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Analyzing soil microbial community diversity using lipoid fatty acid atlas technology

A microbial community and diversity technology, applied in the direction of analysis of materials, preparation of test samples, material separation, etc., can solve the problem of unreported diversity of soil microbial communities

Inactive Publication Date: 2005-03-02
RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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Problems solved by technology

[0006] Although everyone has recognized that PLFA spectrum analysis technology can be used to study soil microbial communities, but the specific use of PLFA spectrum to calculate the diversity of soil microbial communities has not been reported. Based on this aspect, this study proposes a new Calculation method of soil microbial community diversity, that is, calculation method based on PLFA spectrum

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  • Analyzing soil microbial community diversity using lipoid fatty acid atlas technology
  • Analyzing soil microbial community diversity using lipoid fatty acid atlas technology

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Embodiment Construction

[0010] Weigh 4g of soil sample, add 3.6ml of phosphate buffer, 4ml of chloroform and 8ml of methanol, shake for 1 hour in the dark and then centrifuge at 2000rpm for 30min. Transfer the supernatant to a 50ml separating funnel, add 3.6ml phosphate buffer and 4ml chloroform, and separate overnight at room temperature in the dark. The chloroform phase of the lower layer was collected and separated using a silicic acid column, using 5 ml of chloroform, 10 ml of acetone and 5 ml of methanol as eluents. Collect the methanol phase, dry it with high-purity nitrogen, add 1ml methanol:toluene (1:1, V:V) and 1ml 0.56% (w / v) KOH methanol solution, and react at 35°C for 30 minutes. Cool to room temperature, add acetic acid for neutralization, then add 2ml of chloroform:n-hexane (1:4, V:V) and ultrapure water to mix. After static separation, the upper layer liquid was collected, dried with high-purity nitrogen, and then dissolved in chloroform:n-hexane (1:4, V:V) containing a certain amoun...

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Abstract

The invention relates to biochemical method for analysing the diversity of the soil microbiocoenosis,which uses lecithoid fatty acid spectrogram combining technology to analyse the diversity of the soil microbiocoenosis.The steps is: 1)weighing specific quality of soil materials; 2)adding the buffer liquid and organic solution to get organic phase containing liqid; 3)gathering the eluent containing the polar liqid after the organic phase is separated by silicic acid column; 4)gathering the fatty acid produced by the methylesterizing reaction in alkaline environment; 5)using gas phase chromaspectrum-mass spectrum combinmeter to analyse the methylester fatty acid quantitatively to get the content of various kinds of lecithoid fatty acid; 6)calculating the diversity index and judging the diversity of the soil microbiocoenosis. The invention has important practical meanings and applies accurate and quick method for research of the diversity of the microbiocoenosis and the relevant research, especially to assessment and the future developing forecast of the farmland ecology.

Description

Technical field: [0001] Phospholipid fatty acid profile (PLFA) analysis technology is a biochemical method for studying microbial ecology, which is mainly used to study the biomass, community structure, nutritional status and metabolic activities of microbial communities. Furthermore, the method has also been shown to be effective in evolutionary and taxonomic studies. Background technique: [0002] The current description of soil mainly uses some physical and chemical indicators, and rarely involves the status of soil microorganisms. In fact, the biomass of microorganisms in the soil accounts for 80% of the total soil biomass (excluding plant roots), which largely determines the ecological function of the soil. Soil microorganisms play an important role in the metabolism and circulation of almost all plant nutrients, and are closely related to the organic matter content and fertility of the soil. Therefore, the diversity of microorganisms in soil is often used as a biolog...

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Application Information

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IPC IPC(8): G01N1/28G01N30/72G01N30/88G06F19/00
Inventor 齐鸿雁薛凯张洪勋罗海峰
Owner RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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