Filamentous fungus incubator capable of distinguishing absorption of different matrixes and preparation and use method of filamentous fungus incubator

Abstract

The invention provides a filamentous fungus incubator capable of distinguishing absorption of different matrixes and a preparation and use method of the filamentous fungus incubator. The filamentous fungus incubator comprises a circular base plate and a separated top cover, radial straight partition plates are distributed and fixedly connected in the circular base plate at equal intervals, two or three double-layer separation nets are installed in the circular base plate, the separated top cover is installed on the top of the circular base plate, waterproof ventilation holes are formed in one side of the separated top cover, and a movable baffle is arranged in the circular base plate. According to the filamentous fungus incubator capable of distinguishing absorption of different matrixes and the use method and the preparation method of the filamentous fungus incubator, provided by the invention, the growth condition of the same inoculated strain passing through the different matrixes in sequence, the absorption condition of C and N elements in the matrixes, the migration condition of C and N in hyphae and the influence of the matrix arrangement sequence can be observed and measured; people can deeply explore the substrate utilization complexity of the filamentous fungi, and scientific data is provided for research and development of filamentous fungi application technologies.

Description

Technical Field

[0001] The invention relates to the technical field of microbial detection, and in particular to a filamentous fungus culture device capable of distinguishing different matrix absorptions and a preparation and use method thereof. Background Art

[0002] Filamentous fungi are important resource microorganisms, and are particularly important in the production of useful products from organic waste. The culture dish is a traditional device for screening and culturing filamentous fungi. It consists of a flat disc-shaped bottom and a cover, usually made of glass or plastic, with a solid culture medium mainly composed of agar. In the latest research, the culture dish can also be used to study the antagonistic and symbiotic interactions between two filamentous fungi. The method is to inoculate different strains on both sides of the culture dish, observe the growth of the hyphae, or take samples at the junction to test the biochemical components;

[0003] However, there are few studies on how filamentous fungi absorb and utilize different substrates during their growth. Microscopically, filamentous fungi have to pass through different substrates during their growth. During this process, how filamentous fungi absorb elements from different substrates, whether the absorbed elements move within the filamentous fungi, and how filamentous fungi change the properties of different substrates are both rigorous scientific issues and the basis for the development of applied technologies. So far, there are few studies in this area, which to some extent indicates the lack of powerful means to explore such issues, including experimental equipment;

[0004] At present, although some studies have explored the mechanism of mycorrhizal fungi on the transfer of elements C and N and the effects of filamentous fungi on the degradation of different litters, these studies lack complete research equipment. In mycorrhizal fungi research, homemade root boxes and 25μm membranes are often used to isolate fungal hyphae and root hairs, while litter degradation research often uses the net bag replacement method. Although these methods have made some progress, they are difficult to meet the needs of more in-depth and systematic research, especially the lack of equipment for studying the continuous growth of filamentous fungi in different substrates;

[0005] In order to solve the above technical problems, a filamentous fungus culture device capable of distinguishing absorption of different substrates and a preparation and use method thereof are proposed. Summary of the invention

[0006] In view of this, the present invention provides a filamentous fungus culture device for distinguishing absorption of different matrices and a preparation and use method thereof. In view of the fact that there is currently no reliable and established equipment and research means for studying the absorption of elements in different matrices by filamentous fungi and the movement of elements within hyphae, the first purpose of the present invention is to provide a filamentous fungus culture device for distinguishing absorption of different matrices, that is, to allow filamentous fungi to pass through different types of matrices at fixed positions in sequence during growth. By using the filamentous fungus culture device, the growth conditions of the same strain of filamentous fungi in different types of matrices can be clearly determined; the second purpose of the present invention is to provide a test method for quantitatively distinguishing the absorption of elements in different matrices by filamentous fungi and the movement of elements within hyphae based on the combination of isotope labeling technology and subtraction method. This method enables the isotope labeling technology to be supported by physical and chemical equipment, expands the application scope of the isotope technology, and at least provides a beneficial choice.

