Novel low-temperature erythromelastoma sanguineum strain and pennisetum hydridum cultivation method thereof
Through the TsJC206, a new strain of low-temperature hemocytosaccharide, and the fungus grass cultivation method, the high-temperature cultivation restrictions and traditional material consumption problems were solved, and low-cost and sustainable hemocytosaccharide production was achieved.
Patent Information
- Application Number
- CN202510514332.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-04-23
- Publication Date
- 2025-07-25
AI Technical Summary
The existing hemorosomal cultivation technology mostly adopts high temperature conditions, which limits its production time and region. Moreover, traditional cultivation materials consume wood and are costly, making it difficult to achieve sustainable development.
The new strain of low-temperature hemorosum fungus was used as the raw material for culture medium, and low-temperature resistant strains were obtained through ultraviolet mutagenesis and directional screening. Combined with green organic cultivation methods, including the parent species, original species and vegetarian management steps, fresh grass instead of some dry materials.
It has achieved the production of zhizi at 15-20℃, which has reduced production costs, expanded production time and region, and achieved sustainable development and high-quality cultivation of the fungi industry.
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Abstract
Description
Technical Field
[0001] The present invention belongs to the field of microbial cultivation, and particularly relates to a new strain of Trametes sanguinea at low temperature and a cultivation method thereof using Juncao (edible mushroom grass). Background Art
[0002] Trametes sanguinea belongs to Basidiomycotina, Hymenomycetes, Aphyllophorales, Polyporaceae, and Trametes. Through acute toxicology tests on zebrafish models, it was found that its safety is similar to that of Antrodia cinnamomea. This fungus has high medicinal value, can relieve itching, stop bleeding, lower blood sugar, etc., and can also enhance immunity and fight tumors. There is polyporen in its fruiting body, which has an inhibitory effect on Gram-negative and Gram-positive bacteria. The basidiocarp of this fungus is annual, small to medium in size, corky, sessile or nearly sessile, the pileus is semicircular to fan-shaped, reaching 3 - 10 cm in size and 2 - 6 mm in thickness, the surface is smooth or slightly hairy, initially blood red, and gradually becomes orange-red, showing alternating light and dark ring patterns. Trametes sanguinea has a wide range of economic uses and obvious medicinal value, so more and more people are developing and utilizing it. Collecting wild Trametes sanguinea for isolation and purification, ultraviolet mutagenesis, directional screening, identification, and domestication cultivation can provide certain support for large-scale cultivation and development of Trametes sanguinea while protecting germplasm resources.
[0003] Trametes sanguinea is a wood saprophyte, and most of them are cultivated using linden wood, wood chips or cottonseed hulls. Juncao refers to herbaceous plants that contain nutrients suitable for the growth of microorganisms such as edible fungi and medicinal fungi and have comprehensive development and utilization value. Cultivating edible and medicinal fungi with herbaceous plants instead of wood can not only achieve the sustainable development of the mushroom industry, but also reduce the consumption of wood and protect the ecological environment. Experiments show that edible and medicinal fungi cultivated with Juncao as raw materials have higher yields and nutritional values than traditional raw materials such as wood chips and cottonseed hulls, and the heavy metal content of edible and medicinal fungi cultivated with Juncao is lower than that of traditional wood chip formulations.
[0004] The present invention relates to a new strain of Trametes sanguinea with low temperature type. During its growth process, the edge growth area is white, and the white edge disappears after maturity. It is sessile or nearly sessile, the pileus is orange-red, the surface is smooth or slightly hairy, and it is suitable for cultivation with Juncao. The content of the present invention covers the isolation and purification, ultraviolet mutagenesis and directional screening, identification, and cultivation method of this strain. The new strain of Trametes sanguinea with low temperature type is Trametes sanguinea TsJC206, which was deposited at the China Center for Type Culture Collection on June 19, 2023, with the deposit number CCTCC NO: M20231065, and the deposit address is No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province. The GenBank number of the characteristic sequence of Trametes sanguinea in NCBI is OR056420. The artificial cultivation method of Trametes sanguinea TsJC206 includes mother spawn preparation, original spawn preparation, mycelium culture, fruiting body management, harvesting, and preservation. With this strain and the method and formula for artificial cultivation of Trametes sanguinea, green, organic, and sustainable Juncao can be used as raw materials, and Trametes sanguinea can be produced during the relatively low temperature period of 15 - 20 °C, having high economic and ecological value. Summary of the Invention
[0005] The purpose of the present invention is to provide a new strain of Trametes sanguinea with low temperature type and its cultivation method.
