CN115500208B - Method for cultivating poria cocos fruiting bodies - Google Patents
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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
- A01G18/00—Cultivation of mushrooms
- A01G18/20—Culture media, e.g. compost
-
- 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
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- Life Sciences & Earth Sciences (AREA)
- Mycology (AREA)
- Environmental Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Mushroom Cultivation (AREA)
Abstract
The invention discloses a method for cultivating tuckahoe fruiting bodies, which comprises the following steps: (1) preparing a culture medium: mixing pine wood scraps 70 parts, wheat bran powder 15-20 parts, corn cob powder 9.5-14.5 parts and sucrose 0.5 part with water; (2) fungus bag manufacturing: bagging the culture medium obtained in the step (1), sterilizing, cooling, inoculating Poria cocos cultivar, and culturing in dark until mycelia grow over the whole fungus bag; (3) fruiting body cultivation: culturing the fungus bag obtained in the step (2) in a scattered light environment with the temperature of 26-28 ℃ and the air humidity of 70-80% for 15-30 days. The poria cocos sporophore produced by the method has the advantages of high yield, low cost, short period and excellent quality. Compared with the prior art, the method for cultivating the poria cocos sporophore has the advantages of simpler practical operation, no mixed bacteria pollution, high safety, good economic and social benefits and great value for large-scale popularization and use.
Description
Technical Field
The invention belongs to the field of edible fungi, and particularly relates to a method for cultivating poria cocos fruiting bodies.
Background
Poria cocos (Schw.) Wolf is a higher basidiomycete belonging to the genus Poria of the family Polyporaceae, and is originally carried in Shennong Ben Cao Jing, and has effects of promoting diuresis, removing dampness, invigorating spleen and calming heart. The medicinal part of Poria is sclerotium, so that the research is more, and large-scale cultivation is performed at present. However, wild germplasm resources of the poria cocos are deficient, and the cultivated strain is homogenized and seriously degenerated due to long-term asexual propagation, so that the poria cocos is artificially induced to form sexual fruiting bodies, the spore ejected by the sexual fruiting bodies is used for seed production, and the poria cocos strain with strong hypha attraction and high survival rate can be obtained through a sexual propagation mode. Therefore, the cultivation of the poria cocos sporophore is beneficial to the deep research of mating and life history of the poria cocos sporophore, and promotes the development of the poria cocos industry.
At present, less research is conducted on the poria cocos fruiting body, and a better method for culturing the poria cocos fruiting body is lacked in actual production or in a laboratory, so that an effective method for culturing the poria cocos fruiting body needs to be explored. Patent CN202011030917.8 discloses a method for culturing Poria fruiting body, its culture medium (potato, glucose, KH) 2 PO 4 、MgSO 4 .7H 2 O and VB 1 ) The cost is higher, and strict light intensity control is required in the cultivation process, so that the operation difficulty is higher. Patent CN110771429a discloses a method for culturing poria cocos fruiting body by using trichoderma in an induction way, and the method can be used for culturing the poria cocos fruiting body quickly, but trichoderma grows quickly in the culturing process, and can generate gliotoxin and chlorotoxin, so that experiments prove that the trichoderma has the effects of immunosuppression and apoptosis promotion on mammalian cells, and a certain safety risk exists. Meanwhile, trichoderma can be parasitic on tuckahoe mycelia and fruiting bodies, so that the research on obtaining tuckahoe basidiospores from fruiting bodies and obtaining tuckahoe sexual mycelia is greatly influenced, and tuckahoe strains are impure, so that the yield and quality of tuckahoe are influenced.
Disclosure of Invention
In order to solve the technical problems, the invention provides a method for cultivating tuckahoe fruiting bodies, which comprises the following steps:
(1) Preparing a culture medium: mixing pine wood scraps 70 parts, wheat bran powder 15-20 parts, corn cob powder 9.5-14.5 parts and sucrose 0.5 part with water;
(2) And (3) preparing fungus bags: bagging the culture medium obtained in the step (1), sterilizing, cooling, inoculating Poria cocos cultivar, and culturing in dark until mycelia grow over the whole fungus bag;
(3) Culturing fruiting body: culturing the fungus bag obtained in the step (2) in a scattered light environment with the temperature of 26-28 ℃ and the air humidity of 70-80% until the color of the surface of the fruiting body is changed from white to light yellow.
