Soil culture device and use method thereof

Abstract

The invention discloses a soil culture device and a use method thereof, and relates to the technical field of soil culture experiments.The soil culture device comprises a gas cylinder, a vacuum pump, a first metal pipe, a second metal pipe and a culture bottle, and the two ends of the first metal pipe are connected with the gas cylinder and the vacuum pump respectively; a connecting pipe valve and a vacuum valve are respectively arranged on a connecting pipeline between the metal pipe I and the gas cylinder and a connecting pipeline between the metal pipe I and the vacuum pump; a plurality of tee joints are arranged on the first metal tube, the other port of each tee joint is connected with the second metal tube, a two-way valve is arranged on the second metal tube, and the other end of the second metal tube is communicated with the culture bottle through a connecting assembly. According to the soil culture device disclosed by the invention, a plurality of pipelines can be flexibly arranged according to needs, deoxygenization operation of a plurality of soils can be completed at one time, and simultaneous operation of a plurality of persons can also be realized, so that the working efficiency is greatly improved; the device is easy to operate, interference gas in the culture bottle can be removed only by operating several valve switches, and time and labor are saved.

Description

Technical Field

[0001] The present invention relates to the technical field of soil culture experiments, and particularly to a soil culture device and a method for using the same. Background Art

[0002] In experiments in ecology, soil science, etc. when studying topics such as carbon-nitrogen cycling and climate change, for soil (not limited to soil), indoor condition simulation control experiments are often required. Placing the soil in a culture bottle to simulate the natural environment to measure greenhouse gas concentrations such as methane, carbon dioxide, nitrous oxide, etc. is beneficial for studying carbon emission dynamics, nitrogen cycling, and climate change laws, etc., providing data support and theoretical support for climate change prediction.

[0003] Currently, there are various devices for soil anaerobic and aerobic culture experiments. Among them, for soil anaerobic culture for oxygen removal and gas sampling, directly blowing inert gas downward towards the culture bottle is used. This method cannot remove oxygen completely and there will still be residual oxygen. When sampling, directly connecting a gas sampling bag to the culture bottle for sampling, such as in CN202814959U. This method has poor oxygen removal effect, and air will infiltrate when connecting the gas sampling bag and the culture bottle during sampling, which is not precise enough; and the efficiency is not high, not suitable for culturing a large number of soil samples. Some install valves and sampling devices on the culture bottle. This kind of device is difficult, requires cutting and punching holes on the bottle body, and the later stability is not high; such as in CN 210741924 U, during long-term culture, the punching area may become unstable and oxygen may easily infiltrate, affecting the test results; and if culturing a large number of soil samples, a large number of holes need to be punched, with a large installation difficulty and low efficiency. For soil aerobic culture for carbon dioxide removal and gas sampling, generally synthetic air (79% nitrogen and 21% oxygen) is used for purging. This kind of purging is generally carried out at the top of the culture bottle, and the purging effect is not good, and the sampling syringe is not purged either, with a large external interference. Generally speaking, the existing devices and methods do not solve the problem of completely anaerobic or aerobic conditions. Even if complete anaerobic or aerobic conditions are achieved, the device is difficult, with low efficiency, poor stability, not suitable for large-scale culture, and other sampling equipment is easily interfered by the outside world, the experimental results are not accurate enough, and even the experiment fails. Summary of the Invention

[0004] The purpose of the present invention is to provide a soil culture device and a method for using the same, which solve the problems of low efficiency in the test process and inaccurate test results caused by incomplete removal of oxygen or carbon dioxide in anaerobic culture experiments or aerobic culture experiments. The present invention improves the accuracy in the process of soil anaerobic and aerobic culture, maximally eliminates external interference, and improves the success rate of the experiment and the accuracy of the results.

[0005] To achieve the above object, the present invention provides the following solution: The present invention provides a soil cultivation device, including a gas cylinder, a vacuum pump, a first metal tube, a second metal tube, and a cultivation bottle. The two ends of the first metal tube are respectively connected to the gas cylinder and the vacuum pump, and a connecting tube valve and a vacuum valve are respectively arranged on the connecting pipelines of the first metal tube with the gas cylinder and the vacuum pump; A plurality of three-way joints are arranged on the first metal tube, the other port of each three-way joint is connected to the second metal tube, a two-way valve is arranged on the second metal tube, and the other end of the second metal tube is connected to the cultivation bottle through a connecting component.