[0006] To achieve the above purpose, the present invention adopts the following technical solutions: A new strain of Trametes sanguinea with low temperature tolerance. The new strain of Trametes sanguinea is Trametes sanguinea TsJC206, which was deposited at the China Center for Type Culture Collection on June 19, 2023, with the deposit number CCTCC NO: M 20231065, and the deposit address is No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province. The method for obtaining the new strain of Trametes sanguinea is as follows: The fresh fruiting bodies of wild Trametes sanguinea are air-dried. After the scalpel is sterilized by burning on an alcohol lamp and cooled, clean and pollution-free internal tissues are cut from the torn surface and inoculated on the mother culture medium, and then placed in an incubator at a constant temperature of 15-28°C for 5-7 days. The vigorous and fastest-growing mycelia are inoculated into a new mother culture medium for ultraviolet mutagenesis and directional screening, and then continue to be cultured, thereby obtaining the new strain of Trametes sanguinea TsJC206 with vigorous growth, fast growth rate and better low temperature tolerance. The new strain of Trametes sanguinea has the following characteristics: First, one or more tall and fast-growing forage grasses can be used as the main raw material of the culture medium, such as: Pennisetum giganteum, Arundo donax, Pennisetum sinese Roxb., Pennisetum purpureum Schum., etc. Even the fresh grasses of Pennisetum giganteum, Arundo donax, Pennisetum purpureum Schum. can be used for cultivation. The triterpene content of its fruiting bodies can reach 1.15%, the polysaccharide content can reach 10.82%, and the conversion rate is 15%-25%; Second, it is a low temperature type strain, and the fruiting temperature is 15-20°C, which is significantly lower than the fruiting temperature reported in the existing literature and patents, supplementing the fruiting temperature type of Trametes sanguinea; Third, the base at the 182nd site of the ITS sequence in its NCBI database is guanine (G), which is different from the adenine (A) at the 223rd site of the ITS sequence of Trametes sanguinea EOS4 (OR062414.1) strain with the highest similarity in its NCBI database.
[0007] The cultivation method of the above new strain of Trametes sanguinea includes the following steps: S1: Inoculate the new strain of Trametes sanguinea with low temperature tolerance into a test tube containing the mother culture medium, and incubate it in the dark in an incubator at 15-28°C until the mycelia fill the test tube to obtain the mother culture. S2: Inoculate the mother culture into a plastic bag containing the solid original culture medium according to aseptic operation, and incubate it in the dark in an incubator at 15-28°C until the mycelia fill the plastic bag to obtain the original culture; or inoculate the mother culture into a shake flask containing the liquid original culture medium according to aseptic operation, and incubate it in the dark on a shaker at a temperature of 15-28°C and a rotation speed of 120-160 rpm until mycelial pellets appear to obtain the original culture. S3: Inoculate the original culture into a plastic bag containing the cultivation material according to aseptic operation, and incubate it in the dark in a cultivation room at a temperature of 15-28°C and a relative air humidity of 70%-80% until the mycelia fill the plastic bag; Transfer the plastic bag with mature mycelia to the mushroom shed for fruiting management. During fruiting management, the mycelia are in full contact with the air. The temperature in the shed is 15-20°C, the relative air humidity is 90%-110%, and scattered light and sufficient oxygen are provided in the shed, and cultivate until the fruiting bodies are mature.
[0008] Furthermore, the formula of the above mother culture medium is by mass percentage: potato flour 2% - 6%, corn flour 1% - 3%, glucose 0.5% - 2%, agar powder 1.5% - 3%, ferrous sulfate heptahydrate 0.001% - 0.005%, magnesium sulfate 0.001% - 0.005%, calcium sulfate heptahydrate 0.2% - 0.5%, crude sea salt 0.1% - 0.2%, potassium dihydrogen phosphate or dipotassium hydrogen phosphate 0.1% - 0.2%, and the balance is water.
[0009] Furthermore, the formula of the above solid spawn culture medium is by mass percentage: sorghum 15% - 30%, corncob 15% - 30%, wheat bran 5% - 10%, gypsum 0.5% - 1.5%, light calcium carbonate 0.5% - 1.5%, magnesium oxide 0.002% - 0.01%, ferrous sulfate heptahydrate 0.002% - 0.005%, magnesium sulfate 0.001% - 0.01%, crude sea salt 0.1% - 0.2%, potassium dihydrogen phosphate or dipotassium hydrogen phosphate 0.1% - 0.2%, and dried mushroom grass makes up the balance; adjust the water content to 55% - 65%; The mushroom grass is any one or more of Pennisetum giganteum, Arundo donax, Pennisetum purpureum, Neyraudia reynaudiana, Pennisetum purpureum Schum., Pennisetum americanum × P. purpureum, Phragmites australis, Silphium perfoliatum, Cymbopogon citratus, Miscanthus sacchariflorus, Themeda villosa, Sorghum propinquum, Vetiveria zizanioides, Paspalum wettsteinii, Dicranopteris dichotoma; calculated based on the water content of fresh mushroom grass, according to the principle of equal dry weight, the dried mushroom grass can be replaced by fresh mushroom grass.