Further, 70 parts of pine dust, 20 parts of wheat bran powder, 9.5 parts of corn cob powder and 0.5 part of sucrose in the step (1).
Further, the water content of the culture medium in the step (1) is 60% -65%.
Further, the sterilization pressure in the step (2) is 0.13MPa, the temperature is 121 ℃ and the time is 1h.
Further, the inoculation amount of the poria cocos cultivation seeds in the step (2) is 30% of the weight of the culture medium.
Further, the temperature of the light-shielding culture in the step (2) is 27-29 ℃.
Further, the temperature of the light-shielding culture in the step (2) is 28 ℃.
Further, the temperature of step (3) is 26 ℃.
The poria cocos sporophore produced by the method has the advantages of high yield, low cost, short period and excellent quality. Compared with the prior art, the method for cultivating the poria cocos sporophore has the advantages of simpler practical operation, no mixed bacteria pollution, high safety, good economic and social benefits and great value for large-scale popularization and use.
It should be apparent that, in light of the foregoing, various modifications, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.
The above-described aspects of the present invention will be described in further detail below with reference to specific embodiments in the form of examples. It should not be understood that the scope of the above subject matter of the present invention is limited to the following examples only. All techniques implemented based on the above description of the invention are within the scope of the invention.
Drawings
FIG. 1 shows Poria cocos culture medium ((1) number 1-10 is experimental group (1) to (2) number 11-20 is repeated group, and (3) is culture medium of mycelium growing fungus bag
FIG. 2 shows Poria cocos (the pictures are the Poria cocos fruiting bodies obtained by culturing in sequence in test examples (1) to (back))
FIG. 3 Poria cocos fruiting body microstructure
Detailed Description
Example 1 cultivation of Poria cocos fruiting body
(1) Preparing a culture medium: taking 70 parts of pine wood scraps, 20 parts of wheat bran powder, 9.5 parts of corn cob powder and 0.5 part of sucrose, adding water, and uniformly mixing until the water content of a culture medium is 60% -65%;
(2) And (3) preparing fungus bags: placing the culture medium obtained in the step (1) into a 12×24×5 polypropylene bag, sealing with a breathable collar, sterilizing for 1h under the conditions of 0.13MPa and 121 ℃, cooling, inoculating Poria cocos cultivar into the culture medium in an aseptic environment at 30% of the weight of the culture medium, and culturing at 28deg.C in a dark place until mycelia grow over the whole bag;
(3) Culturing fruiting body: culturing the fungus bag obtained in the step (2) in a scattered light environment with the temperature of 26 ℃ and the air humidity of 70% -80% until the color of the surface of the poria cocos fruiting body is changed from white to light yellow, and harvesting.
Example 2 cultivation of Poria cocos fruiting body
(1) Preparing a culture medium: taking 70 parts of pine wood scraps, 15 parts of wheat bran powder, 14.5 parts of corncob powder and 0.5 part of sucrose, adding water, and uniformly mixing until the water content of the culture medium is 60% -65%;
(2) And (3) preparing fungus bags: placing the culture medium obtained in the step (1) into a 12×24×5 polypropylene bag, sealing with a breathable collar, sterilizing for 1h under the conditions of 0.13MPa and 121 ℃, cooling, inoculating Poria cocos cultivar into the culture medium in an aseptic environment at 30% of the weight of the culture medium, and culturing at 28deg.C in a dark place until mycelia grow over the whole bag;
(3) Culturing fruiting body: culturing the fungus bag obtained in the step (2) in a scattered light environment with the temperature of 28 ℃ and the air humidity of 70% -80% until the color of the surface of the poria cocos fruiting body is changed from white to light yellow, and harvesting.
The advantageous effects of the present invention are further illustrated by test examples below.
