High throughput isolation method of fungi

A separation method and technology of pathogenic fungi, applied in the field of separation of strong parasitic pathogenic fungi, can solve problems such as high technical requirements and complicated operations

Inactive Publication Date: 2014-06-25
QINGDAO VLAND BIOTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These single spore isolation methods are complicated to operate and require high technical requirements, which limits their application.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Acquisition of dry rot spores: Pour 10ml of 1% sterile water agar into a sterile petri dish and cool. Dry rot diseased branches with small black spots were collected from the field, and after the surface was wiped and disinfected with 70% alcohol, the diseased skin was cut and pasted on water agar. Invert the petri dish, place a piece of agar for single spore isolation directly under the diseased skin, with the side of the agar facing the diseased skin. Seal the petri dish and keep it moist at room temperature for 2-12 hours. The ascospores in the mature ascus on the diseased skin will be sprayed onto the water agar sheet, and single spores will be picked under a microscope and transferred to PAD for culture.

Embodiment 2

[0016] The conidia of the dry rot fungus were collected from the dry rot diseased branches with small black spots in the field, and the surface was wiped with 70% alcohol for disinfection. Cut off the infected infected skin, soak it with sterile water, place it in a sterile petri dish, and keep it moist at room temperature (20-25 °C) for 12 hours, and the white and elongated conidia horns can be extruded from the sporozoites. Use a capillary needle to pick the spore horns, spread them on the water agar sheet, pick single spores, and transfer them to PDA for culture.

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PUM

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Abstract

The invention belongs to the field of biotechnology, and provides an isolation method for highly parasitic pathogenic fungi, for study in the future. Single spore isolation is an effective method for isolating the highly parasitic pathogenic fungi, and is particularly suitable for isolating the pathogenic fungi that suffer severe pollution caused by isolation with a conventional method and are difficult to isolate successfully, namely, venturia nashicola, marssonina mali and cercospora chionea. A conventional single spore isolation technology is complex in operation and high in technical requirements, thereby restricting applications thereof. In the isolation method provided by the invention, by punching holes with capillary tubes having the diameter of 1 mm, an operation process is greatly simplified; operability of single spore isolation is stronger; and speed is faster. A general technical staff can master the technology through simple training, thereby broadening an application scope of the method. A good effect can be achieved by mainly adopting the method to isolate the highly parasitic pathogenic fungi and ascospore.

Description

technical field [0001] The invention belongs to the field of biological technology, and mainly relates to a method for isolating strong parasitic pathogenic fungi. Background technique [0002] Strong parasitic fungi mainly parasitize on living plants, obtain necessary nutrients from living host tissues, and grow slowly on artificial medium. In addition to pathogenic bacteria, there are also endophytic bacteria, saprophytic bacteria and other miscellaneous bacteria in the host tissue of the disease. Because these miscellaneous bacteria grow quickly on the artificial medium, the separation of strong parasitic fungi by conventional diseased tissue isolation methods is often contaminated by miscellaneous bacteria, and the success rate of isolation is very low. Single spore isolation can avoid contamination caused by miscellaneous bacteria in diseased tissues, and is used for the isolation of strong parasitic fungi with a high success rate. [0003] Single spore isolation is t...

Claims

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Application Information

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IPC IPC(8): C12N1/14
Inventor 王英俊邱梅
Owner QINGDAO VLAND BIOTECH INC
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