Phosphorus-dissolving microorganism enrichment culture and terrace layout integrated device

Abstract

The invention provides a phosphorus-knowing microorganism enrichment culture and terrace layout integrated device, which relates to the technical field of agricultural microorganisms and comprises a selective enrichment culture reactor, a bacterium hanging culture unit and a one-way connecting channel. The enrichment culture reactor is composed of a reactor shell and a top cover, a stirring device and an indissolvable phosphorus particle bed box supported by a supporting plate are arranged in the enrichment culture reactor, an aeration air inlet is formed for introducing sterile air / oxygen, an exhaust port is connected with a sterile filter, and pH, temperature and dissolved oxygen probes can be arranged. The hanging bacteria culture unit is a detachable container, is internally provided with a hollow porous carrier cylinder which is detachably connected with a top cover of the hanging bacteria culture unit, and is provided with a liquid supplementing port and an exhaust port. The one-way connecting channel is composed of a sterile isolating valve, a one-way valve and a quick connector, and a pump body can be arranged according to needs to assist in conveying the culture solution. According to the device, selective enrichment and carrier bacterium hanging can be realized under the closed sterile condition, the pollution risk is reduced, and the culture efficiency and the arrangement convenience are improved.

Description

Technical Field

[0001] This invention relates to the field of agricultural microbial technology, specifically to an integrated device for the enrichment and cultivation of phosphate-solubilizing microorganisms and the layout of terraced fields. Background Technology

[0002] Phosphorus is one of the essential macronutrients for plant growth, but it often exists in soil as insoluble phosphates, making it difficult for crops to directly absorb and utilize. Current agricultural production widely relies on chemical phosphate fertilizers to increase crop phosphorus supply, but long-term or excessive application not only increases planting costs but can also lead to the deterioration of soil physicochemical properties and environmental problems such as eutrophication caused by phosphorus carried by runoff. To reduce reliance on chemical phosphate fertilizers and improve soil phosphorus use efficiency, phosphorus-solubilizing microorganisms have attracted attention because they can convert insoluble inorganic phosphorus into soluble phosphorus that can be absorbed by plants through the production of organic acids and phosphatases.

[0003] The actual application effectiveness of phosphate-solubilizing microorganisms largely depends on the source, activity, and quantity of the microbial community. In existing technologies, the screening and enrichment culture of phosphate-solubilizing microorganisms are mostly conducted under laboratory conditions, typically relying on conventional culture containers, shake flasks, or general fermentation / culture equipment for enrichment and amplification. While this method can obtain a certain amount of microorganisms, it often suffers from problems such as low selective enrichment efficiency, sensitivity to external contamination during the culture process, and difficulty in guaranteeing cell activity and stability. Furthermore, the lack of a smooth connection mechanism between laboratory culture systems and field applications means that the cultured bacterial solutions or agents are prone to contamination, inactivation, or concentration fluctuations during transfer, dispensing, and transportation, leading to unstable actual field application effects. Therefore, how to achieve efficient and selective enrichment culture of phosphate-solubilizing microorganisms under sterile and controllable conditions, and how to reliably transfer the culture system to application units, is one of the key problems that urgently need to be solved in existing technologies.

[0004] For terraced farmland common in mountainous and hilly areas, the uniform application and stratified distribution of phosphate fertilizers and microbial agents are more challenging due to the dispersed plots, significant elevation differences, and complex water flow paths. Current field application methods, such as manual spreading, spraying, or application with irrigation water, struggle to ensure consistent application across different plot levels and maintain a sustained effect. In particular, achieving stratified, quantitative, and continuous application across multiple terrace levels is difficult, resulting in localized enrichment of phosphate-solubilizing microorganisms while other areas are deficient, impacting overall efficacy and stability. If on-site enrichment or expansion of phosphate-solubilizing microorganisms could be achieved in the terraced fields, and the culture system could be organically integrated with the application process in a controllable manner, enabling stratified application and sustained release of phosphate-solubilizing microorganisms across different plot levels, it would significantly improve phosphorus utilization efficiency and reduce fertilizer usage.

