CN113092191A - Method for measuring discharge of mineralized gas of indoor culture soil microorganisms - Google Patents
Method for measuring discharge of mineralized gas of indoor culture soil microorganisms Download PDFInfo
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- CN113092191A CN113092191A CN202110361862.7A CN202110361862A CN113092191A CN 113092191 A CN113092191 A CN 113092191A CN 202110361862 A CN202110361862 A CN 202110361862A CN 113092191 A CN113092191 A CN 113092191A
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- 239000002689 soil Substances 0.000 title claims abstract description 43
- 238000000034 method Methods 0.000 title claims abstract description 42
- 244000005700 microbiome Species 0.000 title claims abstract description 14
- 239000007789 gas Substances 0.000 claims abstract description 102
- 238000012360 testing method Methods 0.000 claims abstract description 7
- 238000005070 sampling Methods 0.000 claims abstract description 6
- 238000007789 sealing Methods 0.000 claims description 22
- 238000004519 manufacturing process Methods 0.000 claims description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- 230000001502 supplementing effect Effects 0.000 claims description 11
- 230000001954 sterilising effect Effects 0.000 claims description 9
- 238000004659 sterilization and disinfection Methods 0.000 claims description 9
- 230000008020 evaporation Effects 0.000 claims description 8
- 238000001704 evaporation Methods 0.000 claims description 8
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 6
- 239000000463 material Substances 0.000 claims description 4
- 229920000742 Cotton Polymers 0.000 claims description 3
- 239000004743 Polypropylene Substances 0.000 claims description 3
- 239000012153 distilled water Substances 0.000 claims description 3
- 229910052757 nitrogen Inorganic materials 0.000 claims description 3
- -1 polypropylene Polymers 0.000 claims description 3
- 229920001155 polypropylene Polymers 0.000 claims description 3
- 238000005303 weighing Methods 0.000 claims description 3
- 238000012258 culturing Methods 0.000 claims 1
- 239000011368 organic material Substances 0.000 claims 1
- 239000005431 greenhouse gas Substances 0.000 abstract description 10
- 238000011160 research Methods 0.000 abstract description 3
- 238000010521 absorption reaction Methods 0.000 abstract description 2
- 238000012544 monitoring process Methods 0.000 abstract description 2
- GQPLMRYTRLFLPF-UHFFFAOYSA-N Nitrous Oxide Chemical compound [O-][N+]#N GQPLMRYTRLFLPF-UHFFFAOYSA-N 0.000 description 4
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 3
- 238000004140 cleaning Methods 0.000 description 3
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 3
- 238000004088 simulation Methods 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 2
- 238000005034 decoration Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 229920001296 polysiloxane Polymers 0.000 description 2
- 238000009423 ventilation Methods 0.000 description 2
- 238000010792 warming Methods 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000001272 nitrous oxide Substances 0.000 description 1
- 239000002680 soil gas Substances 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/02—Devices for withdrawing samples
- G01N1/22—Devices for withdrawing samples in the gaseous state
- G01N1/2294—Sampling soil gases or the like
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/02—Devices for withdrawing samples
- G01N1/22—Devices for withdrawing samples in the gaseous state
- G01N1/2202—Devices for withdrawing samples in the gaseous state involving separation of sample components during sampling
- G01N1/2205—Devices for withdrawing samples in the gaseous state involving separation of sample components during sampling with filters
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/02—Devices for withdrawing samples
- G01N1/22—Devices for withdrawing samples in the gaseous state
- G01N1/2226—Sampling from a closed space, e.g. food package, head space
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/34—Purifying; Cleaning
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/38—Diluting, dispersing or mixing samples
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Physics & Mathematics (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Soil Sciences (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention provides a method for measuring the discharge of mineralized gas of indoor culture soil microorganisms, which belongs to the technical field of greenhouse gas monitoring research and comprises the steps of (1) soil open culture; (2) carrying out closed culture; (3) and (6) collecting gas. The method provided by the invention not only eliminates the influence of external microorganisms on the gas emission of a test system, but also eliminates the influence of gas generated by soil in the early stage on the gas emission of closed culture, realizes closed culture under normal pressure, simultaneously overcomes the problem of absorption of negative pressure on target gas caused by sampling, can finish culture and gas collection of a plurality of culture bottles at one time, ensures the consistency of initial culture concentration, and improves the working efficiency of greenhouse gas determination.
Description
Technical Field
The invention belongs to the technical field of greenhouse gas monitoring research, and particularly relates to a method for measuring discharge of mineralized gas of indoor culture soil microorganisms.
