Preparation method of green fluorescent penicillium

Abstract

The invention discloses a preparation method of green fluorescent penicillium and relates to a method for constructing fluorescent bacteria from wheat black embryo disease pathogenic bacteria. Comprising the following steps: 1, preparing agrobacterium with a green fluorescent GFP gene, and transferring the GFP gene into GV3101 agrobacterium by adopting a freeze-thaw method; 2, propagating agrobacterium which can survive under an antibiotic condition and has a green fluorescent GFP gene to a standard concentration; 3, preparing Sichuan wheat black wild penicillium spore suspension; 4, transferring agrobacterium with a green fluorescent GFP gene into Sichuan wheat black wild penicillium spore to obtain a transformant; and 5, purifying the transformant bacterial liquid by using antibiotics to obtain purified green fluorescent penicillium. The method has the advantages that after the wheat is infected by the green fluorescent penicillium, the green fluorescent mark can be clearly observed at the inoculation part, so that the method can be used for exploring how the single penicillium causes the generation of the wheat black embryo disease and evaluating the pathogenicity of the single penicillium. The method can be used as a powerful tool for researching the generation reason of the wheat black embryo disease and positioning the wheat black embryo disease resistant gene.

Description

technical field [0001] The invention relates to the technical field of pathogenic bacteria in molecular biology and plant pathology, in particular to a method for constructing fluorescent bacteria from pathogenic bacteria of wheat black germ disease. Background technique [0002] wheat( Triticum aestivum L.), belonging to Gramineae (Gramineae), Triticum family, Triticum L. (Triticum L.), originated in western Asia, is one of the earliest cultivated crops, and is also a widely planted food crop. Black point of wheat is a disease of wheat grains. Its typical symptom is that the wheat embryo turns black or brown, and the brown lesions of severe lesions spread to the groin of wheat, also known as black spot. , Black spot disease. The main wheat producing areas in many countries and regions in the world have been affected by the disease, and Canada, Australia, Peru, and China are more seriously affected. Wheat black germ disease is a common wheat grain deterioration disease, ...

AI Technical Summary

Problems solved by technology

At present, the research on wheat black germ disease is still in its infancy, and there are few materials resistant to wheat black germ disease, and the pathogenic bacteria of wheat black germ disease are still unclear