CN113079943A - Cultivation method and extraction method for increasing content of terpenoid in phellinus igniarius sporocarp - Google Patents
Cultivation method and extraction method for increasing content of terpenoid in phellinus igniarius sporocarp Download PDFInfo
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- 150000003505 terpenes Chemical class 0.000 title claims abstract description 24
- 238000012364 cultivation method Methods 0.000 title claims abstract description 18
- 241000123113 Phellinus igniarius Species 0.000 title claims description 30
- 238000000605 extraction Methods 0.000 title abstract description 7
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- MJYQFWSXKFLTAY-OVEQLNGDSA-N (2r,3r)-2,3-bis[(4-hydroxy-3-methoxyphenyl)methyl]butane-1,4-diol;(2r,3r,4s,5s,6r)-6-(hydroxymethyl)oxane-2,3,4,5-tetrol Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O.C1=C(O)C(OC)=CC(C[C@@H](CO)[C@H](CO)CC=2C=C(OC)C(O)=CC=2)=C1 MJYQFWSXKFLTAY-OVEQLNGDSA-N 0.000 claims abstract description 10
- 244000280244 Luffa acutangula Species 0.000 claims abstract description 10
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- 238000005286 illumination Methods 0.000 claims description 10
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- 235000012255 calcium oxide Nutrition 0.000 claims description 7
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- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- 240000001548 Camellia japonica Species 0.000 claims description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 5
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- XBZYWSMVVKYHQN-MYPRUECHSA-N (4as,6as,6br,8ar,9r,10s,12ar,12br,14bs)-10-hydroxy-2,2,6a,6b,9,12a-hexamethyl-9-[(sulfooxy)methyl]-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carboxylic acid Chemical compound C1C[C@H](O)[C@@](C)(COS(O)(=O)=O)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CCC(C)(C)C[C@H]5C4=CC[C@@H]3[C@]21C XBZYWSMVVKYHQN-MYPRUECHSA-N 0.000 abstract description 3
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- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
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- 239000007864 aqueous solution Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000010495 camellia oil Substances 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
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- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
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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
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Mycology (AREA)
- Environmental Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Mushroom Cultivation (AREA)
Abstract
The invention belongs to the technical field of artificial cultivation of edible fungi, and particularly relates to a cultivation method and an extraction method for improving the content of terpenoid substances in phellinus linteus sporocarp. The cultivation method for improving the content of the terpenoid in the phellinus linteus sporocarp is characterized in that the phellinus linteus sporocarp with high content of terpenoid is optimized and developed on the basis of artificial bag material cultivation, mulberry twig sawdust is used as a cultivation substrate, and peel residue, loofah sponge, flaxseed cake pulp, oil tea cake pulp and propolis are added as nutritional raw materials, so that the nutritional requirement of phellinus linteus cultivation can be met, the content of the triterpenoid in the cultivated phellinus linteus sporocarp is effectively improved through induction, and the nutrition and application value of phellinus linteus are effectively improved.
Description
Technical Field
The invention belongs to the technical field of artificial cultivation of edible fungi, and particularly relates to a cultivation method and an extraction method for improving the content of terpenoid substances in phellinus linteus sporocarp.
Background
Phellinus igniarius (Phellinus igniarius) belongs to Basidiomycotina (Basidiomycotina), Hymenomycetes (Hymenomycetes), Aphyllophorales (Aphyllophorales), Hymenochaeyaceae (Hymenochaeyaceae), Phellinus (Phellinus), and is the trade name of Phellinus igniarius (P.igniarius), Phellinus baumii (P.baumii) and Phellinus linteus (P.Linteus), and is generally grown on the stems of withered trees and stumps of broad-leaved trees such as poplar, mulberry, willow, white birch, zelkova, peach, and the like. Phellinus linteus is a precious medicinal fungus developed in recent years for many years and is named as forest gold in the name of America. According to the records of Chinese medicine dictionary, Phellinus can be used for treating gynecological diseases such as metrorrhagia, bloody stranguria, leukorrhagia, amenorrhea, etc.; modern medical research also shows that phellinus igniarius has remarkable effects of resisting tumor, bacteria and fibrosis, resisting oxidation, improving human immunity and the like, is the rare medicinal fungus with the first biological anti-tumor effect internationally acknowledged at present, and is widely applied to the industries of medicines, foods, daily chemicals, health care products and the like.