[0010] Furthermore, the formula of the above liquid spawn culture medium is by mass percentage: potato flour 1% - 3%, glucose 1% - 3%, peptone 0.1% - 0.5%, yeast extract powder 0.1% - 0.5%, ferrous sulfate heptahydrate 0.001% - 0.005%, magnesium sulfate 0.001% - 0.01%, calcium sulfate heptahydrate 0.1% - 0.2%, crude sea salt 0.1% - 0.2%, potassium dihydrogen phosphate or dipotassium hydrogen phosphate 0.1% - 0.2%, and the balance is water.
[0011] Furthermore, the formula of the above cultivation material is by mass percentage: wheat bran or rice bran 10% - 25%, gypsum 0.5% - 1.5%, light calcium carbonate 0.5% - 1.5%, magnesium oxide 0.002% - 0.01%, ferrous sulfate heptahydrate 0.002% - 0.005%, magnesium sulfate 0.001% - 0.01%, crude sea salt 0.1% - 0.2%, potassium dihydrogen phosphate or dipotassium hydrogen phosphate 0.1% - 0.2%, and dried mushroom grass makes up the balance; adjust the water content to 55% - 65%; The said mushroom grass is any one or more of Pennisetum giganteum, Arundo donax, Pennisetum purpureum, Neyraudia reynaudiana, Pennisetum purpureum Schumach. cv. Zixiangcao, Pennisetum americanum×P. purpureum, Phragmites australis, Silphium perfoliatum, Cymbopogon citratus, Miscanthus sacchariflorus, Themeda villosa, Sorghum propinquum, Vetiveria zizanioides, Paspalum wettsteinii, Dicranopteris dichotoma; calculated based on the water content of fresh mushroom grass, according to the principle of equal dry basis weight, the dry mushroom grass can be replaced by fresh mushroom grass.
[0012] The remarkable advantages of the present invention are as follows: (1) It can produce Trametes cinnabarina at 15-20°C. Generally, the fruiting temperature of Trametes cinnabarina is about 28°C, belonging to the high-temperature type mushroom, while Trametes cinnabarina TsJC206 can fruit at 15-20°C, fruiting at a different time from the high-temperature type Trametes cinnabarina, and can cooperate with the general high-temperature type Trametes cinnabarina to produce Trametes cinnabarina for most of the year; it can also produce Trametes cinnabarina in regions and seasons where Trametes cinnabarina could not be produced before due to lower temperatures.
[0013] (2) It uses mushroom grass, a green, organic and sustainable raw material, to cultivate Trametes cinnabarina. Most of the existing cultivation techniques of Trametes cinnabarina use traditional materials such as sawdust. Using mushroom grass, a green, organic and sustainable raw material, is conducive to the high-quality development of the Trametes cinnabarina industry.
[0014] (3) Fresh mushroom grass can be used to replace part of the culture medium for cultivating Trametes cinnabarina. Currently, when cultivating edible and medicinal fungi, basically dried raw materials are used, which is time-consuming and increases costs. Using fresh mushroom grass for cultivation can eliminate the drying step, reduce costs, and can be used in regions and seasons where it is inconvenient to dry the raw materials. Description of the Drawings
[0015] Figure 1 : Mycelial plate diagram of Trametes cinnabarina TsJC206.
[0016] Figure 2 : Microscopic structure diagram of the mycelium of Trametes cinnabarina TsJC206.
[0017] Figure 3 : Microscopic structure diagram of the spores of Trametes cinnabarina TsJC206.
[0018] Figure 4 : Fruit body diagram of wild Trametes cinnabarina (left is the front, right is the back).
[0019] Figure 5 : Mycelial growth of Trametes cinnabarina TsJC206 in solid original seed culture media containing different mushroom grasses.
[0020] Figure 6 : Comparison of fruiting situations of Trametes cinnabarina TsJC206 and common Trametes cinnabarina at 16°C.
[0021] Figure 7 : The fruiting situation of Trametes sanguinea TsJC206 in the cultivation medium containing Miscanthus floridulus and Dicranopteris dichotoma. Detailed implementation manners
[0022] To make the content of the present invention easier to understand, the technical solutions of the present invention will be further described below in conjunction with specific implementation manners. However, the present invention is not limited thereto.