Experimental example 1 study of Poria cocos fruiting body cultivation method
1. Research on influence of culture medium formula on poria cocos fruiting bodies
1. Preparation of substitute culture medium
The culture medium is prepared from pine wood dust, wheat bran powder, corncob powder and sucrose as main raw materials, wherein the proportion of 4 factors in the formula is shown in table 1, a 12×24×5 polypropylene bag is used for preparing the culture medium, the filling amount of each bag is 200g, the water content is 60% -65% of the culture medium matrix, the bags are filled for later use, and a ventilation sleeve ring is used for sealing.
2. Sterilization of poria cocos substitute culture medium
And (3) transferring the substitute culture medium into a sterilizing pot for sterilization, adjusting the pressure to 0.13MPa, sterilizing the substitute culture medium at 121 ℃ and sterilizing the substitute culture medium for 1h.
3. Inoculation of Poria cocos strain
After the culture medium for the substitute is sterilized, the culture medium is placed overnight, the poria cocos culture seeds are inoculated into the culture medium for the substitute according to 30% of the culture medium matrix on an ultra-clean workbench, and the culture medium is placed in a constant-temperature incubator for light-shielding culture at 28 ℃ for 7-9 d until the poria cocos mycelia grow full of the whole fungus bag.
4. Cultivation of Poria fruiting body
After the fungus bag is fully grown by the tuckahoe mycelium, the fungus bag is placed at the temperature of 28 ℃ and the air humidity of 70% -80%, and the color of the surface of the tuckahoe sporophore is changed from white to light yellow by the light scattering (not light-shielding and no light direct irradiation).
Table 1 culture medium formulation
2. Results
The yield of the harvested tuckahoe fruiting bodies, the time for completing one culture period and the culture cost are counted, and are shown in table 2 and fig. 1-2.
TABLE 2 time and yield of Poria cocos fruiting bodies obtained by culturing in each experimental group
As can be seen from table 1: the dry weight of the poria cocos fruiting body obtained by culturing with different culture mediums is greatly different, when the culture medium formula is 70.0% of pine dust, 15% -20.0% of wheat bran powder, 9.5% -14.5% of corncob powder and 0.5% of sucrose, the dry weight of the fruiting body can reach more than 60g/kg, when the pine dust accounts for 70.0% of the formula, the wheat bran powder accounts for 20.0% of the formula, the corncob powder accounts for 9.5% of the formula and the sucrose accounts for 0.5% of the formula, the fruiting body yield is obviously higher than that of other groups, the cost is lowest, and the culture time is short. Thus, the medium formulation of experimental group (1) was the most selected, combining three factors, medium cost, days of culture and fruiting body yield.
At present, most of poria cocos sporophores are obtained by using a flat plate or shake flask culture mode, and glucose, peptone, yeast extract and KH are used for preparing the poria cocos sporophores 2 PO 4 、MgSO 4 The culture medium is adopted, and the method has low yield and high cost; the method uses a small amount of pine wood dust, wheat grains, corn grains and other crops as culture medium to cultivate fruiting bodies with substitute materials, and can effectively improve the fruiting body yield and reduce the cost. The invention uses the substitute material to cultivate fruit bodies, uses the crop waste wheat bran powder to replace wheat grains and uses the corn cob powder to replace corn grains, thereby effectively reducing the cultivation cost. Meanwhile, the specific culture medium in the cultivation method can effectively improve the yield of fruiting bodies, shorten the cultivation time and further reduce the cost.
2. Research on cultivation conditions of poria cocos fruiting bodies
The fruiting body yield of Poria cocos under different culture conditions is also significantly different. Therefore, on the basis of optimizing the formula of the culture medium, the influence of the culture conditions of the poria cocos fruiting bodies is continuously studied.
1) Preparation of substitute culture medium
The culture medium is prepared by using 70% of pine dust, 20% of wheat bran powder, 9.5% of corncob powder and 0.5% of sucrose as raw material formulas of the culture medium, using 12X 24X 5 polypropylene bags, wherein the filling amount of each bag is 200g, the water content is 60% -65% of the culture medium matrix, and the bags are filled for later use and sealed by a breathable sleeve ring.
