CN101271060B - Analytical method for detecting nitrate reductase activity in soil - Google Patents
Analytical method for detecting nitrate reductase activity in soil Download PDFInfo
- Publication number
- CN101271060B CN101271060B CN2007100106805A CN200710010680A CN101271060B CN 101271060 B CN101271060 B CN 101271060B CN 2007100106805 A CN2007100106805 A CN 2007100106805A CN 200710010680 A CN200710010680 A CN 200710010680A CN 101271060 B CN101271060 B CN 101271060B
- Authority
- CN
- China
- Prior art keywords
- distilled water
- solution
- soil
- sample
- test tube
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The invention relates to an analysis method of testing the nitrate reductase activity in the soil, which includes the following steps: 1) n dried and screened soil samples are weighed, adopted and put into test tubes; 2, 4-dinitrophenol solution is added into the test tubes and mixed to be even; and then substrate potassium nitrate solution is added into n-1 test tubes and the same amount of distilled water is added into the other one test tube for sample comparison; glucose solution is respectively added into the test tubes, which is then supplemented by distilled water to be 5 to 10ml and cultivated under constant temperature; 2) the distilled water is added to supplement to 50 to 100ml; 3) potash alum saturated solution is added in to be oscillated and filtered; 4)the distilled water and a chromogenic reagent are added into the filtrate, and a light absorption value A is determined under 520nm; 5) the content of nitrite in the samples is calculated. The analysis method has the advantages of: 1) simplification of operation steps and reduction of the complicacy and uncertainty during the anaerobic culturing process; 2) reduction to dependency on the equipment requirements; 3) high accuracy and easiness in operation; 4) stable and reliably result, and good repeatability.
Description
Technical field
The present invention relates to the mensuration of nitrate reductase activity in the soil, specifically a kind of analytical approach that detects nitrate reductase activity in the soil.
Background technology
Nitrate reductase in the soil can generate nitrite nitrogen with the nitrate-nitrogen reduction that forms in the Soil Nitrogen metabolic process, goes back the donor that the ortho states compound can be used as hydrogen in the soil.
Therefore, the power of nitrate reductase activity has influence on the gaseous loss of nitrogen in the Soil Nitrogen metabolic process in the soil, the utilization ratio of remote effect nitrogenous fertilizer and the polluted by nitrogen of atmosphere.The activity of nitrate reductase is subjected to influencing strongly of edaphic condition such as moisture, temperature, the soil texture in the soil, also is subjected to farming operation system, control measures and different influence of planting plants simultaneously.Therefore, the check to nitrate reductase activity in the soil has very important significance.And up to the present, the mensuration of relevant nitrate reductase activity substantially all is to breathe out now Prokofiev [Soviet Union] work with reference to Φ .X., the method in the books such as " the Methodsin soil biology) " of " soil enzyme activities " that Zheng Hongyuan, Zhou Likai, Zhang Desheng translate and Kandeler (1995).This kind method step is more loaded down with trivial details, need to vacuumize cultivation with the reagent bottle and the suction filtration device of special use, and the difference of cultivating batch has otherness and uncertainty, makes it be difficult to the qualitative comparison and the quantitative examination of nitrate reductase activity in the adapted soil; Simultaneously, handle cumbersome the early stage of this kind method sample, reduced the efficient of experiment, increased the inaccuracy of experimental result.
Summary of the invention
The object of the present invention is to provide a kind of analytical approach of checking nitrate reductase activity in the soil.This method is on the principle basis of using for reference forefathers' assay method, determination step and coloration method to nitrate reductase activity have carried out partly improving (disposal route and assay method to its sample improve), thereby reduce the complicacy of sample preparation and test procedure, make analysis result more accurately reliable, it is better to analyze reappearance.
