A substrate loading and fungal culture observation device with controllable weight and a method of using the same

Abstract

The application relates to a substrate loading and fungus culture observation device with controllable loading weight and a use method thereof, and relates to the technical field of fungus experiment equipment. The material pipe is a transparent quartz pipe body, three quartz branch pipes are welded on the pipe wall as inoculation ports and air inlets, the material pipe holder protects the material pipe, the sealing assembly blocks the material pipe, the first and last end support seats are fixed on one side of the bottom plate to support the material pipe holder, the pressing plate tightly presses the material pipe, the additional support seat top end is provided with a transition pipe which is in communication with the first end of the material pipe, the screw nut mounting seat is slidably mounted on the other side of the bottom plate and can be locked and positioned, the screw lever and the screw nut mounting seat form a screw pair, the inner side end is provided with a loading piston, the outer side end is connected with a driving assembly, and the torque value is monitored in real time through a torque sensor to control the substrate loading weight. The quantitative control and stable maintenance of the loading weight are realized, the inoculation point and the air inlet condition are standardized, the inoculation and culture observation of the fungus culture are facilitated, and the standardization, stability, accuracy and comparability of the experimental data are improved.

Description

Technical Field

[0001] This invention relates to the field of fungal experimental equipment technology, specifically a substrate filling and fungal culture observation device with controllable bulk density and its usage method. Background Technology

[0002] The growth rate and morphology of fungal hyphae are core indicators for evaluating strain viability, substrate compatibility, and environmental adaptability, and are of great guiding significance for optimizing fungal culture and edible / medicinal fungal cultivation processes, strain selection, and production practices. Currently, fungal hyphae culture and observation mainly utilize two types of culture media: PDA medium or production substrate.

[0003] PDA medium has the advantages of simple preparation and good uniformity, which can effectively eliminate the interference caused by differences in the physical properties of the substrate. However, its composition and physical form are significantly different from the solid substrates used in actual production, such as sawdust, wheat bran, straw, and livestock manure. The experimental data on fungal mycelial growth obtained based on PDA medium are difficult to be directly applied to production practice and cannot truly reflect the growth characteristics of fungi in actual production cultivation substrates.

[0004] When using production substrates for cultivation, the particle size, component ratio, water content, and air permeability of the substrate, especially its bulk density (compactness), significantly affect mycelial growth. Current technologies often use culture trays, flasks, bags, or ordinary test tubes to fill with production substrates for fungal culture and growth testing, which has several drawbacks: First, substrate bulk density relies entirely on manual operation experience, making quantitative control and precise repeatability impossible. Even with the same substrate formulation, variations in packing density directly lead to differences in air permeability and water retention, resulting in abnormal fluctuations in mycelial growth rate and morphology, severely interfering with the accuracy and comparability of experimental results. Second, it is difficult to achieve fixed-point, quantitative inoculation and uniform aeration; differences in inoculation location and substrate aeration further amplify experimental errors. Third, frequent operation during observation reduces the airtightness of the cultivation device, easily introducing contaminants, and makes it difficult to accurately mark the mycelial growth front, resulting in significant errors in growth rate measurement.

[0005] In summary, there is currently a lack of a fungal culture experimental device that can precisely control the substrate bulk density (with a wide setting range that is close to actual production conditions) while simultaneously achieving fixed-point inoculation, uniform aeration, and in-situ observation. This makes it impossible to effectively eliminate the interference of substrate bulk density differences, which restricts the precise implementation of fungal growth characteristic research, strain screening, and cultivation process optimization. Summary of the Invention

[0006] To address the shortcomings of the prior art, this invention provides a substrate filling and fungal culture observation device with controllable bulk density and its usage method. It can achieve quantitative control and stable maintenance of bulk density for different solid substrates, while standardizing inoculation sites and ventilation conditions, facilitating inoculation observation of fungal culture, and improving the standardization, stability, accuracy, and comparability of fungal growth experimental data. It is suitable for fungal growth characteristic research and substrate and environmental condition adaptability analysis in scientific research laboratories and industrial testing laboratories.