[0023] Example 1: The wild fresh fruiting bodies of Trametes sanguinea collected were air-dried. After the scalpel was sterilized by burning on an alcohol lamp and cooled, clean and pollution-free internal tissues were cut from the torn surface and inoculated on the mother culture medium, and placed in a constant temperature of 15 - 28°C for 5 - 7 days. The vigorous and fastest-growing mycelia were inoculated into a new mother culture medium for ultraviolet mutagenesis and directional screening, and then continued to be cultured, so as to obtain a new strain of Trametes sanguinea TsJC206 with vigorous growth, fast growth rate and more cold tolerance; the formula of the mother culture medium is calculated by mass percentage as follows: potato whole powder 15%, corn flour 2%, glucose 1%, agar powder 2%, ferrous sulfate heptahydrate 0.002%, magnesium sulfate 0.002%, calcium sulfate heptahydrate 0.2%, crude sea salt 0.1%, potassium dihydrogen phosphate 0.1%, and the balance is water. The new strain of Trametes sanguinea TsJC206 was deposited at the China Center for Type Culture Collection on June 19, 2023. The classification name is named Trametes sanguinea, and the deposit number is CCTCC NO: M 20231065. The deposit address is Wuhan University, China. The GenBank number of the characteristic sequence of the new strain of Trametes sanguinea TsJC206 in NCBI is OR056420.
[0024] The new strain of Trametes sanguinea TsJC206 has the following characteristics: First, TsJC206 can use one or more tall and fast-growing forage grasses as the main raw materials of the culture medium, such as: Pennisetum giganteum, Arundo donax, Pennisetum sinese Roxb., Pennisetum purpureum Schum., etc. Even the fresh grass of Pennisetum giganteum, the fresh grass of Arundo donax, the fresh grass of Pennisetum purpureum Schum. etc. can be used for cultivation. The triterpenoid content of the fruiting body can reach 1.15%, the polysaccharide content can reach 10.82%, and the biological conversion rate can reach 15% - 25%; Second, TsJC206 is a low-temperature type strain, and the fruiting temperature is 15 - 20°C, which is significantly lower than the fruiting temperature reported in the existing literature and patents, supplementing the fruiting temperature type of Trametes sanguinea; Third, the base at the 182nd position of the ITS sequence of TsJC206 in its NCBI database is guanine (G), which is different from the base adenine (A) at the 223rd position of the ITS sequence of the Trametes sanguinea EOS4 (OR062414.1) strain with the highest similarity in its NCBI database.
[0025] Figure 1It is a mycelial plate diagram of Trametes sanguinea TsJC206. Figure 2 It is a microscopic structure diagram of the mycelium of Trametes sanguinea TsJC206. Figure 3 It is a microscopic structure diagram of the spores of Trametes sanguinea TsJC206. Figure 4 It is a fruiting body diagram of wild Trametes sanguinea (the left is the front, and the right is the back). Figure 5 It is the mycelial growth of Trametes sanguinea TsJC206 in solid original seed media containing different forage grasses (from left to right: Pennisetum giganteum, Neyraudia reynaudiana, Phragmites australis, Silphium perfoliatum, Cymbopogon citratus, Miscanthus sacchariflorus, Arundo donax, Themeda villosa, Miscanthus floridulus, Sorghum propinquum, Pennisetum purpureum, Vetiveria zizanioides, Paspalum wettsteinii). Figure 6 It is a comparison of the fruiting body formation of Trametes sanguinea TsJC206 and common Trametes sanguinea under the condition of 16°C. Figure 7 It is the fruiting body formation of Trametes sanguinea TsJC206 in the cultivation material containing Miscanthus floridulus and Dicranopteris dichotoma.
[0026] Example 2: S1: Mother spawn preparation: Inoculate the new strain of Trametes sanguinea TsJC206 into a test tube containing mother spawn medium according to aseptic operation, and incubate it in the dark at 22°C for 8 - 14 days until the mycelium fills the test tube to obtain the mother spawn. Among them, the formula of the mother spawn medium is calculated by mass percentage: potato powder 15%, corn flour 2%, glucose 1%, agar powder 2%, ferrous sulfate heptahydrate 0.002%, magnesium sulfate 0.002%, calcium sulfate heptahydrate 0.2%, crude sea salt 0.1%, potassium dihydrogen phosphate 0.1%, and the balance is water.
