CN109540615B - Slurry culture method for determining soil organic carbon mineralization - Google Patents
Slurry culture method for determining soil organic carbon mineralization Download PDFInfo
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- CN109540615B CN109540615B CN201811332406.4A CN201811332406A CN109540615B CN 109540615 B CN109540615 B CN 109540615B CN 201811332406 A CN201811332406 A CN 201811332406A CN 109540615 B CN109540615 B CN 109540615B
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- soil
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- organic carbon
- mineralization
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- 239000002689 soil Substances 0.000 title claims abstract description 89
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 title claims abstract description 29
- 229910052799 carbon Inorganic materials 0.000 title claims abstract description 29
- 230000033558 biomineral tissue development Effects 0.000 title claims abstract description 28
- 238000012136 culture method Methods 0.000 title claims abstract description 17
- 239000002002 slurry Substances 0.000 title claims abstract description 14
- 239000000243 solution Substances 0.000 claims abstract description 18
- 244000005700 microbiome Species 0.000 claims abstract description 12
- 239000012086 standard solution Substances 0.000 claims abstract description 12
- 239000011521 glass Substances 0.000 claims description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 16
- 239000012153 distilled water Substances 0.000 claims description 8
- 239000011259 mixed solution Substances 0.000 claims description 8
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 6
- JKQOBWVOAYFWKG-UHFFFAOYSA-N molybdenum trioxide Chemical compound O=[Mo](=O)=O JKQOBWVOAYFWKG-UHFFFAOYSA-N 0.000 claims description 6
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 claims description 6
- 238000012258 culturing Methods 0.000 claims description 5
- 239000000565 sealant Substances 0.000 claims description 4
- 238000003892 spreading Methods 0.000 claims description 4
- 239000006228 supernatant Substances 0.000 claims description 4
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 3
- RAOSIAYCXKBGFE-UHFFFAOYSA-K [Cu+3].[O-]P([O-])([O-])=O Chemical compound [Cu+3].[O-]P([O-])([O-])=O RAOSIAYCXKBGFE-UHFFFAOYSA-K 0.000 claims description 3
- 230000000903 blocking effect Effects 0.000 claims description 3
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 claims description 3
- 239000004327 boric acid Substances 0.000 claims description 3
- 239000001110 calcium chloride Substances 0.000 claims description 3
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 3
- 239000011248 coating agent Substances 0.000 claims description 3
- 238000000576 coating method Methods 0.000 claims description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 3
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 3
- 229940099596 manganese sulfate Drugs 0.000 claims description 3
- 239000011702 manganese sulphate Substances 0.000 claims description 3
- 235000007079 manganese sulphate Nutrition 0.000 claims description 3
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 3
- 229910000403 monosodium phosphate Inorganic materials 0.000 claims description 3
- 235000019799 monosodium phosphate Nutrition 0.000 claims description 3
- 239000004323 potassium nitrate Substances 0.000 claims description 3
- 235000010333 potassium nitrate Nutrition 0.000 claims description 3
- 238000004080 punching Methods 0.000 claims description 3
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 claims description 3
- MKWYFZFMAMBPQK-UHFFFAOYSA-J sodium feredetate Chemical compound [Na+].[Fe+3].[O-]C(=O)CN(CC([O-])=O)CCN(CC([O-])=O)CC([O-])=O MKWYFZFMAMBPQK-UHFFFAOYSA-J 0.000 claims description 3
- LRXTYHSAJDENHV-UHFFFAOYSA-H zinc phosphate Chemical compound [Zn+2].[Zn+2].[Zn+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O LRXTYHSAJDENHV-UHFFFAOYSA-H 0.000 claims description 3
- 229910000165 zinc phosphate Inorganic materials 0.000 claims description 3
- 238000000034 method Methods 0.000 abstract description 6
- 238000004519 manufacturing process Methods 0.000 abstract 1
- 230000000087 stabilizing effect Effects 0.000 abstract 1
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 238000005553 drilling Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000001089 mineralizing effect Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Images
Classifications
-
- 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
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
Abstract
A slurry culture method for measuring soil organic carbon mineralization relates to a culture method for soil organic carbon mineralization. The invention aims to solve the technical problems that the existing soil organic carbon mineralization culture method is large in sample consumption and is easily influenced by soil moisture change. The method comprises the following specific steps: manufacturing a culture device; preparing a culture solution without organic carbon; preparing a standard solution containing soil microorganisms; and respectively adding the soil sample and the culture solution into the culture device, covering the cover of the culture device, and shaking up the soil solution in the culture device for later culture. The method is simple to operate, the using amount of the soil sample is small, the soil sample is not influenced by the change of soil moisture, and a new method is provided for deeply researching the stability and the stabilizing mechanism of the organic carbon in the soil.
