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DNA numerator identification method for golden fungus and host epiphyte boreostereum vibrans

A technology of basilar bacteria and host, applied in the field of DNA molecular identification of fungal strains, can solve the problems of inability to judge the accuracy of strains and low yield, and achieve the effect of accurate distinction and identification and reliable technology

Inactive Publication Date: 2008-12-03
江苏同源堂生物工程有限公司
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

This requires that the culture must be accurately identified before cultivation, but the accuracy of the strain cannot be judged solely by morphological characteristics.
Chinese patent (ZL200410022377.3) discloses a method for cultivating yeast-like spores of Auricularia aureus, in order to overcome the weakness of the low yield of Auricularia aureus fruiting body, to find alternative resources of Auricularia auricula, and the yeast-like spores of Auricularia auricula also exist in this patent The problem of bacterial identification

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0019] Example 1 Molecular identification method of the large ribosomal subunit gene D1 / D2 region sequence of Tremella aurantialba Bandoni et Zang

[0020] In this embodiment, Tremella aurantialba Bandoni et Zang bacterial classification CGMCC0179 (China General Microorganism Culture Collection Management Center) is taken for identification by the following steps:

[0021] (1) Genomic DNA extraction, the specific method is as follows:

[0022] 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 of pH 9.0 Tris hydroxymethyl amino methane solution 7mL and pH 8.0 0.05mol / L disodium ethylenediaminetetraacetic acid solution 8mL were mixed and then adjusted to 100mL), ground and mixed; the homogenized liquid was transferred into sterile Add 0.1-0.2mL of 10% sodium dodecylsulfonate solution and 0.3-0.6mL benzyl chloride reagent to the Eppendorf tube...

Embodiment 2

[0031] Example 2 Molecular identification method of the large ribosomal subunit gene D1 / D2 region sequence of the host fungus Stereum hirsutum (Willd.) Fr.

[0032] In this embodiment, the strains to be identified whose thalline morphology is hyphae are taken, and are identified through the following steps:

[0033] (1) Genomic DNA extraction, the specific method is as follows:

[0034] Aseptically collect 0.1 g of artificially cultured mycelia to be identified for 5-7 days in a sterilized mortar, add 0.5-1 mL of extraction buffer (the extraction buffer is 0.15 of pH 9.0 mol / L tris hydroxymethyl aminomethane solution 7mL and pH 8.0 0.05mol / L disodium ethylenediaminetetraacetic acid solution 8mL were mixed, then the volume was adjusted to 100mL), ground and mixed; the homogenized liquid Transfer to a sterile Eppendorf tube, add 0.1-0.2mL of 10% sodium dodecylsulfonate solution and 0.3-0.6mL benzyl chloride reagent, mix well, and incubate at 50°C for 1-2 hours; Add 0.3-0.6mL 3...

Embodiment 3

[0044] In this embodiment, the strains to be identified whose thalli form is yeast-like are taken, and the identification is carried out through the following steps:

[0045] (1) Genomic DNA extraction, the specific method is as follows:

[0046] Aseptically collect 0.1 g of the 3-day cultured thallus of the spore strain to be identified and place it in a sterilized mortar, add 0.5-1 mL of extraction buffer (the extraction buffer is 0.15 mol / L trihydroxy Mix 7 mL of methylaminomethane solution with 8 mL of 0.05 mol / L ethylenediaminetetraacetic acid disodium solution with a pH of 8.0, then dilute to 100 mL), grind and mix; transfer the homogenized liquid into a sterile Eppendorf 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 0.3-0.6mL Mix with 3mol / L NaOAc (pH 5.0), ice-water bath for 15-20min, centrifuge at room temperature at 6000r / min for 15-20min; take the supernatan...

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Abstract

The invention relates to a molecular identification method of a ribosome large-subunit gene D1 / D2 region sequence of a golden tremella and a host fungi crude Stereum hirsutum. The ribosome large-subunit gene sequence of the golden tremella is shown in SEQ ID NO.1, and the ribosome large-subunit gene D1 / D2 region sequence of the host fungi crude Stereum hirsutum is shown in SEQ ID NO.2. The molecular identification method has the steps as follows: the ribosome large-subunit gene D1 / D2 region sequence of the strains to be identified is obtained after extraction, and is judged through being compared with the sequence in the SEQ ID NO.1 and the SEQ ID NO.2, so as to judge whether the strains to be identified is the golden tremella strains or host fungi crude Stereum hirsutum strains. By utilizing the molecular identification method, the golden tremella strains and the host fungi crude Stereum hirsutum strains which are cultured artificially can be identified accurately; the reliable technology is provided for strains identification, breed conservation, strain quality control, effective strain propagation, and artificial cultivation of the golden tremella.

Description

technical field [0001] The invention relates to a DNA molecular identification method of fungal strains, more specifically to a molecular identification method of the large ribosomal subunit gene D1 / D2 region sequence of golden ear and host fungus P. Background technique [0002] Golden ear Tremella aurantialba Bandoni et Zang is rare in the wild and endemic to China. Due to the particularity of life, golden ear fungus must parasitize on the surface of Stereum hirsutum (Willd.) Fr. and form a heterogeneous complex together. Therefore, the existence of a small amount of host fungi is the normal growth of golden ear fungus. basis of development. Due to the strong ability of Bacillus pilosae to decompose and utilize carbohydrates such as lignocellulose, it is very easy to be in a growth advantage in the growth and development history of golden ear. Mycelial culture, but it is mistaken for golden ear mycelia, and it is difficult to obtain a real golden ear product by using thi...

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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