Background
An increase in the concentration of greenhouse gases in the atmosphere is a major contributor to global warming, carbon dioxide (CO)2) Methane (CH)4) And nitrous oxide (N)2O) are the 3 most important greenhouse gases in the atmosphere, their contribution to global warming accounts for nearly 80%, and their atmospheric concentrations increase at annual rates of 0.5%, 0.8%, and 0.3%, respectively. Of all greenhouse gas emissions, soil is the most important emission source, and it is estimated that atmospheric CO is present at 5% -20% per year215% -30% of CH4And 80% -90% of N2O is derived from soil. Therefore, scholars at home and abroad carry out relevant observation and research on the emission of 3 main greenhouse gases of different soil of ecological systems.
However, no method for measuring the discharge of mineralized greenhouse gases of the soil microorganisms cultured in the room, which is closer to the actual culture, exists at present. Therefore, there is a need to establish a scientific method for measuring the mineralized greenhouse gas emission of indoor culture soil microorganisms.
Disclosure of Invention
In view of the above, the present invention provides a method for measuring the discharge of mineralized gas from indoor cultured soil microorganisms.
In order to achieve the above purpose, the invention provides the following technical scheme:
the invention provides a method for measuring the discharge of mineralized gas of indoor culture soil microorganisms, which comprises the following steps:
(1) soil open culture
Placing the processed soil sample into a culture tank of a culture gas production device, covering and sealing a sealing cover with a super filter, wherein the sealing cover is provided with a vent hole to keep the inside and the outside of the bottle to be freely ventilated, and performing aerobic culture, and supplementing water lost due to evaporation in the culture process to reach the total weight consistent with that before the culture;
(2) closed culture
Connecting and sealing the gas collection tank and the sealing cover, closing the gas collection two-way valve at the same time to enable the culture bottle to be in a sealed state, and collecting external gas at the same time, wherein the concentration of the external gas is the initial concentration before culture; according to the test requirements, the whole device is placed under corresponding conditions for closed culture for 1-24 h;
(3) gas collection
And when the closed culture is finished, connecting the gas collection needle tube with the gas collection two-way valve, opening the gas collection two-way valve, and repeatedly pushing and pulling the gas collection needle tube to fully mix the gas in the culture bottle and immediately collect the gas.
Preferably, the culture tank, the super filter and the gas collecting tank in the steps (1) to (3) are prepared by adopting high-permeability polypropylene high-temperature and high-pressure resistant materials.
Preferably, the soil sample treated in the step (1) comprises soil obtained by different treatment methods of water content, nitrogen concentration, pH value, culture temperature, sterilization and addition of special strains.
Preferably, the method for achieving the consistency with the total weight before culture in the step (1) comprises the following steps: weighing and recording the total weight of the soil sample and the culture bottle before culture, supplementing water lost due to evaporation every 2-4d by a gravimetric method in the culture process, and supplementing distilled water into the culture bottle through a rubber stopper by adopting an injector with a slender needle head.
Preferably, after the gas collection in the step (3) is finished, the gas collection tank is taken down, open culture is continued according to the step (1), and the steps (2) to (3) are repeated to realize sampling in different time periods.
Preferably, the super filter in the step (1) comprises a bottom cover, filter cotton and an upper cover, and a rubber plug is arranged in the center.
Preferably, the apparatus of steps (1) to (3) is sterilized before the soil open culture of step (1).
Preferably, the temperature of the sterilization is 121 ℃ and the time is 30 min.
The invention has the beneficial effects that:
the method provided by the invention not only eliminates the influence of external microorganisms on the gas emission of a test system, but also eliminates the influence of gas generated by soil in the early stage on the gas emission of closed culture, realizes closed culture under normal pressure, simultaneously overcomes the problem of absorption of negative pressure on target gas caused by sampling, can finish culture and gas collection of a plurality of culture bottles at one time, ensures the consistency of initial culture concentration, and improves the working efficiency of greenhouse gas determination.
Detailed Description
The invention provides a method for measuring the discharge of mineralized gas of indoor culture soil microorganisms, which comprises the following steps:
(1) soil open culture
Placing the processed soil sample into a culture tank of a culture gas production device, covering and sealing a sealing cover with a super filter, wherein the sealing cover is provided with a vent hole to keep the inside and the outside of the bottle to be freely ventilated, and performing aerobic culture, and supplementing water lost due to evaporation in the culture process to reach the total weight consistent with that before the culture;
(2) closed culture
Connecting and sealing the gas collection tank and the sealing cover, closing the gas collection two-way valve at the same time to enable the culture bottle to be in a sealed state, and collecting external gas at the same time, wherein the concentration of the external gas is the initial concentration before culture; according to the test requirements, the whole device is placed under corresponding conditions for closed culture for 1-24 h;
(3) gas collection
And when the closed culture is finished, connecting the gas collection needle tube with the gas collection two-way valve, opening the gas collection two-way valve, and repeatedly pushing and pulling the gas collection needle tube to fully mix the gas in the culture bottle and immediately collect the gas.