[0007] The technical solution of the present invention is achieved as follows: a filamentous fungus culture device for distinguishing different substrate absorption comprises a circular bottom plate and a detachable top cover;

[0008] The interior of the circular chassis is evenly distributed and fixedly connected with radial straight partitions, the interior of the circular chassis is installed with two or three double-layer isolation nets, the separate top cover is installed on the top of the circular chassis, one side of the separate top cover is provided with a waterproof vent hole, and the interior of the circular chassis is provided with a movable baffle.

[0009] Further preferably, the height of the circular chassis is 1-3 cm, and the bottom of the circular chassis is made of transparent material.

[0010] Further preferably, the pore size of the double-layer isolation net is 200 μm, and the distance between the two layers of the double-layer isolation net is 5 mm.

[0011] Further preferably, the height of the radial straight partition is the same as the height of the circular chassis.

[0012] Further preferably, the movable baffle is arc-shaped, and its direction is opposite to the concentric circles formed by the double-layer isolation net.

[0013] Further preferably, the detachable top cover material is a light-proof material, and the waterproof ventilation hole has a diameter of 5-10 mm.

[0014] The method for preparing a filamentous fungus culture device for distinguishing different substrate absorption comprises the following steps:

[0015] S1. Manufacturing a circular chassis;

[0016] S2. Use a circular double-layer isolation net to divide the circular chassis into 3-4 concentric circles;

[0017] S3, use radial straight partitions to divide each concentric circle into 6-9 small chambers;

[0018] S4. A movable baffle can be added to each small room as needed;

[0019] S5. Each chamber is equipped with a detachable top cover with waterproof vents.

[0020] Further preferably, in S2, the height of the circular double-layer isolation net is the same as the height of the circular bottom plate, and among the multiple concentric circles, the ratio of the radius of the first concentric circle to the radial width of the other circles is 1:1, specifically 2-4 cm.

[0021] A method for using a filamentous fungus incubator for distinguishing different substrate absorptions comprises the following steps:

[0022] (1) Each concentric circle is filled with the same matrix, including ordinary matrix and labeled matrix;

[0023] (2) In the 3 or 4 chambers in the same radial direction, ensure that the 3 or 4 substrates to be studied are present simultaneously and without duplication, and there is only one labeled substrate;

[0024] (3) At the end of the culture, the growth of hyphae in each chamber can be observed, or hyphae can be separated from each chamber independently to test the isotope abundance;

[0025] (4) Based on the isotope labeling and subtraction methods, the growth conditions of filamentous fungi in different substrates, the absorption of different substrates, the migration of elements such as C and N in the hyphae, and the influence of the substrate arrangement order were quantified.

[0026] More preferably, the influence of the arrangement order of the substrates on the growth and absorption of filamentous fungi is clarified, the growth status of filamentous fungi in each substrate can be quantified, and the amount of C, N and other elements in the filamentous fungi in each cell that come from the substrates of other cells can be distinguished.

[0027] The embodiment of the present invention has the following advantages due to the adoption of the above technical solution:

[0028] 1. The filamentous fungus incubator for distinguishing different substrate absorption provided by the present invention and its use method and preparation method can observe and measure the growth status of the same inoculated strain when it sequentially passes through different substrates, the absorption of C and N elements in the substrate, the migration of C and N in the hyphae, and the influence of the substrate arrangement sequence, which helps people to explore the complexity of filamentous fungi in using substrates and provide scientific data for the research and development of filamentous fungi application technology;

[0029] Second, the present invention uses a filamentous fungus incubator to make it possible to accurately and quantitatively evaluate the utilization degree of different substrates by filamentous fungi, and can more accurately evaluate the utilization value of organic waste, making the isotope labeling technology materialized, more intuitive, and more applicable.

[0030] The above summary is for illustrative purposes only and is not intended to be limiting in any way. In addition to the illustrative aspects, embodiments and features described above, further aspects, embodiments and features of the present invention will be readily apparent by reference to the accompanying drawings and the following detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0032] Figure 1 It is a structural diagram of the filamentous fungus culture device of the present invention;

[0033] Figure 2 It is a structural diagram of a circular bottom plate in the filamentous fungus incubator of the present invention;

[0034] Figure 3 is a flow chart of the preparation method of the filamentous fungus culture device of the present invention;

[0035] Figure 4 The present invention is a flowchart of the method for using the filamentous fungus culture device.

