Method for determining content of Thelephora ganbajun mycelia in soil
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A method of determination, the technology of dried bacteria, applied in the field of determination of mycelium content, to achieve the effect of rapid detection and simple method
Inactive Publication Date: 2020-04-17
YUNNAN UNIV
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However, the selection of single-passage genes requires professio
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Embodiment 1
[0023] To determine the standard curve, the specific steps are as follows:
[0024] 1. Materials
[0025] Ganba bacteria soil samples were collected in Wuding County, Yimen County and Lufeng County, Yunnan Province (see Table 1). Collection method: in each Ganba bacteria growth place, use the 5-point method to take the fruiting body as the center radius 5cm , the soil with a depth of 14cm, every 2cm is a soil depth, a total of 7 soil samples with different depths are put into the prepared sterile sampling bags, numbered, and the soil samples are stored at -80°C (soil samples contain dry bacteria fungal mixed samples);
[0026] Table 1 Collection information of soil samples of D.
[0027]
[0028] 2. Method
[0029] 2.1 DNA extraction:
[0030] Use QIAGEN's The DNA Isolation Kit was used for DNA extraction, and 300 mg of dry fungus soil samples were taken at each depth, and the extracted soil fungal DNA mixed liquid was placed at -20°C for later use;
[0031] 2.2 Prim...
Embodiment 2
[0061] Determination of the mycelium content of Drosophila in soil:
[0062] 1. Materials
[0063] Ganba bacteria soil samples were collected in Wuding County, Yimen County and Lufeng County, Yunnan Province (see Table 2). Collection method: in each Ganba bacteria growth place, use the 5-point method to take the fruiting body as the center radius 5cm , the soil with a depth of 14cm, every 2cm is a soil depth, a total of 7 soil samples with different depths are put into the prepared sterile sampling bags, numbered, and the soil samples are stored at -80°C (soil samples contain dry bacteria fungal mixed samples);
[0064] Table 2 Collection information of soil samples of D.
[0065]
[0066] 2. Method
[0067] 2.1 DNA extraction:
[0068] Use QIAGEN's The DNA Isolation Kit was used for DNA extraction, and 300 mg of dry fungus soil samples were taken at each depth, and the extracted soil fungal DNA mixed liquid was placed at -20°C for later use;
[0069] 2.2 Primer synt...
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Abstract
The invention discloses a method for determining the content of Thelephora ganbajun mycelia in soil. The method comprises the following steps: in a Thelephora ganbajun growing field, taking seven soilsamples at different depths from soil in a circular area where sporocarp is taken as a center, a radius is 5 cm and a depth is 14 cm through a five-point method, wherein a depth of 2 cm is taken as one soil depth, putting the seven soil samples into prepared sterile sampling bags, numbering the sterile sampling bags, and storing the soil samples at a temperature of -80 DEG C; carrying out DNA extraction on the soil sample to obtain a liquid which is a mixed liquid of DNAs of soil fungi, and naturally cooling the mixed liquid of the DNAs of the soil fungi at -20 DEG C for later use; and carrying out a qPCR test by adopting a primer Tg, and substituting a detected Ct value into a standard curve equation to calculate the mycelium content of the soil. The method can achieve the purpose of rapidly detecting the content of Thelephora ganbajun mycelia in soil, and is simple and efficient.
Description
technical field [0001] The invention relates to a method for determining the content of mycelium, in particular to a method for determining the content of mycelia in soil of D. Background technique [0002] The determination of mycelium content plays a very important role in exploring the growth mechanism and metabolic activities of fungi, and mycelium content can be used as a parameter to measure the growth status of fungi. The method for determining the mycelium content of soil fungi is generally an indirect measurement method, and the indirect measurement method is to calculate the content of the mycelium by measuring the content of some special substances (such as chitin, etc.) in the fungal mycelium. The main methods are: chitin method, ergosterol method, protein method and nucleic acid method. Different measurement methods have certain limitations, and a more suitable measurement method should be selected according to actual needs during actual measurement. [0003] ...
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