[0006] In one embodiment, the gas cylinder is a nitrogen cylinder or a synthetic air cylinder, and a pressure reducing valve and a gas cylinder valve are arranged at the gas outlet end of the gas cylinder.

[0007] In one embodiment, the first metal tube is a horizontally placed metal tube, and the horizontally placed metal tube is formed by connecting one section of metal tube and a three-way joint, and the lower interface of the three-way joint is connected to the second metal tube.

[0008] In one embodiment, the second metal tube is a vertically placed metal tube, and the vertically placed metal tube is composed of two sections of metal tubes and a two-way valve. The two-way valve is connected in the middle of the two sections of metal tubes and controls the input and output of gas.

[0009] In one embodiment, the connecting component includes a Luer three-way valve, a main spiral hose, and a needle. The lower end of the second metal tube is connected to the Luer three-way valve, and the cultivation bottle and the Luer three-way valve are connected by a main spiral hose with a needle, and the needle is inserted into the cultivation bottle.

[0010] In one embodiment, the cultivation bottle includes a bottle body, a silica gel plug, and a perforated cover. The silica gel plug is hermetically arranged at the opening of the bottle body, and the perforated cover is buckled on the silica gel plug and is threadedly connected to the bottle body.

[0011] In one embodiment, it further includes a gas sampling device, and the gas sampling device includes a syringe, a gas sampling Luer three-way valve, and a gas sampling needle. The injection port of the syringe is connected to the gas sampling needle through the gas sampling Luer three-way valve.

[0012] In one embodiment, it further includes a dilution device, and the dilution device includes a dilution syringe, a dilution Luer three-way valve, and a dilution needle. The injection port of the dilution syringe is connected to the dilution needle through the dilution Luer three-way valve.

[0013] The present invention also provides a usage method of the soil cultivation device, which is applied to the above-mentioned soil cultivation device, and includes the following steps:

[0014] When performing anaerobic cultivation of soil, the gas cylinder uses a nitrogen cylinder, and high-purity nitrogen is filled in the gas cylinder. All valves are closed, and the vacuum pump is powered on;

[0015] Step 1: Perform the pumping and blowing cycle three times

[0016] Place the culture bottle filled with soil in place, insert the needle of the connection component into the culture bottle. Next, turn on the vacuum pump, the vacuum pump valve, the two-way valve, and the Luer three-way valve in the connection component in sequence to evacuate the air. When the silicone stopper of the culture bottle deforms and sinks, close the vacuum pump valve. Then, turn on the gas cylinder valve, the pressure reducing valve, and the gas cylinder connection pipe valve of the gas cylinder in sequence, and introduce nitrogen into the culture bottle. When the silicone stopper bulges, immediately close the gas cylinder connection pipe valve. Subsequently, turn on the vacuum pump valve to evacuate the air. When the silicone stopper deforms and sinks, close the vacuum pump valve. Take this as one cycle, and repeat the operations of evacuating the air and introducing nitrogen three times. At this time, the vacuum pump valve is in the closed state, and the culture bottle is in a vacuum state;

[0017] Step 2: Nitrogen blow for 10 minutes

[0018] Open the gas cylinder connection pipe valve and introduce nitrogen. When the silicone stopper bulges, insert the needle of the auxiliary threaded hose with the auxiliary threaded hose into the culture bottle to continuously input nitrogen. The other end of the auxiliary threaded hose is inserted into a water bottle filled with water. After 10 minutes, turn off the gas cylinder connection pipe valve, the two-way valve, and the Luer three-way valve in sequence. When there are no more bubbles in the water bottle, pull out the main threaded hose needle and the auxiliary threaded hose needle on the culture bottle. At this time, the inside of the culture bottle is in an anaerobic state and the pressure is the atmospheric pressure, and it can be cultured;