At present, due to the restriction of the particularity and complexity of physiological states and external environment, the formation of the fruit body of phellinus igniarius in nature is difficult, so that the natural phellinus igniarius is rare in quantity, the resource available for medicine in nature is limited, and the demand of domestic and foreign markets for phellinus igniarius is increased, so that medicine farmers in domestic places take away to collect the phellinus igniarius, the fruit body cannot be formed in large quantity, the phellinus igniarius cannot become a stable industrial product source, and the development of the phellinus igniarius industry is very unfavorable, so that the defect of wild resources is made up through artificial cultivation, and the method is very important. However, artificial cultivation of Phellinus linteus is extremely difficult, and has problems of harsh culture conditions and long growth cycle.
At present, two methods are mainly used for artificially cultivating phellinus igniarius, one of the methods is mainly the cut-log cultivation of mulberry trees, poplar trees and oak trees, but the cut-log cultivation needs a large amount of wood resources, the resource waste is serious, and the large-scale cultivation is difficult; the other is bag cultivation, but has the problems of insufficient nutrient supply of the culture medium, small fruit body development, easy contamination of mixed bacteria and the like, and particularly, the content of active substances is not ideal.
Disclosure of Invention
Therefore, the technical problem to be solved by the invention is to provide a cultivation method for increasing the content of terpenoid in phellinus linteus sporocarp, so as to solve the problem that the content of terpenoid active substances in phellinus linteus sporocarp cultivated by artificial bagged materials in the prior art is low;
the second technical problem to be solved by the invention is to provide a method for extracting terpenoid active substances from phellinus igniarius sporocarp.
In order to solve the technical problems, the cultivation method for improving the content of terpenoids in the phellinus linteus fruiting body comprises the following steps:
(1) preparing a cultivation bag material: taking 20-30 parts by weight of mulberry twig sawdust, 10-20 parts by weight of peel residues, 5-12 parts by weight of loofah sponge, 3-8 parts by weight of flaxseed cake dregs, 3-8 parts by weight of oil-tea camellia cake dregs, 1-3 parts by weight of propolis, 1-5 parts by weight of quick lime, KH2PO40.3-0.8 weight part, and uniformly mixing to obtain the required culture medium; adding water into the culture medium, mixing uniformly, and bagging for later use;
(2) mycelium culture: inoculating a phellinus igniarius mother seed into the cultivation bag material, and carrying out constant-temperature cultivation on the inoculated cultivation bag material until the cultivation bag material is full of mycelia;
(3) and (3) fruiting body culture: cutting the bag material full of mycelia to produce mushroom, continuously performing constant temperature culture until the fruiting body is mature, and harvesting.
Specifically, in the step (1), the cultivation bag material comprises the following components: 25 parts of mulberry twig sawdust, 15 parts of peel residues, 8 parts of loofah sponge, 5 parts of flaxseed cake meal and oil5 parts of tea cake dregs, 2 parts of propolis, 3 parts of quicklime and KH2PO40.5 part by weight.
Specifically, in the step (1), the peel dregs comprise apple peel dregs, orange peel dregs and/or pear peel dregs.
Specifically, in the step (1), the mass ratio of the culture medium to water is 40-50%: 50 to 60 percent.
Specifically, in the step (2), the inoculation amount of the phellinus igniarius mother seeds is 5-20 wt% of the culture medium.
Specifically, in the step (2), the process conditions of the mycelium culturing step include: the culture temperature is controlled at 20-28 deg.C, air humidity is 50-70%, and illumination intensity is 10-20 lux.
