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A method for preparing n2o with definite 15n abundance and measuring nitrogen cycle based on 15n isotope tracer method
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An isotope tracer, 15NO2--N technology, applied in the biological field, can solve the problems of being easily oxidized into nitrate, affecting the content and abundance, and the instability of nitrite, so as to reduce gas storage and transportation costs, and operation Simple and low-cost effect
Active Publication Date: 2021-10-22
NINGBO URBAN ENVIRONMENT OBSERVATION & RES STATION NUEORS CHINESE ACADEMY OF SCI +1
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However, there are big problems in this method: first, nitrite is unstable, and it is easily oxidized into nitrate during storage, which affects 15 N 2 The content and abundance of O; the second is the production 15 N 2 O gas is not easy to store, so it can only be prepared and used immediately
At present, most studies on fungal denitrification have focused on the effect of fungi on soil N. 2 The contribution of O release has not yet been produced by fungi with a defined abundance 15 N 2 O's research
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Embodiment 1
[0029] (1), according to the following components to prepare the Cha formula solid medium: NaNO 3 2g, K 2 HPO 4 1g, KCl 0.5g, MgSO 4 0.5g, FeSO 4 0.01g, 30g sucrose, 18g agar, 1000ml water, sterilize at 121°C for 20min after preparation;
[0030] Prepare Chase Liquid Medium according to the following composition: NaNO 3 2g, K 2 HPO 4 1g, KCl 0.5g, MgSO 4 0.5g, FeSO 4 0.01g, 30g sucrose, 1000ml water, sterilized at 121°C for 20min after preparation.
[0031] (2) Strain screening:
[0032] Collect 5 g of soil samples from Hangzhou Longjing Tea Garden into a sterilized 100 mL Erlenmeyer flask, add 45 mL of sterilized deionized water, and vortex for 1 min to obtain a soil suspension that is 10 -1 two-fold dilution solution; and then respectively put 4 test tubes containing 9mL sterilized deionized water into -2 to 10 -5 Numbered in sequence, use a pipette to draw 1ml of 10 -1 Double dilution to number 10 -2 In the test tube, blow gently three times to make the...
Embodiment 2
[0042] The only difference between this embodiment and Example 1 is that the composition of the Chase solid medium is: NaNO 3 1.8g, K 2 HPO 4 0.8g, KCl 0.4g, MgSO 4 0.4g, FeSO 4 0.01g, 27g sucrose, 20g agar, 1000ml water, sterilize at 126°C for 17min after preparation;
[0043] The composition of the Chase liquid medium is: NaNO 3 1.8g, K 2 HPO 4 0.8g, KCl 0.4g, MgSO 4 0.4g, FeSO 4 0.01g, 27g sucrose, 1000ml water, sterilized at 126°C for 17min after preparation.
Embodiment 3
[0045] The only difference between this embodiment and Example 1 is that the composition of the Chase solid medium is: NaNO 3 2.2g, K 2 HPO 4 1.3g, KCl 0.6g, MgSO 4 0.6g, FeSO 4 0.01g, 32g sucrose, 15g agar, 1000ml water, sterilize at 115°C for 23min after preparation;
[0046] The composition of the Chase liquid medium is: NaNO 3 2.2g, K 2 HPO 4 1.3g, KCl 0.6g, MgSO 4 0.6g, FeSO 4 0.01g, 32g sucrose, 1000ml water, sterilized at 115°C for 23min after preparation.
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Abstract
The present invention relates to a kind of preparation that utilizes Penicillium violet 15 N Abundance N 2 O and based on 15 The invention relates to a method for measuring nitrogen cycle by an N isotope tracing method, which belongs to the field of biotechnology. The Penicillium violet in the present invention has been preserved in the General Microorganism Center of China Microbial Strain Preservation Management Committee on August 7, 2017, and the preservation number is CGMCCNo.14146, which has a definite 15 N Abundance N 2 The preparation method of O comprises the steps of: collecting soil samples, carrying out strain cultivation, gas measurement screening and identification after dilution; 15 N abundance NaNO 3 Configure Chapei's liquid medium, inoculate Penicilliumlavender, and collect the N produced after shaking culture. 2 O, add lye to absorb and remove CO 2 , that is obtained with a certain 15 N Abundance N 2 O. The method of the invention produces gas quickly and accurately, and is easy to operate and low in cost; the prepared gas is used as a standard gas for nitrogen cycle research, and the result has high accuracy and precision.
Description
technical field [0001] The invention belongs to the field of biological technology, and relates to a kind of 15 N Abundance N 2 O and based on 15 N isotope tracer method for measuring the nitrogen cycle. Background technique [0002] 15 N isotope tracer technology is the most effective method for us to carry out the study of nitrogen cycle. It can distinguish the rate and flux of nitrogen turnover processes (mineralization, nitrification, denitrification, etc.). Currently for 15 NO 3 - -N and 15 NO 2 - -N is detected by adding chemical reagents (such as hydroxylaminehydrochloride) to reduce nitrate nitrogen and nitrite nitrogen to 15 N 2 O gas was measured. In this method, the standard 15 N 2 O gas for calibration. And currently 15 N 2 The preparation method of O is mainly to 15 N-labeled nitrite is produced by chemical reduction reaction as raw material. However, there are big problems in this method: first, nitrite is unstable, and it is easily oxidized...
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