[0019] Step 3: Take a gas sample

[0020] First, a transfer bottle needs to be made. The structure of the transfer bottle is the same as that of the culture bottle. The transfer bottle is made in the same way as in Step 1 and Step 2, and the transfer bottle is always kept full of gas. Take out the gas sampling device, insert the gas sampling device into the transfer bottle, open the sampling Luer three-way valve to connect the transfer bottle with the syringe, and the gas enters the syringe. Rotate the sampling Luer three-way valve to connect the syringe with the outside, and at the same time apply pressure with your finger to release the gas in the syringe. After repeating the operation of the gas sampling device 5 times, insert the gas sampling device into the cultured culture bottle and make an up-and-down pumping action to mix the gas in the culture bottle evenly. After pumping 10 times, extract the required volume of gas, close the sampling Luer three-way valve, and transfer it to the gas sampling bag to complete the gas sampling operation;

[0021] Step 4: Dilute the gas sample

[0022] Insert the dilution device into the transfer bottle, open the dilution Luer three-way valve to connect the transfer bottle with the dilution syringe, and the gas enters the dilution syringe. Rotate the dilution Luer three-way valve to connect the dilution syringe with the outside, and at the same time apply pressure with your finger to release the gas in the dilution syringe. After repeating the operation of the dilution device 5 times, transfer the required nitrogen to the gas sampling bag to complete the dilution operation, and the gas can be tested on the machine.

[0023] The present invention also provides a method for using a soil cultivation device, which is applied to the above-mentioned soil cultivation device and includes the following steps:

[0024] When performing aerobic soil cultivation, a synthetic air bottle is used for the gas cylinder. The gas in the synthetic air bottle is 79% nitrogen + 21% oxygen. All valves are closed, and the vacuum pump is powered on.

[0025] Step 1: Perform the pumping and blowing cycle three times

[0026] Place the culture bottle filled with soil in place, insert the needle of the connection component into the culture bottle. Next, sequentially open the vacuum pump, the vacuum pump valve, the two-way valve, and the Luer three-way valve in the connection component to evacuate the air. When the silicone stopper of the culture bottle is deformed and sunken, close the vacuum pump valve. Then, sequentially open the gas cylinder valve, the pressure reducing valve, and the gas cylinder connection pipe valve of the gas cylinder to introduce synthetic air into the culture bottle. When the silicone stopper bulges, immediately close the gas cylinder connection pipe valve. Subsequently, open the vacuum pump valve to evacuate the air. When the silicone stopper is deformed and sunken, close the vacuum pump valve. Taking this as one cycle, repeat the operations of evacuating the air and introducing synthetic air three times. At this time, the vacuum pump valve is in the closed state, and the culture bottle is in a vacuum state.

[0027] Step 2: Nitrogen blow for 10 minutes

[0028] Open the gas cylinder connection pipe valve to introduce synthetic air. When the silicone stopper bulges, insert the needle of the sub-threaded hose with the sub-threaded hose into the culture bottle, and insert the other end of the sub-threaded hose into the water bottle filled with water to continuously input synthetic air. After 10 minutes, sequentially close the gas cylinder connection pipe valve, the two-way valve, and the Luer three-way valve. Wait until there are no more bubbles in the water bottle, then pull out the main threaded hose needle and the sub-threaded hose needle on the culture bottle. At this time, the inside of the culture bottle is in an anaerobic state, and the pressure is the atmospheric pressure, and cultivation can be carried out.

[0029] Step 3: Take a gas sample

[0030] First, a transfer bottle needs to be made. The structure of the transfer bottle is the same as that of the culture bottle. The transfer bottle is made in the same way as in Step 1 and Step 2, and the transfer bottle is always kept full of gas. Take out the gas sampling device, insert the gas sampling device into the transfer bottle, open the gas sampling Luer three-way valve to connect the transfer bottle with the syringe, and the gas enters the syringe. Rotate the gas sampling Luer three-way valve to connect the syringe with the outside, and at the same time, apply pressure with your finger to release the gas in the syringe. Repeat the operation of the gas sampling device 5 times, then insert the gas sampling device into the cultured culture bottle and perform the up and down pumping and pressing actions to make the gas in the culture bottle mix evenly. After pumping and pressing 10 times, extract the required volume of gas, close the gas sampling Luer three-way valve, and transfer it to the gas sampling bag to complete the gas sampling operation.

