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ITS sequence of Tremella aurantiablba Bandoni and Stereum hirsutum and method for identifying golden fungus using the same

A technology for the identification of Bacillus pilosa and gold, which is applied to the DNA sequence of fungal species and the use of the sequence to identify its species, the ITS region sequence of the fungus Auricularia auricosa and the use of the sequence to identify its species, which can solve the problem of complex sources of Auricularia aureus, Issues affecting the clinical efficacy of golden ear, consumer interests, comprehensive and objective identification of golden ear varieties, etc.

Inactive Publication Date: 2009-01-21
江苏同源堂生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The sources of commercially available golden ears are complex, involving species with similar colors and shapes such as Tremellaaurantia Schw.ex Fr., Tremella encephala Pers., Tremellamesenterica Retz ex Fr. or Tremella lutescens Fr., or even Some artificially dyed white fungus directly pretends to be golden fungus, which directly affects the clinical efficacy of golden fungus and the interests of consumers
The current identification methods of golden ear include trait identification and microscopic anatomy, but these methods have not yet made a comprehensive and objective identification of the golden ear species

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Using the sequence of the ITS region of Tremella aurantialba Bandoni et Zang to identify the fruiting bodies of Auricularia aurantii

[0031] (1) get the fungus fruiting body similar to golden ear in appearance, extract the genome DNA of fruiting body, concrete method is as follows:

[0032]Aseptically collect and sterilize 0.1 g of the fruiting body to be identified and put it in a sterilized mortar, add 0.5-1 mL of extraction buffer, this extraction buffer is 0.15 mol / L tris hydroxymethyl aminomethane solution with pH 9.0 and 7 mL It is prepared by mixing 8 mL of 0.05 mol / L disodium ethylenediamine tetraacetic acid solution with a pH of 8.0, then distilling to 100 mL, grinding and mixing; transfer the homogenized liquid into a sterile centrifuge tube, and add 0.1-0.2 mL of mass 10% sodium dodecylsulfonate solution and 0.3-0.6mL benzyl chloride reagent, mix well, keep warm at 50°C for 1-2 hours; add 0.3mL 3mol / L NaOAc (pH 5.2) and mix well , ice-water bath f...

Embodiment 2

[0050] Example 2 Using the sequence of the ITS region of Tremella aurantialba Bandoni et Zang to identify Auricularia species CGMCC0179

[0051] (1) Extract the genomic DNA of the existing spore strain CGMCC0179 (China General Microorganism Culture Collection and Management Center) to be identified, the specific method is as follows:

[0052] Aseptically collect 0.1 g of artificially cultured spore strains to be identified for 3 days in a sterilized mortar, add 0.5-1 mL of extraction buffer, the extraction buffer is 0.15 mol / L at pH 9.0 It is prepared by mixing 7 mL of tris hydroxymethyl aminomethane solution and 8 mL of 0.05 mol / L disodium ethylenediamine tetraacetic acid solution with a pH of 8.0, then distilling to 100 mL, grinding and mixing; transfer the homogenized liquid into a sterile centrifuge In the tube, add 0.1-0.2mL of 10% sodium dodecylsulfonate solution and 0.3-0.6mL benzyl chloride reagent, mix thoroughly, and keep warm at 50°C for 1-2 hours; add 0.3mL 3mol / ...

Embodiment 3

[0070] (1) Extract the mycelium genome DNA of the existing bacterial species to be identified, the specific method is as follows:

[0071] Aseptically collect 0.1 g of the mycelium of the strain to be identified that has been cultured for 5 days and place it in a sterilized mortar, add 0.5-1 mL of extraction buffer (the extraction buffer is 0.15 mol / L trimethylol at pH 9.0 7mL of diaminomethane solution and 8mL of 0.05mol / L disodium edetate solution with a pH of 8.0 were mixed, and the volume was adjusted to 100mL) and ground and mixed. Transfer the homogenized liquid into a sterile centrifuge tube, add 0.1-0.2mL of 10% by mass sodium dodecylsulfonate solution and 0.3-0.6mL of benzyl chloride reagent, mix well, and store at 50°C Keep warm for 1-2 hours. Add 0.3mL 3mol / L NaOAc (pH 5.2) and mix well, put in ice-water bath for 15-20min, and centrifuge at room temperature at 6000r / min for 15-20min. Take the supernatant and add an equal volume of absolute ethanol to precipitate a...

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Abstract

The invention relates to a DNA sequence of a fungal strain and a method for identifying variety by means of the sequence. An ITS zone sequence of tremella aurantiabla is shown in SEQ ID NO.1, and an ITS zone sequence of host fungus stereum hirsutum is shown in SEQ ID NO.2. During identification, an ITS zone sequence of a to-be-identified sample is extracted at first and then is respectively compared with the SEQ ID NO.1 sequence and the SEQ ID NO.2 sequence to obtain a result. The method can accurately identify the authenticity of a to-be-identified fungus, and can also carry out accurate identification to artificially cultured yeast-like spore strain of tremella aurantiabla and host fungus stereum hirsutum strain; moreover, the method can accurately distinguish and identify the authenticity and purity of tremella aurantiabla variety or strain, thereby providing a reliable technique for variety identification, protection, strain quality control and effective strain breeding.

Description

technical field [0001] The present invention relates to a DNA sequence of a fungus variety and a method for identifying its variety by using the sequence. More specifically, the present invention relates to the sequence of the ITS region of the fungus Auricularia auricosa and a method for identifying its variety by using the sequence, which belongs to molecular biology methods The technical field of identification of fungal species and detection of fungal purity. Background technique [0002] Tremella aurantialba Bandoni et Zang is a precious edible and medicinal fungus of the genus Tremella. It is rich in nutrients and can provide a variety of amino acids, vitamins and mineral elements necessary for the human body. It is a good biological nutrition resource. Golden ear has a long history of medicinal use, and its shape, medicinal parts and properties, flavors and functions are described in detail in "Famous Doctors" and "Compendium of Materia Medica". "Chinese Medicinal Fu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/31
Inventor 刘春卉瞿伟箐俞建国马维新
Owner 江苏同源堂生物工程有限公司
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