Description
Technical Field
The invention relates to a method for culturing slurry for mineralizing soil organic carbon.
Background
The existing soil organic carbon mineralization culture method is that about 40-60 g of soil sample is placed into a culture bottle of 500ml, and then the soil moisture content in the culture bottle is increased to 60% of the field water capacity for constant temperature culture.
Disclosure of Invention
The invention aims to solve the technical problems that the existing soil organic carbon mineralization culture method is large in sample consumption and is easily influenced by soil moisture change, and provides a slurry culture method for measuring soil organic carbon mineralization.
The invention relates to a slurry culture method for measuring soil organic carbon mineralization, which comprises the following steps:
taking a centrifugal tube, taking down a cover of the centrifugal tube, punching a hole in the center of the cover, blocking the punched hole by using a rubber plug, coating a sealant around the rubber plug on the inner side of a bottle cover, airing the cover, and covering the cover on the centrifugal tube to obtain a culture device;
step two, preparing a Hoagland culture solution, wherein the Hoagland culture solution consists of 1M calcium chloride, 1M potassium nitrate, 1M magnesium sulfate, 0.1M sodium ferric ethylenediamine tetraacetate, 1M sodium dihydrogen phosphate, 0.04M boric acid, 0.008M manganese sulfate, 0.008M zinc phosphate, 0.0003M copper phosphate, 0.0003M molybdenum trioxide and distilled water;
step three, preparing a standard solution containing soil microorganisms: uniformly spreading farmland soil in a glass bottle, adding distilled water into the glass bottle to adjust the soil water content to 10-12%, then putting the glass bottle into an incubator for culturing for 3 days, taking a soil sample from the glass bottle after the culture is finished, adding the soil sample into a centrifugal tube, vibrating, centrifuging, and taking out supernatant to obtain standard solution containing soil microorganisms;
and step four, putting 1-2 g of a soil sample to be cultured into a culture device, then adding 19.8mL of Hoagland's culture solution obtained in the step two and 0.2mL of standard solution containing soil microorganisms, covering a cover, shaking the mixed solution uniformly, and putting the mixed solution on a constant-temperature shaking table for culture.
The invention has the following beneficial effects:
in the process of measuring the mineralization rate of the soil sample, the amount of the used soil sample is very small, only 1-2 g is enough, the whole culture process is carried out in a sealed environment with sufficient moisture, the mineralization rate of the soil sample is not influenced by moisture change, and the mineralization rate of organic carbon in the soil can be measured more accurately.
Description of the drawings:
FIG. 1 shows the mineralization rate of organic carbon in black soil of farmland in northeast region;
FIG. 2 shows the mineralization accumulation of organic carbon in black soil of farmland in northeast region.