[0027] S2: Original spawn preparation: Inoculate 1 cm 2 of the mother spawn into a plastic bag containing solid original spawn medium according to aseptic operation, and incubate it in the dark at 22°C until the mycelium fills the plastic bag to obtain the original spawn. Among them, the formula of the solid original spawn medium is calculated by mass percentage: sorghum 15%, corn cob 15%, wheat bran 10%, gypsum 1%, light calcium carbonate 1%, magnesium oxide 0.002%, ferrous sulfate heptahydrate 0.002%, magnesium sulfate 0.001%, crude sea salt 0.1%, potassium dihydrogen phosphate 0.2%, and Pennisetum giganteum (specifically dry Pennisetum giganteum) makes up the balance; adjust the water content to 65%.
[0028] S3: Cultivation and fruiting body management: Inoculate 1 - 2 g of stock culture into the mushroom bag containing cultivation medium according to aseptic operation. Transfer the inoculated mushroom bag to the culture rack in the disinfected culture room and conduct dark cultivation at a temperature of 22°C and a relative air humidity of 70% - 80% until the mycelium fills the bag. Transfer the mushroom bag with mature mycelium to the mushroom shed for fruiting body management. During fruiting body management, the mycelium is in full contact with the air. The temperature in the shed is 18°C, and the relative air humidity is 90% - 110%. Provide scattered light and sufficient oxygen in the shed. Cultivate for 15 - 25 days until the fruiting body matures, stops growing, the white edges of the ganoderma all turn dark red, and all the ganoderma slices have a relatively consistent color, then pick. Among them, the formula of the cultivation medium is by mass percentage: wheat bran 10%, gypsum 1%, light calcium carbonate 1%, magnesium oxide 0.002%, ferrous sulfate heptahydrate 0.002%, magnesium sulfate 0.002%, crude sea salt 0.2%, potassium dihydrogen phosphate 0.2%, Pennisetum giganteum (specifically dry Pennisetum giganteum) to make up the balance; adjust the water content to 62%.
[0029] Taking a mushroom bag with a wet weight of 600 g of cultivation medium as an example, the fresh weight of the first - flush ganoderma is 20 - 30 g / bag, and the fresh weight of the second - flush ganoderma is 15 - 20 g / bag.
[0030] Example 3: S1: Mother culture preparation: Inoculate the new strain of Trametes cinnabarina TsJC206 into the test tube containing mother culture medium according to aseptic operation. Conduct dark cultivation in an incubator at 24°C for 8 - 14 days until the mycelium fills the test tube to obtain the mother culture. Among them, the formula of the mother culture medium is by mass percentage: potato powder 10%, corn flour 1%, glucose 2%, agar powder 2%, ferrous sulfate heptahydrate 0.003%, magnesium sulfate 0.001%, calcium sulfate heptahydrate 0.3%, crude sea salt 0.2%, potassium dihydrogen phosphate 0.1%, and the balance is water.
[0031] S2: Stock culture preparation: Inoculate 1 cm 2 of the mother culture into the mushroom bag containing solid stock culture medium. Conduct dark cultivation in an incubator at 24°C until the mycelium fills the bag to obtain the stock culture. Among them, the formula of the solid stock culture medium is by mass percentage: sorghum 20%, corncob 20%, wheat bran 8%, gypsum 1%, light calcium carbonate 0.5%, magnesium oxide 0.002%, ferrous sulfate heptahydrate 0.002%, magnesium sulfate 0.001%, crude sea salt 0.1%, potassium dihydrogen phosphate 0.1%, and Arundo donax makes up the balance (specifically, first use dry Arundo donax to make up the balance, and then calculate according to the water content of fresh Arundo donax, and replace dry Arundo donax with fresh Arundo donax according to the principle of equal dry - basis weight); adjust the water content to 65%.
[0032] S3: Cultivation and fruiting management: Inoculate 1 - 2 g of original strain into the fungus bags containing cultivation materials according to aseptic operation, transfer the inoculated fungus bags onto the culture racks in the disinfected culture room, and conduct dark cultivation at a temperature of 24°C and a relative air humidity of 70% - 80% until the mycelium fills the fungus bags; transfer the fungus bags with mature mycelium into the mushroom shed for fruiting management. During fruiting management, the mycelium is in full contact with the air, the temperature in the shed is 19°C, and the relative air humidity is 90% - 110%. Provide scattered light and sufficient oxygen in the shed, and cultivate for 15 - 25 days until the fruiting bodies mature, stop growing, the white edges of the ganoderma turn dark red completely, and all the ganoderma slices have a relatively consistent color, then pick them. Among them, the formula of the cultivation materials by mass percentage is: rice bran 10%, gypsum 1%, light calcium carbonate 0.5%, magnesium oxide 0.003%, ferrous sulfate heptahydrate 0.003%, magnesium sulfate 0.003%, crude sea salt 0.1%, potassium dihydrogen phosphate 0.2%, and Arundo donax fills the balance (specifically, first fill the balance with dry Arundo donax, then calculate according to the water content of fresh Arundo donax, and replace the dry Arundo donax with fresh Arundo donax according to the principle of equal dry basis weight); adjust the water content to 65%.
