, turn the material 10-20cm above the surface of the pile to the middle of the pile, and turn the middle material to the bottom and surface of the pile. After turning the pile, the shape of the pile is the same as the initial pile; when the temperature of the material rises to above 65℃ again for 24 hours, turn the pile for the second time, and when the temperature of the material rises to above 65℃ again for 24 hours, the fermentation ends. When the material temperature drops to below 30℃, the fermented edible fungus residue is obtained;

[0048] (3) The edible fungus residue fermented in step (2) is mixed with vermiculite (2-4 mm) and perlite (3-6 mm) in a ratio of 2:2:1, and the pH value is adjusted to 7.4-7.6 with lime powder to prepare a cultivation medium synthesized by truffle mycorrhiza.

[0049] Example 4

[0050] Control group (peat soil: vermiculite: perlite = 2:2:1)

[0051] Embodiment 1 (fermented edible fungus residue: vermiculite: perlite=2:2:1)

[0052] Embodiment 2 (fermented edible fungus residue: vermiculite: perlite=2:2:1)

[0053] Example 3 (Fermented edible fungus residue: vermiculite: perlite = 2:2:1)

[0054] The cultivation substrates prepared in Example 1, Example 2 and Example 3 were used as the experimental group, and a control group in which traditional peat soil was used instead of the edible fungus residue of the present invention was set. The sterile seedlings of Pinus sylvestris were cultivated. After 2-3 months of cultivation, the mycorrhizal formation was observed. After the mycorrhizal formation was determined, the mycorrhizal morphology was recorded, and then 4 mycorrhizal seedlings were randomly taken out for each treatment to count the mycorrhizal roots. The mycorrhizal seedlings were taken out from the substrate bag, rinsed, and placed in a statistical container. The mycorrhizae wrapped by the same hyphae sheath were counted as one mycorrhizae, and the root system was divided into long and short ones that were easy to count, the number of mycorrhizae was recorded, and the mycorrhizal infection rate was calculated (Table 1).

[0055] Table 1 Comparative experiment on truffle mycorrhizal infection rate of different fermentation cultivation matrix formulas

[0056]

[0057]

[0058] Note: Capital letters indicate significant differences among treatments at the P 0.01 level.

[0059] Result analysis: When other ingredients (vermiculite, perlite) remain unchanged, the mycorrhizal infection rate can be significantly increased by replacing the peat soil in the traditional formula with the fermentation material of Example 1, and the infection rate is increased from 80.2% to 95.8%. The infection rate is increased to 95.1% by replacing the peat soil in the traditional formula with the fermentation material of Example 2, and the infection rate is increased to 95.7% by replacing the peat soil in the traditional formula with the fermentation material of Example 3. It can be seen that the synthetic infection rate of truffle mycorrhizae can be significantly affected by replacing the peat soil in the traditional formula with the fermentation material. And within the scope of the present invention, the infection rates of Examples 1, 2 and 3 are all higher than the infection rate of the control group.

[0060] Example 5

[0061] Control group (peat soil: vermiculite: perlite = 2:2:1)

[0062] Treatment group 1 (fermented edible fungus residue: vermiculite: perlite = 2:2:1)

[0063] Treatment group 2 (fermented edible fungus residue: vermiculite: perlite = 2:1:1)

[0064] Treatment group 3 (fermented edible fungus residue: vermiculite: perlite = 2:2:0.5)

[0065] The fermented edible fungi residue prepared by Example 1 was mixed with vermiculite and perlite in different proportions to obtain the cultivation substrates (treatment groups 1, 2, and 3) with different treatments. Traditional peat soil was set as a control group. After 2-3 months of inoculation of truffle inoculants in the cultivation of aseptic seedlings of Pinus sylvestris var. mongolica, the mycorrhizal formation was observed. After determining the mycorrhizal formation, 4 mycorrhizal seedlings were randomly taken out for each treatment to count mycorrhizal roots. The mycorrhizal seedlings were taken out from the substrate bag, rinsed clean, and put into a statistical container. The mycorrhizal roots wrapped by the same hyphae cover were used as mycorrhizal roots during counting. The root system was divided into the length that was easy to count, and the number of mycorrhizal roots was recorded. After obtaining the data, the method of average variance and significant difference analysis was used to analyze the influence of different treatments of the substrate formula on the mycorrhizal infection rate (Table 2).

[0066] Table 2 Comparative experiment on truffle mycorrhizal infection rate of different fermentation medium formulas

[0067]

[0068] Note: Capital letters indicate significant differences among treatments at the P 0.01 level.

[0069] Results: When other ingredients (vermiculite and perlite) are the same, fermented material replaces the peat soil in the traditional formula, which can significantly increase the mycorrhizal infection rate, from 80% to 96% (treatment 1). Reducing the proportion of vermiculite in the matrix formula (treatment 2) resulted in an infection rate of 94.9%. Reducing the proportion of perlite in the matrix formula (treatment 3) resulted in an infection rate of 94.8%. It can be seen that vermiculite and perlite, as common substrates for water retention and air permeability, mainly affect the growth environment of mycorrhizae, while fermented material affects the synthetic infection rate of truffle mycorrhizae. And within the scope of the present invention, the infection rates of treatments 1, 2 and 3 are all higher than those of the control group.