[0007] To achieve the above objectives, the present invention adopts the following technical solution: A substrate filling and fungal culture observation device with controllable bulk density consists of two parts: a fungal culture observation mechanism and a substrate filling mechanism with controllable bulk density. The fungal culture observation device includes a material tube, a material tube holder, and a sealing assembly. The material tube is a transparent quartz tube with three quartz branch tubes fused to its upper wall. The branch tube at the midpoint serves as the inoculation port, and the branch tubes on both sides serve as ventilation ports. The material tube holder is an integrally formed support structure, including an arc-shaped support groove at the bottom, a circular baffle at the end, and an annular baffle with external threads at the front end. The annular baffle is screwed with a matching nut. The sealing assembly includes plugs for sealing the two ends of the material tube, filter plugs for sealing the inoculation port, and filter breathable membranes installed at the ventilation ports on both sides. The controllable bulk density substrate filling mechanism includes a base plate, an end support, a head support, an additional support, a pressure plate, a transition tube, a nut mounting base, a lever, a drive assembly, and a torque sensor. The end support and the head support are fixed to one side of the upper surface of the base plate, respectively supporting the end and head of the material tube support. A limiting plate is provided on the outer side of the top of the end support to restrict the axial displacement of the material tube support. The pressure plate is fixed to the top of the end support to press and fix the material tube. The additional support is fixed to the middle of the upper surface of the base plate. The transition tube is detachably fixed to the top of the additional support and coaxially connected to the head of the material tube. The nut mounting base is slidably installed on the other side of the upper surface of the base plate and is provided with a positioning screw. The lever is coaxially arranged with the material tube and screwed to the top of the nut mounting base to form a lead screw pair. A loading piston is provided on the inner end of the lever, and the outer end is connected to the drive assembly. The torque sensor is connected in series in the transmission path of the lever to monitor the torque value of the substrate compaction process in real time to control the bulk density of the substrate.

[0008] Furthermore, the material pipe has an inner diameter of 30mm, a length of 300mm, and a wall thickness of 3mm. The three quartz branch pipes have a diameter of 16mm and a length of 30mm. The center of the two vents is 30mm away from the corresponding edge of the material pipe.

[0009] Furthermore, the arc-shaped groove of the material pipe support is adapted to the outer wall of the material pipe, and the annular baffle can axially compress the plugs at both ends of the material pipe through the screw-on nut, thereby achieving the functions of sealing the ends of the material pipe and protecting the pipe body.

[0010] Furthermore, the drive assembly uses a handle, which is manually rotated in both directions to drive the lever to rotate, thereby realizing the axial advancement and retraction of the loading piston.

[0011] Furthermore, the drive component adopts a geared motor, which is slidably connected to a slide rail fixed to the bottom of the outer side of the nut mounting base to achieve guidance. By switching the forward and reverse directions of the geared motor, the wire lever is driven to rotate, thereby realizing the axial advancement and retraction of the loading piston.

[0012] A method for using a substrate filling and fungal culture observation device with controllable bulk density includes the following steps: S1. Preparation of production matrix: Prepare the production matrix according to the experimental target, mix it evenly, add water according to the set material-to-water ratio and stir until fully mixed before use. S2. Assembly and positioning of the device: Insert a plug into the end of the material pipe. After the material pipe is placed in the material pipe support, place it together on the top of the end support and the head support. Adjust the position so that the end of the material pipe support abuts against the limit plate. Then install the pressure plate to press and position the material pipe. Install a fixed additional support in front of the head support. S3. Segmented filling and bulk density control: Segmented filling is adopted and the single filling amount of the production matrix is ​​pre-calculated. The corresponding number of transition tubes are added with a single quantitative amount of production matrix. One transition tube is installed and fixed on the top of the auxiliary support base and ensured to be coaxially attached to the first end of the material tube support. The sliding nut mounting base allows the loading piston to enter the transition tube opening. The positioning screw is tightened to complete the locking. The drive component drives the wire lever to rotate, controlling the loading piston to push the production matrix in the transition tube into the material tube for compaction. The torque value is monitored in real time using a torque sensor. After reaching the preset threshold, the loading piston is retracted. The next transition tube is replaced and the compaction operation is repeated until all the production matrix is ​​filled and the bulk density meets the preset index. The material tube support and material tube are then removed. S4. Sealing and Sterilization: Install a plug at the beginning of the material tube, screw the nut onto the external thread at the beginning of the material tube support and tighten it to seal both ends of the material tube. At the same time, use a filter gel plug to seal the inoculation port. Install filter breathable membranes at the two vents respectively. Then put the material tube together with the material tube support into the sterilization equipment for sterilization. S5. Aseptic inoculation and culture: Under aseptic conditions, quantitatively inoculate the target fungal strain into the midpoint of the substrate through the inoculation port. After inoculation, seal the inoculation port with a filter gel plug and transfer the material tube along with the material tube holder into the experimental environment for static culture. S6. Growth observation and data recording: During the cultivation process, the mycelial growth status is observed regularly through the material tube. By marking the leading edge of the mycelial growth, the straight-line growth distance between the two ends of the mycelium is measured, and the mycelial growth rate is calculated in combination with the cultivation time.