Specifically, in the step (3), the process conditions of the fruiting body cultivation step include: the culture temperature is controlled at 20-28 deg.C, air humidity is 50-70%, and illumination intensity is 20-80 lux.
The invention also discloses a phellinus igniarius sporocarp cultivated by the method.
The invention also discloses a method for extracting terpenoids in the phellinus igniarius sporocarp, which comprises the step of carrying out alcohol extraction by taking the phellinus igniarius sporocarp as a raw material.
The cultivation method for improving the content of the terpenoid in the phellinus linteus sporocarp is characterized in that the phellinus linteus sporocarp with high content of terpenoid is optimized and developed on the basis of artificial bag material cultivation, mulberry twig sawdust is used as a cultivation substrate, and peel residue, loofah sponge, flaxseed cake pulp, oil tea cake pulp and propolis are added as nutritional raw materials, so that the nutritional requirement of phellinus linteus cultivation can be met, the content of the triterpenoid in the cultivated phellinus linteus sporocarp is effectively improved through induction, and the nutrition and application value of phellinus linteus are effectively improved.
Detailed Description
In the following examples of the present invention, the phellinus linteus mother strain is obtained by separation and purification by a conventional method, activated by a conventional PDA plate medium, and inoculated into a conventional PD mother strain medium for culture, thereby obtaining the phellinus linteus mother strain.
Example 1
The cultivation method for increasing the content of terpenoids in phellinus linteus fruiting bodies comprises the following steps:
(1) preparing a cultivation bag material: taking 20 parts by weight of mulberry twig sawdust, 20 parts by weight of apple peel residues, 5 parts by weight of loofah sponge, 8 parts by weight of flaxseed cake pulp, 3 parts by weight of oil tea cake pulp, 3 parts by weight of propolis, 1 part by weight of quick lime and KH2PO40.8 part by weight, and uniformly mixing to obtain the required culture medium; taking the culture medium according to the ratio of 50%: adding 50% of water by mass, mixing uniformly, and bagging for later use;
(2) mycelium culture: inoculating a phellinus igniarius mother seed liquid material into the cultivation bag material obtained in the step (1) according to the inoculation amount of 10%, placing the inoculated cultivation bag material at the temperature of 25 ℃ for constant-temperature cultivation, controlling the air humidity to be 60% and the illumination condition to be 15lux, keeping the cultivation time in a spaced ventilation state, and observing the cultivation condition of the bag material at any time until the cultivation bag material is full of mycelia;
(3) and (3) fruiting body culture: cutting a cut on the bag material full of mycelia for fruiting, continuously controlling the temperature to be 25 ℃, the air humidity to be 60% and the illumination intensity to be 50lux for constant-temperature culture, keeping the interval ventilation state between cultures, observing the fruiting condition of the bag material at any time, and harvesting in time.
Example 2
The cultivation method for increasing the content of terpenoids in phellinus linteus fruiting bodies comprises the following steps:
(1) preparing a cultivation bag material: 30 parts of mulberry twig sawdust, 10 parts of orange peel residues, 12 parts of loofah sponge, 3 parts of flaxseed cake pulp, 8 parts of oil tea cake pulp, 1 part of propolis, 5 parts of quick lime and KH2PO40.3 part by weight, and uniformly mixing to obtain the required culture medium; taking the culture medium according to the ratio of 50%: adding 50% of water by mass, mixing uniformly, and bagging for later use;
(2) mycelium culture: inoculating a phellinus igniarius mother seed liquid material into the cultivation bag material obtained in the step (1) according to the inoculation amount of 10%, placing the inoculated cultivation bag material at the temperature of 25 ℃ for constant-temperature cultivation, controlling the air humidity to be 60% and the illumination condition to be 15lux, keeping the cultivation time in a spaced ventilation state, and observing the cultivation condition of the bag material at any time until the cultivation bag material is full of mycelia;
(3) and (3) fruiting body culture: cutting a cut on the bag material full of mycelia for fruiting, continuously controlling the temperature to be 25 ℃, the air humidity to be 60% and the illumination intensity to be 50lux for constant-temperature culture, keeping the interval ventilation state between cultures, observing the fruiting condition of the bag material at any time, and harvesting in time.