[0033] Taking the fungus bag with a wet weight of 600 g of cultivation materials as an example, the fresh weight of the first flush of ganoderma is 15 - 25 g / bag, and the fresh weight of the second flush of ganoderma is 10 - 20 g / bag.
[0034] Example 4: S1: Mother strain preparation: Inoculate the new strain of Trametes sanguinea TsJC206 into the test tubes containing mother strain medium according to aseptic operation, and conduct dark cultivation in an incubator at 25°C for 8 - 14 days until the mycelium fills the test tubes to obtain the mother strain. Among them, the formula of the mother strain medium by mass percentage is: potato powder 18%, corn flour 3%, glucose 2%, agar powder 3%, ferrous sulfate heptahydrate 0.002%, magnesium sulfate 0.002%, calcium sulfate heptahydrate 0.5%, crude sea salt 0.2%, potassium dihydrogen phosphate 0.2%, and the balance is water.
[0035] S2: Original strain preparation: Inoculate 1 - 3 g of mother strain into 150 ml of liquid original strain medium according to aseptic operation, and conduct dark cultivation in a shaker at 25°C and 130 rpm for 3 - 7 days until mycelial pellets appear to obtain the original strain. Among them, the formula of the liquid original strain medium by mass percentage is: potato powder 2%, glucose 2%, peptone 0.3%, yeast extract powder 0.3%, ferrous sulfate heptahydrate 0.003%, magnesium sulfate 0.003%, calcium sulfate heptahydrate 0.1%, crude sea salt 0.2%, potassium dihydrogen phosphate 0.2%, and the balance is water.
[0036] S3: Cultivation and Ganoderma lucidum fruiting management: Under aseptic operation, inoculate 3 - 5 ml of original strain into the plastic bags containing cultivation materials. Move the inoculated plastic bags onto the culture racks in the disinfected culture room and conduct dark cultivation at a temperature of 25°C and a relative air humidity of 70% - 80% until the mycelium fills the plastic bags. Move the plastic bags with mature mycelium into the mushroom shed for fruiting management. During fruiting management, the mycelium is in full contact with the air. The temperature in the shed is 18°C, and the relative air humidity is 90% - 110%. Provide scattered light and sufficient oxygen in the shed. Cultivate for 15 - 25 days until the fruiting bodies mature, stop growing, the edges of the white Ganoderma lucidum turn dark red completely, and all Ganoderma lucidum slices have a relatively consistent color, then pick them. Among them, the formula of the cultivation materials by mass percentage is: wheat bran 20%, gypsum 1%, light calcium carbonate 1%, magnesium oxide 0.003%, ferrous sulfate heptahydrate 0.002%, magnesium sulfate 0.002%, crude sea salt 0.1%, potassium dihydrogen phosphate 0.2%, Arundo donax (specifically dry Arundo donax) to make up the balance; adjust the water content to 65%.
[0037] Taking the plastic bag with 600 g of wet cultivation materials as an example, the fresh weight of the first - flush Ganoderma lucidum is 25 - 35 g / bag, and the fresh weight of the second - flush Ganoderma lucidum is 15 - 20 g / bag.
[0038] Example 5: S1: Mother strain preparation: Under aseptic operation, inoculate the new strain of Trametes coccinea TsJC206 into the test tubes containing mother strain medium, and conduct dark cultivation in an incubator at 25°C for 8 - 14 days until the mycelium fills the test tubes to obtain the mother strain. Among them, the formula of the mother strain medium by mass percentage is: potato powder 15%, corn flour 2%, glucose 2%, agar powder 3%, ferrous sulfate heptahydrate 0.003%, magnesium sulfate 0.002%, calcium sulfate heptahydrate 0.3%, crude sea salt 0.2%, potassium dihydrogen phosphate 0.2%, and the balance is water.
[0039] S2: Original strain preparation: Under aseptic operation, inoculate 1 - 3 g of mother strain into 150 ml of liquid original strain medium, and conduct dark cultivation in a shaker at 25°C and 140 rpm for 3 - 7 days until mycelial pellets appear to obtain the original strain. Among them, the formula of the liquid original strain medium by mass percentage is: potato powder 3%, glucose 2%, peptone 0.3%, yeast extract powder 0.4%, ferrous sulfate heptahydrate 0.002%, magnesium sulfate 0.003%, calcium sulfate heptahydrate 0.2%, crude sea salt 0.2%, potassium dihydrogen phosphate 0.2%, and the balance is water.
