Method for testing high-pollution background fungi by separation and purification

A technology for separation, purification and inspection method, which is applied in the field of separation, purification and inspection of highly polluting background fungi, which can solve the problems of not being able to meet the requirements of separation purity, and cannot ensure the reduction of background interfering bacteria pollution, so as to achieve good fungal inspection adaptability and improve Biosafety, the effect of improving experimental efficiency

Pending Publication Date: 2019-12-10
郑萍
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Problems solved by technology

[0003] Because the background of the isolated Trichoderma is rich in various microorganisms, such as spores, actinomycetes, penicillium, aspergillus, phage, etc., the content is extremely high. Therefore, the liquid dilution method cannot ensure that t

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1 uses the isolation, screening and purification of Trichoderma species. It is used as follows:

[0061] The water agar with 1.5% agar content was sterilized at 121° C. for 15 minutes, spread on a plate with a thickness of about 3 mm, and was set aside after solidification.

[0062] The separation layer medium was based on PDA and then added with VB1: 8mg / L, 5ml of Eupatoria violet extract, and 5% cellulose. Lay flat plates with a thickness of 3mm, and set aside after cooling.

[0063] Weigh 2g of the soil and spread it into a circle with a diameter of 20mm with a sterile applicator stick. Remove the outer water agar to prepare a sample separation dish.

[0064] After sterilizing the glass ring, paste it on the medium of the separation layer to prepare a separation dish, buckle it with the sample separation dish, and place it upright in a 22°C incubator for cultivation.

[0065] When the aerial hyphae of filamentous fungi produced by the soil on the feeder m...

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PUM

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Abstract

The invention discloses a method for testing high-pollution background fungi by separation and purification. A double-sided forward fungi purification and separation culture method is adopted, according to a double-sided forward separation method, two culture media are arranged in one culture dish, air is taken as a separation medium, aerial hyphae are induced to grow towards a target culture layer through allelochemicals, or spores are induced to be produced through a feeding layer, and fungi cysts are used for cracking and ejecting spores to separate and purify single spores. The biologicalsafety can be improved, the experiment time can be shortened, the experiment efficiency can be improved, and used consumables and added toxic substances can be reduced; the method has high flexibility, one or more inhibitors or growth stimulants can be added to any culture layer, growth of the fungi can be interfered through the allelochemicals serving as signals, and the purpose of separation andpurification is achieved; the phage pollution of industrial fungi can be well separated, and a bacterial strain polluted by the phage is favorably purified; and the method is simple, convenient and easy to popularize, and has good fungus detection adaptability.

Description

technical field [0001] The invention relates to a biological detection technology, in particular to a method for separation and purification of high-contamination background fungi. Background technique [0002] As we all know, Trichoderma is difficult to purify and isolate due to the growth of many symbiotic fungi and bacteria in the habitat, and due to the competitive growth among fungi and spore scattering, the isolation effect is very poor and there are biosafety risks. Moreover, the traditional fungal culture experiment period is long, which greatly reduces the experimental efficiency. Trichoderma is one of the most widely used microbial species in industry. [0003] Because the background of the isolated Trichoderma is rich in various microorganisms, such as spores, actinomycetes, penicillium, aspergillus, phage, etc., the content is extremely high. Therefore, the liquid dilution method cannot ensure that the pollution of the background interfering bacteria is reduced....

Claims

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Application Information

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IPC IPC(8): C12N1/14C12N1/02C12R1/645
CPCC12N1/14C12N1/02
Inventor 郑萍周翔
Owner 郑萍
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