Method for fast extracting AM epiphyte environment DNA in plant rhizosphere soil

A technology of plant roots and fungi, applied in the fields of molecular biology and applied microbiology, can solve the problems of undiscovered public reports, achieve the effect of reducing degradation and promoting the release of DNA

Inactive Publication Date: 2008-03-26
YUNNAN UNIV
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  • Abstract
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Problems solved by technology

[0005] Through document search, do not find the public report of the same document as the content of the present invention

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Extraction of AMF genomic DNA from the rhizosphere soil of Dodonaea viscose:

[0040] Take by weighing 100g of willow plant rhizosphere soil, collect AM fungus-related structures (AMF spores, extraroot hyphae, etc.) DNA was extracted with 20% Chelex-100 resin, and nested-PCR was performed, and the AMF target gene was obtained by cloning and sequencing. And submit the obtained gene to GenBank, the sequence accession number is EU118739.

Embodiment 2

[0042] Genomic DNA extraction of AM fungi in rhizosphere soil of Heteropogon contortus;

[0043] It is basically the same as in Example 1, and the obtained gene is submitted to GenBank, and the sequence accession number is EU118750.

Embodiment 3

[0045] Genomic DNA Extraction of AM Fungus from D. viscose Root:

[0046] Weigh 2 g of fresh willow root system, sterilize the surface with 75% ethanol, cool and grind into powder with liquid nitrogen, add 1000 μl CTAB solution and 20% Chelex-100 resin for DNA extraction according to the method of the present invention, and perform nested-PCR , the objective gene of AM fungus was obtained by cloning and sequencing. And submit the obtained gene to GenBank, the sequence accession number is EU118692.

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Abstract

The present invention relates to process of fast extracting AM fungal genome DNA in plant rhizosphere soil environment, and belongs to the field of molecular biology and applied microbiology. The process includes the first wet screening to eliminate great amount of PCR proliferation limiting factors from rhizosphere soil, collecting AM fungal related structures of rhizosphere soil for DNA extraction, mechanically breaking wall and adding CTAB to promote DNA release, and further purifying with Chelex-100 resin. The process is easy, simple and fast, and has great DNA extracting amount, capacity of being concentrated and purified and other advantages. The present invention has excellent application foreground.

Description

Technical field: [0001] The invention relates to a method for quickly extracting environmental DNA of AM fungi in plant rhizosphere soil, belonging to the fields of molecular biology and applied microbiology. Background technique: [0002] Arbuscular mycorrhizal fungus (Arbuscular mycorrhizal fungus, AMF) is a class of soil microorganisms widely distributed in nature, it can form a mutualistic symbiont with the roots of more than 90% of terrestrial vascular plants - arbuscular mycorrhizal fungi Mycorrhiza, AM). More and more studies have shown that a large number of AMF extrarhizomycelia in the rhizosphere play important ecological functions in the soil ecosystem. It is an important factor in the composition of flora, and it is also an important factor affecting the allocation of resources in the ecosystem, the stability of the ecosystem, species diversity and productivity. Arbuscular mycorrhizae appear to affect the processes and functions of living systems at different o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31C12N15/10
Inventor 李涛赵之伟
Owner YUNNAN UNIV
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