A Composite Fluorescent Staining Method for Identification of Clubroot in Cruciferous Plants

Abstract

The invention discloses a composite fluorescent dyeing method for identifying clubroot disease of cruciferous plants, comprising the following operation steps: (1) inoculating the cabbage two weeks after sowing the cabbage; (2) controlling the cabbage within 2 days after inoculation (3) Three weeks after inoculation, take cabbage roots to make paraffin sections, first stain with aniline blue for 2-4min, and then sequentially use ddH 2 O, 70% alcohol and 100% alcohol were cleaned, then stained with Nile red for 50-70s, and finally washed with 100% alcohol for 30 seconds, dried, and observed under the ultraviolet excitation light of a fluorescence microscope. The invention provides a composite fluorescent dyeing method for identifying clubroot disease of Decidaceae plants, which is simple to operate, environmentally friendly, and low in cost, and can specifically dye the cell wall of cabbage root cells into red, and dye starch granules into bright green or Blue-green, while Plasma rhizogenes are stained blue.

Description

technical field [0001] The invention belongs to the technical field of fluorescent counterstaining and has Involvement of a compound for the identification of clubroot in cruciferous plants Fluorescence staining method . Background technique [0002] Cruciferous clubroot is a soil-borne disease caused by Plasmodiophora brassicae Woron., which seriously endangers the safe production of Brassicaceae plants. Plasmodium brassicae is divided into two stages in the process of infecting plants. The initial infection stage (or root hair infection, that is, the dormant spores in the soil become primary zoospores reach the surface of the root hair, penetrate the cell wall and invade the interior of the root hair. , this stage is called the initial infection stage) and the secondary infection stage (or cortical infection, that is, the pathogenic bacteria form a secondary protoplasm mass inside the root hair, a part of the nucleus is cleaved to form multinucleated zoosporangia, and e...

AI Technical Summary

Problems solved by technology

Although there are many staining methods, the commonly used staining methods for the infection process of clubroot are limited to aniline blue, basic fuchsin, methylene blue, toluidine blue, saffron, astra blue, and Nile red. It is a staining agent with poor specificity. It cannot distinguish plant cells and Plasmodium in cells well, which is not conducive to the cytological observation of Plasmodium infecting hosts.

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