Method for constructing chroogomphis rutillus fruiting body surrounding soil fungus community internet testing services (ITS) library
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A technology of rivet mushroom seeds and soil, which is applied in the field of constructing the ITS library of soil fungal communities around the fruiting body of blood red rivet mushroom, and achieves the effect of important application value
Active Publication Date: 2012-07-11
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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However, the research on the microbial population in its growth environment has not been reported
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Embodiment 1
[0052] Example 1. Construction of the ITS library of the soil fungal community around the fruiting body of the blood-red rivet mushroom
[0053] 1. Collection of samples
[0054] Soil samples were collected at an altitude of 790 meters in Beijing (GPS coordinates N4057.186 E11629.923) at a place 10 cm below the fruiting body of the rivet mushroom, and placed in a sterile kraft paper bag.
[0055] 2. Extraction of metagenomic DNA from soil samples
[0056] (1) Weigh 5g of the soil sample collected in step 1, suspend in 3ml TENP buffer, vortex and shake for 10min, centrifuge at 12000rpm for 10min, and take the precipitate.
[0057] (2) Wash the precipitate in step (1) three times with TENP buffer, and then add 3 ml of PBS buffer to wash once.
[0058] (3) The precipitation of step (2) is carried out the extraction of metagenomic group, the extraction of metagenomic group adopts Mega Preps PowerMax Soil DNA Isolation Kit (Mo Bio, Carlsbad, CA, USA) to carry out according to ins...
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Abstract
The invention discloses a method for constructing a chroogomphis rutillus fruiting body surrounding soil fungus community internet testing services (ITS) library. The method comprises the following steps of: extracting a metagenome of chroogomphis rutillus fruiting body surrounding soil; performing polymerase chain reaction (PCR) amplification by using the metagenome as a template and using ITS1 and ITS4, and reclaiming DNA fragments of 700bp; connecting each DNA fragment and a pMD19-T vector, performing PCR amplification by using RV-M and M13-47, performing enzyme digestion on each PCR product by using HinfI, Hae III, Msp I, Taq I and Mbo I respectively, and performing electrophoresis to obtain a molecular map. By using the method, the chroogomphis rutillus fruiting body surrounding soil fungus community ITS library can be constructed; and the method has significance for determining the growth environment microbial fungus community structure, the heredity and the functional diversity of chroogomphis rutillus fruiting bodies, provides important information for deeply discussing artificial acclimation and culture of chroogomphis rutillus, and has important application value.
Description
technical field [0001] The invention relates to a method for constructing an ITS library of a soil fungal community around the fruit body of the rivet mushroom. Background technique [0002] Blood red rivet mushroom (Chroogomphis rutillus), Basidiomycotina, Phytomycetes, Agaricaceae, Rivetaceae, Chroogomphis genus, commonly known as red mushroom, pine umbrella, pine mushroom, meat mushroom, shaped like a rivet, the color is dark Red, rare wild mycorrhizal edible fungus, is an important ectophytic large mycorrhizal fungus of coniferous trees, growing in groups or scattered in summer and autumn. The mushroom is thick, smooth, delicate, delicious, high in nutritional value, rich in protein, contains 8 kinds of essential amino acids, calcium, iron and other minerals and a large amount of carbohydrates needed by the human body, and is also a dietary fiber and natural antioxidant An important source of and loved by the people. [0003] Blood red rivet mushroom is an ectomycorrhi...
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