In the invention, the method is carried out by utilizing an integrated ventilation, culture and gas production device, wherein the integrated ventilation, culture and gas production device comprises a culture tank, a super filter and a gas collector; the device is made of high-permeability polypropylene high-temperature and high-pressure resistant materials, can be sterilized by high-temperature steam, and has the advantages that the bottle body is not deformed after the temperature is 121 ℃ for 30 minutes. The super filter comprises a bottom cover, filter cotton and an upper cover, wherein the center of the super filter is provided with a rubber plug for supplementing water lost due to evaporation in the culture process, and a vent hole is formed in the super filter. The culture gas production device comprises a culture bottle with an opening at the upper end, a sealing plug is arranged at the opening at the upper end, a gas production hole is formed in the sealing plug, a gas production pipe is fixed in the gas production hole, one end of the gas production pipe is inserted into the culture bottle, the other end of the gas production pipe is connected with a gas production two-way valve, and the upper port of the gas production two-way valve is connected with a gas production needle tube. The invention has no special requirements on the device and the connecting structure thereof, and can achieve the aim of the invention.
In the invention, the soil sample treated in the step (1) comprises soil obtained by different treatment methods of water content, nitrogen concentration, pH value, culture temperature, sterilization and special strain addition.
In the present invention, the method for achieving the total weight before culture in the step (1) comprises: weighing and recording the total weight of the soil sample and the culture bottle before culture, supplementing water lost due to evaporation every 2-4d by a gravimetric method in the culture process, and supplementing distilled water into the culture bottle through a rubber stopper by adopting an injector with a slender needle head.
In the invention, after the gas collection in the step (3) is finished, the gas collection tank is taken down, the open culture is continued according to the step (1), and the steps (2) to (3) are repeated to realize the sampling in different time periods.
In the present invention, the apparatus in steps (1) to (3) is sterilized before the soil open culture in step (1). The temperature of the sterilization is 121 ℃, and the time is 30 min.
The device capable of measuring the discharge of the mineralized gas of the indoor cultured soil microorganisms has the advantages of simple structure, easiness in manufacturing, easiness in obtaining materials, low price, convenience in disassembly and assembly, easiness in cleaning, convenience in operation and good simulation effect, and is suitable for culture and gas collection of indoor simulated soil. By utilizing the technical scheme of the invention, the gas is directly produced by the gas production needle tube through the two-way valve, so that the problems of pinhole blockage caused by inserting a slender needle into the silicone tube for gas production and air leakage of the silicone plug caused by inserting the needle are avoided to a great extent; the problem that the emission of target gas during the closed culture period is influenced by the increase of the concentration of the gas released by the soil in the early stage in the culture bottle can be solved, the air near the culture bottle is collected as the initial culture gas, the concentration of the initial gas is consistent, and the workload is reduced by half; the method can realize no mixed bacteria pollution in the closed culture process, and can avoid the problem of the mixed bacteria pollution to the target gas in the sampling process, thereby obtaining the soil gas emission which is relatively close to the actual condition.
The soil culture and gas collection device has the characteristics of simple manufacture, low cost, convenience in cleaning, convenience and quickness in operation and the like, can keep normal-pressure closed culture in the using process, avoids negative pressure from influencing target gas emission, has the advantage of improving the working efficiency, and ensures the quality of indoor simulation experiments.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
A method for measuring the discharge of mineralized gas from indoor cultured soil microorganisms comprises device sterilization, soil open culture, closed culture and gas collection, and comprises the following steps:
(1) device sterilization
Before the test treatment, the device was placed in an autoclave for sterilization at 121 ℃ for 30 min.
(2) Soil open culture
Placing the processed soil sample into a culture tank of a culture gas production device, covering and sealing a sealing cover with a super filter, wherein the sealing cover is provided with a vent hole to keep the inside and the outside of the bottle to be freely ventilated, and performing aerobic culture, and supplementing water lost due to evaporation in the culture process to reach the total weight consistent with that before the culture;
(3) closed culture
Connecting and sealing the gas collection tank and the sealing cover, closing the gas collection two-way valve at the same time to enable the culture bottle to be in a sealed state, and collecting external gas at the same time, wherein the concentration of the external gas is the initial concentration before culture; according to the test requirements, the whole device is placed under corresponding conditions for closed culture for 1-24 h;
(4) gas collection
And when the closed culture is finished, connecting the gas collection needle tube with the gas collection two-way valve, opening the gas collection two-way valve, and repeatedly pushing and pulling the gas collection needle tube to fully mix the gas in the culture bottle and immediately collect the gas.
