Method for simply and quickly separating cercospora monospores

Abstract

The invention relates to a method for simply and quickly separating cercospora monospores. A specially-made spore taking device is adopted, one end of the spore taking device is made into a flat round spore taking ball, and the other end of the spore taking device is made into a spore taking shovel which is slightly sharp at the front end and is in a flat shovel shape. The spore taking device can glue pathogen spores of a small amount, a culture medium is evenly coated with the pathogen spores, it can be guaranteed that one spore arises in a view of 10*10 times, the spore density is low, a target spore is easy to find, blade residual bodies do not adhere easily, the pathogen spores can be separated easily, and the separation rate and separation speed are improved effectively. The chosen spore is a spore which is cultured for 24 h and is germinated, the result that germination cannot occur after single spores are chosen in other methods is effectively avoided, and the success rate is high. The spore is chosen in a mode of spore taking through adhesion, the operation is quick, the success rate is high, the picking efficiency is improved, the operation technique difficulty of the method is not high, the method can be mastered skillfully after several trainings, and the method is easy to handle, simple and easy to learn.

Description

technical field [0001] The invention relates to a simple and rapid method for isolating single spores of the genus Cercospora, which belongs to the technical field of microorganism culture. Background technique [0002] Cercospora fungi are plant pathogenic bacteria that can parasitize on the stems, leaves, flowers or fruits of plants and cause diseases. Maize gray spot disease (Cercosporazeina) (MeiselB, KorsmanJ, KloppersFJ, et al. Cercosporazeina is the causala gent of greyleaf spot disease of maize in southern Africa [J]. European journal of plant pathology, 2009, 124 (4): 577-583.), sugar beet brown spot disease (Cercospollarde, Assolabeticola) etal.Cercosporabeticola-worldwidedistributionandincidence[J].CercosporabeticolaSacc.biology,agronomicinfluenceandcontrolmeasuresinsugarbeet.,2000:5-16.)、辣椒褐斑病(Cercosporacapsici)(KawagoeH.InfluenceoftemperatureandrelativehumidityondiseaseoccurrenceofsweetpeppercausedbyCercosporacapsiciinaplastichouse[J].AnnalsofthePhytopathologica...

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Problems solved by technology

[0004] Wherein, " single spore separator " is a kind of single spore isolation device (ConstantinescuO.Aninstrumentandprocedureforsingle-sporeisolation[J].TransactionoftheBritishMycologicalSociety, 1988,91(4):700-702.), but this device is installed comparatively complicated, needs microscope Equipment transformation, and there is no widespread use of this device in China

In contrast, manual separation is more common, but there are also some shortcomings in different methods, for example, the cutter separation method (HoWC, KoWH. Asimple method for obtaining single-spore isolates offungi [J]. Botanical Bulletin of Academia Sinica, 1997, 38.), needs to be examined under a microscope. Mark the single spore, and then cut the single spore agar block with a cutter, often the cutting range is inaccurate, and the single spore cannot be cut; directly pick the agar plate coated with spores under the microscope, but the petri dish cannot be fixed. Can not move at will, causing separation rate to reduce (Fang Zhongda. Plant Disease Research Methods [M]. Beijing: Agricultural Press, 1998.); 29-38.), eyebrow acupuncture method (Liu Changhua, Wang Zhenhua. Research on the relationship between inoculation concentration and incidence of corn head smut in field[J]. Maize Science, 2008,16(1):119-121.), these methods The spores are basically picked out by picking needles. Most of the tools for making picking needles are insect needles or glass picking needles. The insect needles are relatively thin, so it is easy to determine the location of the spores under a microscope, but the insect needles are soft and difficult to pick out spores Agar blocks, and the hardness of the glass pick is moderate, but the diameter is relatively thick, and the observation under the microscope often occupies the entire field of view, and the position of the spores cannot be determined. At the same time, the tip of the glass pick is fragile, and extra care must be taken during operation

These factors reduce the success rate and operational efficiency of single spore isolation, and limit the application of single spore isolation method to a certain extent.

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