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Method for preparing N2O with determined <15>N abundance and measuring nitrogen cycle based on <15>N isotope tracer technology

A strain and gas measurement technology, applied in the biological field, can solve problems such as being easily oxidized to nitrate, affecting content and abundance, and instability of nitrite, so as to reduce gas storage and transportation costs, easy operation and low cost low effect

Active Publication Date: 2018-07-20
NINGBO URBAN ENVIRONMENT OBSERVATION & RES STATION NUEORS CHINESE ACADEMY OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

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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  • Method for preparing N2O with determined &lt;15&gt;N abundance and measuring nitrogen cycle based on &lt;15&gt;N isotope tracer technology

Examples

Experimental program
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Effect test

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; then respectively press 10 -2 to 10 -5 Numbered in sequence, use a pipette to draw 1ml of 10 -1 Double dilution to number 10 -2 Into the test tube, blow gently three times to make the suspension fully mix, and get 10 -2 double dilution; and...

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 invention relates to a method for preparing N2O with a determined <15>N abundance by using Penicillium janthinillum and measuring nitrogen cycle based on a <15>N isotope tracer technology, and belongs to the biotechnical field. The Penicillium janthinillum is preserved in China General Microbiological Culture Collection Center on August 7, 2017 with the storage number of CGMCC No. 14146. The method for preparing N2O with the determined <15>N abundance comprises the following steps: collecting a soil sample, diluting the soil sample, and carrying out culturing, gas measurement screening andidentification on the above strain; and preparing a Czapek Dox liquid medium by using NaNO3 with the determined <15>N abundance, inoculating the medium with the Penicillium janthinillum strain, performing shaking culture, collecting produced N2O, and adding an alkaline solution to absorb and remove CO2 in the N2O in order to obtain the N2O with the determined <15>N abundance. The method has the advantages of rapidness and accuracy in gas production, simplicity in operation and low cost; and the prepared gas achieves highly accurate and highly precise results when used in nitrogen cycle studying as a standard gas.

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 hydroxylamine hydrochloride) 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 based on 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 ox...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12Q1/04G01N27/62C12R1/80
CPCC12N1/14C12Q1/04C12N1/145C12R2001/80G01N27/62G01N2333/385
Inventor 李雅颖郑宁国姚槐应吴愉萍
Owner NINGBO URBAN ENVIRONMENT OBSERVATION & RES STATION NUEORS CHINESE ACADEMY OF SCI
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