A kind of single spore separation method of wheat powdery mildew

Abstract

The invention provides a single spore isolation method of Blumeria graminis, and belongs to the technical field of plant pathogenic obligate fungus parasite isolation method. The single spore isolation method comprises following steps: 1, field Blumeria graminis infected samples are collected, and indoor spore production is carried out so as to obtain samples for further processing; 2, an electrically fluorinated liquid is adopted to prepare a spore suspension liquid; 3, suspension liquid coating is carried out to prepare single spore fungus dishes; 4, the single spore fungus dishes are selected and are inverted placed on wheat detached leaf segments; 5, the inoculated leaf segments are delivered into an illumination incubator for 10 days of culturing so as to obtain pure Blumeria graminis fungus colonies. The single spore isolation method is simple and convenient, and is capable of obtaining pure cultured Blumeria graminis at a relatively high success rate via separation, and solving a problem that conventional fungus separation method is not suitable for Blumeria graminis single spore isolation.

Description

technical field [0001] The invention belongs to the application technical field of a method for isolating plant pathogenic obligate parasites, and in particular relates to a method for isolating a single spore of wheat powdery mildew. Background technique [0002] The single spore isolation of pathogenic bacteria is the basic technology of fungal disease research, and it is widely used in the genetic variation, physiological and biochemical characteristics of fungi, as well as the purification and preservation of strains. Currently commonly used single spore isolation methods are: agar plate dilution method (Fang Zhongda. Plant Disease Research Methods (3rd Edition). Beijing: China Agricultural Press, 1998:137-139. Choi Y W, Hyde K D, HoW H.Single spore isolation of fungi.Fungal Diversity,1999,3:29-38.); imprint pipetting single spore isolation method (Zhao Ning. imprint pipetting single spore isolation method. Plant Protection, 1988,14(1):48-49 .); small slide single spore...

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Problems solved by technology

[0003] Since wheat powdery mildew is an obligate parasite, it needs to obtain necessary nutrients from living host tissues and cannot grow on synthetic media, the above method is not suitable for the isolation of single spores of wheat powdery mildew

And in the existing literature, it is generally to obtain its "single spore heap isolate", this kind of single spore heap visible to the naked eye cannot discharge the "single" colony formed by the germination of two or more single spores, thus affecting Pathogenic mechanism, single spore sequencing, etc.

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