Black fungus standard bead strain and application method
By preparing fixed-size bead-like mycelium strains, the instability of black fungus strains during the inoculation process and the challenges of mechanized inoculation were solved, achieving efficient and stable mycelial growth and automated inoculation.
Patent Information
- Application Number
- CN202511820380.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-04
- Publication Date
- 2026-03-03
AI Technical Summary
Existing black fungus strains suffer from problems such as unstable yield, high contamination rate of miscellaneous bacteria, and weak mycelial vitality. In particular, it is difficult to achieve uniformity in specifications, control nutrition, and achieve mechanized inoculation when using ordinary sawdust, branches, or liquid strains.
Using standard black fungus granules, a new culture medium ratio is used to prepare a standardized granule spawn, which includes a main culture medium and a secondary culture medium. The framework is made of plastic or metal materials and is spherical, cubic or cylindrical in shape, which is conducive to mechanical and automated identification and inoculation.
It improved the viability and shape certainty of the strain, enabled mechanized and precise inoculation, shortened the germination and colonization time, and improved the growth rate and stability of mycelium.
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Abstract
Description
Technical Field
[0001] This invention relates to the field of edible fungi cultivation technology, and in particular to a method for the production and application of a standard spore strain of black fungus. Background Technology
[0002] Black fungus is a specialty of China and a famous delicacy. my country is a major producer of black fungus, accounting for over 96% of the world's total output, and its products are exported to more than 80 countries and regions worldwide, becoming a pillar industry in many cities and counties. However, many technical challenges remain in black fungus production across the country, especially the quality of spawn and spawn bags, which are often overlooked or ignored. Spawn bag production suffers from unstable yield, high contamination rates by other microorganisms, and weak mycelial vitality. There are many reasons for this, but the influence of the spawn is a significant one. With ordinary sawdust spawn, the mycelium tears and particles disperse during inoculation, making it difficult to control the inoculation amount, resulting in slow germination and colonization, and making mechanical inoculation difficult. With spawn from branches and sticks, the nutrients within the branches and sticks are difficult to regulate uniformly, and the shape of the pores in the spawn bag differs, preventing sufficient adhesion after inoculation. Liquid spawn suffers from too many quality-affecting factors and overly refined nutrients in the culture medium, leading to unstable spawn characteristics and vitality. In conclusion, it is essential to develop a new strain of fungus that is nutritionally complete and controllable, has a fixed shape and size, is easy to inoculate precisely by mechanization, and exhibits rapid germination and colonization, strong mycelial vigor, and stable species characteristics. Summary of the Invention
[0003] This invention, through a novel culture medium formulation and the use of new materials to shape the granules, produces a granulated spawn that not only enhances the viability of black fungus spawn but also facilitates automated mechanical identification and handling of the granular spawn. This overcomes various problems inherent in previous spawns made from sawdust, branches, wood blocks, grains, and liquids, such as inconsistent sizes, nutritional deviations, poor mycelial viability, serotype degeneration, and susceptibility to infection in the spawn bags. Furthermore, it promotes mechanized and automated inoculation. This invention solves the problems associated with using sawdust, branches, grains, and liquids as substrates for spawn production and application in black fungus.
[0004] To address the existing problems, this invention provides a standard black fungus bead spawn that can be shaped into fixed beads, promotes the growth of black fungus mycelium, and enhances the vitality of the spawn. The bead spawn has a defined shape and consistent size, making it suitable for automated inoculation. It is beneficial for the growth and storage of black fungus mycelium, and can be accurately identified and grasped during inoculation, and accurately inserted into the designated position on the black fungus spawn bag. After inoculation, it germinates and grows rapidly.
[0005] The present invention provides a standard spore strain of black fungus, comprising a standard spore strain framework and a main culture medium filled within the framework.
[0006] The main culture medium contains the following components by weight: 39-41 parts fine hardwood sawdust, 38-40 parts white glutinous corn flour, 3.8-4.2 parts elm bark powder, 1.9-2.1 parts konjac powder, 3.8-4.2 parts natural calcium-based bentonite, and 0.02-0.03 parts chitosan.