[0031] Step 4, Dilute the gas sample

[0032] Insert the dilution device into the transfer flask. Open the dilution Luer three-way valve to connect the transfer flask with the dilution syringe, so that the gas enters the dilution syringe. Rotate the dilution Luer three-way valve to connect the dilution syringe with the outside, and at the same time apply pressure with fingers to release the gas in the dilution syringe. Repeat the operation of the dilution device 5 times, then transfer the required synthetic gas to the gas sampling bag to complete the dilution operation, and the gas can be tested on the machine.

[0033] The present invention has achieved the following beneficial technical effects compared with the prior art:

[0034] In the soil cultivation device and its using method of the present invention, the soil cultivation device includes a gas cylinder, a vacuum pump, a first metal pipe, a second metal pipe and a cultivation flask. The two ends of the first metal pipe are respectively connected to the gas cylinder and the vacuum pump, and a connecting pipe valve and a vacuum valve are respectively arranged on the connecting pipelines of the first metal pipe with the gas cylinder and the vacuum pump; a plurality of three-ways are arranged on the first metal pipe, and the other ports of each three-way are connected to the second metal pipe. A two-way valve is arranged on the second metal pipe, and the other end of the second metal pipe is connected to the cultivation flask through a connecting component. The soil cultivation device of the present invention can flexibly set multiple pipelines according to needs, can complete the deoxidation operation of multiple soils at one time, or can be operated by multiple personnel at the same time, greatly improving the work efficiency; the operation is simple, and only a few valve switches need to be manipulated to remove the interfering gas in the cultivation flask, saving time and effort. A threaded hose is provided, which can be flexibly moved according to the position of the cultivation flask, facilitating the operation of personnel. All the interfaces of this device are tightened by screwing, and liquid and solid raw tapes are added, with good firmness, good sealing performance and strong stability. 90% of the fixed parts are made of metal materials, which are firm and durable and have good stability. 10% of the moving parts are made of plastic materials, which have good flexibility, light weight, are convenient to disassemble, and have low replacement cost. Description of the Drawings

[0035] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required to be used in the embodiments. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained based on these drawings without creative efforts.

[0036] Figure 1 It is a schematic diagram of the overall structure of the soil cultivation device;

[0037] Figure 2 It is a schematic diagram of the state of connecting the water bottle;

[0038] Figure 3 It is an operation diagram of taking a gas sample;

[0039] Figure 4 Operation diagram for diluting gas samples

[0040] Figure 5 Structural diagram of gas sampling device

[0041] Figure 6 Structural diagram of culture flask

[0042] Figure 7 Structural diagram of gas cylinder

[0043] Among them, 1. Nitrogen gas cylinder (synthetic air cylinder); 2. Gas cylinder connecting pipe; 3. Gas cylinder connecting pipe valve; 4. Three-way joint; 5. Vacuum pump valve; 6. Vacuum pump connecting pipe; 7. Vacuum pump; 8. Two-way valve; 9. Luer three-way valve; 10. Main threaded hose; 11. Needle; 12. Culture flask; 13. Horizontally placed metal pipe; 14. Vertically placed metal pipe; 15. Sub-threaded hose; 16. Water bottle; 17. Gas sampling device; 18. Intermediate bottle; 19. Gas sampling bag; 20. Syringe; 21. Gas sampling Luer three-way valve; 22. Gas sampling needle; 23. Punching cover; 24. Silicone plug; 25. Bottle body; 27. Gas cylinder valve; 28. Pressure reducing valve; 29. Gas cylinder body Specific implementation manners

[0044] The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention

[0045] The purpose of the present invention is to provide a soil culture device and its use method, which solves the problems of low efficiency and inaccurate test results caused by incomplete removal of oxygen or carbon dioxide in anaerobic culture experiments or aerobic culture experiments. The present invention improves the accuracy in the processes of soil anaerobic and aerobic culture, maximally eliminates external interference, and improves the success rate of the experiment and the accuracy of the results

[0046] In order to make the above objects, features, and advantages of the present invention more obvious and understandable, the present invention will be further described in detail below with reference to the accompanying drawings and specific implementation manners

[0047] As Figures 1 - 7As shown in the figure, the present invention provides a soil culture device, which is generally divided into a metal part (gas cylinder 1), a gas cylinder connecting pipe valve 3, a tee 4, a vacuum pump valve 5, a vacuum pump 7, a two-way valve 8, a horizontally placed metal pipe 13, and a vertically placed metal pipe 14), a plastic part (gas cylinder connecting pipe 2, vacuum pump connecting pipe 6, Luer tee valve 9, main threaded hose 10, needle 11), and an experimental part (culture bottle 12).