[0005] However, current technologies still lack a comprehensive, modular solution for terraced fields that integrates the selective enrichment and cultivation of phosphate-solubilizing microorganisms with tiered deployment in terraced environments. In particular, there is a lack of a suitable microbial culture device and its supporting structure for on-site use, enabling the enrichment and cultivation process to be conducted under controlled, aseptic conditions. This would allow for a more convenient and controllable connection of the culture system to on-site deployment units, achieving stable transfer and quantitative deployment of the culture products to meet the deployment needs of multi-level terraced fields. Therefore, it is necessary to provide a novel device that organically combines the enrichment and cultivation of phosphate-solubilizing microorganisms with their deployment in terraced environments, improving the efficiency and stability of on-site preparation and deployment of phosphate-solubilizing microorganisms, thereby enhancing sustainable agricultural productivity. Summary of the Invention

[0006] An integrated device for phosphate-solubilizing microbial enrichment culture and terraced field layout includes a selective enrichment culture reactor, a microbial culture unit, and a one-way connection channel. The enrichment culture reactor includes a reactor shell and a top cover, with the top cover enclosing the top of the reactor shell. Inside the reactor shell are a stirring device and a bed of insoluble phosphate particles. The stirring device includes a stirring motor mounted above the top cover, a stirring shaft driven by the stirring motor, and an impeller connected to the stirring shaft. The bed of insoluble phosphate particles is located at the bottom of the reactor shell and supported by a particle bed support plate, and contains insoluble phosphate particles. The reactor shell also has an aeration inlet for introducing sterile air. The microbial culture unit is a container detachably connected to the reactor shell, and contains at least one hollow porous carrier tube connected to the top cover of the microbial culture unit and is detachable. The one-way connection channel includes a sterile isolation valve, a one-way valve, and a quick connector, which together form a one-way flow connection channel from the reactor shell to the microbial culture unit.

[0007] Furthermore, the reactor shell is equipped with a pH probe, a temperature probe, and a dissolved oxygen probe.

[0008] Furthermore, the reactor shell has a sterile filter at the exhaust port.

[0009] Furthermore, an aeration inlet is provided at the bottom of the reactor shell, through which sterile air or oxygen is introduced into the reactor shell.

[0010] Furthermore, a culture medium inlet and a metering pump are provided above the reactor shell. The metering pump is connected to a culture medium storage container to quantitatively add culture medium or reagents into the reactor shell.

[0011] Furthermore, the top cover of the bacterial culture unit is provided with a liquid replenishment port, which can be detached or sealed with a cap.

[0012] Furthermore, the bacterial culture unit is equipped with an exhaust port so that gas is discharged when the culture medium enters the bacterial culture unit.

[0013] Furthermore, the unidirectional flow connection channel can be equipped with a pump to assist in transporting the culture medium from the reactor shell to the bacterial culture unit as needed.

