SU1370134A1 - Method of determining nitrogen monoxide separated by soil microorganisms - Google Patents

Abstract

The invention relates to agriculture, in particular to methods for the determination of nitrous oxide released by soil microorganisms. The aim of the invention is to improve the accuracy of determination. The method is as follows: a sample of the soil sample is taken, incubated on a nutrient medium under anaerobic conditions in the presence of acetylene, the gas phase is removed, carbon dioxide is removed from it by passing through filtering material from granulated ascarite and in the gas phase nitrous oxide is determined by gas chromatography chesky. The method allows to increase the accuracy of the analysis and determine the nitrous oxide produced by the yeast cultures. 2 tab.

Description

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The invention relates to agriculture, in particular to the assessment of the biological activity of soil microorganisms, and in particular to methods for the determination of nitrous oxide.

The purpose of the invention is to improve the accuracy of determination.

The method is carried out as follows.

Example. Pure cultures of soil microorganisms are grown in 15 ml penicillin vials containing 5 ml of the corresponding medium (Gilta, wort). After sowing, the vials are closed with rubber stoppers, the air is displaced above the medium surface with argon, and acetylene is introduced (10% by volume). The vials are incubated at 28 ° C for 3 days. Gas sampling (1 ml) was carried out with a standard medical syringe (prototype) and analyzed on a gas chromatograph.

For comparison, in the same cultures, the selection of the gas sample for content was carried out by the proposed method. To this end, pure cultures of soil microorganisms are grown in 15 ml penicillin vials containing 5 ml of the corresponding meal (Gilta, wort). After seeding, the vials are sealed with rubber stoppers, the air is forced out above the medium surface with argon and acetylene is introduced (1 ° C medium with argon and acetyl (10% vol.) Is introduced. The vials are incubated for 1 day. The gas sample (1 ml) is taken with a syringe Ascarite gasket with a thickness of 1 mm, weight O, A g (fraction 0.3-0.5 mm) and gas chromatographic determination of the content in it.

The results of analyzes of gas samples taken in two ways are presented in Table. 1 and 2.

As follows from the table. 1 and 2 in the samples taken by the standard method (I), nitrous oxide was not detected, whereas the proposed method for the selection of gas samples (II) reveals a significant release of nitrous oxide by bacterial and yeast cultures.

Soil yeast pp. Cryptococcus and Lipomyces nitrous oxide did not release the maximum production of N 0 from the pp. Hansenula and Pichia yeast, which possess active enzymes katase and pro-oxidase; (tab. 1). Of

FOR 2

tab. 2 it follows that the activity of de-nitrification in soil bacteria Bacillus sp. decreases during incubation, therefore, it is advisable to determine it no later than 2-3 days of growing the culture on a nutrient medium.

Claims (1)

Thus, the proposed method of determining the amount of nitrous oxide released by microorganisms allows to significantly improve the accuracy of NjO determination, as well as to study this process over time. Invention Formula The method of determining nitrous oxide secreted by soil microorganisms, which involves incubating the sample on a nutrient medium under anaerobic conditions in the presence of acetylene, followed by gas phase selection and gas chromatographic analysis of nitrous oxide in it, and tl and h that, in order to increase the accuracy of the determination, pure culture of soil microorganisms is subjected to incubation; carbon dioxide is removed directly during the gas phase selection process by passing granules through filtering material ized ascarite. ,Table 1 Hansenula polymorpha 2733 ABOUT 24.0 The same 273A O 26.2 Hansenula sp. 847. About 4.8 Pichia gui lermondii  3 137013A Table 2 8.4 12.1 10.9 9.0 Continued table. 2 N, 0, mg / mp, medium A 1 (G ten LTD oh oh 17.0 13.7 .2 6,6 3.8