[0070] Example 6

[0071] Traditional substrate formula (peat soil: vermiculite: perlite = 2:2:1)

[0072] Formula of fermentation material matrix (fermented edible fungus residue: vermiculite: perlite = 2:2:1)

[0073] Inoculum dosage (spores / strain): 1.2×10 4 , 1.2×10 5 , 1.2×10 6 , 1.2×10 7 , 1.2×10 8

[0074] The fermented edible fungus residue prepared in Example 1 was mixed with vermiculite and perlite to obtain a cultivation matrix, and a traditional peat soil matrix formula was set at the same time. The spores of different inoculation amounts of the microbial agent were inoculated into the sterile seedlings of Pinus sylvestris var. mongolica. After 2-3 months of cultivation, the mycorrhizal formation was observed. After confirming the mycorrhizal formation, 4 mycorrhizal seedlings were randomly taken out from each treatment, and the mycorrhizal counts were performed to calculate the infection rate. After obtaining the data, the average variance and significant difference analysis methods were used to analyze the effects of different inoculation amounts of the microbial agent on the mycorrhizal infection rate (Table 3).

[0075] Table 3 Effects of different inoculation amounts of bacterial agents on mycorrhizal infection rate

[0076]

[0077]

[0078] Note: Capital letters indicate significant differences among treatments at the P 0.01 level.

[0079] Results: The traditional substrate formula and fermented substrate formula were used to plant sterile Pinus sylvestris seedlings, and inoculated with truffle spores of different bacterial doses. 7 spores / strain and 1.2×10 8The mycorrhizal infection rates of the three treatments were 79.9% and 80.1%, respectively, with no significant difference between the two, but significantly higher than the three treatments with low concentration inoculation. Therefore, the optimal truffle inoculation amount for cultivation with traditional substrate was 1.2×10 7 spores / plant. When cultivated with fermented material substrate, the spore inoculation amount was 1.2×10 6 , 1.2×10 7 and 1.2×10 8 When the number of spores / strain was 1.2×10 4 and 1.2×10 5 Therefore, the optimal truffle inoculum size was 1.2×10 6 And within the scope of the present invention, the infection rate of the culture using the fermentation medium is higher than that of the traditional medium.

[0080] The various embodiments in the specification are described in a progressive manner, and each embodiment focuses on the differences from other embodiments, and the same and similar parts between the various embodiments can be referred to each other. The above description of the disclosed embodiments enables professionals and technicians in the field to implement or use the present invention. Various modifications to these embodiments will be obvious to professionals and technicians in the field, and the general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but will conform to the widest range consistent with the principles and novel features disclosed herein.

Claims

1. A cultivation medium synthesized by truffle mycorrhiza, characterized in that The product is prepared by uniformly mixing fermented edible fungus residue, vermiculite and perlite in a volume ratio of 1-2:1-2:0.5-1, and adjusting the pH value to 7.4-7.

6.

2. The synthetic cultivation medium of truffle mycorrhiza according to claim 1, characterized in that The edible fungus residue is one or more of golden ear, black fungus, white ear, shiitake mushroom, mulberry linterus and / or ganoderma lucidum.

3. The synthetic cultivation medium of truffle mycorrhiza according to claim 1, characterized in that The edible fungus residue is one or more of golden fungus, black fungus or mulberry linterus.

4. The synthetic cultivation medium of truffle mycorrhiza according to claim 3, characterized in that The edible mushroom residue is the waste residue generated after the mushroom sticks have finished fruiting or have been contaminated by mold.

5. The synthetic cultivation medium of truffle mycorrhiza according to claim 1, characterized in that, The fermentation process of the edible fungus residue is as follows: (1) taking edible fungus residue, spraying lime water until the water content is 65-70%, and preparing a pre-treated base material; (2) The base material of step (1) is used to build a pile. When the temperature of the pile reaches 55-65° C., the timing is started. After fermentation for 24 hours, the pile is turned once. After turning the pile, the fermentation is continued until the material temperature reaches 55-65° C., the timing is started. After fermentation for 24 hours, the pile is turned a second time. After turning the pile, the fermentation is continued until the material temperature reaches 55-65° C., the timing is started. After fermentation for 24 hours, the fermentation is terminated. When the material temperature drops to below 30° C., a fermented edible fungus residue is obtained.

6. The synthetic cultivation medium of truffle mycorrhiza according to claim 5, characterized in that The concentration of the lime water in step (1) is 1-2%.

7. The synthetic cultivation medium of truffle mycorrhiza according to claim 5, characterized in that The pile body in step (2) is a trapezoidal pile body with a width of 2.0-2.5m, a height of 1.2-1.5m and an unlimited length; the turning process is as follows: turning the pile body 10-20cm on the surface of the pile body to the middle of the pile body, turning the middle pile body to the bottom and surface of the pile body, and the shape of the pile body after turning the pile body is the same as the initial pile body.

8. Application of any cultivation substrate described in claim 1-7 in truffle mycorrhiza production.

9. A method for producing truffle mycorrhizae, characterized in that: The truffle spores are mixed with the cultivation substrate according to any one of claims 1 to 7, and the cultured sterile seedlings are planted in the cultivation substrate and co-cultured with the truffle spores for 2 to 3 months to obtain mycorrhizal seedlings, which can be transplanted in the field for afforestation and truffle production.

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