[0040] S3: Cultivation and fruiting body management: Inoculate 3 - 5 ml of original strain into the mushroom bag containing cultivation material according to aseptic operation. Transfer the inoculated mushroom bag to the culture rack in the disinfected culture room, and conduct dark cultivation at a temperature of 25°C and a relative air humidity of 70% - 80% until the mycelium fills the mushroom bag. Transfer the mushroom bag with mature mycelium to the mushroom shed for fruiting body management. During fruiting body management, the mycelium is in full contact with the air. The temperature in the shed is 18°C, and the relative air humidity is 90% - 110%. Scattered light and sufficient oxygen are provided in the shed. After cultivating until the fruiting body matures, pick it. Among them, the formula of the cultivation material by mass percentage is: wheat bran 20%, gypsum 1.5%, light calcium carbonate 1.5%, magnesium oxide 0.002%, ferrous sulfate heptahydrate 0.002%, magnesium sulfate 0.003%, crude sea salt 0.2%, potassium dihydrogen phosphate 0.2%, and Pennisetum giganteum makes up the balance (specifically, first make up the balance with dry Pennisetum giganteum, and then calculate according to the water content of fresh Pennisetum giganteum, and replace dry Pennisetum giganteum with fresh Pennisetum giganteum according to the principle of equal dry basis weight); adjust the water content to 65%.
[0041] Taking the mushroom bag with a wet weight of 600 g of cultivation material as an example, the fresh weight of the first flush of fruiting bodies is 15 - 20 g / bag, and the fresh weight of the second flush of fruiting bodies is 10 - 15 g / bag.
[0042] Figure 1 It is a mycelium plate map of Trametes sanguinea TsJC206.
[0043] Figure 2 It is a microscopic structure map of the mycelium of Trametes sanguinea TsJC206.
[0044] Figure 3 It is a microscopic structure map of the spores of Trametes sanguinea TsJC206.
[0045] Figure 4 It is a fruiting body map of wild Trametes sanguinea (the left is the front view, and the right is the back view).
[0046] Figure 5 It shows the mycelium growth of Trametes sanguinea TsJC206 in solid original strain culture media containing different Pennisetum. From left to right, the Pennisetum in the culture media are Pennisetum giganteum, Neyraudia reynaudiana, Phragmites australis, Silphium perfoliatum, Cymbopogon citratus, Miscanthus sacchariflorus, Arundo donax, Themeda villosa, Miscanthus floridulus, Sorghum propinquum, Pennisetum purpureum, Vetiveria zizanioides, and Paspalum wettsteinii.
[0047] Figure 6 It shows the fruiting body formation of Trametes sanguinea TsJC206 and common Trametes sanguinea under sawdust cultivation conditions (the cultivation material consists of sawdust, wheat bran, and gypsum in a mass ratio of 78:20:2, and the water content is adjusted to 55% - 65%) at 16°C. The left is Trametes sanguinea TsJC206, and the right is common Trametes sanguinea.
[0048] Figure 7The fruiting situation of Trametes sanguinea TsJC206 at 16°C under the condition of cultivation with Juncao (the cultivation material consists of Miscanthus floridulus, Dicranopteris dichotoma, wheat bran and gypsum in a mass ratio of 40:38:20:2, and the water content is adjusted to 55% - 65%).
[0049] The above are only the preferred embodiments of the present invention, and all equivalent changes and modifications made according to the scope of the patent application of the present invention shall fall within the scope covered by the present invention.
Claims
1. A new strain of Trametes sanguinea with low temperature type, characterized in that: The new strain of Trametes sanguinea is Trametes sanguinea ( Trametes sanguinea ) TsJC206, which was deposited at the China Center for Type Culture Collection on June 19, 2023, with the deposit number CCTCC NO: M 20231065 and the deposit address at No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province.