Detailed Description
The first embodiment is as follows: the slurry culture method for determining soil organic carbon mineralization of the embodiment comprises the following steps:
taking a centrifugal tube, taking down a cover of the centrifugal tube, punching a hole in the center of the cover, blocking the punched hole by using a rubber plug, coating a sealant around the rubber plug on the inner side of a bottle cover, airing the cover, and covering the cover on the centrifugal tube to obtain a culture device;
step two, preparing a Hoagland culture solution, wherein the Hoagland culture solution consists of 1M calcium chloride, 1M potassium nitrate, 1M magnesium sulfate, 0.1M sodium ferric ethylenediamine tetraacetate, 1M sodium dihydrogen phosphate, 0.04M boric acid, 0.008M manganese sulfate, 0.008M zinc phosphate, 0.0003M copper phosphate, 0.0003M molybdenum trioxide and distilled water;
step three, preparing a standard solution containing soil microorganisms: uniformly spreading farmland soil in a glass bottle, adding distilled water into the glass bottle to adjust the soil water content to 10-12%, then putting the glass bottle into an incubator for culturing for 3 days, taking a soil sample from the glass bottle after the culture is finished, adding the soil sample into a centrifugal tube, vibrating, centrifuging, and taking out supernatant to obtain standard solution containing soil microorganisms;
and step four, putting 1-2 g of a soil sample to be cultured into a culture device, then adding 19.8mL of Hoagland's culture solution obtained in the step two and 0.2mL of standard solution containing soil microorganisms, covering a cover, shaking the mixed solution uniformly, and putting the mixed solution on a constant-temperature shaking table for culture.
The second embodiment is as follows: the first difference between the present embodiment and the specific embodiment is: the farmland soil in the third step is black soil or soil with the organic carbon content of more than 2 percent. The rest is the same as the first embodiment.
The third concrete implementation mode: the first difference between the present embodiment and the specific embodiment is: the mass of farmland soil taken in the third step is 50 g. The rest is the same as the first embodiment.
The fourth concrete implementation mode: the first difference between the present embodiment and the specific embodiment is: and in the third step, the water content of the soil is adjusted to 11-12%. The rest is the same as the first embodiment.
The fifth concrete implementation mode: the first difference between the present embodiment and the specific embodiment is: the culture temperature of the incubator in the third step is 25 ℃. The rest is the same as the first embodiment.
The sixth specific implementation mode: the first difference between the present embodiment and the specific embodiment is: 50/1 soil samples of farmland soil quality were taken from the glass bottles in step three. The rest is the same as the first embodiment.
The seventh embodiment: the first difference between the present embodiment and the specific embodiment is: in the fourth step, the swing speed of the constant temperature shaking table is 80-100 r/min. The rest is the same as the first embodiment.
The beneficial effects of the present invention are demonstrated by the following examples:
example 1
In this embodiment, taking black soil in northeast as an example, the specific implementation scheme is as follows:
step one, taking one 50mL plastic centrifuge tube, taking down a cover of the centrifuge tube, drilling a hole in the center of the cover by using an electric drill, and then plugging the drilled hole by using a rubber plug, wherein in order to ensure the sealing effect, a sealant needs to be coated around the rubber plug on the inner side of the bottle cover. Then the cover is dried and covered on a centrifuge tube to prepare the culture device.
Step two, the culture solution is prepared by adopting an improved Hoagland culture solution preparation method. The culture solution was prepared with distilled water, and the amounts of various nutrients in the culture solution were shown in Table 1.
TABLE 1 recipe ratio of improved Hoagland culture solution
Step three, weighing 50g of farmland black soil in northeast regions, uniformly spreading the farmland black soil in a 500ml glass bottle, adding 6g of distilled water into the glass bottle, adjusting the water content of the soil to 12%, and then putting the glass bottle into an incubator for culturing for 3 days. After the culture, 1g of soil sample is taken from the glass bottle, added into a 50ml centrifuge tube, shaken and centrifuged. Taking out the supernatant to obtain the standard solution containing the soil microorganisms.
And step four, putting 1-2 g of a farmland black soil sample to be cultured in the northeast region into a culture device, then adding 19.8ml of improved Hoagland culture solution and 0.2ml of standard solution containing soil microorganisms, covering a cover, shaking the mixed solution uniformly, putting the mixed solution into a constant-temperature shaking table for culture, and periodically measuring the amount of carbon dioxide mineralized and released by the soil.