[0036] Figure numerals: 11, circular chassis; 12, detachable top cover; 13, waterproof vent; 14, radial straight partition; 15, double-layer isolation net; 16, movable baffle. DETAILED DESCRIPTION

[0037] In the following, only some exemplary embodiments are briefly described. As those skilled in the art will appreciate, the described embodiments may be modified in various ways without departing from the spirit or scope of the present invention. Therefore, the drawings and descriptions are considered to be exemplary and non-restrictive in nature.

[0038] The embodiments of the present invention are described in detail below with reference to the accompanying drawings.

[0039] Embodiment 1

[0040] like Figure 1 and Figure 2 As shown, the embodiment of the present invention provides a filamentous fungus incubator for distinguishing absorption of different substrates, which is composed of a circular bottom plate 11 and a detachable top cover 12;

[0041] The circular chassis 11 has radial straight partitions 14 evenly distributed and fixedly connected inside, two double-layer isolation nets 15 installed inside, a separate top cover 12 installed on the top of the circular chassis 11, a waterproof vent 13 is opened on one side of the separate top cover 12, and a movable baffle 16 is provided inside the circular chassis 11.

[0042] In one embodiment, the height of the circular bottom plate 11 is 3 cm, and the bottom of the circular bottom plate 11 is made of a transparent material. The height of the circular bottom plate 11 is specified to clarify the volume of each chamber, that is, the volume of each matrix, to ensure that the filamentous fungi have sufficient growth space in each matrix. The bottom is made of a transparent material to facilitate the observation of the growth of the mycelial fungi.

[0043] In one embodiment, the pore size of the double-layer isolation net 15 is 200 μm, and the distance between the two layers of the double-layer isolation net 15 is 5 mm, which can isolate different substrates and prevent substrates from contacting each other, thereby ensuring the purity of the substrate in each chamber.

[0044] In one embodiment, the height of the radial straight partition 14 is the same as that of the circular bottom plate 11, which can ensure that each chamber is closed and prevent filamentous fungi from entering other chambers during the growth process.

[0045] In one embodiment, the movable baffle 16 is arc-shaped and its direction is opposite to the concentric circle formed by the double-layer isolation net 15. The movable baffle 16 can prolong the growth time of mycelium in the chamber and can be installed or removed as needed.

[0046] In one embodiment, the separate top cover 12 is made of opaque material, the waterproof vent hole 13 has a diameter of 10 mm, and the separate top cover 12 of each small chamber opens and closes independently without affecting other small chambers, while preventing water vapor from overflowing. The 6 or 9 small chambers in the center circle use 1 independent separate top cover 12.

[0047] like Figure 3 As shown, the embodiment of the present invention also provides a method for preparing a filamentous fungus culture device for distinguishing different matrix absorption, comprising the following steps:

[0048] S1, manufacturing a circular chassis 11;

[0049] S2, using a circular double-layer isolation net 15 to divide the circular chassis 11 into three concentric circles;

[0050] S3, using radial straight partitions 14 to equally divide each concentric circle into 6 small chambers;

[0051] S4. A movable baffle 16 can be added to each chamber as needed;

[0052] S5. Equip each small room with a separate top cover 12 with a waterproof vent 13.

[0053] In one embodiment, in S2, the height of the circular double-layer isolation net 15 is the same as the height of the circular bottom plate 11, which can ensure that each chamber is closed and prevent the filamentous fungi from entering other chambers during the growth process. In addition, among the multiple concentric circles, the ratio of the radius of the first concentric circle to the radial width of the other circular rings is 1:1, which can ensure that the radial growth distance of the filamentous fungi in each chamber is the same, specifically 3 cm.