Example 3
The cultivation method for increasing the content of terpenoids in phellinus linteus fruiting bodies comprises the following steps:
(1) preparing a cultivation bag material: 25 parts of mulberry twig sawdust, 5 parts of apple peel residues, 5 parts of orange peel residues, 5 parts of white pear peel residues, 8 parts of loofah sponge, 5 parts of flaxseed cake residues, 5 parts of oil-tea camellia cake residues, 2 parts of propolis, 2 parts of quicklime and KH2PO40.5 part by weight, and uniformly mixing to obtain the required culture medium; taking the culture medium according to the ratio of 50%: adding 50% of water by mass, mixing uniformly, and bagging for later use;
(2) mycelium culture: inoculating a phellinus igniarius mother seed liquid material into the cultivation bag material obtained in the step (1) according to the inoculation amount of 10%, placing the inoculated cultivation bag material at the temperature of 25 ℃ for constant-temperature cultivation, controlling the air humidity to be 60% and the illumination condition to be 15lux, keeping the cultivation time in a spaced ventilation state, and observing the cultivation condition of the bag material at any time until the cultivation bag material is full of mycelia;
(3) and (3) fruiting body culture: cutting a cut on the bag material full of mycelia for fruiting, continuously controlling the temperature to be 25 ℃, the air humidity to be 60% and the illumination intensity to be 50lux for constant-temperature culture, keeping the interval ventilation state between cultures, observing the fruiting condition of the bag material at any time, and harvesting in time.
Comparative example 1
The method for cultivating phellinus linteus in the comparative example is the same as example 3, and is different from the method in the embodiment in that the same amount of sucrose is used to replace the peel residue in the cultivation bag material.
Comparative example 2
The method for cultivating phellinus linteus in the comparative example is the same as that in example 3, only the loofah sponge is not added in the cultivation bag material.
Comparative example 3
The method for cultivating phellinus linteus in the comparative example is the same as example 3, and is different from the method in the embodiment only in that the flaxseed cake is not added to the cultivation bag material.
Comparative example 4
The method for cultivating phellinus linteus in the comparative example is the same as example 3, and is different from the method in the embodiment only in that the tea-oil camellia cake is not added into the cultivation bag material.
Examples of the experiments
1. Phellinus igniarius sporophore weight and mass
The weight and mass of the fruiting bodies of Phellinus linteus harvested in example 3 and comparative examples 1-4 were recorded, and the results are shown in Table 1 below.
TABLE 1 Phellinus linteus fruiting body culture
| Numbering | Weight of fruiting body/g | Quality and shape of fruiting body |
| Example 3 | 45g | Preferably, it is close to wild |
| Comparative example 1 | 38g | Preferably, it is close to wild |
| Comparative example 2 | 43g | Preferably, it is close to wild |
| Comparative example 3 | 41g | Preferably, it is close to wild |
| Comparative example 4 | 42g | Preferably, it is close to wild |
2. The content of triterpenes in Phellinus Linteus fruiting body
The phellinus linteus fruiting bodies harvested in the above example 3 and comparative examples 1-4 are respectively cleaned, dried and crushed, 3 times of 95% ethanol aqueous solution is added into the crushed phellinus linteus material, extraction is carried out for 1h at 50 ℃ under the assistance of ultrasound, extraction is carried out for 2 times in total, the extract is collected and combined, the content of triterpenoids in the extract is detected after concentration, meanwhile, the content of triterpenoids is detected by taking commercially available wild phellinus linteus fruiting bodies as a control, and the detection results are recorded in the following table 2.
The analysis and determination of the content of the triterpenoids adopt a conventional ultraviolet colorimetric method, ursolic acid is taken as a standard substance, and the content of the triterpenoids in the extracting solution is determined.