[0048] The gas cylinder 1 is connected to the gas cylinder connecting pipe 2, and the gas cylinder connecting pipe 2 is connected to the horizontally placed metal pipe 13. A gas cylinder connecting pipe valve 3 is installed in the middle to control the input of gas. The horizontally placed metal pipe 13 is connected by a section of shorter metal pipes and a tee 4. The lower interface of the tee 4 is connected to the vertically placed metal pipe 14. The vertically placed metal pipe 14 is composed of two sections of shorter metal pipes connected by a two-way valve 8 in the middle to control the input and output of gas. The lower end of the vertically placed metal pipe 14 is connected to a Luer tee valve 9. The Luer tee valve 9 serves as the control switch at the end. The culture bottle 12 is connected to the Luer tee valve 9 by a main threaded hose 10 with a needle 11. Finally, the needle 11 is inserted into the culture bottle 12 during operation. At the rightmost end of the device is a vacuum pump 7, which is responsible for evacuating the air, connected by a vacuum pump connecting pipe 6, and the output of gas is controlled by a vacuum pump valve 5. The culture bottle 12 is composed of a punched lid 23, a silica gel plug 24, and a bottle body 25; the gas sampling device 17 is composed of a syringe 20, a gas sampling Luer tee valve 21, and a gas sampling needle 22; the gas cylinder 1 is composed of a gas cylinder valve 27, a pressure reducing valve 28, and a gas cylinder body 29; the dilution device is composed of a dilution syringe, a dilution Luer tee valve, and a dilution needle.

[0049] The following describes the usage method and sampling method of this device in the culture experiment, taking anaerobic culture as an example.

[0050] Before using this device, ensure that the gas cylinder 1 uses a nitrogen gas cylinder, the nitrogen gas cylinder contains high-purity nitrogen and the gas is sufficient, all valves are closed, and the vacuum pump is powered on.

[0051] Step 1, perform three pumping and blowing cycles. Place the culture bottle filled with soil in front of this device and insert the needle 11 into the culture bottle 12. Next, turn on the vacuum pump 7, the vacuum pump valve 5, the two-way valve 8, and the Luer tee valve 9 in sequence to evacuate the air. When the silica gel plug 24 deforms and depresses and no longer depresses, close the vacuum pump valve 5. Then turn on the gas cylinder valve 27, the pressure reducing valve 28, and the gas cylinder connecting pipe valve 3 in sequence to introduce nitrogen into the culture bottle 12. When the silica gel plug 24 bulges, immediately close the gas cylinder connecting pipe valve 3. Then open the vacuum pump valve 5 to evacuate the air. When the silica gel plug 24 deforms and depresses and no longer depresses, close the vacuum pump valve 5. This is one cycle. Repeat the operations of evacuating the air and introducing nitrogen three times. At this time, the vacuum pump valve 5 is in the closed state, and the culture bottle 12 is in a vacuum state.

[0052] Step 2: Nitrogen blow for 10 minutes. Open the valve 3 of the gas cylinder connecting pipe, introduce nitrogen. When the silica gel plug 24 protrudes, insert the needle of the secondary threaded hose 15 with the secondary threaded hose into the culture bottle 12, and insert the other end of the secondary threaded hose 15 into the water bottle 16 filled with water to make nitrogen continuously input. After 10 minutes, turn off the valve 3 of the gas cylinder connecting pipe, the two-way valve 8, and the Luer three-way valve 9 in sequence. Wait until there are no more bubbles in the water bottle 16, then pull out the needle 11 on the culture bottle 12 and the needle of the secondary threaded hose. At this time, the inside of the culture bottle 12 is in an anaerobic state and the pressure is the atmospheric pressure, and cultivation can be carried out.

