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Analytical method for detecting nitrate reductase activity in soil

An analysis method and reductase technology, which are applied in the preparation of test samples, biochemical equipment and methods, and the determination/inspection of microorganisms, etc., can solve the problems that are difficult to adapt to the qualitative comparison and quantitative research of nitrate reductase activity, and increase the number of experimental results. Accuracy, troublesome pre-processing of samples, etc., to achieve stable and reliable analysis results, reduce complexity and uncertainty, and reduce test costs

Inactive Publication Date: 2008-09-24
SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
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  • Claims
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Problems solved by technology

The steps of this method are cumbersome, and special reagent bottles and suction filtration devices are required for vacuum cultivation, and the differences and uncertainties of the cultivation batches make it difficult to adapt to the qualitative comparison and analysis of nitrate reductase activity in soil. Quantitative research; at the same time, the pre-processing of samples in this method is cumbersome, which reduces the efficiency of the experiment and increases the inaccuracy of the experimental results

Method used

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  • Analytical method for detecting nitrate reductase activity in soil
  • Analytical method for detecting nitrate reductase activity in soil
  • Analytical method for detecting nitrate reductase activity in soil

Examples

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preparation example Construction

[0041] 8. Preparation of standard curve: draw 1ml of the above-mentioned standard stock solution, and set the volume to 100ml, which is 2.5mgNO 2 - -N / L solution. Then draw 0, 1, 2, 3, 4, and 5ml of the solution into a 50ml volumetric flask, add a small amount of distilled water and 4ml of developer, and distill to volume. The amount of nitrite nitrogen contained in this standard series is 0, 0.05, 0.1, 0.15, 0.2, 0.25mgNO 2 - -N / L. Colorimetric determination of a range of known concentrations of NaNO at 520nm 2 The absorbance value A' of the solution is expressed as NaNO 2 The concentration of the solution and the measured absorbance value A' prepare a standard curve to obtain the slope b;

[0042] Steps:

[0043] 1) Weigh 4 parts of 1g soil air-dried samples passed through a 1mm sieve into 4 test tubes, add 20mg CaCO 3 , after mixing, add 1 mL of 0.8mM / L 2,4-dinitrophenol solution to the test tubes, and mix well; then add 1-1.5 mL of 0.05%-0.15% substrate KNO to thre...

Embodiment 1

[0053] The black soil used in this example was collected from the Hailun Ecological Experimental Station of the Chinese Academy of Sciences, and three different treatments were set up: No. 1 control, ordinary soil that was cultivated at 25°C for more than 24 hours without any treatment or adding any soil additives; No. 3 is the soil added with the nitrification inhibitor DCD and incubated at 25°C for more than 24 hours; No. 3 is the soil added with the nitrification inhibitor DMPP and incubated at 25°C for more than 24 hours. The soil moisture content is 20% (air-dried soil weight). In No. 2 and No. 3 soils where nitrification inhibitors were added, the addition amount of the two inhibitors was 50 ppm, and the activities of the three nitrate reductases were detected by the above method.

[0054] The specific analysis steps for each soil are as follows:

[0055] 1) Weigh 4 parts of 1g soil air-dried samples passed through a 1mm sieve into 4 test tubes, add 20mg CaCO 3 , after...

Embodiment 2

[0066] The brown soil of three different planting crops used in this example was collected at the Shenyang Ecological Experiment Station of the Chinese Academy of Sciences: wherein the previous crop of No. 4 soil was rice, the previous crop of No. 5 soil was corn, and the previous crop of No. 6 soil was soybeans. The soil of three kinds of different tillage systems all is 20% (air-dried soil weight) in water content, and temperature is cultivated more than 24h under the condition of 25 ℃, measures its nitrate reductase activity, and concrete steps are as follows: substrate potassium nitrate concentration is 0.1%, Add 0.50mL, the glucose concentration is 1%, add 1mL, the rest of the steps are the same as Example 1.

[0067] The test results are as follows:

[0068]

[0069]

[0070] The data in the table also shows the stability and high precision of the analysis results.

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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 [0001] The invention relates to the determination of the activity of nitrate reductase in soil, in particular to an analysis method for detecting the activity of nitrate reductase in soil. Background technique [0002] Nitrate reductase in the soil can reduce the nitrate nitrogen formed in the process of soil nitrogen metabolism to nitrite nitrogen, and the reduced compounds in the soil can be used as hydrogen donors. [0003] [0004] Therefore, the activity of nitrate reductase in soil affects the gaseous loss of nitrogen in the process of soil nitrogen metabolism, and indirectly affects the use efficiency of nitrogen fertilizer and nitrogen pollution in the atmosphere. The activity of nitrate reductase in soil is strongly affected by soil conditions such as moisture, temperature, and soil texture, as well as by farming systems, management practices, and different planting crops. Therefore, it is of great significance to test the activity of nitrate red...

Claims

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Application Information

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IPC IPC(8): G01N21/31G01N1/28C12Q1/26
Inventor 武志杰隽英华陈利军史云峰
Owner SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
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