TABLE 2 content of triterpenes in Phellinus Linteus
| Numbering | Phellinus igniarius sporophore with triterpenoid content mg/g |
| Example 3 | 10.6 |
| Comparative example 1 | 10.1 |
| Comparative example 2 | 9.1 |
| Comparative example 3 | 8.5 |
| Comparative example 4 | 8.3 |
| Comparative example | 5.2 |
From the above results, it can be seen that the quality of the phellinus igniarius sporocarp is improved, the content of triterpenes in the phellinus igniarius sporocarp is effectively increased through adjustment of the nutritional ingredients in the phellinus igniarius artificial cultivation bag material, and the nutritional and medicinal values of the phellinus igniarius sporocarp are improved.
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are within the scope of the invention.
Claims (9)
1. A cultivation method for increasing the content of terpenoids in phellinus linteus sporocarp is characterized by comprising the following steps:
(1) preparing a cultivation bag material: taking 20-30 parts by weight of mulberry twig sawdust, 10-20 parts by weight of peel residues, 5-12 parts by weight of loofah sponge, 3-8 parts by weight of flaxseed cake dregs, 3-8 parts by weight of oil-tea camellia cake dregs, 1-3 parts by weight of propolis, 1-5 parts by weight of quick lime, KH2PO40.3 to 0.8 weight portion, and mixing evenlyObtaining a required culture medium; adding water into the culture medium, mixing uniformly, and bagging for later use;
(2) mycelium culture: inoculating a phellinus igniarius mother seed into the cultivation bag material, and carrying out constant-temperature cultivation on the inoculated cultivation bag material until the cultivation bag material is full of mycelia;
(3) and (3) fruiting body culture: cutting the bag material full of mycelia to produce mushroom, continuously performing constant temperature culture until the fruiting body is mature, and harvesting.
2. The cultivation method for increasing the content of terpenoids in phellinus linteus fruiting body according to claim 1, wherein in the step (1), the cultivation bag material comprises the following components: 25 parts of mulberry twig sawdust, 15 parts of peel residues, 8 parts of loofah sponge, 5 parts of flaxseed cake pulp, 5 parts of oil-tea camellia cake pulp, 2 parts of propolis, 3 parts of quick lime and KH2PO40.5 part by weight.
3. The cultivation method for increasing the content of terpenoids in phellinus linteus fruiting body according to claim 1 or 2, wherein in the step (1), the peel dregs comprise apple peel dregs, orange peel dregs and/or pear peel dregs.
4. The cultivation method for improving the content of terpenoids in phellinus linteus fruiting body according to any one of claims 1 to 3, wherein in the step (1), the mass ratio of the cultivation substrate to water is 40-50%: 50 to 60 percent.
5. The cultivation method for increasing the content of terpenoids in phellinus linteus fruiting body according to any one of claims 1 to 4, wherein in the step (2), the inoculation amount of phellinus linteus mother species is 5-20 wt% of the cultivation medium.
6. The cultivation method for improving the content of terpenoids in phellinus linteus fruiting body according to any one of claims 1 to 5, wherein in the step (2), the process conditions of the mycelium cultivation step include: the culture temperature is controlled at 20-28 deg.C, air humidity is 50-70%, and illumination intensity is 10-20 lux.
7. The cultivation method for improving the content of terpenoids in phellinus linteus fruiting body according to any one of claims 1 to 6, wherein in the step (3), the process conditions of the fruiting body cultivation step include: the culture temperature is controlled at 20-28 deg.C, air humidity is 50-70%, and illumination intensity is 20-80 lux.
8. Phellinus linteus fruiting body cultivated by the method according to any one of claims 1 to 7.
9. A method for extracting terpenoids from Phellinus linteus fruiting body, which comprises extracting with ethanol the Phellinus linteus fruiting body of claim 8 as raw material.
Priority Applications (1)
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