2. A cultivation method of the new strain of Trametes sanguinea as described in claim 1, characterized in that: It includes the following steps: S1: Inoculate a new strain of Trametes sanguinea (low-temperature type) into a test tube containing a slant culture medium, and incubate it in the dark at 15-28 °C in an incubator until the mycelium fills the test tube to obtain the slant culture; S2: Inoculate the slant culture into a plastic bag containing a solid spawn culture medium according to aseptic operation, and incubate it in the dark at 15-28 °C in an incubator until the mycelium fills the plastic bag to obtain the spawn; or inoculate the slant culture into a shake flask containing a liquid spawn culture medium according to aseptic operation, and incubate it in the dark at 15-28 °C and 120-160 rpm on a shaker until hyphal pellets appear to obtain the spawn; S3: Inoculate the spawn into a plastic bag containing the cultivation substrate according to aseptic operation, and incubate it in the dark at 15-28 °C and 70%-80% relative air humidity in a cultivation room until the mycelium fills the plastic bag; transfer the plastic bag with mature mycelium to a mushroom shed for fruiting management. During fruiting management, the mycelium is in full contact with the air, the temperature in the shed is 15-20 °C, and the relative air humidity is 90%-110%. Provide diffused light and sufficient oxygen in the shed, and cultivate until the fruiting body matures.
3. The cultivation method according to claim 2, characterized in that: The formula of the slant culture medium is as follows by mass percentage: potato flour 2%-6%, corn flour 1%-3%, glucose 0.5%-2%, agar powder 1.5%-3%, ferrous sulfate heptahydrate 0.001%-0.005%, magnesium sulfate 0.001%-0.005%, calcium sulfate heptahydrate 0.2%-0.5%, crude sea salt 0.1%-0.2%, potassium dihydrogen phosphate or dipotassium hydrogen phosphate 0.1%-0.2%, and the balance is water.
4. The cultivation method according to claim 2, characterized in that: The formula of the solid spawn culture medium is as follows by mass percentage: sorghum 15%-30%, corncob 15%-30%, wheat bran 5%-10%, gypsum 0.5%-1.5%, light calcium carbonate 0.5%-1.5%, magnesium oxide 0.002%-0.01%, ferrous sulfate heptahydrate 0.002%-0.005%, magnesium sulfate 0.001%-0.01%, crude sea salt 0.1%-0.2%, potassium dihydrogen phosphate or dipotassium hydrogen phosphate 0.1%-0.2%, and dried mushroom grass makes up the balance; adjust the water content to 55%-65%.
5. The cultivation method according to claim 4, characterized in that: The mushroom grass is any one or more of Pennisetum giganteum, Arundo donax, Pennisetum purpureum, Neyraudia reynaudiana, Pennisetum purpureum × Pennisetum americanum, Pennisetum americanum, Phragmites australis, Silphium perfoliatum, Cymbopogon citratus, Miscanthus sacchariflorus, Themeda japonica, Sorghum propinquum, Vetiveria zizanioides, Paspalum wettsteinii, Dicranopteris dichotoma; calculated based on the water content of fresh mushroom grass, and according to the principle of equal dry weight, the dried mushroom grass can be replaced by fresh mushroom grass.
6. The cultivation method according to claim 2, wherein: The formula of the liquid spawn culture medium is as follows by mass percentage: potato flour 1%-3%, glucose 1%-3%, peptone 0.1%-0.5%, yeast extract powder 0.1%-0.5%, ferrous sulfate heptahydrate 0.001%-0.005%, magnesium sulfate 0.001%-0.01%, calcium sulfate heptahydrate 0.1%-0.2%, crude sea salt 0.1%-0.2%, potassium dihydrogen phosphate or dipotassium hydrogen phosphate 0.1%-0.2%, and the balance is water.
7. The cultivation method according to claim 2, characterized in that: The formula of the cultivation material is as follows by mass percentage: wheat bran or rice bran 10% - 25%, gypsum 0.5% - 1.5%, light calcium carbonate 0.5% - 1.5%, magnesium oxide 0.002% - 0.01%, ferrous sulfate heptahydrate 0.002% - 0.005%, magnesium sulfate 0.001% - 0.01%, crude sea salt 0.1% - 0.2%, potassium dihydrogen phosphate or dipotassium hydrogen phosphate 0.1% - 0.2%, and dried mushroom grass makes up the balance; adjust the water content to 55% - 65%.
8. The cultivation method according to claim 7, characterized in that: The mushroom grass is any one or more of Pennisetum giganteum, Arundo donax, Pennisetum purpureum, Neyraudia reynaudiana, Pennisetum purpureum × Pennisetum americanum, Pennisetum americanum, Phragmites australis, Silphium perfoliatum, Cymbopogon citratus, Triarrhena sacchariflora, Themeda japonica, Sorghum propinquum, Vetiveria zizanioides, Paspalum wettsteinii, Dicranopteris dichotoma; calculated according to the water content of fresh mushroom grass, and in accordance with the principle of equal dry basis weight, the dried mushroom grass can be replaced by fresh mushroom grass.