[0053] Step 3: Take a gas sample. First, a transfer bottle 18 needs to be made. The making of the transfer bottle 18 is the same as that in Step 1 and Step 2. Just replace the culture bottle 12 with an empty bottle. During the making process, close the unused two-way valve 8 and the Luer three-way valve 9, and always keep the transfer bottle 18 filled with gas. Note that the pressure of the pressure reducing valve 28 should not be opened too large, just 0.1 Mpa. Take out the gas sampling device 17, insert it into the transfer bottle 18, open the gas sampling Luer three-way valve 21 to connect the transfer bottle 18 with the syringe 20, and the gas enters the syringe 20. Rotate the gas sampling Luer three-way valve 21 to connect the syringe 20 with the outside, and at the same time apply pressure with fingers to release the gas in the syringe 20. The purpose of this operation is to flush the residual air in the gas sampling device 17 and keep no miscellaneous gas interference in the gas sampling device 17. After repeating this 5 times, insert the gas sampling device 17 into the cultured culture bottle 12 and make an up-and-down pumping action to make the gas in the culture bottle 12 evenly mixed. After pumping 10 times, extract the required volume of gas, close the gas sampling Luer three-way valve 21, and transfer it to the gas sampling bag 19 to complete the gas sampling operation.

[0054] Step 4: Dilute the gas sample. Insert the dilution device (the composition of the dilution device is the same as that of the gas sampling device 17) into the transfer bottle 18, open the dilution Luer three-way valve to connect the transfer bottle 18 with the dilution syringe, and the gas enters the dilution syringe. Rotate the dilution Luer three-way valve to connect the dilution syringe with the outside, and at the same time apply pressure with fingers to release the gas in the dilution syringe. After repeating this 5 times, transfer the required nitrogen to the gas sampling bag 19 to complete the dilution operation. The gas can be tested on the machine.

[0055] For the aerobic cultivation step, just replace the nitrogen gas cylinder with a synthetic air cylinder (79% nitrogen + 21% oxygen), and the other steps are the same as those for anaerobic cultivation.

[0056] This method combines a suction cycle and nitrogen purging, with good deoxygenation effect, high experimental accuracy and success rate. This method sets up a transfer bottle 18, which facilitates the cleaning of the gas sampling device 17 and the dilution device, reduces experimental errors, and at the same time is conducive to the collection of dilution gas, improving the operation efficiency. The culture bottle 12 used in this method is provided with a perforated lid 23 and a silicone stopper 24, which facilitates the insertion of the needle for gas sampling, and the silicone material will not leak even if inserted multiple times, with good sealing performance; this method uses a water bottle to observe the pressure inside the culture bottle, with low cost and good effect.

[0057] It should be noted that for those skilled in the art, it is obvious that the present invention is not limited to the details of the above exemplary embodiments, and the present invention can be implemented in other specific forms without departing from the spirit or basic characteristics of the present invention. Therefore, from any point of view, the embodiments should be regarded as exemplary and non-limiting. The scope of the present invention is defined by the appended claims rather than the above description. Therefore, all changes falling within the meaning and scope of the equivalent elements of the claims are intended to be included in the present invention, and any reference signs in the claims should not be regarded as limiting the claims involved.

[0058] Specific examples are used in the present invention to elaborate on the principles and implementation manners of the present invention. The description of the above embodiments is only used to help understand the method and its core idea of the present invention; at the same time, for those of ordinary skill in the art, according to the idea of the present invention, there will be changes in the specific implementation manners and application scopes. In summary, the content of this specification should not be construed as a limitation of the present invention.

Claims

1. A soil culture device, characterized in that: It includes a gas cylinder, a vacuum pump, a metal tube 1, a metal tube 2 and a culture bottle, wherein the two ends of the metal tube 1 are respectively connected to the gas cylinder and the vacuum pump, and a connecting pipe valve and a vacuum valve are respectively provided on the connecting pipelines between the metal tube 1 and the gas cylinder and the vacuum pump; the metal tube 1 is provided with a plurality of tees, and the other end of each of the tees is connected to the metal tube 2; the metal tube 2 is provided with a two-way valve, and the other end of the metal tube 2 is connected to the culture bottle through a connecting component.