[0054] like Figure 4 As shown, the embodiment of the present invention also provides a method for using a filamentous fungus incubator for distinguishing different matrix absorptions, comprising the following steps:

[0055] (1) Each concentric circle is filled with the same matrix, including ordinary matrix and labeled matrix;

[0056] (2) In the three chambers in the same radial direction, ensure that the three matrices to be studied exist simultaneously and without duplication, and that there is only one labeled matrix;

[0057] (3) At the end of the culture, the growth of hyphae in each chamber can be observed, or hyphae can be separated from each chamber independently to test the isotope abundance;

[0058] (4) Based on the isotope labeling and subtraction methods, the growth conditions of filamentous fungi in different substrates, the absorption of different substrates, the migration of elements such as C and N in the hyphae, and the influence of the substrate arrangement order were quantified.

[0059] Embodiment 2

[0060] like Figure 1 and Figure 2 As shown, the embodiment of the present invention provides a filamentous fungus incubator for distinguishing absorption of different substrates, which is composed of a circular bottom plate 11 and a detachable top cover 12;

[0061] The circular chassis 11 has radial straight partitions 14 evenly distributed and fixedly connected inside, three double-layer isolation nets 15 installed inside, a separate top cover 12 installed on the top of the circular chassis 11, a waterproof vent 13 is opened on one side of the separate top cover 12, and a movable baffle 16 is provided inside the circular chassis 11.

[0062] In one embodiment, the height of the circular bottom plate 11 is 2 cm, and the bottom of the circular bottom plate 11 is made of a transparent material. The height of the circular bottom plate 11 is specified to clarify the volume of each chamber, that is, the volume of each matrix, to ensure that the filamentous fungi have sufficient growth space in each matrix. The bottom is made of a transparent material to facilitate the observation of the growth of the mycelial fungi.

[0063] In one embodiment, the pore size of the double-layer isolation net 15 is 180 μm, and the distance between the two layers of the double-layer isolation net 15 is 5 mm, which can isolate different substrates and prevent substrates from contacting each other, thereby ensuring the purity of the substrate in each chamber.

[0064] In one embodiment, the height of the radial straight partition 14 is the same as that of the circular bottom plate 11, which can ensure that each chamber is closed and prevent filamentous fungi from entering other chambers during the growth process.

[0065] In one embodiment, the movable baffle 16 is arc-shaped and its direction is opposite to the concentric circle formed by the double-layer isolation net 15. The movable baffle 16 can prolong the growth time of mycelium in the chamber and can be installed or removed as needed.

[0066] In one embodiment, the separate top cover 12 is made of opaque material, the waterproof vent hole 13 has a diameter of 5 mm, and the separate top cover 12 of each small chamber opens and closes independently without affecting other small chambers, while preventing water vapor from overflowing. The 6 or 9 small chambers in the central circle use 1 independent separate top cover 12.

[0067] like Figure 3 As shown, the embodiment of the present invention also provides a method for preparing a filamentous fungus culture device for distinguishing different matrix absorption, comprising the following steps:

[0068] S1, manufacturing a circular chassis 11;

[0069] S2, using a circular double-layer isolation net 15 to divide the circular chassis 11 into 4 concentric circles;

[0070] S3, using radial straight partitions 14 to equally divide each concentric circle into 9 small chambers;

[0071] S4. A movable baffle 16 can be added to each chamber as needed;

[0072] S5. Equip each small room with a separate top cover 12 with a waterproof vent 13.

[0073] In one embodiment, in S2, the height of the circular double-layer isolation net 15 is the same as the height of the circular bottom plate 11, which can ensure that each chamber is closed and prevent the filamentous fungi from entering other chambers during the growth process. In addition, among the multiple concentric circles, the ratio of the radius of the first concentric circle to the radial width of the other circular rings is 1:1, which can ensure that the radial growth distance of the filamentous fungi in each chamber is the same, specifically 3 cm.

[0074] like Figure 4 As shown, the embodiment of the present invention also provides a method for using a filamentous fungus incubator for distinguishing different matrix absorptions, comprising the following steps:

[0075] (1) Each concentric circle is filled with the same matrix, including ordinary matrix and labeled matrix;

[0076] (2) In the four chambers in the same radial direction, ensure that the four substrates to be studied exist simultaneously and without duplication, and there is only one labeled substrate;

[0077] (3) At the end of the culture, the growth of hyphae in each chamber can be observed, or hyphae can be separated from each chamber independently to test the isotope abundance;

[0078] (4) Based on the isotope labeling and subtraction methods, the growth conditions of filamentous fungi in different substrates, the absorption of different substrates, the migration of elements such as C and N in the hyphae, and the influence of the substrate arrangement order were quantified.

