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Molecular marker for identifying white Pleurotus pulmonarius strain Qian side 02 and specific primer pair

A lung-shaped Pleurotus and molecular marker technology, applied in the direction of microorganism-based methods, microorganisms, biochemical equipment and methods, etc., can solve the problem of undiscovered SCAR markers, and achieve real-time and on-site detection, strong specificity, and identification accurate effect

Active Publication Date: 2022-02-18
GUIZHOU INST OF SOIL & FERTILIZER
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] After searching, no report was found on the use of SCAR markers for the identification of Pleurotus sinensis 02 species

Method used

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  • Molecular marker for identifying white Pleurotus pulmonarius strain Qian side 02 and specific primer pair
  • Molecular marker for identifying white Pleurotus pulmonarius strain Qian side 02 and specific primer pair
  • Molecular marker for identifying white Pleurotus pulmonarius strain Qian side 02 and specific primer pair

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Specific Random Primer Screening Test of Example 1 Pleurotus pulmonica Strain Qianfang 02

[0027] (1) Test strains: Qianfang 02 (white Pleurotus pleurotus domesticated and cultivated by Guizhou Institute of Soil and Fertilizer, preserved in China Center for Typical Cultures, and its preservation number is CCTCC NO: 2021259); Jingxiu 1 (Beijing Pleurotus pleurotus strain bred by Jiji Garden); Pteris cerevisiae (light gray Pleurotus pleurotus strain bred by Jijiyuan in Beijing); Xiaxiu 004 (brown lung-shaped Pleurotus strain bred by Jiangsu Tianlong Fungus Industry) Pleurotus strain); Xiu 03 (grey lung-shaped Pleurotus strain from Chuangjian Taijiang, Qiandongnan).

[0028] (2) Test method

[0029] (1) Mycelia culture:

[0030] Qianbian 02, Jingxiu No. 1, Pteris edodes, Xiaxiu 004 and Xiu 03 preserved in low temperature were respectively inoculated into different PDA medium (prepared PDA medium according to the usual method; 200g of potatoes in every 1L medium, glucose...

Embodiment 2

[0041] Example 2 The design of the Guizhou side 02 specific primer pair and the screening test of SCAR molecular markers

[0042] Proceed as follows:

[0043] (1) Test strain: with embodiment 1.

[0044] (2) Test method:

[0045] (1) Design of specific primer pairs

[0046] According to the nucleotide sequences of 6 specific DNA fragments of Guizhou side 02 shown in Table 2, adopt PRIMER5.0 software to design specific primer pairs; Described specific primer pairs are shown in Table 3:

[0047] Table 3 Specific primer pairs designed for Guizhou side 02

[0048]

[0049]

[0050] (2) Genomic DNA extraction. According to the method described in step (2) of Example 1, the genomic DNAs of Qianbian 02, Jingxiu No. 1, Pteris praecox, Xiaxiu 004 and Xiu 03 were extracted respectively.

[0051] (3) SCAR-PCR amplification

[0052] The genomic DNA of the different strains described in (2) was used as a template, and the 6 pairs of primers shown in Table 3 were used for SCAR-PCR ...

Embodiment 3

[0057] Embodiment 3 Utilizes specific primer pair of the present invention to the comparative identification test of bacterial strain Qianbian 02

[0058] (1) Test strain: with embodiment 1.

[0059] (2) Test method:

[0060] (1) Genomic DNA extraction. According to the method described in step (2) of Example 1, the genomic DNAs of Qianbian 02, Jingxiu No. 1, Pteris praecox, Xiaxiu 004 and Xiu 03 were extracted respectively.

[0061] (2) Perform PCR amplification using the genomic DNA extracted in step (1) as a template and MM16-F and MM16-R as primers; see step (3) of Example 2 for the reaction system and reaction conditions of PCR amplification. Take 4 μL of each of the above five PCR products and mix them with 0.5 times the volume of 6×Loadding Buffer respectively. The amplified products are detected by 1.5% agarose gel electrophoresis, placed in 1×TBE buffer, the voltage is 120V, and the electrophoresis time is 30min. After electrophoresis, stain with EB for 10 min, and...

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Abstract

The invention belongs to the field of edible mushroom strain molecular identification and in particular, discloses a molecular marker for identifying white Pleurotus pulmonarius strain Qian side 02, the molecular marker is a DNA fragment, and the nucleotide sequence of the DNA fragment is shown as SEQ ID NO: 1. The invention also discloses a specific primer pair for amplifying the molecular marker and a detection kit containing the primer pair. The SCAR molecular marker and the specific primer have strong specificity to the Qian side 02 of the strain, are accurate and reliable to identify the Qian-side 02 of the strain, and can provide a sufficient and reliable basis for infringement processing of the Qian side 02; and secondly, the SCAR molecular marker detection method is simple and rapid, and is suitable for real-time and field detection.

Description

technical field [0001] The invention belongs to the field of molecular identification of edible fungus strains, and in particular relates to a molecular marker for identifying the Pleurotus pleurotus strain Qianfang 02, and also relates to a specific primer pair for amplifying the molecular marker. Background technique [0002] Lung-shaped Pleurotus (also known as Xiuzhen mushroom. Alias: Pleurotus pleurotus, Pleurotus phoenix, Indian oyster mushroom or Himalayan oyster mushroom, etc.; its Latin name: Pleurotus pulmonarius (Fr.) Quél.) belongs to the genus Pleurotus of the family Pleurotus. The lung-shaped Pleurotus is small and beautiful in appearance, not only delicate in texture and delicious in taste, but also rich in nutrition, rich in protein, sugar, unsaturated fatty acid, vitamins, folic acid and various trace elements; the polysaccharide contained in it has anti-tumor function, It can also be used to improve human immunity. Therefore, Pleurotus aeruginosa is regard...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11C12R1/645
CPCC12Q1/6895C12Q1/6858C12Q2600/156C12Q2531/113C12Q2565/125Y02A50/30
Inventor 王晓敏杜慕云杨珍陈旭张邦喜朱森林陈波张钦语唐玲杨仁德
Owner GUIZHOU INST OF SOIL & FERTILIZER
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