2. The soil cultivation device according to claim 1, characterized in that: The gas cylinder is a nitrogen cylinder or a synthetic air cylinder, and a pressure reducing valve and a gas cylinder valve are arranged at the gas outlet end of the gas cylinder.

3. The soil cultivation device according to claim 1, characterized in that: The metal tube 1 is a transverse metal tube, which is formed by connecting metal tubes in sections and tees, and the lower interface of the tee is connected to the metal tube 2.

4. The soil cultivation device according to claim 1, characterized in that: The second metal pipe is a longitudinal metal pipe, which is composed of two sections of metal pipes and a two-way valve. The two-way valve is connected in the middle of the two sections of metal pipes and controls the input and output of gas.

5. The soil cultivation device according to claim 1, characterized in that: The connecting assembly includes a Luer three-way valve, a main spiral hose and a needle. The lower end of the metal tube 2 is connected to the Luer three-way valve. The culture bottle and the Luer three-way valve are connected by a main spiral hose with a needle, and the needle is inserted into the culture bottle.

6. The soil cultivation device according to claim 1, characterized in that: The culture bottle comprises a bottle body, a silicone plug and a perforated cover. The silicone plug is sealed at the opening of the bottle body, and the perforated cover is buckled on the silicone plug and threadedly connected to the bottle body.

7. The soil cultivation device according to claim 1, characterized in that: It also includes an air extraction device, which includes a syringe, an air extraction Luer three-way valve and an air extraction needle. The injection port of the syringe is connected to the air extraction needle through the air extraction Luer three-way valve.

8. The soil cultivation device according to claim 1, characterized in that: A dilution device is also included, which includes a dilution syringe, a dilution Luer three-way valve and a dilution needle. The injection port of the dilution syringe is connected to the dilution needle through the dilution Luer three-way valve.

9. A method for using a soil culture device, applied to the soil culture device according to any one of claims 1 to 8, characterized in that: The following steps are involved: When conducting soil anaerobic culture, a nitrogen cylinder is used as the gas cylinder, and the cylinder is filled with high-purity nitrogen, all valves are closed, and the vacuum pump is powered on; Step 1: Pump and blow three times Put the culture bottle filled with soil in place, insert the needle of the connecting assembly into the culture bottle, then open the vacuum pump, the vacuum pump valve, the two-way valve and the Luer three-way valve in the connecting assembly in sequence to evacuate, when the silicone plug of the culture bottle is deformed and sunken, close the vacuum pump valve, open the gas cylinder valve, the pressure reducing valve and the gas cylinder connecting pipe valve of the gas cylinder in sequence, pass nitrogen into the culture bottle, and immediately close the gas cylinder connecting pipe valve when the silicone plug protrudes, then open the vacuum pump valve to evacuate, when the silicone plug is deformed and sunken, close the vacuum pump valve; this is a cycle, after repeating the vacuum evacuation and nitrogen passing operation for three cycles, the vacuum pump valve is now in a closed state, and the culture bottle is in a vacuum state; Step 2: Nitrogen blowing for 10 minutes Open the valve of the gas cylinder connecting pipe and introduce nitrogen. When the silicone plug protrudes, insert the needle of the secondary threaded hose with the secondary threaded hose into the culture bottle, and insert the other end of the secondary threaded hose into the water bottle filled with water to continuously input nitrogen. After 10 minutes, turn off the valve of the gas cylinder connecting pipe, the two-way valve, and the Luer three-way valve in turn. When no more bubbles emerge in the water bottle, unplug the needle of the main threaded hose and the needle of the secondary threaded hose on the culture bottle. At this time, the culture bottle is in an anaerobic state and the pressure is atmospheric pressure, and culture can be carried out; Step 3: Take gas sample Make a transfer bottle, and the making of the transfer bottle is the same as step one and step two; the structure of the transfer bottle is consistent with that of the culture bottle, and no culture soil is placed in the transfer bottle, and the transfer bottle is always kept full of gas; take out the gas extraction device, insert the gas extraction device into the transfer bottle, open the gas extraction Luer three-way valve to connect the transfer bottle with the syringe, and the gas enters the syringe, and the gas extraction Luer three-way valve is rotated to connect the syringe with the outside world, and at the same time, the finger applies pressure to release the gas in the syringe, and the operation of the gas extraction device is repeated 5 times, and the gas extraction device is inserted into the culture bottle after culture to perform up and down pumping and pressing actions to mix the gas in the culture bottle evenly, and the required volume of gas is extracted after 10 pumping and pressing, and the gas extraction Luer three-way valve is closed, and the gas is transferred to the gas collection bag to complete the gas extraction operation; Step 4: Dilute the gas sample Take the dilution device and insert it into the transfer bottle, open the dilution Luer three-way valve to connect the transfer bottle with the dilution syringe, and the gas enters the dilution syringe. Rotate the dilution Luer three-way valve to connect the dilution syringe with the outside world. At the same time, apply pressure with your fingers to release the gas in the dilution syringe. After repeating the operation of the dilution device 5 times, take the required nitrogen and transfer it to the gas collection bag to complete the dilution operation. The gas can be tested on the machine.