For achieving the above object, the technical solution used in the present invention is:
A kind of analytical approach that detects nitrate reductase activity in the soil,
1) take by weighing soil air-dry sample n part that 0.5-1g crosses 0.5-1.5mm sieve and be respectively charged into n and prop up in the test tube, n 〉=2 add 2 of 1-3mL 0.5-1mM/L respectively in test tube, 4-dinitrophenol dinitrophenolate solution, mixing; Prop up the substrate KNO that adds 1-1.5mL weight concentration 0.05-0.15% in the test tube to n-1 wherein then
3Solution, the distilled water that adds equivalent in other 1 test tube contrasts as sample; In test tube, add the donor of the glucose solution of 1-2mL weight concentration 0.5%-1% more respectively, be supplemented to 5-10mL with distilled water again, shake up, 29-31 ℃ of constant temperature culture 24h behind air-locked beyond the Great Wall rubber plug as hydrogen; Do reagent blank contrast (do not add soil, all the other are handled with sample) simultaneously;
2) after cultivation finishes, invisible spectro native liquid mixture is transferred in the triangular flask of 100mL fully, is supplemented to 50-100mL with distilled water again with 10-40mL distilled water;
3) add potassium alum saturated solution 1-2mL, vibration is filtered;
4) get filtrate 1mL in the 50mL volumetric flask, add 1-3mL distilled water and 4-8mL developer (excessive), be settled to scale with distilled water then with respect to nitrite ion.After 15 minutes, under 520nm, use spectrophotometer colorimetric estimation light absorption value A;
5) preparation of standard working curve: draw nitrite anions standard reserving solution 1mL, be settled to 100mL, this is 2.5mgNO
2 -The solution of-N/L.Draw this liquid 0,1,2,3,4,5mL more respectively in the 50mL volumetric flask, add a small amount of distilled water and 4mL developer, and use the distilled water constant volume.The amount that this standard series contains nitrite nitrogen is respectively 0,0.05,0.1,0.15,0.2,0.25mgNO
2 --N/L.The NO of a series of concentration known of colorimetric estimation under 520nm
2 -The light absorption value A ' of-N standard solution is with NO
2 -The concentration of-N solution and light absorption value A ' make the working stamndard curve, obtain slope b;
6), calculate the concentration S of the nitrite anions in the solution of measuring, S=A*b according to light absorption value A and slope b;
7) calculation sample Central Asia nitrate radical content;
8) calculate nitrate reductase activity:
Computing formula is: [(S
1-S
2) * 50*V
1/ 1000*V
2]/W.T=mgNO
2-N/g soil .24h
In the formula: S
1Be the NO in the sample
2The content of-N (mg/L); S
2Be the NO in the sample contrast
2The content of-N (mg/L); 50 is the liquor capacity (mL) of constant volume; V
1Be extract cumulative volume (mL); V
2For drawing the volume (mL) of filtrate; W is for claiming soil sample heavy (g); T is for cultivating the time (h) of soil sample; 1000 is the unit conversion coefficient.
For the sour earth sample, should reconcile its pH before analyzing is 7-8.5, can adopt the method that adds lime carbonate to reconcile, and adds 15-25mgCaCO in every test tube respectively
3, in test tube, add 2 of 1-3mL 0.5-1mM/L, 4-dinitrophenol dinitrophenolate solution, mixing behind the mixing more respectively.(for calacareous soil, lime carbonate need not to add, because the pH value of soil itself promptly can satisfy the optimal pH of microorganism).
Anaerobism is cultivated employed distilled water, before the use, need be heated to the O in the boiling expeling water
2, sealing is cooled to room temperature then.
Described developer is the alpha-naphthylamine solution of weight concentration 0.1% of acetic acid preparation of weight concentration 35% and the isopyknic mixed liquor of sulfanilic acid solution of weight concentration 0.5%, and their adding is excessive with respect to nitrite anions.
The improved foundation of the inventive method:
Soil nitrate reductase and nitrite reductase, soil hydroxyl amine reductase is the same all is the reduction enzyme that participates in nitrogen metabolism in the soil.They all need the anaerobism condition of culture in the mensuration process.The assay method of activity of nitrous acid reductase then is with sodium nitrite (NaNO
2) be substrate, cultivate through 24 hours anaerobism, by NO in the unit interval
2 -The reduction of N characterizes; The mensuration of hydroxyl amine reducing ferment activity then is with azanol (NH
2OH) be substrate, cultivate, weigh the activity of hydroxylamine reductase by the reduction of azanol in the unit interval through 5 hours anaerobism.Identical therewith, measuring nitric acid reductase activity then is with potassium nitrate (KNO in this method
3) be substrate, by adding 2, the 4-dinitrophenol dinitrophenolate suppresses the activity of nitrite reductase, cultivates through 24 hours anaerobism, by detecting NO in the unit interval
2 -The growing amount of-N characterizes the activity of nitrate reductase.The fluid-tight that adds 5-10mL eliminating dissolved oxygen DO, purpose are exactly to create the good foster environment of detesting for culture experiment.
