Method for separating single fungus spore

Abstract

The invention discloses a method for separating a single fungus spore. According to the technical scheme of the invention, the method comprises the following steps of: inserting a glass capillary tube into a Pasteur pipet from a tip; enclosing the tip pipe orifice of the Pasteur pipet with a Parafilm sealing film; picking a fungus material to be separated by using a dissecting needle; positioning the center of the pipe orifice of the capillary tube over against and close to the spore; and transferring the spore to be upper side of a culture medium to finish fungus and spore separation, wherein the culture medium is a PDA (Potato Dextrose Agar) culture medium or SDA (Sahli Dextrose Agar) or MEA (Malt Extract Agar) culture medium. The Pasteur pipet is combined with glass capillary tubes of different specifications, so that sucking and transferring operation of various fungus spores is realized. The method has the characteristics of low preparation cost, easiness of manufacturing and operation, convenience for detaching and sterilizing, reusability and the like. Meanwhile, the defect that various conventional methods are unsuitable for separating fungus spores from a liquid environment is made up.

Description

technical field [0001] The invention relates to a separation method, in particular to a separation method for single spores of fungi, and belongs to the technical field of microorganism cultivation. Background technique [0002] The pure culture obtained from single spore culture is the basis for the isolation and identification of fungi and basic physiological and biochemical research, so it is particularly important to isolate fungal single spores. At present, there are many methods for single spore isolation, such as using tools such as fixed insect needles, eyebrow needles, glass capillaries, and handmade glass needles for single spore isolation. American scientist Hansen disclosed a method of separating single-spores using glass capillary in "Science" 1926 64:384 "A simple method of obtaining single-spore cultures". After the medium is solidified, the capillary fragments with single spores are intercepted under the microscope for subsequent cultivation. This method re...

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Problems solved by technology

This method requires high experimental skills. On the one hand, it is difficult to suck the medium into the capillary and intercept the capillary fragment of a single spore; Fungaldiversity" 1999 2: 47-63 "Single-spore isolation using a hand-made glassneedle" reported a hand-made glass needle for single-spore isolation, which can realize the separation and transfer of various fungal spores, However, the production and use of glass needles in this method are relatively difficult, and they are easily broken and inconvenient to preserve; Cai Rongrong and Jin Minzhong introduced a single-spore direct picking to obtain a single-spore strain in "Plant Protection" at 3:53 in 1991. In the "method", the isolation of single spores of pathogenic bacteria was realized with the help of insect needles fixed on the microscope stage, but the operation of this method was cumbersome, and because the insect needles were thick and fixed, they lacked flexibility and were very difficult to obtain. It is difficult to actually pick and transfer individual spores; Sun Guangyu and Zhang Tianyu used a bent tip fixed on a glass rod in "Plant Protection" 1996, 10(1): 40-41 "A Simple Single Spore Separation Technique". Insect Needle made a spore-picking needle. Although this tool is more convenient to operate, it is still limited by the fact that the insect needle is too thick compared to the spores, which is not conducive to the realization of single spore picking, and is not suitable for high humidity and liquid environments. single spore picking operation; Lin Daifu proposed a special eyebrow picking method in "Mountain Agricultural Biology Journal" 2003 22 (5): 462-463 "eyebrow picking method for single spore isolation", which can be very good The separation of single spores can be achieved, but this method is still not suitable for picking spores from a liquid environment, and its production is cumbersome and not conducive to preservation

So far, although there are many methods for single spore isolation, they all have certain limitations in operation, application and storage, and cannot successfully isolate single spores for various operating environments.

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