[0007] Furthermore, in the above technical solution, the surface of the strain is adhered with a layer of auxiliary culture medium, which, by weight, contains 84-86 parts of fine hardwood sawdust, 9.5-10.5 parts of white glutinous corn flour, 1.9-2.1 parts of elm bark powder, 1.9-2.1 parts of konjac powder, 0.02-0.03 parts of chitosan, and 0.8-1 parts of gypsum powder.
[0008] Furthermore, in the above technical solution, the weight ratio of the main culture medium to the auxiliary culture medium is in the range of 10-12:1.
[0009] Furthermore, in the above technical solution, the standard bead-like microorganism framework is a sphere with a diameter of 10-12 mm, or a cube with a side length of 9-10 mm, or a cylinder with a diameter and height of 9-10 mm, and the framework is made of plastic or metal materials.
[0010] This invention provides a method for producing standard granule strains of black fungus, comprising the following steps: A standard granule fungal culture framework is prepared, wherein the framework is a sphere with a diameter of 10-12 mm, a cube with a side length of 9-10 mm, or a cylinder with a diameter and height of 9-10 mm. To prepare the main culture medium, weigh out 39-41 parts of fine hardwood sawdust, 38-40 parts of white glutinous corn flour, 3.8-4.2 parts of elm bark powder, 1.9-2.1 parts of konjac powder, 3.8-4.2 parts of natural calcium-based bentonite, and 0.02-0.03 parts of chitosan by weight percentage. After dry mixing, add hot water at 60-80℃ and stir. Mix the materials at a ratio of 1:1 until the mixture can be formed into a ball by hand.
[0011] The main culture medium is filled into the standard granular mycelium frame; Prepare the auxiliary culture medium by weighing out 84-86 parts of fine hardwood sawdust, 9.5-10.5 parts of white glutinous corn flour, 1.9-2.1 parts of elm bark powder, 1.9-2.1 parts of konjac powder, 0.02-0.03 parts of chitosan, and 0.8-1 parts of gypsum powder by weight percentage. After mixing thoroughly with dry ingredients, add water and stir; mix according to a material-to-water ratio of 1:1. The frame filled with the main culture medium is rolled in the auxiliary culture medium to make a layer of auxiliary culture medium adhere to the surface, forming beads. The beads are placed into spawn bottles or bags, sterilized, inoculated with black fungus spawn, and cultured in a ventilated environment at 25-30℃.
[0012] This invention discloses an inoculation method for black fungus spawn bags, using the aforementioned standard black fungus spawn, comprising the following steps: To prepare the mycelium bags, a central hole is punched in the middle of the culture medium. The central hole is wider at the top and narrower at the bottom, with the diameter of the lower part of the central hole slightly smaller than the outer diameter of the standard bead-like mycelium strain. Under aseptic conditions, the standard bead-like bacterial strain is identified and picked up by an automated inoculation machine and inoculated into the central well, so that the bacterial strain is located in the middle of the depth of the central well.
[0013] Furthermore, in the above technical solution, the standard mycelium strain is placed at 20-30℃ for 12-24 hours before inoculation to allow the mycelium to regain its vitality, which is conducive to the germination and growth of the strain. Attached Figure Description
[0014] Figure 1 This is a circular frame diagram of the standard *Vallisneria natans* strain.
[0015] Figure 2 This is a standard cube framework diagram of the *Vallisneria* strain.
[0016] Figure 3 This is a standard cylindrical framework diagram of the *Vallisneria natans* strain.
[0017] Figure 4 This is a schematic diagram of the structure of a special culture bottle. Detailed Implementation
[0018] Example 1 A standard granule strain for black fungus includes a standard granule strain framework and a main culture medium filled within the framework; The main culture medium contains the following components by weight: 40 parts fine hardwood sawdust, 39 parts white glutinous corn flour, 4 parts elm bark powder, 2 parts konjac powder, 4 parts natural calcium-based bentonite, and 0.02 parts chitosan.
[0019] The surface of the strain is adhered with a layer of auxiliary culture medium, which, by weight, contains 85 parts of fine hardwood sawdust, 10 parts of white glutinous corn flour, 2 parts of elm bark powder, 2 parts of konjac powder, 0.02 parts of chitosan, and 0.8 parts of gypsum powder.