[0079] The above is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art can easily think of various changes or substitutions within the technical scope disclosed by the present invention, which should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

Claims

1. A filamentous fungus incubator for distinguishing different matrix absorptions, characterized in that: It comprises a circular bottom plate (11) and a detachable top cover (12); The circular chassis (11) is evenly distributed and fixedly connected with radial straight partitions (14), the circular chassis (11) is installed with two or three double-layer isolation nets (15), the detachable top cover (12) is installed on the top of the circular chassis (11), a waterproof vent hole (13) is opened on one side of the detachable top cover (12), and the circular chassis (11) is provided with a movable baffle (16) inside.

2. The filamentous fungus incubator for distinguishing different matrix absorption according to claim 1, characterized in that: The height of the circular bottom plate (11) is 1-3 cm, and the bottom of the circular bottom plate (11) is made of a transparent material.

3. The filamentous fungus incubator for distinguishing different matrix absorption according to claim 1, characterized in that: The pore size of the double-layer isolation net (15) is 200 μm, and the distance between the two layers of the double-layer isolation net (15) is 5 mm.

4. The filamentous fungus incubator for distinguishing different matrix absorption according to claim 1, characterized in that: The height of the radial straight partition (14) is the same as the height of the circular bottom plate (11).

5. The filamentous fungus incubator for distinguishing different matrix absorption according to claim 1, characterized in that: The movable baffle (16) is arc-shaped, and its direction is opposite to the concentric circle formed by the double-layer isolation net (15).

6. The filamentous fungus incubator for distinguishing different matrix absorption according to claim 1, characterized in that: The material of the detachable top cover (12) is a light-proof material, and the diameter of the waterproof vent hole (13) is 5-10 mm.

7. A method for preparing a filamentous fungus culture device for distinguishing different matrix absorptions according to any one of claims 1 to 6, characterized in that: The following steps are involved: S1, manufacturing a circular chassis (11); S2, using a circular double-layer isolation net (15) to divide the circular chassis (11) into 3-4 concentric circles; S3, using radial straight partitions (14) to equally divide each concentric circle into 6-9 small chambers; S4. A movable baffle (16) may be added to each chamber as required; S5. Each chamber is equipped with a separate top cover (12) with a waterproof vent (13).

8. The method for preparing a filamentous fungus culture device for distinguishing different substrate absorption according to claim 7, characterized in that: In S2, the height of the circular double-layer isolation net (15) is the same as the height of the circular bottom plate (11), and among the multiple concentric circles, the ratio of the radius of the first concentric circle to the radial width of the other circles is 1:1, preferably 2-4 cm.

9. A method for using a filamentous fungus incubator for distinguishing different substrate absorptions, characterized in that: The following steps are involved: (1) Each concentric circle is filled with the same matrix, including ordinary matrix and labeled matrix; (2) In the 3 or 4 chambers in the same radial direction, ensure that the 3 or 4 substrates to be studied are present simultaneously and without duplication, and there is only one labeled substrate; (3) At the end of the culture, the growth of hyphae in each chamber can be observed, or hyphae can be separated from each chamber independently to test the isotope abundance; (4) Based on the isotope labeling and subtraction methods, the growth conditions of filamentous fungi in different substrates, the absorption of different substrates, the migration of elements such as C and N in the hyphae, and the influence of the substrate arrangement order were quantified.

10. The method for using the filamentous fungus culture device for distinguishing different matrix absorption according to claim 9, characterized in that: By clarifying the effect of matrix arrangement order on the growth and absorption and utilization of filamentous fungi, the growth status of filamentous fungi in each matrix can be quantified, and the amount of C, N and other elements in the filamentous fungi in each chamber that come from the matrix of other chambers can be distinguished.

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