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Method for identifying predator nematophagous hyphomycete arthrobotrys through DNA bar codes

A technique for Arthrospora and Mycelia, which is applied in the field of fungal species identification, and achieves the effects of low completeness requirements, easy comparison, and improved identification efficiency.

Inactive Publication Date: 2015-11-18
YUNNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, it has not been reported that a specific DNA barcode can be applied to the identification of all fungi, so for specific fungal groups, experiments are still needed to find a suitable gene sequence to use as a DNA barcode

Method used

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  • Method for identifying predator nematophagous hyphomycete arthrobotrys through DNA bar codes
  • Method for identifying predator nematophagous hyphomycete arthrobotrys through DNA bar codes
  • Method for identifying predator nematophagous hyphomycete arthrobotrys through DNA bar codes

Examples

Experimental program
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Effect test

Embodiment 1

[0018] Embodiment 1: The establishment of the gene bank of the RPB2 gene sequence fragment that preys on the nematode hyphomycetes Arthrobush sp. DNA barcode

[0019] (1) There are 32 species and 54 strains of the selected strains that prey on the nematode hyphomycete Arthrobush, all of which are recorded in "Arthroxa and related genera" in Volume 33 of "Mycology of China" Species with a clear concept of spores, and each common species contains at least 3 strains to meet the needs of the subsequent calculation of the maximum genetic variation value within the species. See Table 1 for details.

[0020] Table 1 Sample information sheet

[0021]

[0022]

[0023]

[0024] (2) Inoculate the pure culture strains of the samples listed in Table 1 on a CMA plate [corn flour medium: 30 g of corn flour, 16 g of agar, 1000 ml of water, natural pH, corn flour and an appropriate amount of water and boil for 30 minutes, filter with four layers of gauze, and the filtrate Add water...

Embodiment 2

[0028] Example 2: Identification of unknown species of Arthrobush fungus

[0029] (1) The suspected but unidentified Arthroplexia pure culture strain SP1 was selected and divided into two parts, one was identified by traditional taxonomic methods, and the other was identified by the method used in the present invention.

[0030] (2) Using the steps described in Example 1 to extract the genomic DNA in the strain, using SEQ ID NO.55 and SEQ ID NO.56 as primers to amplify the RPB2 gene sequence fragment. As shown in Example 1, PCR product detection, sequencing, and DNA barcode sequence acquisition were carried out, and then the following results were judged.

[0031] (3) By comparing the sequence of the fungus to be identified with the existing sequence in the gene bank, the genetic distance method based on Kimura-2-parameter probability and the phylogenetic tree method based on cluster analysis are used to establish identification rules. Identify species and obtain the similari...

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Abstract

The invention provides a method for identifying predator nematophagous hyphomycete arthrobotrys through DNA bar codes and belongs to the field of fungus species identification. According to the method, RPB2 genes serve as target DNA bar code genes for identifying the predator nematophagous hyphomycete arthrobotrys, combined arthrobotrys sample experiment data and target fungus data merged strategy is adopted to establish a high-cavity database, meanwhile, RPB2 genes to be identified are compared with a gene library, an identifying rule is established based on a system generation tree method of genetic distance method and clustering analysis of the Kimura-2-parameter probability to identify species. The method has the advantages that the RPB2 genes serve as the DNA bar codes most suitable for identifying the nematophagous hyphomycete arthrobotrys, and the method is universal and easy to amplify and compare. The identifying efficiency and the reliability and accuracy of the identifying method are greatly superior to those of a conventional DNA bar code method, and the method makes up for the blank of the nematophagous hyphomycete arthrobotrys DNA bar code molecular markers and the method provides a useful research tool for researches on germplasm resource excavation, biocontrol application and genetic diversity of nematophagous hyphomycete.

Description

Technical field: [0001] The invention belongs to the field of identification of fungal species, in particular, the invention relates to a method for identifying nematode-predating Hyphomycetes Arthrobush by using a DNA barcode. Background technique: [0002] Plant parasitic nematodes are a kind of plant diseases that commonly occur in the world. At present, there are more than 70 known species of root-knot nematodes, which harm more than 3,000 plants. The global annual losses due to nematodes are as high as 158 billion US dollars. . In my country, nematodes harm almost all crops such as vegetables, tobacco, flowers, forest trees, Chinese medicinal materials, cotton, soybeans, etc. Pine wood nematodes are called smokeless forest fires, which occur widely in many provinces and regions in China, and become a major cause of agricultural and forestry production. One of the important limiting factors. Faced with such serious nematode infestation, a variety of nematicidal reagents...

Claims

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Application Information

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IPC IPC(8): C40B50/06G06F19/24
Inventor 张颖张克勤张云润董建勇和晓霞徐建平
Owner YUNNAN UNIV
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