Method for carrying out spilling separation on ralstonia solanacearum of casuarina

Abstract

The invention discloses a method for carrying out spilling separation on ralstonia solanacearum of casuarina. The method comprises the following steps of digging a casuarina disease root which is 8 to 15 cm in length, washing cleanly, and cutting two ends of the casuarina disease root into flat new sections; then soaking one part, which is 3 to 5 cm in length, of one end into sterile water for 4 to 12 hours, and enabling milky bacterial ooze to flow out from the section on the other end of the casuarina disease root; picking the bacterial ooze by using an inoculating needle, carrying out streak cultivation on a bacteria culture medium, and carrying out amplification culture by selecting a separated typical single colony. The method for carrying out spilling separation on the ralstonia solanacearum of the casuarina, disclosed by the invention, has the advantages of easiness in operation, simpleness, convenience and quickness, high reliability and low content of infectious microbes.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for spilling and separating casuarina solanacearum. Background technique [0002] At present, the isolation of Ralstonia solanacearum from Casuarina generally adopts the conventional method of plant bacteria isolation—dilution separation method. The dilution separation method is a routine method for the isolation of plant bacteria and has long been used in the isolation of Casuarina R. solanacearum. Its technical principle: Under natural conditions, Ralstonia solanacearum invades from the root cap of the root system, invades the vascular bundles of the xylem of the root system through the intercellular space, reproduces and spreads in a large number in the root vascular bundles, and finally blocks the liquid flow in the vascular bundles Cause the death of the plant. After the diseased root is taken by dilution and separation method, it is cleaned and peeled off ...

AI Technical Summary

Problems solved by technology

[0006] 2. The operation is difficult

Because R. solanacearum grows and reproduces in the vascular tissue, it needs to cut the root system to dilute and isolate the pathogen. However, the root system of casuarina has a large wood density and dense cells, so it is laborious and time-consuming to cut and chop the root system; secondly, the outer bark of the root system contains impurities. There are many bacteria. To ensure the separation effect, the root bark needs to be peeled off. The bark is closely connected with the root xylem, and the peeling process is difficult to operate.

[0007] 3. The content of miscellaneous bacteria is high, and secondary separation is usually required

[0008] 4. The reliability of the method of judging virulent strains by traditional visual observation using TTC medium is poor

Due to the mixed strains of Casuarina solanacearum, rapid strain variation and serious pathogenic differentiation, TTC medium is used as a semi-selective medium to specifically distinguish R. There are errors in the strength of pathogenicity

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