10. A method for using a soil culture device, applied to the soil culture device according to any one of claims 1 to 8, characterized in that: The following steps are involved: When aerobic soil cultivation is carried out, the gas cylinder uses a synthetic air cylinder, the gas in the synthetic air cylinder is 79% nitrogen + 21% oxygen, all valves are closed, and the vacuum pump is powered on; Step 1: Pump and blow three times Put the culture bottle filled with soil in place, insert the needle of the connecting assembly into the culture bottle, then open the vacuum pump, the vacuum pump valve, the two-way valve and the Luer three-way valve in the connecting assembly in sequence to evacuate, when the silicone plug of the culture bottle is deformed and sunken, close the vacuum pump valve, open the gas cylinder valve, the pressure reducing valve and the gas cylinder connecting pipe valve of the gas cylinder in sequence, pass synthetic air into the culture bottle, and immediately close the gas cylinder connecting pipe valve when the silicone plug protrudes, then open the vacuum pump valve to evacuate, when the silicone plug is deformed and sunken, close the vacuum pump valve; this is a cycle, after repeating the vacuum extraction and synthetic air passing operation for three cycles, the vacuum pump valve is now in a closed state, and the culture bottle is in a vacuum state; Step 2: Nitrogen blowing for 10 minutes Open the valve of the gas cylinder connecting pipe and introduce synthetic air. When the silicone plug protrudes, insert the needle of the secondary threaded hose with the secondary threaded hose into the culture bottle, and insert the other end of the secondary threaded hose into the water bottle filled with water to continuously input synthetic air. After 10 minutes, turn off the valve of the gas cylinder connecting pipe, the two-way valve, and the Luer three-way valve in turn. When no more bubbles emerge from the water bottle, unplug the needle of the main threaded hose and the needle of the secondary threaded hose on the culture bottle. At this time, the culture bottle is in an anaerobic state and the pressure is atmospheric pressure, and culture can be carried out; Step 3: Take gas sample Make a transfer bottle, and the making of the transfer bottle is the same as step one and step two; the structure of the transfer bottle is consistent with that of the culture bottle, and no culture soil is placed in the transfer bottle, and the transfer bottle is always kept full of gas; take out the gas extraction device, insert the gas extraction device into the transfer bottle, open the gas extraction Luer three-way valve to connect the transfer bottle with the syringe, and the gas enters the syringe, and the gas extraction Luer three-way valve is rotated to connect the syringe with the outside world, and at the same time, the finger applies pressure to release the gas in the syringe, and the operation of the gas extraction device is repeated 5 times, and the gas extraction device is inserted into the culture bottle after culture to perform up and down pumping and pressing actions to mix the gas in the culture bottle evenly, and the required volume of gas is extracted after 10 pumping and pressing, and the gas extraction Luer three-way valve is closed, and the gas is transferred to the gas collection bag to complete the gas extraction operation; Step 4: Dilute the gas sample Insert the dilution device into the transfer bottle, open the dilution Luer three-way valve to connect the transfer bottle with the dilution syringe, and the gas enters the dilution syringe. Rotate the dilution Luer three-way valve to connect the dilution syringe with the outside world. At the same time, apply pressure with your fingers to release the gas in the dilution syringe. Repeat the operation of the dilution device 5 times, take the required synthetic gas and transfer it to the gas collection bag to complete the dilution operation. The gas can be tested on the machine.

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