Reaction principle used in the present invention is:
Utilize potassium nitrate to be substrate, soil sample under anaerobic, 30 ℃ of constant temperature culture 24h.Nitrite reductase is by adding 2, and the 4-dinitrophenol dinitrophenolate suppresses.Under the effect of nitrate reductase, be nitrite nitrogen with nitrate-nitrogen reduction, the nitrite anions of generation and the colour developing of Γ р и с с reagent reacting, the amount of the nitrite nitrogen that 520nm colorimetric estimation enzymatic reaction back generates characterizes the activity of nitrate reductase.
Compare with the analytical approach in past, advantage of the present invention mainly contains:
1) equipment needed thereby simple, reduced experimentation cost.Culture experiment among the present invention is at diameter 10-15mm, long carries out in the simple glass test tube of 100-150mm, does not need to realize with being used for of mentioning in the original method the special-purpose Dewar bottle and the vacuum pump of the band grinding port plug that vacuumizes.
2) compare with traditional Pbenoldisulfonic Acid colourimetry, this kind coloration method is simpler, and coloration method sensitivity used in the present invention is higher, has improved the accuracy of analyzing greatly.
3) simple to operate, analysis result is reliable and stable, favorable reproducibility.Originally method need be provided with sterile soil in contrast, and this method only needs a solution that does not add substrate in contrast; Simultaneously and since involved in the present invention to cultural method reduced the complicacy and the uncertainty of anaerobism incubation, so reproducibility of analysis results is very good.
4) simplified operation steps.It is exactly to cultivate needed anaerobic environment in order to create that the soil face is positioned at liquid level following, reduces the dependence to air-extractor and vacuum environment.
Embodiment
The reagent preparation:
1.0.05%-0.15% KNO
3Solution: take by weighing 0.05-0.15gKNO
3Be settled to 100mL with distilled water.
2.0.5%-1% glucose solution: accurately take by weighing 0.50-1.00g glucose, be settled to 100mL with distilled water.
3. lime carbonate: analytical reagent.
4. potassium alum saturated solution: aluminium potassium sulfate is dissolved in the distilled water, until supersaturation.
5. developer 1 (Γ р и с с reagent): with proportion is that 1.04 acetic acid is prepared 0.1% alpha-naphthylamine solution and 0.5% sulfanilic acid solution (b) respectively, with preceding equal-volume a, b is mixed.
The acetic acid of proportion 1.04 (acetic acid that is equivalent to percent by weight 35%) configuration: get the acetic acid of 175m1100%, be settled to 500ml, be 35%, the acetic acid of proportion 1.04.
Take by weighing 0.2g alpha-naphthylamine percent by weight 35%, the acetate dissolution of proportion 1.04 also is settled to 200ml, and this is (a) liquid.
Take by weighing 1.0g sulfanilic acid percent by weight 35%, the acetate dissolution of proportion 1.04 also is settled to 200ml, and this is (b) liquid.
6.2,4-dinitrophenol dinitrophenolate solution (2,4-DNP, 0.8mM): take by weighing 0.1481g 2,4-DNP is settled to 1000mL with distilled water in distilled water.Heating makes it whole dissolvings.At 5-300 μ g 2, measure the suitableeest inhibitor consumption of each great soil group in the 4-DNP scope.
7. standard stock solution (250mgNO
2 --N/L): dissolving 1.2314gNaNO
2And be settled to 1000ml with distilled water, and being put in 4 ℃ of refrigerators and preserving, the holding time should not be of a specified duration excessively, is advisable to be no more than several weeks.