[0020] The weight ratio of the main culture medium to the auxiliary culture medium is 10:1.
[0021] The standard bead-like microorganism framework is a spherical body with a diameter of 10-12 mm, and the framework is made of plastic or metal materials; A method for producing a standard granule strain of black fungus includes the following steps: A standard granular mycelium framework was prepared, wherein the framework was a spherical shape with a diameter of 10-12 mm; To prepare the main culture medium, weigh out 40 parts of fine hard hardwood sawdust, 39 parts of white glutinous corn flour, 4 parts of elm bark powder, 2 parts of konjac powder, 4 parts of natural calcium-based bentonite, and 0.02 parts of chitosan by weight percentage. After dry mixing, add hot water at 60-80℃ and stir. Mix the materials at a ratio of 1:1 until the mixture can be formed into a ball by hand.
[0022] The main culture medium is filled into the standard granular mycelium frame; Prepare the auxiliary culture medium by weighing 85 parts of fine hardwood sawdust, 10 parts of white glutinous corn flour, 2 parts of elm bark powder, 2 parts of konjac powder, 0.02 parts of chitosan, and 0.8 parts of gypsum powder by weight percentage.
[0023] After mixing thoroughly with dry ingredients, add water and stir; mix according to a material-to-water ratio of 1:1. The frame filled with the main culture medium is rolled in the auxiliary culture medium to make a layer of auxiliary culture medium adhere to the surface, forming beads. The beads are placed into spawn bottles or bags, sterilized, inoculated with black fungus spawn, and cultured in a ventilated environment at 25-30℃.
[0024] A method for inoculating black fungus spawn bags includes the following steps: To prepare the mycelium packs, a central hole is punched in the middle of the culture medium. The central hole is wider at the top and narrower at the bottom, with the lower part of the hole slightly smaller than the outer diameter of the standard bead-like mycelium strain. Under aseptic conditions, the standard bead-like bacterial strain is identified and picked up by an automated inoculation machine and inoculated into the central well, so that the bacterial strain is located in the middle of the depth of the central well.
[0025] The standard mycelium strain is placed at 20-30℃ for 12-24 hours before inoculation to allow the mycelium to regain its vitality, which is conducive to the germination and growth of the strain.
[0026] Example 2 The difference from Example 1 is as follows: The standard bead-like microorganism framework is a cube with a side length of 9-10 mm, and the framework is made of plastic or metal. The main culture medium contains the following components by weight: 39 parts fine hardwood sawdust, 38 parts white glutinous corn flour, 3.8 parts elm bark powder, 1.9 parts konjac powder, 3.8 parts natural calcium-based bentonite, and 0.02 parts chitosan. The supplementary culture medium, by weight, contains 84 parts fine hardwood sawdust, 9.5 parts white glutinous corn flour, 1.9 parts elm bark powder, 1.9 parts konjac powder, 0.02 parts chitosan, and 0.8 parts gypsum powder. Example 3 The difference from Example 1 is as follows: The standard bead-like microorganism framework is a cylinder with a diameter and height of 9-10 mm, and the framework is made of plastic or metal. The main culture medium contains the following components by weight: 41 parts fine hard hardwood sawdust, 40 parts white glutinous corn flour, 4.2 parts elm bark powder, 2.1 parts konjac powder, 4.2 parts natural calcium-based bentonite, and 0.03 parts chitosan.
[0027] The supplementary culture medium, by weight, contains 86 parts fine hardwood sawdust, 10.5 parts white glutinous corn flour, 2.1 parts elm bark powder, 2.1 parts konjac powder, 0.03 parts chitosan, and 1 part gypsum powder. Example 4 The difference from Example 1 is as follows: The standard bead-like microorganism framework is a cube with a side length of 9-10 mm, and the framework is made of plastic or metal. The main culture medium contains the following components by weight: 39 parts fine hardwood sawdust, 40 parts white glutinous corn flour, 4 parts elm bark powder, 2 parts konjac powder, 3.8 parts natural calcium-based bentonite, and 0.02 parts chitosan.