[0013] Furthermore, in step S3, the total loading mass is calculated in advance based on the inner volume of the material pipe and the target bulk density. Then, the single loading amount and corresponding compaction length of the production matrix are divided according to the volume of the transition pipe. The uniformity of the matrix density of the whole pipe is ensured by segmented loading.

[0014] Furthermore, in step S4, the sterilization operation is performed at 121°C for 60-120 minutes under moist heat sterilization, and after sterilization, it is naturally cooled to below 28°C.

[0015] Furthermore, in step S6, by setting up multiple sets of parallel material tubes, comparative experiments are conducted on different substrate formulations, different bulk densities, different strains, or different culture conditions to obtain experimental data on the culture of corresponding fungal mycelia.

[0016] Compared with the prior art, the beneficial effects of the present invention are: 1. Quantitative control and stable production substrate bulk density throughout the process: Through the screw drive compaction and torque sensor feedback, the compaction of the substrate filling can be precisely set and controlled, adapting to different types, ratios, moisture contents and particle sizes of production substrates. This helps to maintain the stability of substrate bulk density throughout the sterilization and cultivation process, solving the industry pain point of solid substrates being difficult to shape and prone to rebound, and eliminating the interference of bulk density differences on mycelial growth experiments from the root. 2. Standardization of the entire process of fungal culture conditions: The structure design of central fixed-point inoculation and bilateral symmetrical ventilation ensures the consistency of inoculation position and ventilation conditions. The hyphae grow bidirectionally along the material tube, which can offset the minor errors caused by local unevenness of the substrate and significantly improve the repeatability and data reliability of parallel experiments. 3. Modular design with strong adaptability and practicality: Multiple transition tubes are set up for segmented filling, which facilitates precise batch filling of production matrix into the material tube. The transition tubes make the filling amount of each batch precise and controllable, and maximize the uniformity of matrix bulk density. At the same time, it is equipped with two optional modes: manual drive and motor drive, which are suitable for small-batch flexible experiments and high-throughput batch testing scenarios, respectively. The transparent quartz material tube supports in-situ non-destructive observation, and the mycelial growth front can be marked and the growth rate measured without opening the cap. When used with the plug, filter gel plug and material tube holder, the operation is simple and can effectively prevent contamination by other bacteria, while also providing protection to avoid breakage. 4. Possesses high scientific research and production application value: It helps to accurately conduct comparative studies on the effects of nutritional factors such as substrate type, ratio, particle size, water content, and pH, as well as environmental factors such as temperature, humidity, and gas concentration (oxygen / carbon dioxide) on fungal growth, providing reliable experimental support for strain selection and cultivation process optimization, and filling the gap in the field of standardized cultivation and observation equipment for production substrates. Attached Figure Description

[0017] Figure 1 This is a schematic diagram of the main structure of the device of the present invention in manual drive mode; Figure 2 This is a three-dimensional schematic diagram of the device of the present invention in manual drive mode; Figure 3 This is a schematic diagram of the main structure of the device of the present invention in motor drive mode; Figure 4 This is a three-dimensional schematic diagram of the device of the present invention in motor drive mode; Figure 5 This is a schematic diagram of the disassembled structure of the material pipe and the material pipe support in the device of the present invention.