8. do the preparation of typical curve: draw above-mentioned standard stock solution 1ml, be settled to 100ml, this is 2.5mgNO
2 -The solution of-N/L.Draw this liquid 0,1,2,3,4,5ml more respectively in the 50ml volumetric flask, add a small amount of distilled water and 4ml developer, and use the distilled water constant volume.The amount that this standard series contains nitrite nitrogen is respectively 0,0.05,0.1,0.15,0.2,0.25mgNO
2 --N/L.The NaNO of a series of concentration known of colorimetric estimation under 520nm
2The light absorption value A ' of solution is with NaNO
2The concentration of solution and the light absorption value of mensuration A ' preparation standard curve obtain slope b;
Operation steps:
1) takes by weighing 4 parts of soil air-dry samples that 1g crosses 1mm sieve in 4 test tubes, add 20mgCaCO respectively
3, in test tube, add 2 of 1mL 0.8mM/L behind the mixing respectively, 4-dinitrophenol dinitrophenolate solution, mixing; The substrate KNO that in 3 test tubes wherein, adds 1-1.5mL 0.05%-0.15% then
3Solution, the distilled water that adds equivalent in other 1 test tube contrasts as sample; In test tube, add the donor of the glucose solution of 1-2mL 0.5%-1% more respectively, be supplemented to 5-10mL with distilled water again, shake up, 30 ℃ of (± 1 ℃) constant temperature culture 24h behind air-locked beyond the Great Wall rubber plug as hydrogen; Do reagent blank contrast (do not add soil, all the other are handled with sample) simultaneously;
2) after cultivation finishes, invisible spectro native liquid mixture is transferred in the triangular flask of 100mL fully, is supplemented to 50mL with distilled water again with 30mL distilled water;
3) add potassium alum saturated solution 1mL, fine and close filter paper filtering is used in vibration;
4) get filtrate 1mL in the 50mL volumetric flask, add the developer (excessive) of a small amount of distilled water and 4mL, be settled to scale with distilled water then with respect to nitrite anions.After 15 minutes, under 520nm, use spectrophotometer colorimetric estimation light absorption value A;
5), calculate the concentration S of the nitrite anions in the solution of measuring, S=A*b according to light absorption value A and slope b;
6) calculation sample Central Asia nitrate radical content;
7) calculate nitrate reductase activity:
Computing formula is: [(S
1-S
2) * 50*V
1/ 1000*V
2]/W.T=mgNO
2-N/g soil .24h
In the formula: S
1Be the NO in the sample
2The content of-N (mg/L); S
2Be the NO in the sample contrast
2The content of-N (mg/L); 50 is the liquor capacity (mL) of constant volume; V
1For soaking body fluid cumulative volume (mL); V
2For drawing the volume (mL) of filtrate; W is for claiming soil sample heavy (g); 1000 is the unit conversion coefficient; T is the time (h) that soil is cultivated.
Embodiment 1
The employed black earth of present embodiment picks up from the ecological testing station of the Helen of the Chinese Academy of Sciences, establishes three different processing: No. 1 position contrast, both without any processing with add any soil additive at 25 ℃ of normal soils of cultivating more than the 24h; No. 2 for adding nitrification inhibitor DCD and the soil more than 25 ℃ of cultivation 24h; No. 3 for adding nitrification inhibitor DMPP and the soil more than 25 ℃ of cultivation 24h.Soil moisture content is 20% (wind desiceted soil is heavy).Add the soil of nitrification inhibitor No. 2 and No. 3, wherein the addition of two kinds of inhibitor all is 50ppm, detects the activity of three kinds of nitrate reductases with said method.
It is as follows that every kind of soil must be made a concrete analysis of step:
1) takes by weighing 4 parts of soil air-dry samples that 1g crosses 1mm sieve in 4 test tubes, add 20mgCaCO respectively
3, in test tube, add 2 of 1mL 0.8mM/L behind the mixing respectively, 4-dinitrophenol dinitrophenolate solution, mixing; The KNO that in 3 test tubes wherein, adds 1mL 0.05% then
3Solution, the distilled water that adds equivalent in other 1 test tube contrasts as sample; In test tube, add the donor of the glucose solution of 1mL 1% more respectively, be supplemented to 10mL with distilled water again, shake up, 30 ℃ of (± 1 ℃) constant temperature culture 24h behind air-locked beyond the Great Wall rubber plug as hydrogen; Do reagent blank contrast (do not add soil, all the other are handled with sample) simultaneously;
2) after cultivation finishes, invisible spectro native liquid mixture is transferred in the triangular flask of 100mL fully, is supplemented to 50mL with distilled water again with 30mL distilled water;
3) add potassium alum saturated solution 1mL, fine and close filter paper filtering is used in vibration;
4) get filtrate 1mL in the 50mL volumetric flask, add the developer (excessive) of a small amount of distilled water and 4mL, be settled to scale with distilled water then with respect to nitrite anions.After 15 minutes, under 520nm, use spectrophotometer colorimetric estimation light absorption value A;
5), calculate the concentration S of the nitrite anions in the solution of measuring, S=A*b according to light absorption value A and slope b;
6) calculation sample Central Asia nitrate radical content;
7) calculate nitrate reductase activity.