[0028] The auxiliary culture medium contains, by weight, 84 parts of fine hardwood sawdust, 10.5 parts of white glutinous corn flour, 1.9 parts of elm bark powder, 2.1 parts of konjac powder, 0.03 parts of chitosan, and 0.9 parts of gypsum powder.
[0029] Comparative Example 1 The conventional culture medium for microbial strains is prepared by mixing 85 parts hardwood sawdust, 12 parts wheat bran, 2 parts soybean meal, and 1 part gypsum powder, and then proceeding with the standard mixing, bagging, sterilization, inoculation, and cultivation.
[0030] The production and application effects of the black fungus granule strain in Example 1 above were compared with those of the conventional black fungus sawdust strain in Comparative Example 1: The culture medium and production process of the *Varicella spp.* strain are as described in Example 1 above.
[0031] In Example 1, the *Varicella spp.* strain and in Comparative Example 1 were both produced and packaged into 15cm × 28cm plastic bags, with a filling height of 12cm. Sterilization was performed using a centrally located indented stomata. Test-tube mother cultures were used for all cultures, and the bags were incubated at 22–15℃. Inoculation into the bags was performed using an automated inoculation machine.
[0032] The experimental data and results are shown in Table 1.
[0033] Table 1. Comparison of Production and Use of *Varicella granulata* and Conventional Wood Chip Fungi.
[0034] Comparative studies show that: *Mycorrhizal* spawn grows faster than conventional spawn due to its better air permeability, with a cultivation time 79.19% shorter; its mycorrhizal structure is denser and stronger due to the more balanced nutrient content of the culture medium; it is easier to pick up with an automatic inoculation machine, eliminating the need for mechanical removal, and its inoculation speed is 1.5 times faster than conventional spawn; when inoculating the spawn bags, each spawn bead falls precisely into the center of the bag's central hole, while with conventional spawn, the inoculation amount is uneven, with most accumulating at the top of the central hole; after inoculating the bags with spawn, the bags fully colonize in 80% of the time compared to conventional spawn due to the robust spawn and better air permeability; and the mycorrhizal structure of spawn within the bags is significantly stronger than that of conventional spawn.
[0035] Comparative Examples 2-6 The differences from the embodiments are shown in Tables 2 and 3.
[0036] Table 2. Formulation of the main culture medium for comparative examples
[0037] Table 3 Comparative Supplementary Culture Medium Formulation
[0038] Table 4 shows the statistical results of Example 1 and the comparative example.
[0039] Example 1: The culture medium forms a normal clump, and there is no water accumulation at the bottom of the bag (bottle) after sterilization. The mycelium grows rapidly, and the mycelium is white and dense. During inoculation, the automatic inoculation machine accurately picks up the mycelium and inoculates it into the middle of the cultivation bag. The mycelium germinates and grows quickly in the culture bag.
[0040] In Comparative Example 2, white glutinous corn flour is the main component for the formation of beads in the *Mycorrhiza regia* strain; without it, the beads are difficult to form. When white glutinous corn flour was replaced with yellow glutinous corn flour, the bead formation on the culture medium was poor. During mycorrhizal cultivation, the mycelium grew rapidly and was white in color, but it was weak. This may be because many yellow glutinous corn varieties contain less sticky substances and have a higher sugar content. When white glutinous corn flour was replaced with glutinous rice flour, although the bead formation on the culture medium was better, the subsequent cultivation results were similar to those with yellow glutinous corn, again due to the high sugar content of glutinous rice.
[0041] Compared with Example 3, elm bark powder was removed and an equal amount of white glutinous corn flour was added. In addition to the decrease in the granulation of the culture medium, the mycelium of black fungus grew too fast during the cultivation of fungi because the sugar content of corn flour was too high.
[0042] Compared to Example 4, omitting konjac powder has several drawbacks: firstly, it affects the shaping of the beads; secondly, water tends to accumulate at the bottom of the spawn bags (bottles), shortening the shelf life; and thirdly, the mycelium appears slightly thinner during spawn cultivation. Konjac powder's main component, glucomannan, is a hemicellulose that not only retains water, gels, binds, and shapes the beads, but also helps retain water in the culture medium, alleviating water accumulation at the bottom of the spawn bags (bottles). Furthermore, glucomannan is readily decomposed and absorbed by the black fungus mycelium and synthesized by the black fungus into glucomannan, the main component of black fungus polysaccharides.