[0018] In the diagram: 1. Base plate; 2. End support seat; 3. Head support seat; 4. Additional support seat; 5. Material pipe support; 6. Material pipe; 6-1. Plug; 6-2. Filter plug; 6-3. Filter membrane; 7. Pressure plate; 8. Transition pipe; 9. Slide rail; 10. Wire nut mounting seat; 11. Wire lever; 12. Drive assembly. Detailed Implementation

[0019] The technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the invention, not all embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[0020] like Figures 1-5 As shown, a substrate filling and fungal culture observation device with controllable bulk density comprises two parts: a fungal culture observation mechanism and a substrate filling mechanism with controllable bulk density. The fungal culture observation mechanism is used to complete the entire process of substrate filling, sealing sterilization, inoculation culture, and growth observation under controllable bulk density conditions. The substrate filling mechanism with controllable bulk density is used to quantitatively fill the production substrate into the fungal culture observation mechanism and precisely control the bulk density index. The specific scheme is as follows: Combination Figure 1 , Figure 3 , Figure 5 As shown, the fungal culture observation device includes a material tube 6, a material tube holder 5, and a sealing assembly.

[0021] Material tube 6 is made of high-temperature and moisture-heat sterilizable quartz glass, with an inner diameter of 30mm, a length of 300mm, and a wall thickness of 3mm. The tube is completely transparent, allowing direct observation of the internal substrate and mycelial growth. Material tube 6 is arranged horizontally, with three quartz branch tubes perpendicular to the tube body fused evenly along its axial direction above the tube wall. The three quartz branch tubes have a diameter of 16mm and a length of 30mm. The quartz branch tube located at the midpoint of the axial direction serves as the inoculation port for fungal inoculation. The quartz branch tubes located on both sides serve as vents, with the center of each vent 30mm from the corresponding edge of material tube 6, ensuring uniform aeration within the tube during cultivation.

[0022] The material tube support 5 is an integrally molded support structure, including an arc-shaped groove at the bottom, a circular baffle at the end, and an annular baffle at the beginning. The outer wall of the annular baffle is fitted with a threaded screw-on nut. The arc of the arc-shaped groove is adapted to the outer wall of the material tube 6 to support the material tube 6; the circular baffle is used to limit the axial displacement of the end of the material tube 6; the annular baffle, through the screw-on nut, can axially compress the plugs 6-1 at both ends of the material tube 6 to achieve end sealing of the material tube 6. The material tube support 5 has a dual function: first, to protect the material tube 6 during filling, sterilization, and handling, reducing the risk of tube breakage; second, to provide radial and axial constraints on the material tube 6, and to achieve precise positioning during the filling process in conjunction with the support base.

[0023] The sealing assembly includes two plugs 6-1, one filter gel plug 6-2, and two filter breathable membranes 6-3. During the filling process, only the end of the material tube 6 is sealed with one plug 6-1. The open end of the material tube 6 is connected to the annular baffle of the material tube support 5 for substrate filling. After filling, both ends of the material tube 6 are sealed with one plug 6-1. The two plugs 6-1 are then tightened at the beginning of the material tube support 5 to provide an elastic seal to both ends of the material tube 6. The filter gel plug 6-2 is used to seal the inoculation port. It is removed during the inoculation operation and then resealed after inoculation. The two filter breathable membranes 6-3 are sealed on the vent and remain stationary for ventilation of the material tube 6 during fungal culture.

[0024] Combination Figures 1-4 As shown, the substrate filling mechanism with controllable bulk density includes a base plate 1, an end support seat 2, a head support seat 3, an additional support seat 4, a pressure plate 7, a transition tube 8, a slide rail 9, a nut mounting seat 10, a nut lever 11, a drive assembly 12, and a torque sensor.

[0025] The base plate 1 is a horizontally set rectangular plate, which serves as the mounting base for the entire device and ensures the coaxiality and structural stability of each component.

[0026] The end support 2 and the first support 3 are arranged in a straight line and fixed at intervals on one side of the upper surface of the base plate 1. The end support 2 has an arc-shaped groove on the inner side of its top end to support the end of the material pipe support 5. The end support 2 has a limiting plate reserved on the outer side of its top end to limit the axial displacement of the material pipe support 5 and prevent the pipe from moving backward during the filling process. The first support 3 has an arc-shaped through groove on its top end to support the first end of the material pipe support 5.