[0014] Compared with the prior art, the present invention has the following advantages: This device addresses the problems in the prior art, such as low enrichment efficiency, easy contamination during the cultivation process, disconnect between cultivation and field application, and difficulties in coordinating with terraced field layout. Based on the structure defined in the claims, it provides a systematic solution: Firstly, regarding selective enrichment cultivation, the enrichment cultivation reactor consists of a reactor shell and a top cover forming a closed cultivation space. The reactor shell contains a stirring device and a bed of sparingly soluble phosphate particles. The stirring device includes a stirring motor mounted above the top cover, a stirring shaft driven by the stirring motor, and a stirring impeller connected to the stirring shaft. The stirring impeller enhances mixing and mass transfer of the cultivation system, ensuring sufficient contact and maintaining homogeneity between the microorganisms and the sparingly soluble phosphate particles in the bed of sparingly soluble phosphate particles. The reactor provides uniform and stable enrichment conditions. Simultaneously, a sparingly soluble phosphorus microparticle bed is positioned at the bottom of the reactor shell and supported by a microparticle bed support plate. An aeration inlet is located at the bottom of the reactor shell, allowing the introduction of sterile air or oxygen. A culture medium inlet with a metering pump is located at the top of the reactor shell, enabling the quantitative addition of culture medium or reagents, thereby improving the controllability and stability of the enrichment culture process. Secondly, regarding sterility reliability and anti-contamination, a sterile filter is installed at the exhaust port of the reactor shell to control the exhaust process with a sterile barrier. A unidirectional flow connection channel, consisting of a sterile isolation valve, a one-way valve, and a quick connector, is provided between the reactor shell and the bacterial culture unit. By utilizing the isolation function of the aseptic isolation valve, the one-way check valve, and the quick-sealing connection characteristics of the quick connector, the risk of cross-contamination from open operation and backflow is reduced. Furthermore, the one-way flow connection channel can be equipped with a pump to assist in transporting the culture medium from the reactor shell to the bacterial culture unit, further improving the stability of closed transfer. Thirdly, regarding the connection between culture and application, the bacterial culture unit is a container detachably connected to the reactor shell. The bacterial culture unit contains at least one hollow porous carrier tube, which is connected to the top cover of the bacterial culture unit and is detachable. This allows the enriched culture product to attach and be carried within the bacterial culture unit, facilitating subsequent transfer and application. The top cover of the incubation unit is equipped with a liquid replenishment port, and the unit itself is equipped with an exhaust port. These ports are used to expel gas when the culture medium enters the incubation unit and facilitate liquid replenishment and maintenance, thereby reducing contamination and activity fluctuations caused by unstable transfer, gas resistance, or frequent opening and closing. Fourthly, in terms of process monitoring and stable operation, the reactor shell is equipped with a pH probe, a temperature probe, and a dissolved oxygen probe, providing basic support for online monitoring and process control of enrichment culture conditions. This makes the enrichment culture of phosphate-solubilizing microorganisms more stable. Furthermore, through the modular docking of the incubation unit with the unidirectional flow connection channel, a more convenient, controllable, and aseptically compliant path is provided for connection and delivery at dispersed locations in terraced fields. Attached Figure Description

[0015] Figure 1 This is a schematic diagram of the overall structure of the present invention; Figure 2 This is a top view of the present invention; Figure 3 This is a cross-sectional view of the present invention; Figure 4 This is a front view of the present invention; Figure 5 This is a rear view of the present invention; The components include: 1. Reactor shell; 2. Inoculum culture unit; 3. Top cover; 4. Culture medium inlet; 5. Exhaust port; 6. Stirring motor; 7. Metering pump; 8. Sterile filter; 9. Stirring shaft; 10. Stirring impeller; 11. Insoluble phosphorus microparticle bed box; 12. Microparticle bed support plate; 13. Hollow porous carrier cylinder; 14. Circulation pump; 15. Circulation pipe; 16. Sterile isolation valve; 17. One-way valve; 18. Quick connector; 19. Liquid replenishment port; 20. Exhaust port; 21. pH probe; 22. Temperature probe; 23. Dissolved oxygen probe; 24. Aeration inlet. Detailed Implementation

[0016] The subject matter described herein will now be discussed with reference to exemplary embodiments. It should be understood that these embodiments are discussed merely to enable those skilled in the art to better understand and implement the subject matter described herein, and are not intended to limit the scope, applicability, or examples set forth in the claims. The function and arrangement of the elements discussed may be changed without departing from the scope of this specification. Various processes or components may be omitted, substituted, or added as needed in the various examples. For example, the described methods may be performed in a different order than described, and steps may be added, omitted, or combined. Furthermore, features described in some examples may be combined in other examples.

[0017] As used herein, the term "comprising" and its variations are open terms meaning "including but not limited to". The term "based on" means "at least partially based on". The terms "one embodiment" and "an embodiment" mean "at least one embodiment". The term "another embodiment" means "at least one other embodiment". The terms "first", "second", etc., may refer to different or the same objects. Other definitions, whether explicit or implicit, may be included below. Unless explicitly indicated by the context, the definition of a term shall remain consistent throughout the specification.

