Method and system for detecting maize rhizosphere soil microorganisms based on deep sequencing

Abstract

The invention belongs to the technical field of soil microbiology, and discloses a method and a system for detecting maize rhizosphere soil microorganisms based on deep sequencing. The method comprises the following steps: 1, collecting rhizosphere soil; 2, extracting high-quality microorganism genome DNA from the soil; 3, performing PCR to amplify a 16S rDNA V4 region in the DNA by using a double-tag primer, and constructing a library; 4, sequencing a qualified library to obtain pure READS; 5, splicing, wherein each sample produces at least 50 thousand effective TAG for clustering into operable taxons; 6, analyzing species composition, structure, diversity and relative abundance difference significance 7, determining the composition, the structure, the diversity and the relative abundanceof transgenic saline-alkaline-resistant maize rhizosphere soil prokaryotic microflorae. The method and the system for detecting the maize rhizosphere soil microorganisms based on deep sequencing aresupported by a major national genetic modification specific subject, namely, a national genetically modified corn wheat soybean environmental safety assessment technology (2016ZX08011003).

Description

technical field [0001] The invention belongs to the technical field of soil microbiology, and in particular relates to a method and system for detecting corn rhizosphere soil microorganisms based on deep sequencing. Background technique [0002] The rhizosphere (Rhizosphere) is defined as the biologically active area closely surrounding the viable root system, which contains a large number of microorganisms, such as bacteria, actinomycetes, and fungi, as well as some algae and viruses, generally within 1cm from the root surface . Different plants or even different growth stages of the same plant, their root exudates will affect the growth and development of rhizosphere microorganisms; and rhizosphere microorganisms participate in various physiological and biochemical processes such as the decomposition of soil organic matter, the formation of humus, the transformation and circulation of nutrients, etc. The process, especially the nitrogen-fixing bacteria group, converts nit...

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Problems solved by technology

[0006] Existing technologies have low detection throughput, small scale, and low coverage of non-culturable microorganisms; they cannot accurately detect differences in the composition, structure, diversity, and relative abundance of microbial communities in the rhizosphere of plants of different genotypes, including transgenic crops. The rhizosphere soil obtained by shaking off method actually contains a large amount of non-rhizosphere soil, which leads to inaccurate subsequent identification and detection results

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