[0027] The pressure plate 7 is fixed to the top of the end support 2 by screws. The end of the pressure plate 7 can be pre-made with an arc-shaped notch to match the shape of the vent at the end of the material pipe 6, which is used to press and fix the material pipe 6, further improving the positioning stability during the filling process.

[0028] The auxiliary support 4 is fixed in the middle of the upper surface of the base plate 1. It is located in front of the first support 3 and has a sleeve integrally installed at the top horizontally. The sleeve is used to detachably insert the transition tube 8 and has locking screws on the side wall for positioning. There are multiple transition tubes 8 for pre-filling the production matrix to be loaded into the material tube 6. The outer diameter of the transition tube 8 is the same as the outer diameter of the material tube 6, and the inner diameter of the transition tube 8 should be slightly smaller than the inner diameter of the material tube 6 to facilitate the entry of the production matrix. One end of the transition tube 8 can be chamfered to facilitate the introduction of the production matrix. In the filling state, the transition tube 8 is coaxially attached to the first end of the material tube support 5. Since the first end of the material tube support 5 is set as an annular baffle, the transition tube 8 and the material tube 6 are coaxially connected. The transition tube 8 serves as a storage cavity for single filling, used to realize segmented filling and segmented compaction, and to ensure the uniformity of the density of the matrix in the whole tube.

[0029] The above components are assembled to form a basic support and positioning module, which, in conjunction with the transmission-drive module, enables the batch compaction and filling of the substrate. The device is characterized by its miniaturization and lightweight design. During the cultivation stage, a three-dimensional tray can be used, facilitating the stacking of multiple material tubes, improving the utilization of cultivation space, and also facilitating experimental comparisons. The transmission-drive module can be operated manually or by a motor. In manual drive mode, combined with Figures 1-2As shown, a nut mounting base 10 is slidably mounted on the other side of the base plate 1. A slide rail 9 is provided on the surface of the base plate 1 along the direction of the material tube 6 for guidance, so that the nut mounting base 10 can only move horizontally along the direction of the material tube 6. A positioning screw is provided at the bottom of the nut mounting base 10 for locking and positioning. The nut lever 11 is located on the axis of the material tube 6 and is screwed to the top of the nut mounting base 10 to form a lead screw pair, which converts the rotational motion of the nut lever 11 into linear motion along the axis of the material tube 6. A loading piston is installed on the inner end of the nut lever 11 and is clearance-fitted with the transition tube 8 and the material tube 6 to insert the matrix material in the transition tube 8 into the material tube 6. A handle is coaxially fixed on the outer end of the nut lever 11 as a drive component 12. The nut lever 11 is rotated by manually rotating the handle in both directions to achieve the axial advancement and retraction of the loading piston. This mode has a simple structure, low cost, and does not require an external power supply, making it suitable for flexible experimental scenarios with small batches and multiple parameters.

[0030] In motor drive mode, combined Figures 3-4 As shown, a nut mounting base 10 is slidably mounted on the other side of the base plate 1. The nut mounting base 10 can only move horizontally along the material tube 6 and is locked in place by a positioning screw. The nut lever 11 is located on the axis of the material tube 6 and is screwed into the top of the nut mounting base 10 to form a lead screw pair, which can convert the rotational motion of the nut lever 11 into linear motion along the axis of the material tube 6. A loading piston is installed on the inner end of the nut lever 11 and is clearance-fitted with the transition tube 8 and the material tube 6 to push the material in the transition tube 8 into the material tube 6. The difference is that the outer end of the nut lever 11 is connected to a geared motor as a drive component 12, while the slide rail 9 is placed on the surface of the base plate 1 and fixed to the outer bottom of the nut mounting base 10. The geared motor and the slide rail 9 are slidably connected to provide guidance. By switching the forward and reverse rotation of the geared motor, the nut lever 11 is driven to rotate, realizing the axial advancement and retraction of the loading piston. This mode can be combined with the control system to automatically complete the predetermined advancement speed and torque, with low operation intensity and higher consistency of bulk density control, and is suitable for high-throughput experimental scenarios with batch samples.

