Identification method for existence of fungus in ganoderma lucidum tissue and assessment method for necessity of fungus for formation of ganoderma lucidum

An identification method, Taisui technology, applied in the direction of biochemical equipment and methods, microbial determination/inspection, etc., can solve the problem of not being able to obtain all the genetic information of various types of bacteria, avoid the interference of polysaccharides, and simplify the operation. Effect

Active Publication Date: 2019-08-02
河北省农林科学院经济作物研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] One of the purposes of the present invention is to provide an identification method for the presence of fungi in Tai Sui tissues,

Method used

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  • Identification method for existence of fungus in ganoderma lucidum tissue and assessment method for necessity of fungus for formation of ganoderma lucidum
  • Identification method for existence of fungus in ganoderma lucidum tissue and assessment method for necessity of fungus for formation of ganoderma lucidum
  • Identification method for existence of fungus in ganoderma lucidum tissue and assessment method for necessity of fungus for formation of ganoderma lucidum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Determination of Tai Sui Tissue Melting Temperature

[0052] 1. Determination of melting temperature

[0053] The test tube water bath method is used for determination. The test tube is used to hold Tai Sui tissue pieces and water, and the water bath provides the melting heat source.

[0054] The size of the test tube was 18*18 mm, and the volume of water was set at 10 mL to match the capacity of the disposable syringe filter in the subsequent step. After filling this amount of water, the water level will account for 1 / 3 of the length of the test tube.

[0055] The water bath is an electric heating automatic temperature control type, the temperature control accuracy is 0.5°C, and its water level should be slightly higher than the water level of the test tube.

[0056] Taisui tissue is cut into squares with a side length of 3-5 mm, and the weight of each tube is 0.4 g (wet weight, the same below).

[0057] According to observation, the Tai Sui is solid at na...

Embodiment 2

[0064] Example 2, Extraction of Fungi Information in Tai Sui Tissue

[0065] 1. Preparation of Tai Sui tissue solution

[0066] Weigh 0.6 g of Tai Sui's internal tissue, cut into pieces or debris with a side length of about 1.5 mm, and put it into a test tube with 10 mL of sterile distilled water. Under the dissolution temperature measured in Example 1, that is, in a water bath at 85°C, the test tube was shaken before and after bathing, and it could completely dissolve into a light red viscous solution in about 6 minutes.

[0067] 2. Filter out the polysaccharide in the solution

[0068] Immediately pour the dissolved Tai Sui solution into a 10 mL syringe with a 0.22 µm disposable syringe filter, and press to filter. Reserve the filtrate in a 10 mL sterile centrifuge tube.

[0069] After filtering, pull out the piston rod of the syringe, add sterile water preheated to the melting temperature, and then push the water through the membrane.

[0070] 3. Extraction of 4 major b...

Embodiment 3

[0087] Example 3 Identification and inspection of 5 major fungioids

[0088] 1. Selection and treatment of positive control bacteria

[0089] The known bacteria of bacteria, archaea, fungi, myxomycetes and mycoplasma were selected as the respective positive control bacteria for the identification of the five major fungi, wherein the positive control bacteria for the identification of mycoplasma were Mycoplasma hyorhinois ( M. Hyorhinis ); Pseudomonas was selected as the positive control bacteria for bacterial identification ( Pseudomonas sp. ); the positive control bacteria for the identification of archaea were selected from the inner lake archaea of ​​the salt field ( H. salinus MCCC 4K00021); Fusarium was selected as the positive control bacteria for fungal identification ( Fusarium sp. ); the positive control bacteria for the identification of slime molds were Calderma sp. ( Didymium sp. ).

[0090] Put ≥ 0.1 g of wet bacteria of each positive control bacteria (10...

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Abstract

The invention provides an identification method for existence of fungus in ganoderma lucidum tissue and an assessment method for necessity of fungus for formation of ganoderma lucidum. The identification method comprises the steps of a, measuring the dissolving temperature of a ganoderma lucidum tissue block; b, preparing a ganoderma lucidum tissue dissolving solution; c, filtering the ganoderma lucidum tissue dissolving solution with a disposable needle filter, so as to enrich the fungus that cannot pass through a filter membrane on the filter membrane, and mycoplasma is retained in the filter liquor by passing through the filter membrane; d, taking out the filter membrane in step c and cutting up the filter membrane, then extracting DNA by using a CTAB method; e, selecting respective positive control fungus of five fungi, and processing by the same method as steps b to d; f, identification of the existence of the five fungi: identifying the mycoplasma by taking the filter liquor as an amplification template, and identifying the other four fungi by taking the DNA extracted from the thallus on the filter membrane as an amplification template. By applying the identification method provided by the invention, the existence of the five fungi can be measured, the fungus constitution information in the ganoderma lucidum can be accurately acquired, and a foundation is laid for quicklyand more accurately verifying the biological attributes.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for identifying the presence of fungi in Tai Sui tissues and an evaluation method for the necessity of fungi for the formation of Tai Sui. Background technique [0002] Tai Sui is a mysterious object that is recorded in ancient Chinese books and is being discovered continuously in modern times. Its status in the biological world has been being explored. Since Tai Sui is found in the soil layer without photosynthetic pigments such as chlorophyll, revealing its biological taxonomy basically starts with studying its fungal composition. [0003] Since the first modern Tai Sui discovered in 1992 was identified as a slime mold, the Tai Sui discovered later has been interpreted as a slime mold complex, but so far there has been no confirmatory report of cultivating Tai Sui or its analogues with isolated slime molds , more researchers turned to pay more attention to the existence o...

Claims

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Application Information

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IPC IPC(8): C12Q1/686C12Q1/6806
CPCC12Q1/686C12Q1/6806C12Q2521/107C12Q2527/107
Inventor 王朝江
Owner 河北省农林科学院经济作物研究所
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