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Detection method of soil anammox microbial community structure

A technology of microbial community and anammox, which is applied in the field of soil microbial community structure detection, can solve the problems of high cost, inability to quickly analyze the community structure of anammox bacteria, and heavy workload of cloning libraries

Active Publication Date: 2019-01-25
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method avoids the requirement for the pure culture of anammox bacteria, but the construction of clone library is heavy workload and high cost, and it cannot realize the rapid analysis of the community structure of anammox bacteria
[0003] Cloning and DGGE are commonly used methods to study the structure of microbial communities, but these two methods can only reflect the similarity and difference between community structures. If you want to accurately analyze the content of each microorganism, you need to combine other techniques

Method used

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  • Detection method of soil anammox microbial community structure
  • Detection method of soil anammox microbial community structure
  • Detection method of soil anammox microbial community structure

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Embodiment 1, the detection method of soil anammox microbial community structure in vegetable garden soil, carries out following steps successively:

[0055] Prepare 6 100mL culture flasks, and each culture flask carries out the following steps 1) to 4):

[0056] 1), add 30g of fresh soil to be tested in each culture bottle, absorb freshly prepared 1.2mL concentration of 5g / L 13 C-urea solution is evenly sprinkled in the soil of each culture bottle and stirred rapidly to make the soil 13 The concentration of C-urea-N reaches about 200mg / kg;

[0057] 2), absorb 1.2mL of KNO with a concentration of 5g / L 3 Sprinkle the solution evenly in the culture bottle soil obtained in step 1), and stir to make the NO 3 － -N concentration is about 200mg / kg;

[0058] 3) Quickly add 6ml of water to adjust to a muddy state and seal the 100mL culture bottle with a rubber stopper;

[0059] 4) Fill the culture bottle with inert gas and drive away the air with a syringe, and put it into...

Embodiment 3

[0097] Example 3. Combined application of organic fertilizer and inorganic fertilizer to treat paddy soil soil anammox microbial community structure diversity.

[0098] The test soil was collected from Jianchun Village, Zhiqian Town, Jintan City, Jiangsu Province, and the amount of pig manure composted was 6000kg hm -2 +50% NPK treatment; wherein, pure NPK (urea, superphosphate, potassium sulfate) treatment fertilizer dosage is: N 300kg hm -2 , P 2 o 5 120kg·hm -2 , K 2 O 100kg·hm -2 ; The nutrient content of pig manure compost is: organic matter 45.4%, N 2.3%, P 2 o 5 2.9%, K 2 O1.2%, water content 29.1%. Take the same process parameters and process steps as in Example 1.

[0099] Measured NO 3 - -N, NH 4 + -N concentration results as Figure 5 Shown, calculated, NO 3 - -N, NH 4 + -N concentration decreased the fastest at 7 days and 14 days after culture.

[0100] The obtained data takes the 15:0iso of 14d as an example:

[0101] P FAME =16.273,P ISTD ...

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Abstract

The invention discloses a soil anaerobic ammonium oxidation microbial flora structure detection method. The method comprises the following steps: arranging six containers, wherein the following content is respectively and successively carried out for each container: loading soil to be detected, 13C-urea solution and KNO3 solution into the container, uniformly stirring, adding water, adjusting to a slurry state, and sealing the container; replacing air in the container by using inert gases, placing the container into an anaerobic culture box, and carrying out anaerobic culture; after the six containers are respectively cultured for different times, taking out the soil, measuring the soil, selecting the soil in the container corresponding to a culture phase with the highest change rate according to a content change rate of NO3<->N and NH4<+>-N, and extracting PLFA in the soil; and determining abundance D13C of 13C in the extracted PLFA by virtue of gas chromatography-combustion-isotopic ratio mass spectrometry, so that a soil anaerobic ammonium oxidation microbial floral structure can be acquired.

Description

technical field [0001] The invention belongs to the technical field of soil microbial community structure detection, in particular to a method for detecting soil anammox microbial community structure, in particular to a method for measuring soil anammox microbial community structure by using 13C-PLFA. Background technique [0002] Anaerobic ammonium oxidation (ANAMMOX-Anaerobic ammonium oxidation) means that under anaerobic conditions, microorganisms convert NO 2 - is the electron acceptor, NH 4 + As the electron donor, the NH 4 + , NO 2 - Biochemical process of conversion to nitrogen gas. In 1977, starting from chemical thermodynamics, Broda boldly predicted the existence of Anammox process when calculating its evolutionary relationship with organisms. The first named anammox strain was physically isolated from a laboratory enrichment culture in a bioreactor using Percoll gradient centrifugation. Subsequently, Vande Graaf et al. confirmed the existence of the anamm...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/24G01N30/02
CPCG01N30/02G01N33/24G01N2030/025
Inventor 张奇春周慧芳顾超
Owner ZHEJIANG UNIV