[0031] In addition, both modes are equipped with torque sensors, which are connected in series in the transmission path of the wire lever 11 to monitor the torque value during the matrix compaction process in real time. By preset torque threshold, the degree of matrix compaction can be precisely controlled. Combined with the loading quality and material tube volume, an accurate matrix bulk density index can be obtained, ensuring the accuracy and repeatability of bulk density control.

[0032] like Figures 1-5 As shown, based on the above device, a method for using a substrate filling and fungal culture observation device with controllable bulk density is proposed. In practical applications, it can be used to study the mycelial growth of large fungi such as Auricularia auricula-judae, Ganoderma lucidum, and Lentinus edodes in a growth substrate mainly composed of sawdust and wheat bran. The method includes the following steps: S1. Preparation of production matrix: Prepare the production matrix according to the experimental objectives. For example, use a mixed matrix of 88% wood chips, 11% wheat bran, 0.5% lime, and 0.5% gypsum. After mixing evenly, add water according to the set material-to-water ratio and stir. In this embodiment, the material-to-water ratio is 1:1.2. Mix thoroughly and set aside.

[0033] S2. Device assembly and positioning: First, insert the plug 6-1 into the end of the material pipe 6. Then, place the material pipe 6 as a whole into the arc-shaped groove of the material pipe support 5. Place the material pipe support 5 together with the material pipe 6 on the top of the end support 2 and the beginning support 3. Adjust the position so that the end of the material pipe support 5 abuts against the limiting plate of the end support 2. Then, install the pressure plate 7 and lock it in place to fix the position of the material pipe 6 and the material pipe support 5. Place the additional support 4 in a suitable position in front of the beginning support 3 and fix it to the base plate 1.

[0034] S3. Segmented filling and bulk density control: The total loading mass is calculated in advance based on the inner volume of the material pipe 6 and the target bulk density. Then, the single loading amount and corresponding compaction length of the production matrix are determined based on the volume of the transition pipe 8.

[0035] Take the corresponding number of transition tubes 8 and add a single quantitative amount of production matrix. First, insert one transition tube 8 into the sleeve at the top of the auxiliary support 4 and lock it to ensure that the transition tube 8 is coaxially attached to the first end of the material tube support 5. Slide the nut mounting seat 10 to make the loading piston enter the tube opening of the transition tube 8. Tighten the positioning screw to complete the locking. Then, drive the wire lever 11 to rotate through the drive assembly 12 to control the loading piston to advance axially and push the production matrix in the transition tube 8 into the material tube 6 for compaction. During the compaction process, the torque value is monitored in real time by the torque sensor. When the torque reaches the preset threshold, the advancement is stopped to complete the single-segment matrix filling. Then, the loading piston is retracted, and the next transition tube 8 is replaced and fixed on the sleeve of the auxiliary support 4. The advancement and compaction operation is repeated until all the production matrix is ​​filled.

[0036] The production matrix is ​​filled into the material tube 6 in sections to ensure uniform density and consistent bulk density throughout the tube. In this embodiment, the net capacity of the material tube 6 is 198ml, and the target bulk density is 0.8g / ml, so the total mass of material to be added in batches is 158g.

[0037] After all the production substrates are filled, loosen the positioning screws, retract the nut mounting base 10 and the nut lever 11, remove the pressure plate 7, and take out the material tube support 5 and the material tube 6.

[0038] S4. Sealing and sterilization: Insert another plug 6-1 into the first end of the material pipe 6, screw the matching nut onto the external thread of the first end of the material pipe support 5, tighten the nut to axially compress the plugs 6-1 at both ends, so that the two ends of the material pipe 6 are sealed tightly; at the same time, use the filter gel plug 6-2 to seal the inoculation port, and seal the filter breathable membrane 6-3 on the two vents respectively.

[0039] The material tube 6, together with the material tube holder 5, is placed into the sterilization equipment for sterilization. In this embodiment, the material tube is placed in the sterilizer and sterilized by moist heat at 121°C for 60-120 minutes. After sterilization, the material tube is allowed to cool naturally to below 28°C.