2) Sterilizing Poria substitute culture medium
And (3) transferring the substitute culture medium into a sterilizing pot for sterilization, adjusting the pressure to 0.13MPa, sterilizing the substitute culture medium at 121 ℃ and sterilizing the substitute culture medium for 1h.
3) Inoculation of Poria cocos strain
After the culture medium for the substitute is sterilized, the culture medium is placed overnight, the poria cocos culture seeds are inoculated into the culture medium for the substitute according to 30% of the culture medium matrix on an ultra-clean workbench, and the culture medium is placed in a constant-temperature incubator for light-shielding culture at 28 ℃ for 7-9 d until the poria cocos mycelia grow full of the whole fungus bag.
4) Cultivation of Poria fruiting body
Culturing Poria mycelia in scattered light (not light-shielding and directly irradiating without light) until the surface color of Poria fruiting body is changed from white to light yellow. Wherein the culture temperature and humidity and the yield of Poria cocos fruiting body at the time of harvesting are shown in Table 3.
TABLE 3 yields of Poria cocos fruiting bodies under different culture conditions
Annotation: "-" means no fruiting body formation
As can be seen from table 3: the yield of the poria cocos fruiting bodies is obviously different under different temperatures, different humidity and different shading degrees, and the fruiting bodies are higher in yield at 26-28 ℃ and highest in yield at 26 ℃; the yield of the poria cocos fruiting bodies is higher under the condition of 70-80% of humidity; no fruiting body is generated under the experimental conditions of complete light shielding or direct light irradiation. Thus, the culture conditions selected in the present invention are: the temperature is 26 ℃, the air humidity is 70-80 percent, and the culture is carried out by scattered light (not light-proof and directly irradiated without light). Compared with the existing published conditions, the culture conditions are completely different, strict illumination control is not needed, other microorganism spores are not needed to be inoculated, and the culture process is simpler and more controllable.
3. Verification
According to the optimal formula of the culture medium: preparing a fungus bag by using pine wood dust with a ratio of 70.0%, wheat bran powder with a ratio of 20.0%, corn cob powder with a ratio of 9.5% and sucrose with a ratio of 0.5%; optimal culture conditions: the temperature is 26 ℃, the air humidity is 70-80 percent, and the culture is carried out by scattered light (not light-proof and directly irradiated without light). Verification tests were performed according to the above optimal culture technique, 3 times in parallel, and the results are shown in table 4.
TABLE 4 verification of stability of optimal culture Process for Poria cocos
From the results, it can be seen that: the average dry weight of the poria cocos fruiting body is 107.28 +/-4.65 g, the dry weight of the fruiting body is increased by about 17% compared with that of the fruiting body after only optimizing the culture medium, and the RSD value of 3 parallel verification tests is 4.33%, so that the process stability is good.
4. Discussion of the invention
The current research reports on the poria cocos fruiting bodies are few, and the poria cocos fruiting bodies are mostly concentrated on the culture process. The invention cultures the poria cocos fruiting body, prepares seeds by using the ejected basidiospores, and obtains the poria cocos strain by a sexual propagation mode. The invention observes the microscopic structure of the poria cocos fruiting body obtained by the culture in the optimal process, and discovers that the poria cocos fruiting body obtained by the invention is rich in basidiospores (see figure 3), and the poria cocos strain can be obtained by a sexual propagation mode.
In summary, the invention has the following advantages: (1) The formula of the poria cocos sporophore culture medium is optimized through a mixing design method, and on the basis, the culture conditions are continuously optimized, so that the optimal formula of the culture medium is obtained: pine wood dust accounts for 70.0%, wheat bran powder accounts for 20.0%, corn cob powder accounts for 9.5%, and sucrose accounts for 0.5%; the optimal culture conditions are as follows: the temperature is 26 ℃, the air humidity is 70-80 percent, and the culture is carried out by scattered light (not light-proof and directly irradiated without light). The formation rate of the poria cocos fruiting body can reach 100% by performing 3 parallel verification tests under the culture medium and the culture conditions, the fruiting body yield (dry weight) is 107.28 +/-4.65 g/kg of the culture medium, and the RSD value of 3 tests is 4.33%, which indicates that the process stability is good.