[0043] Compared to Example 5, without the addition of natural calcium-based bentonite, the beads had low hardness and were easily broken. Without the addition of natural calcium-based bentonite, the growth rate of the microorganisms was slower, resulting in a shorter shelf life.
[0044] Comparative Example 6, without the addition of chitosan, resulted in reduced mycelial density and shorter culture time for the strain. This is related to chitosan's ability to stimulate enzyme activity and promote metabolism.
Claims
1. A standard mycelium strain for black fungus, characterized in that: Includes a standard granular culture frame and a master culture medium filled within the frame; The main culture medium contains the following components by weight: 39-41 parts fine hardwood sawdust, 38-40 parts white glutinous corn flour, 3.8-4.2 parts elm bark powder, 1.9-2.1 parts konjac powder, 3.8-4.2 parts natural calcium-based bentonite, and 0.02-0.03 parts chitosan.
2. The standard mycelium strain for black fungus according to claim 1, characterized in that: The surface of the fungal strain is adhered with a layer of auxiliary culture medium, which, by weight, contains 84-86 parts of fine hardwood sawdust, 9.5-10.5 parts of white glutinous corn flour, 1.9-2.1 parts of elm bark powder, 1.9-2.1 parts of konjac powder, 0.02-0.03 parts of chitosan, and 0.8-1 parts of gypsum powder.
3. The standard mycelium strain for black fungus according to claim 1, characterized in that: The weight ratio of the main culture medium to the auxiliary culture medium is 10-12:
1.
4. The standard mycelium strain for black fungus according to claim 1, characterized in that: The standard bead-like microorganism framework is shaped as a sphere with a diameter of 10-12 mm, or a cube with a side length of 9-10 mm, or a cylinder with a diameter and height of 9-10 mm. The framework is made of plastic or metal.
5. A method for producing a standard granule strain of black fungus, characterized in that... Includes the following steps: To prepare the main culture medium, weigh out 39-41 parts of fine hard hardwood sawdust, 38-40 parts of white glutinous corn flour, 3.8-4.2 parts of elm bark powder, 1.9-2.1 parts of konjac powder, 3.8-4.2 parts of natural calcium-based bentonite, and 0.02-0.03 parts of chitosan by weight percentage. After dry mixing evenly, add hot water at 60-80℃ and stir, mixing the materials at a ratio of 1:
1. The main culture medium is filled into the standard granular mycelium frame; Prepare the auxiliary culture medium by weighing out 84-86 parts of fine hardwood sawdust, 9.5-10.5 parts of white glutinous corn flour, 1.9-2.1 parts of elm bark powder, 1.9-2.1 parts of konjac powder, 0.02-0.03 parts of chitosan, and 0.8-1 parts of gypsum powder by weight percentage. After mixing thoroughly with dry ingredients, add water and stir; mix according to a material-to-water ratio of 1:
1. The frame filled with the main culture medium is rolled in the auxiliary culture medium to make a layer of auxiliary culture medium adhere to the surface, forming beads. The beads are placed into spawn bottles or bags, sterilized, inoculated with black fungus spawn, and cultured in a ventilated environment at 25-30℃.
6. The method for producing the standard black fungus granule strain according to claim 5, characterized in that: This includes preparing a standard granular microorganism framework, wherein the framework is a sphere with a diameter of 10-12 mm, a cube with a side length of 9-10 mm, or a cylinder with a diameter and height of 9-10 mm.
7. A method for inoculating black fungus spawn bags, characterized in that... Using the black fungus standard granule strain as described in claim 1 or 2, the following steps are included: To prepare the mycelium packs, a central hole is punched in the middle of the culture medium. The central hole is wider at the top and narrower at the bottom, with the lower part of the hole slightly smaller than the outer diameter of the standard bead-like mycelium strain. Under aseptic conditions, the standard bead-like bacterial strain is identified and picked up by an automated inoculation machine and inoculated into the central well, so that the bacterial strain is located in the middle of the depth of the central well.