[0040] S5. Aseptic inoculation and culture: Under aseptic conditions, open the filter plug 6-2 of the inoculation port and quantitatively inoculate the target fungal strain into the midpoint of the production substrate through the inoculation port. After inoculation, re-tighten the filter plug 6-2 of the inoculation port.

[0041] The entire set of material tubes 6 and material tube holders 5 were moved into a constant temperature incubator and statically incubated according to the temperature, humidity and gas concentration conditions set in the experiment.

[0042] S6. Growth Observation and Data Recording: During the cultivation process, the mycelial growth status is observed regularly through the transparent material tube 6, and indicators such as mycelial color and morphology are recorded; a mark is made at the leading edge of mycelial growth, the straight-line growth distance between the two ends of the mycelium is measured, and the mycelial growth rate is calculated in combination with the cultivation time.

[0043] By setting up multiple sets of parallel material tubes 6, comparative experiments can be conducted on different substrate formulations, different bulk densities, different strains, and different culture conditions. Based on stable and consistent bulk densities and culture conditions, accurate and comparable experimental data on fungal mycelial culture can be obtained.

[0044] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the invention can be implemented in other forms without departing from its spirit or essential characteristics. Therefore, the embodiments should be considered illustrative and non-limiting in all respects, and the scope of the invention is defined by the appended claims rather than the foregoing description. Thus, all variations falling within the meaning and scope of the equivalents of the claims are intended to be included within the present invention. No reference numerals in the claims should be construed as limiting the scope of the claims.

[0045] Furthermore, it should be understood that although this specification describes embodiments, not every embodiment contains only one independent technical solution. This narrative style is merely for clarity. Those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can also be appropriately combined to form other embodiments that can be understood by those skilled in the art.

Claims

1. A substrate filling and fungal culture observation device with controllable bulk density, characterized in that: It consists of two parts: a fungal culture and observation mechanism and a substrate filling mechanism with controllable bulk density; The fungal culture observation device includes a material tube, a material tube holder, and a sealing assembly. The material tube is a transparent quartz tube with three quartz branch tubes fused to its upper wall. The branch tube at the midpoint serves as the inoculation port, and the branch tubes on both sides serve as ventilation ports. The material tube holder is an integrally formed support structure, including an arc-shaped support groove at the bottom, a circular baffle at the end, and an annular baffle with external threads at the front end. The annular baffle is screwed with a matching nut. The sealing assembly includes plugs for sealing the two ends of the material tube, filter plugs for sealing the inoculation port, and filter breathable membranes installed at the ventilation ports on both sides. The controllable bulk density substrate filling mechanism includes a base plate, an end support, a head support, an additional support, a pressure plate, a transition tube, a nut mounting base, a lever, a drive assembly, and a torque sensor. The end support and the head support are fixed to one side of the upper surface of the base plate, respectively supporting the end and head of the material tube support. A limiting plate is provided on the outer side of the top of the end support to restrict the axial displacement of the material tube support. The pressure plate is fixed to the top of the end support to press and fix the material tube. The additional support is fixed to the middle of the upper surface of the base plate. The transition tube is detachably fixed to the top of the additional support and coaxially connected to the head of the material tube. The nut mounting base is slidably installed on the other side of the upper surface of the base plate and is provided with a positioning screw. The lever is coaxially arranged with the material tube and screwed to the top of the nut mounting base to form a lead screw pair. A loading piston is provided on the inner end of the lever, and the outer end is connected to the drive assembly. The torque sensor is connected in series in the transmission path of the lever to monitor the torque value of the substrate compaction process in real time to control the bulk density of the substrate.

2. The substrate filling and fungal culture observation device with controllable bulk density according to claim 1, characterized in that: The material pipe has an inner diameter of 30mm, a length of 300mm, and a wall thickness of 3mm. The three quartz branch pipes have a diameter of 16mm and a length of 30mm. The center of the two vents is 30mm away from the corresponding edge of the material pipe.