The soil culture and gas collection device has the characteristics of simple manufacture, low cost, convenience in cleaning, convenience and quickness in operation and the like, can keep normal-pressure closed culture in the using process, avoids negative pressure from influencing target gas emission, has the advantage of improving the working efficiency, and ensures the quality of indoor simulation experiments.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (8)
1. A method for measuring the discharge of mineralized gas from indoor culture soil microorganisms is characterized by comprising the following steps:
(1) soil open culture
Placing the processed soil sample into a culture tank of a culture gas production device, covering and sealing a sealing cover with a super filter, wherein the sealing cover is provided with a vent hole to keep the inside and the outside of the bottle to be freely ventilated, and performing aerobic culture, and supplementing water lost due to evaporation in the culture process to reach the total weight consistent with that before the culture;
(2) closed culture
Connecting and sealing the gas collection tank and the sealing cover, closing the gas collection two-way valve at the same time to enable the culture bottle to be in a sealed state, and collecting external gas at the same time, wherein the concentration of the external gas is the initial concentration before culture; according to the test requirements, the whole device is placed under corresponding conditions for closed culture for 1-24 h;
(3) gas collection
And when the closed culture is finished, connecting the gas collection needle tube with the gas collection two-way valve, opening the gas collection two-way valve, and repeatedly pushing and pulling the gas collection needle tube to fully mix the gas in the culture bottle and immediately collect the gas.
2. The method according to claim 1, wherein the culture tank, the super filter and the gas collecting tank in the steps (1) to (3) are prepared by using a high-permeability polypropylene high-temperature and high-pressure resistant material.
3. The method as claimed in claim 1, wherein the soil sample treated in step (1) comprises soil obtained by different treatment methods of water content, pH value, culture temperature, organic material addition, nitrogen concentration, sterilization, and special strain addition.
4. The method according to claim 1, wherein the step (1) of achieving the weight corresponding to the total weight before culturing comprises: weighing and recording the total weight of the soil sample and the culture bottle before culture, supplementing water lost due to evaporation every 2-4d by a gravimetric method in the culture process, and supplementing distilled water into the culture bottle through a rubber stopper by adopting an injector with a slender needle head.
5. The method as claimed in claim 1, wherein after the gas collection in step (3) is completed, the gas collection tank is removed, the open culture is continued according to step (1), and steps (2) to (3) are repeated to realize sampling in different time periods.
6. The method according to claim 1, wherein the super filter in the step (1) comprises a bottom cover, filter cotton and an upper cover, and the center of the super filter is provided with a rubber plug.
7. The method according to claim 1, wherein the apparatus of steps (1) to (3) is sterilized before the soil is cultured in the open state in step (1).
8. The method of claim 7, wherein the sterilization is performed at a temperature of 121 ℃ for a period of 30 min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110361862.7A CN113092191A (en) | 2021-04-02 | 2021-04-02 | Method for measuring discharge of mineralized gas of indoor culture soil microorganisms |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202110361862.7A CN113092191A (en) | 2021-04-02 | 2021-04-02 | Method for measuring discharge of mineralized gas of indoor culture soil microorganisms |
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| CN113092191A true CN113092191A (en) | 2021-07-09 |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090301234A1 (en) * | 2006-04-27 | 2009-12-10 | St. Francis Xavier University | Apparatus and method for measuring the surface flux of a soil gas component |
| CN105973656A (en) * | 2016-05-18 | 2016-09-28 | 沈阳大学 | Soil respiration measurement culture and gas collection device and measurement method thereof |
| CN106932241A (en) * | 2017-02-09 | 2017-07-07 | 山东省农业科学院农业资源与环境研究所 | A kind of method of culture soil greenhouse gas emission in measuring cell |
| CN106940365A (en) * | 2017-02-09 | 2017-07-11 | 山东省农业科学院农业资源与环境研究所 | A kind of indoor soil incubation device for integrating ventilation culture with gas collecting and application |
-
2021
- 2021-04-02 CN CN202110361862.7A patent/CN113092191A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090301234A1 (en) * | 2006-04-27 | 2009-12-10 | St. Francis Xavier University | Apparatus and method for measuring the surface flux of a soil gas component |
| CN105973656A (en) * | 2016-05-18 | 2016-09-28 | 沈阳大学 | Soil respiration measurement culture and gas collection device and measurement method thereof |
| CN106932241A (en) * | 2017-02-09 | 2017-07-07 | 山东省农业科学院农业资源与环境研究所 | A kind of method of culture soil greenhouse gas emission in measuring cell |
| CN106940365A (en) * | 2017-02-09 | 2017-07-11 | 山东省农业科学院农业资源与环境研究所 | A kind of indoor soil incubation device for integrating ventilation culture with gas collecting and application |
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Application publication date: 20210709 |