Test findings is as follows:
By data in the table as can be seen, result difference reached the level of signifiance between each was handled, and less standard difference shows that the deviation between this analytical approach result is less, and degree of accuracy is higher, favorable reproducibility.
Embodiment 2
The employed three kinds of different brown earth of planting plant of present embodiment are all adopted in ecological experiment station, Chinese Academy of Sciences Shenyang: wherein No. 4 soil preceding crops are paddy rice, and No. 5 soil preceding crops are corn, and No. 6 soil preceding crops are soybean.The soil of three kinds of different cropping systems is 20% (wind desiceted soil is heavy) in water cut all, temperature is being cultivated under 25 ℃ of conditions more than the 24h, measure its nitrate reductase activity, concrete steps are as follows: substrate potassium nitrate concentration is 0.1%, add 0.50mL, concentration of glucose is 1%, adds 1mL, and all the other steps are with example 1.
Test findings is as follows:
Data have shown the stability and the higher precision of analysis result equally in the table.
Claims (4)
1. analytical approach that detects nitrate reductase activity in the soil is characterized in that:
1) take by weighing soil air-dry sample n part that 0.5-1g crosses 0.5-1.5mm sieve and be respectively charged into n and prop up in the test tube, n 〉=2 add 2 of 1-3mL 0.5-1mM/L respectively in test tube, 4-dinitrophenol dinitrophenolate solution, mixing; Prop up the substrate KNO that adds 1-1.5mL weight concentration 0.05-0.15% in the test tube to n-1 wherein then
3Solution, the distilled water that adds equivalent in other 1 test tube contrasts as sample; In test tube, add the donor of the glucose solution of 1-2mL weight concentration 0.5%-1% more respectively, be supplemented to 5-10mL with distilled water again, shake up, 29-31 ℃ of constant temperature culture 24h behind air-locked beyond the Great Wall rubber plug as hydrogen; Do the reagent blank contrast simultaneously;
2) after cultivation finishes, invisible spectro native liquid mixture is transferred in the triangular flask of 100mL fully, is supplemented to 50-100mL with distilled water again with 10-40mL distilled water;
3) add potassium alum saturated solution 1-2mL, vibration is filtered;
4) get filtrate 1mL in the 50mL volumetric flask, add 1-3mL distilled water and 4-8mL developer, be settled to scale with distilled water then, after 15 minutes, under 520nm, use spectrophotometer colorimetric estimation light absorption value A;
5) the light absorption value A ' of the nitrite anions standard solution of a series of concentration known of colorimetric estimation under 520nm makes the working stamndard curve with the concentration of nitrite anions and the light absorption value A ' of mensuration, obtains slope b;
6), calculate the concentration S of the nitrite anions in the solution of measuring, S=A*b according to light absorption value A and slope b;
7) calculation sample Central Asia nitrate radical content;
8) calculate nitrate reductase activity:
Computing formula is: [(S
1-S
2) * 50 * V
1/ (1000 * V
2)]/(W * T), mgNO
2 --Ng
-1Soil24h
-1
In the formula: S
1Be the NO in the sample
2 -The content of-N (mg/L); S
2Be the NO in the sample contrast
2 -The content of-N (mg/L); 50 is the liquor capacity (mL) of constant volume; V
1Be extract cumulative volume (mL); V
2For drawing the volume (mL) of filtrate; W is for claiming soil sample heavy (g); T is for cultivating the time (h) of soil sample; 1000 is the unit conversion coefficient.
2. analytical approach according to claim 1 is characterized in that: for the sour earth sample, should reconcile its pH before analyzing is 7-8.5, adopts the method that adds lime carbonate to reconcile, and adds 15-25mgCaCO in every test tube respectively
3, in test tube, add 2 of 1-3mL 0.5-1mM/L, 4-dinitrophenol dinitrophenolate solution, mixing behind the mixing more respectively.
3. analytical approach according to claim 1 is characterized in that: anaerobism is cultivated employed distilled water, before the use, need be heated to the O in the boiling expeling water
2, sealing is cooled to room temperature then.