[0018] Example An integrated device for phosphorus-solubilizing microbial enrichment culture and terraced field layout includes a selective enrichment culture reactor, a microbial culture unit, and a one-way connecting channel. The enrichment culture reactor includes a reactor shell 1 and a top cover 3, with the top cover 3 enclosing the top of the reactor shell 1. Inside the reactor shell 1, there is a stirring device and a bed of sparingly soluble phosphorus microparticles 11. The stirring device includes a stirring motor 6 mounted above the top cover 3, a stirring shaft 9 driven by the stirring motor 6, and a stirring impeller 10 connected to the stirring shaft 9. The bed of sparingly soluble microparticles 11 is located at the bottom of the reactor shell 1 and is composed of microorganisms... The particle bed support plate 12 supports the reactor shell 1, which contains insoluble phosphate particles. The reactor shell 1 is also provided with an aeration inlet 24 for introducing sterile air. The bacterial culture unit 2 is a container that is detachably connected to the reactor shell 1. The bacterial culture unit 2 is provided with at least one hollow porous carrier cylinder 13. The hollow porous carrier cylinder 13 is connected to the top cover of the bacterial culture unit 2 and is detachable. The one-way connection channel includes a sterile isolation valve 16, a one-way valve 17, and a quick connector 18. The three of them constitute a one-way flow connection channel from the reactor shell 1 to the bacterial culture unit 2.

[0019] Furthermore, the reactor shell 1 is equipped with a pH probe 21, a temperature probe 22, and a dissolved oxygen probe 23.

[0020] Furthermore, a sterile filter 8 is provided at the exhaust port 5 of the reactor shell 1.

[0021] Furthermore, an aeration inlet 24 is provided at the bottom of the reactor shell 1, through which sterile air or oxygen is introduced into the reactor shell 1.

[0022] Furthermore, a culture medium inlet 4 is provided above the reactor shell 1, and a metering pump 7 is provided. The metering pump 7 is connected to a culture medium storage container to quantitatively add culture medium or reagents into the reactor shell 1.

[0023] Furthermore, the top cover of the bacterial culture unit 2 is provided with a liquid replenishment port 19, which can be detached or sealed with a cap.

[0024] Furthermore, the bacterial culture unit 2 is provided with an exhaust port 20, which discharges gas when the culture medium enters the bacterial culture unit 2.

[0025] Furthermore, the unidirectional flow connection channel can be equipped with a pump to assist in transporting the culture medium from the reactor shell 1 to the bacterial culture unit 2 as needed.

[0026] like Figure 1As shown, the device of the present invention comprises two parts: a selective enrichment culture reactor and a microbial culture unit 2, connected by a unidirectional connection channel. First, insoluble phosphate particles (such as phosphate rock powder) are filled into the insoluble phosphate particle bed box 11 at the bottom of the reactor shell 1, and the particle bed support plate 12 is installed to fix the position of the insoluble phosphate particle bed box 11. Then, pre-prepared liquid culture medium is added to the reactor shell 1 to an appropriate level. The amount of liquid added can be precisely controlled through the culture medium inlet 4 and the metering pump 7, and culture medium or inducing agents can be added as needed. Next, the top cover 3 is opened, and the required phosphate-solubilizing microbial strains are inoculated into the culture medium, or environmental samples are inoculated to screen for enrichment of phosphate-solubilizing microorganisms. After inoculation, the top cover 3 is closed tightly, and the stirring motor 6 is started, driving the stirring shaft 9 and the stirring impeller 10 to stir at an appropriate speed, so that the culture medium and the insoluble phosphate particles are fully mixed. At the same time, the aeration inlet 24 is opened to introduce sterile air for aeration. A sterile filter 8 is installed at the exhaust port 5 of the reactor shell 1 to maintain the sterility of the gas exchange process. With the combined action of stirring and aeration, the dissolved oxygen level in the culture medium can be monitored by the dissolved oxygen probe 23 and used for process control. The pH of the culture medium can be monitored in real time by the pH probe 21 and adjusted to the suitable range for microorganisms by adding acid or alkali solutions. The temperature probe 22 is used to monitor the culture temperature and ensure stable culture conditions.

[0027] After culturing under the above conditions for a period of time, the insoluble phosphates are gradually dissolved by the phosphate-solubilizing microorganisms, and the concentration of soluble phosphorus in the culture medium increases. This indicates that the phosphate-solubilizing microorganisms have been successfully enriched and proliferated. The cultivation process can be judged by combining the monitoring results of pH probe 21, temperature probe 22, and dissolved oxygen probe 23. When the cell density in the culture medium reaches the predetermined value or the phosphorus dissolution meets the requirements, the inoculation operation can be carried out.

