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Complete ITS sequence of dictyophora indusiata, dictyophora duplicate and dictyophora phalloidea and application in identifying variety of dictyophora

A technology of Dictyophora with short skirt and Dictyophora with long skirt, which is applied in the field of molecular identification methods of its varieties, can solve the problems of species confusion, inability to comprehensively and objectively identify Dictyophora varieties, and affect the use value of Dictyophora and the interests of consumers , to achieve the effect of reliable technology

Inactive Publication Date: 2018-07-27
周培富 +4
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the extremely similar morphological characteristics of the three, it leads to confusion among species in terms of selection and breeding, management and protection of germplasm resources, experimental research, and product development of Dictyophora; and due to regional differences, different regions The nutritional value of Dictyophora itself will also have certain differences. If Dictyophora strains are mixed, Dictyophora will be negatively affected in terms of seed selection and breeding, germplasm resource management, experimental research, and Dictyophora product development. It will also directly affect the utilization value of bamboo fungus itself and the interests of consumers
Therefore, it is necessary to accurately identify the species of Dictyophora before conducting relevant research, but it is not possible to make a comprehensive and objective identification of Dictyophora species simply by using morphological characteristics and microscopic anatomy

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 The ITS1-5.8S-ITS2 region sequence of wild Dictyophora strain ZST was used to identify Dictyophora species.

[0030] In this example, the wild bamboo fungus strain ZST (collected by the patent declaration laboratory) was taken, and the identification was carried out through the following steps.

[0031] (1) Preparation of wild Dictyophora strain ZST mycelium.

[0032] Inoculate the wild bamboo fungus strain ZST strain into PDA liquid medium, and cultivate it at a constant temperature of 25°C. When the mycelium grows to an appropriate amount, collect the mycelium and wash the liquid medium contained in the liquid on the mycelium with sterile distilled water. Suck the water in the mycelium with sterile filter paper, put it into a 1.5 mL centrifuge tube, and freeze it at -20°C for later use.

[0033] (2) Extraction of genomic DNA.

[0034] Weigh 1.5 g of mycelia stored at -20°C into a sterilized and frozen mortar, add an equal volume of quartz sand to grind; tr...

Embodiment 2

[0045] Example 2 The sequence of the ITS1-5.8S-ITS2 region of the wild Dictyophora strain YZS was used to identify Dictyophora species.

[0046] In this example, the wild bamboo fungus strain YZS (collected by the patent declaration laboratory) is taken, and the method is the same as in Example 1. The ITS1-5.8S-ITS2 region sequence (SEQ NO.5) of the strain was compared with the homologous sequences of SEQ NO.1, SEQ NO.2 and SEQ NO.3 using DNAMAN software, and it was found that SEQ NO. .5 and SEQ NO.1, SEQ NO.2 and SEQ NO.3 sequence similarities are 97.35%, 99.81% and 99.62%, respectively, so the wild Dictyophora strain YZS is identified as Dictyophora kiltica.

Embodiment 3

[0047] Example 3 The sequence of the ITS1-5.8S-ITS2 region of the wild Dictyophora strain ZZC2 was used to identify Dictyophora species.

[0048] In this example, the wild bamboo fungus strain ZZC2 (collected by the laboratory for patent application) is used, and the method is the same as in Example 1. The ITS1-5.8S-ITS2 region sequence (SEQ NO.6) of the strain was compared with the homologous sequences of SEQ NO.1, SEQ NO.2 and SEQ NO.3 using DNAMAN software, and it was found that SEQ NO. .6 The sequence similarities with SEQ NO.1, SEQ NO.2 and SEQ NO.3 are 97.17%, 99.62% and 99.81%, respectively, so the wild Dictyophora strain ZZC2 is identified as Dictyophora clavium.

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PUM

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Abstract

The invention relates to a DNA sequence of a fungus variety as well as a molecular identification method for identifying a variety of the fungus by using the DNA sequence. An ITS1-5.8S-ITS2 sequence of the dictyophora indusiata is as shown by SEQ NO.1; an ITS1-5.8S-ITS2 sequence of dictyophora duplicate is as shown by SEQ NO.2; and an ITS1-5.8S-ITS2 sequence of the dictyophora phalloidea is as shown by SEQ NO.3. When in identification, a genome of a to-be-identified sample is firstly extracted, an ITS1-5.8S-ITS2 fragment is amplified in a PCR manner, then cloning and sequencing are performed,the obtained sequence is compared separately with the sequences shown by SEQ NO.1, SEQ NO.2 and SEQ NO. 3 so as to determine the variety of the dictyophora indusiata, dictyophora duplicate or dictyophora phalloidea, the variety of the dictyophora indusiata, dictyophora duplicate and dictyophora phalloidea can be accurately distinguished and determined, so that a reliable technology can be providedfor identifying the species, protecting the varieties, controlling the quality of the strain, effectively breeding the strain and artificially cultivating the dictyophora.

Description

technical field [0001] The present invention relates to a DNA sequence of a fungal species and a method for identifying the species by using the sequence, more specifically, the present invention relates to a DNA sequence of a fungal species, a species of Dictyophora longum, a species of Dictyophora kiltus and a species of Dictyophora clavula The ITS1-5.8S-ITS2 sequence and the molecular identification method for identifying its species by using the sequence belong to the technical field of identifying fungal species by molecular biological methods. Background technique [0002] Dictyophora( Dictyophora spp. ) belongs to Basidiomycotina, Abdomycetes, Phalloides, Phalloidaceae, and Dictyophora. It is one of the world's rare large edible fungi, with a total of 12 species. In my country, there are 7 species of Dictyophora with long skirt, Dictyophora with short skirt, Dictyophora with yellow skirt, Dictyophora with scarlet, Dictyophora with red support, Dictyophora with wrinkl...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6869C12Q1/04C12N15/11
CPCC12Q1/6869C12Q1/6895C12Q2531/113
Inventor 周培富汤洪敏丁邦菊张静黎璐
Owner 周培富
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