Method for AMF spore surface disinfection

Abstract

The invention discloses a method for AMF spore surface disinfection, with the technical scheme comprising: putting filter paper and absorbent paper respectively inside several culture dishes; preparing spore surface antiseptic solution; putting the filter paper carrying AMF spores between the double-layer filter paper inside the culture dish; absorbing the antiseptic solution for dropping on the upper filer paper in a dropwise manner; shifting the double-layer filter paper inside the culture dish with filter paper for absorbing the adsorbed antiseptic solution via the filter paper; taking outthe upper filter paper and putting the upper filter paper inversely inside another culture dish, and transferring the AMF spores on the filter paper, thereby completing the surface disinfection. The invention realizes the surface disinfection of AMF spores through the combination of double-layer filter paper and culture dishes. The method is easy and mild to operate, consumes relatively low amount of antiseptic solution, and causes no loss of spores before or after disinfection, without affecting the vitality of the spores and without the assistance of other apparatus.

Description

technical field [0001] The invention relates to a surface disinfection method, in particular to a surface disinfection method for arbuscular mycorrhizal fungus spores, which belongs to the field of microorganism cultivation. Background technique [0002] Arbuscular mycorrhizal fungi (AMF) are a type of plant symbiotic bacteria that cannot be purely cultured. In the study of the non-symbiotic stage and physiology and biochemistry of AMF, it is often necessary to disinfect the surface of its spores to exclude the interference of other microorganisms. Because the spores of AMF have the characteristic of sticking to the wall, in general utensils, they tend to be attached to the wall of the utensil, and it is difficult to make it completely submerged in the disinfectant solution, so that it cannot be well sterilized. [0003] The most commonly used AMF spore surface disinfection methods are: "Kitasato beaker" method and "Eppendorf surface sterilized" method. The "Kitasato beaker...

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Problems solved by technology

The "Kitasato beaker" method was proposed by the British scientist Mosse in "Transactions of the British Mycological Society" 1959 42: 273-286 "The regular germination of resting spores and some observations on the growth requirements of an Endogone sp.causing vesicular-arbuscular mycorrhiza" , is a method of disinfection based on a suction filtration system. The purpose of surface disinfection of spores is achieved by continuously adding disinfectant so that the spores are always immersed in the disinfectant. Although this method has a good disinfection effect, its operation is complicated and requires Auxiliary of special instruments, such as vacuum pump and special aseptic hood, in addition, this method will lose a large number of spores when the operating area is large; the "Eppendorf surface sterilized" method was published by Spanish scientist Cano et al. Cultures open new prospects for basic research in arbuscular mycorrhizas" proposes a method of disinfecting the surface of spores by centrifugation. This method is to transfer the spores to an Eppendorf tube containing a disinfectant, and use centrifugal force to sterilize the spores. The spores are immersed in the disinfectant to achieve surface disinfection. This disinfection method requires the assistance of a centrifuge. Although the surface disinfection of AMF spores can be achieved to a certain extent, due to the intensive operation, the activity of the spores will decrease after surface disinfection.

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