[0028] Before incubation begins, the culture unit 2 can be connected to the reactor shell 1 via quick connector 18, but the aseptic isolation valve 16 is closed to temporarily isolate the reactor shell 1 from the culture unit 2 to maintain sterility. The hollow porous carrier cylinder 13 inside the culture unit 2 is pre-sterilized and loaded, and kept ready for use. When incubation is required, the aseptic isolation valve 16 and check valve 17 are opened, and the culture medium in the reactor shell 1 is pumped into the culture unit 2 in batches via gravity or by starting the circulation pump 14 and circulation pipe 15 until the culture medium submerges the hollow porous carrier cylinder 13. During the culture medium filling process, the vent 20 on the top cover of the culture unit 2 should be open to expel air. After the liquid is full, the vent 20 should be closed, or a sterile filter 8 should be connected as needed to maintain aseptic ventilation. To ensure sufficient adhesion of microorganisms, the culture medium can be left to stand or slowly circulate in the inoculum culture unit 2, allowing the phosphate-solubilizing microorganisms to gradually adhere to the surface and inner wall of the hollow porous carrier cylinder 13, forming a stable biofilm.

[0029] After the predetermined incubation period, the bacterial film formation on the hollow porous carrier cylinder 13 is basically complete. At this time, the aseptic isolation valve 16 is closed, the quick connector 18 is disconnected, and the incubation unit 2 is removed from the reactor shell 1, thus obtaining an independent incubation unit 2 module carrying highly active phosphate-solubilizing microorganisms.

[0030] Next, the inoculated culture unit 2 can be applied to terraced farmland. For example, in terraced fields where soil phosphorus nutrition needs to be improved, select several representative plot levels and place an inoculated hollow porous carrier tube 13 in the irrigation ditch or soil of each terrace level. Specifically, the hollow porous carrier tube 13 can be removed from the top cover of the inoculated culture unit 2 and partially buried in the soil root zone or fixed at the water flow point on the field ridge; or it can be placed together with the inoculated culture unit 2 in a suitable location in the field. Because the hollow porous carrier tube 13 can retain water and nutrients, the attached phosphorus-solubilizing microorganisms can continue to survive in the field environment and continue to play the role of converting insoluble phosphorus into soluble phosphorus; when irrigating, the water flowing through each level of the field will also wash the hollow porous carrier tube 13, promoting the diffusion of the microorganisms and their metabolites to the surrounding soil, thereby promoting the activation of phosphorus in the entire terraced soil layer by layer. If necessary, the above culture and inoculation process can be repeated periodically, and new hollow porous carrier tubes 13 can be replaced to ensure the quantity and activity of microorganisms.

[0031] In this embodiment, the cultivation and application of phosphate-solubilizing microorganisms are integrated through this device: the cultivation process is completed within a closed and controllable system consisting of reactor shell 1 and top cover 3, and stable operation is achieved through culture medium inlet 4, metering pump 7, aeration inlet 24, exhaust port 5, and sterile filter 8; subsequently, the culture medium is introduced into the inoculation unit 2 through a one-way connection channel consisting of sterile isolation valve 16, one-way valve 17, and quick connector 18 (which can be combined with circulation pump 14 and circulation pipe 15), and the hollow porous carrier cylinder 13 is inoculated with microorganisms. Finally, the inoculated hollow porous carrier cylinder 13 is used for field deployment, thereby reducing the risk of contamination and activity loss in the traditional culture transfer, packaging, and transportation process, and improving the convenience of application in terraced fields.

[0032] Those skilled in the art will understand that the various embodiments disclosed above can be modified and altered in various ways without departing from the spirit of the invention. Therefore, the scope of protection of this invention should be defined by the appended claims.

[0033] It should be noted that not all steps and units in the above processes are necessary; some steps or units can be omitted as needed. The execution order of each step is not fixed and can be determined as required. The device structure described in the above embodiments can be a physical structure or a logical structure. That is, some units may be implemented by the same physical entity, or some units may be implemented by multiple physical entities, or they may be jointly implemented by certain components in multiple independent devices.