3. The substrate filling and fungal culture observation device with controllable bulk density according to claim 1, characterized in that: The arc-shaped groove of the material pipe support is adapted to the outer wall of the material pipe, and the annular baffle can axially compress the plugs at both ends of the material pipe through the screw-on nut, so as to achieve the sealing of the material pipe end and the protection of the pipe body.

4. The substrate filling and fungal culture observation device with controllable bulk density according to claim 1, characterized in that: The drive assembly uses a handle. By manually rotating the handle in both directions, the lever is driven to rotate, thereby achieving the axial advancement and retraction of the loading piston.

5. The substrate filling and fungal culture observation device with controllable bulk density according to claim 1, characterized in that: The drive component uses a geared motor, which is slidably connected to a slide rail fixed to the bottom of the outer side of the nut mounting base for guidance. The forward and reverse switching of the geared motor drives the wire lever to rotate, thereby realizing the axial advancement and retraction of the loading piston.

6. A method for using a substrate filling and fungal culture observation device with controllable bulk density, characterized in that: The substrate filling and fungal culture observation device with controllable bulk density according to any one of claims 1-5, the method of using it includes the following steps: S1. Preparation of production matrix: Prepare the production matrix according to the experimental target, mix it evenly, add water according to the set material-to-water ratio and stir until fully mixed before use. S2. Assembly and positioning of the device: Insert a plug into the end of the material pipe. After the material pipe is placed in the material pipe support, place it together on the top of the end support and the head support. Adjust the position so that the end of the material pipe support abuts against the limit plate. Then install the pressure plate to press and position the material pipe. Install a fixed additional support in front of the head support. S3. Segmented filling and bulk density control: Segmented filling is adopted and the single filling amount of the production matrix is ​​pre-calculated. The corresponding number of transition tubes are added with a single quantitative amount of production matrix. One transition tube is installed and fixed on the top of the auxiliary support base and ensured to be coaxially attached to the first end of the material tube support. The sliding nut mounting base allows the loading piston to enter the transition tube opening. The positioning screw is tightened to complete the locking. The drive component drives the wire lever to rotate, controlling the loading piston to push the production matrix in the transition tube into the material tube for compaction. The torque value is monitored in real time using a torque sensor. After reaching the preset threshold, the loading piston is retracted. The next transition tube is replaced and the compaction operation is repeated until all the production matrix is ​​filled and the bulk density meets the preset index. The material tube support and material tube are then removed. S4. Sealing and Sterilization: Install a plug at the beginning of the material tube, screw the nut onto the external thread at the beginning of the material tube support and tighten it to seal both ends of the material tube. At the same time, use a filter gel plug to seal the inoculation port. Install filter breathable membranes at the two vents respectively. Then put the material tube together with the material tube support into the sterilization equipment for sterilization. S5. Aseptic inoculation and culture: Under aseptic conditions, quantitatively inoculate the target fungal strain into the midpoint of the substrate through the inoculation port. After inoculation, seal the inoculation port with a filter gel plug and transfer the material tube along with the material tube holder into the experimental environment for static culture. S6. Growth observation and data recording: During the cultivation process, the mycelial growth status is observed regularly through the material tube. By marking the leading edge of the mycelial growth, the straight-line growth distance between the two ends of the mycelium is measured, and the mycelial growth rate is calculated in combination with the cultivation time.

7. The method of using the substrate filling and fungal culture observation device with controllable bulk density according to claim 6, characterized in that: In step S3, the total loading mass is calculated in advance based on the inner volume of the material pipe and the target bulk density. Then, the single loading amount and corresponding compaction length of the production matrix are divided according to the volume of the transition pipe. The uniformity of the matrix density of the whole pipe is ensured by segmented loading.

8. The method of using the substrate filling and fungal culture observation device with controllable bulk density according to claim 6, characterized in that: In step S4, the sterilization operation is performed at 121°C for 60-120 minutes under moist heat sterilization, and after sterilization, it is naturally cooled to below 28°C.

9. The method of using the substrate filling and fungal culture observation device with controllable bulk density according to claim 6, characterized in that: In step S6, by setting up multiple sets of parallel material tubes, comparative experiments are conducted on different substrate formulations, different bulk densities, different strains, or different culture conditions to obtain experimental data on the culture of corresponding fungal mycelia.

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