4. analytical approach according to claim 1, it is characterized in that: described developer is the alpha-naphthylamine solution of weight concentration 0.1% of acetic acid preparation of weight concentration 35% and the isopyknic mixed liquor of sulfanilic acid solution of weight concentration 0.5%, and the adding of mixed liquor is excessive with respect to nitrite anions.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007100106805A CN101271060B (en) | 2007-03-21 | 2007-03-21 | Analytical method for detecting nitrate reductase activity in soil |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007100106805A CN101271060B (en) | 2007-03-21 | 2007-03-21 | Analytical method for detecting nitrate reductase activity in soil |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101271060A CN101271060A (en) | 2008-09-24 |
| CN101271060B true CN101271060B (en) | 2011-06-08 |
Family
ID=40005160
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2007100106805A Expired - Fee Related CN101271060B (en) | 2007-03-21 | 2007-03-21 | Analytical method for detecting nitrate reductase activity in soil |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101271060B (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102128736B (en) * | 2010-11-26 | 2012-12-19 | 中国科学院南京土壤研究所 | Dry land nitrification and denitrification field in-situ measuring device and method for testing by using same |
| CN103913450A (en) * | 2014-03-21 | 2014-07-09 | 中山市南方新元食品生物工程有限公司 | Accurate quantitative detection method for activity of lipoxygenase |
| CN105543330A (en) * | 2016-01-25 | 2016-05-04 | 东北农业大学 | Nitrate reductase measuring kit and application |
| CN110643676A (en) * | 2019-10-11 | 2020-01-03 | 石河子大学 | Novel method for measuring activity of soil nitrate reductase |
| CN110907445A (en) * | 2019-12-06 | 2020-03-24 | 西安外事学院 | Microfluidic biochip detection device, preparation method and detection method |
| CN116908128B (en) * | 2023-08-22 | 2024-04-19 | 中国科学院东北地理与农业生态研究所 | Kit for detecting nitrate reductase activity and detection method thereof |
-
2007
- 2007-03-21 CN CN2007100106805A patent/CN101271060B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN101271060A (en) | 2008-09-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101271060B (en) | Analytical method for detecting nitrate reductase activity in soil | |
| Kaiser et al. | Evaluation of methods to estimate the soil microbial biomass and the relationship with soil texture and organic matter | |
| Voroney et al. | Determination of kC and kNin situ for calibration of the chloroform fumigation-incubation method | |
| Berríos et al. | Spectrophotometric method for determining gibberellic acid in fermentation broths | |
| CN101586145A (en) | Analyzing method for detecting activity of soil xylanase | |
| CN100494994C (en) | Analysis method for detecting activity of nitrous acid reductase in soil | |
| CN101625310A (en) | Analysis method for detecting arylsulfatase activity in soil | |
| CN103525896A (en) | Quantitative high-vitality yeast cell screening method based on TTC (2,3,5-triphenyltetrazolium chloride) staining method | |
| Solaiman | Measurement of microbial biomass and activity in soil | |
| CN102564979A (en) | Method for determining alcohol concentration by using enzyme cycling method and alcohol determination kit | |
| CN100494979C (en) | Analytical method for detecting hydroxyl amine reducing enzyme activity in soil | |
| Lehtokari et al. | Determination of ATP from compost using the firefly bioluminescence technique | |
| CN101270385A (en) | Analysis method for testing soil denitrification enzyme liveness | |
| Egami et al. | Nitrate | |
| Bostick et al. | Methodologies for the determination of adenosine phosphates | |
| CN113655006B (en) | Urinary system knot Dan Chengdan risk factor detection and test system | |
| CN102495011B (en) | Method for determining activity of bacterial nitrite reductase | |
| CN115639333A (en) | Method for detecting ammonia nitrogen content by using nitration biological reaction | |
| CN102108377A (en) | Method for determining activity of soil arginine desaminase | |
| CN104034721A (en) | Detection method of ammoniacal nitrogen in fermenting liquid | |
| CN101294191A (en) | Analysis method for testing ammonium oxidizing enzyme activity in soil | |
| CN101251451B (en) | Method for digesting available nutrient from acid soil as well as combined digestion agent thereof | |
| CN101294192A (en) | Analysis method for testing catalase activity in soil | |
| CN110643676A (en) | Novel method for measuring activity of soil nitrate reductase | |
| CN101294193A (en) | Analysis method for testing desaturase activity in soil |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20110608 Termination date: 20120321 |