[0034] The specific embodiments described above are exemplary embodiments, but do not represent all embodiments that can be implemented or fall within the scope of the claims. The term "exemplary" as used throughout this specification means "serving as an example, instance, or illustration" and does not imply that it is "preferred" or "advantageous" compared to other embodiments. Specific details are included to provide an understanding of the described techniques. However, these techniques can be practiced without these specific details. In some instances, well-known structures and apparatuses are shown in block diagram form to avoid obscuring the concepts of the described embodiments.

[0035] The foregoing description of this disclosure is provided to enable any person skilled in the art to implement or use this disclosure. Various modifications to this disclosure will be apparent to those skilled in the art, and the general principles defined herein can be applied to other variations without departing from the scope of this disclosure. Therefore, this disclosure is not limited to the examples and designs described herein, but is consistent with the widest scope of the principles and novel features disclosed herein.

Claims

1. An integrated device for the enrichment and cultivation of phosphate-solubilizing microorganisms and the layout of terraced fields, comprising a selective enrichment culture reactor, a microbial culture unit, and a unidirectional connecting channel, characterized in that: The enrichment culture reactor includes a reactor shell (1) and a top cover (3). The top cover (3) is closed on the top of the reactor shell (1). The reactor shell (1) is equipped with a stirring device and a sparingly soluble phosphate microparticle bed box (11). The stirring device includes a stirring motor (6) installed above the top cover (3), a stirring shaft (9) driven by the stirring motor (6), and a stirring impeller (10) connected to the stirring shaft (9). The sparingly soluble microparticle bed box (11) is located at the bottom of the reactor shell (1) and supported by a microparticle bed support plate (12). It contains sparingly soluble phosphate microparticles. The reactor shell (1) is also provided with an aeration inlet (24) for introducing sterile air. The bacterial culture unit (2) is a container that can be detachably connected to the reactor shell (1). The bacterial culture unit (2) is provided with at least one hollow porous carrier cylinder (13). The hollow porous carrier cylinder (13) is connected to the top cover of the bacterial culture unit (2) and is a detachable structure. The one-way connection channel includes a sterile isolation valve (16), a one-way valve (17), and a quick connector (18). The three of them constitute a one-way flow connection channel from the reactor shell (1) to the bacterial culture unit (2).

2. The integrated device for phosphorus-solubilizing microbial enrichment culture and terraced field layout according to claim 1, characterized in that: The reactor shell (1) is equipped with a pH probe (21), a temperature probe (22) and a dissolved oxygen probe (23).

3. The integrated device for phosphorus-solubilizing microbial enrichment culture and terraced field layout according to claim 1, characterized in that: The reactor shell (1) has a sterile filter (8) at the exhaust port (5).

4. The integrated device for enriching and cultivating phosphorus-solubilizing microorganisms and terracing according to claim 1, characterized in that: The bottom of the reactor shell (1) is provided with an aeration inlet (24) through which sterile air or oxygen is introduced into the reactor shell (1).

5. The integrated device for phosphorus-solubilizing microbial enrichment culture and terraced field layout according to claim 1, characterized in that: The reactor shell (1) is provided with a culture medium inlet (4) and a metering pump (7) is provided. The metering pump (7) is connected to the culture medium storage container to add culture medium or reagents into the reactor shell (1) in a quantitative manner.

6. The integrated device for phosphorus-solubilizing microbial enrichment culture and terraced field layout according to claim 1, characterized in that: The top cover of the bacterial culture unit (2) is provided with a liquid replenishment port (19), which can be detached or sealed with a cap.

7. The integrated device for enriching and cultivating phosphorus-solubilizing microorganisms and terracing according to claim 1, characterized in that: The bacterial culture unit (2) is equipped with an exhaust port (20) so that gas is discharged when the culture medium enters the bacterial culture unit (2).

8. The integrated device for enriching and cultivating phosphorus-solubilizing microorganisms and terracing according to claim 1, characterized in that: The unidirectional flow connection channel can be equipped with a pump as needed to assist in transporting the culture medium from the reactor shell (1) to the bacterial culture unit (2).

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