(2) The substitute culture medium uses wheat bran powder to replace wheat grains as a nitrogen source and corn cob powder to replace corn as a carbon source, so that the cost of the culture medium can be effectively reduced by about 40 percent, and the cost of each bag of culture medium is only 0.23 yuan. All materials of the culture medium are pine scraps and crop wastes, are cheap and easy to obtain, promote comprehensive utilization of resources, and have good economic and social benefits.
(3) Compared with other methods at present, the culture condition is easier to operate, strict illumination intensity control is not needed, other microorganism spores are not needed to be supplemented, and the culture process is simple and controllable to operate.
(4) The method for culturing the poria cocos fruiting body by using the substitute material has the advantages of high yield, low cost, simplicity and controllability in operation, good repeatability and the like. Meanwhile, the poria cocos sporophore basidiospore obtained by culture is rich, the poria cocos strain is obtained by a sexual propagation mode, deep research on sexual propagation of poria cocos can be promoted, and the poria cocos sporophore basidiospore has reference value for improving yield and quality stability of poria cocos.
Claims (6)
1. A method for cultivating tuckahoe fruiting bodies, which is characterized by comprising the following steps:
(1) Preparing a culture medium: taking 70 parts of pine wood scraps, 15-20 parts of wheat bran powder, 9.5-14.5 parts of corncob powder and 0.5 part of sucrose, and adding water to mix uniformly to obtain a culture medium with the water content of 60% -65%;
(2) And (3) preparing fungus bags: bagging the culture medium obtained in the step (1), sterilizing, cooling, inoculating poria cocos cultivars, and culturing in a dark place at the temperature of 27-29 ℃ until mycelia grow over the whole fungus bag;
(3) Culturing fruiting body: culturing the fungus bag obtained in the step (2) in a scattered light environment with the temperature of 26-28 ℃ and the air humidity of 70% -80% until the color of the surface of the fruiting body is changed from white to light yellow.
2. The method according to claim 1, characterized in that: 70 parts of pine dust, 20 parts of wheat bran powder, 9.5 parts of corn cob powder and 0.5 part of sucrose in the step (1).
3. The method according to claim 1, characterized in that: the sterilization pressure in the step (2) is 0.13MPa, the temperature is 121 ℃ and the time is 1h.
4. The method according to claim 1, characterized in that: the inoculation amount of the poria cocos cultivar in the step (2) is 30% of the weight of the culture medium.
5. The method according to claim 1, characterized in that: the temperature of the light-shielding culture in the step (2) is 28 ℃.
6. The method according to claim 1, characterized in that: the temperature in the step (3) is 26 ℃.
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| CN1076825A (en) * | 1993-03-29 | 1993-10-06 | 王汉培 | Method with living pine parasitic cultivation Poria cocos |
| KR20180112149A (en) * | 2017-03-30 | 2018-10-12 | 경상북도(농업기술원) | Mass production method of wolfiporia cocos fruitbody |
| CN112136598A (en) * | 2020-09-23 | 2020-12-29 | 重庆市中药研究院 | Efficient poria cocos planting method |
| CN112449992A (en) * | 2020-11-11 | 2021-03-09 | 江西省林业科学院 | Poria cocos production seed culture medium and preparation method of poria cocos production seeds |
| CN113383674A (en) * | 2021-07-07 | 2021-09-14 | 云茯苓普洱有限公司 | Planting method of poria cocos |
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| CN1076825A (en) * | 1993-03-29 | 1993-10-06 | 王汉培 | Method with living pine parasitic cultivation Poria cocos |
| KR20180112149A (en) * | 2017-03-30 | 2018-10-12 | 경상북도(농업기술원) | Mass production method of wolfiporia cocos fruitbody |
| CN112136598A (en) * | 2020-09-23 | 2020-12-29 | 重庆市中药研究院 | Efficient poria cocos planting method |
| CN112449992A (en) * | 2020-11-11 | 2021-03-09 | 江西省林业科学院 | Poria cocos production seed culture medium and preparation method of poria cocos production seeds |
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