According to this example, the black soil of the northeast farmland was cultured for 67 days, and the mineralization rate (fig. 1) and the cumulative mineralization amount (fig. 2) of the organic carbon in the black soil of the northeast farmland were obtained by measuring the mineralization amount of the organic carbon in the soil 15 times in total. In the embodiment, the requirement of test determination can be met only by using 1-2 g of soil samples, the influence of the change of the water content of the soil on the mineralization of organic carbon in the soil is avoided, and the mineralization rate and the accumulated mineralization quantity of the organic carbon in the black soil of the farmland in the northeast region are accurately determined.
Claims (7)
1. A slurry culture method for measuring soil organic carbon mineralization is characterized by comprising the following steps;
taking a centrifugal tube, taking down a cover of the centrifugal tube, punching a hole in the center of the cover, blocking the punched hole by using a rubber plug, coating a sealant around the rubber plug on the inner side of a bottle cover, airing the cover, and covering the cover on the centrifugal tube to obtain a culture device;
step two, preparing a Hoagland culture solution, wherein the Hoagland culture solution consists of 1M calcium chloride, 1M potassium nitrate, 1M magnesium sulfate, 0.1M sodium ferric ethylenediamine tetraacetate, 1M sodium dihydrogen phosphate, 0.04M boric acid, 0.008M manganese sulfate, 0.008M zinc phosphate, 0.0003M copper phosphate, 0.0003M molybdenum trioxide and distilled water;
step three, preparing a standard solution containing soil microorganisms: uniformly spreading farmland soil in a glass bottle, adding distilled water into the glass bottle to adjust the soil water content to 10-12%, then putting the glass bottle into an incubator for culturing for 3 days, taking a soil sample from the glass bottle after the culture is finished, adding the soil sample into a centrifugal tube, vibrating, centrifuging, and taking out supernatant to obtain standard solution containing soil microorganisms;
and step four, putting 1-2 g of a soil sample to be cultured into a culture device, then adding 19.8mL of Hoagland's culture solution obtained in the step two and 0.2mL of standard solution containing soil microorganisms, covering a cover, shaking the mixed solution uniformly, and putting the mixed solution on a constant-temperature shaking table for culture.
2. The slurry culture method for detecting the mineralization of organic carbon in soil according to claim 1, wherein the farmland soil in the third step is black soil or soil with organic carbon content more than 2%.
3. The slurry culture method for determining the organic carbon mineralization of soil according to claim 1, wherein the mass of the farmland soil in the third step is 50 g.
4. The slurry culture method for detecting the mineralization of organic carbon in soil according to claim 1, wherein the water content of soil is adjusted to 11-12% in the third step.
5. The slurry culture method for measuring the mineralization of organic carbon in soil according to claim 1, wherein the culture temperature of the culture box in the third step is 25 ℃.
6. The slurry culture method for determining the organic carbon mineralization of soil as claimed in claim 1, wherein 1/50 soil samples of farmland soil quality are taken from the glass bottles in the third step.
7. The slurry culture method for detecting the mineralization of organic carbon in soil according to claim 1, wherein the swing speed of the constant temperature shaking table in the fourth step is 80-100 r/min.
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| US4016065A (en) * | 1975-10-16 | 1977-04-05 | Idaho Research Foundation, Inc. | Electrolytic field respirometer |
| CN102778551B (en) * | 2012-07-26 | 2014-09-03 | 中国科学院东北地理与农业生态研究所 | Organic carbon mineralized culture method for determining soil organic carbon mineralized rate by utilizing apparatus without soil |
| CN103760119A (en) * | 2014-01-14 | 2014-04-30 | 沈阳建筑大学 | Method for determining inhibition effect of plant sample on soil nitrification |
| BR112017010955A2 (en) * | 2014-11-25 | 2018-02-14 | Univ Colorado State Res Found | composition for improving the rate at which organic soil phosphorus is converted to orthophosphate, method for improving the rate at which organic soil phosphorus is converted to orthophosphate, and method of increasing a plant's growth rate |
| CN206096125U (en) * | 2016-10-26 | 2017-04-12 | 贵州大学 | Device of organic carbon mineralize mineralization of indoor survey soil |
| CN108130304A (en) * | 2018-02-10 | 2018-06-08 | 厦门昶青生物科技有限公司 | Bacillus composite bacterial liquid and preparation method thereof |
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