Method for in-vitro propagation of grape downy mildew pathogens

Abstract

The invention discloses a method for in-vitro propagation of grape downy mildew pathogens. The method includes the steps: selecting grape varieties sensitive to downy mildews, collecting third grape leaves to fifth grape leaves of annual new shoots from top to bottom, drilling leaf discs from the middles of the sterilized grape leaves by the aid of a hole puncher, horizontally placing the leaf discs into a moisturizing culture device, enabling the backs of the leaf discs to be upward, and dripping sterilized distilled water on each round leaf dish; preparing grape downy mildew pathogen sporangia into grape downy mildew pathogen sporangium suspension liquid, uniformly coating water agar slabs with the suspension liquid, cutting the slabs coated with the pathogen sporangia into 2mm*2mm agar blocks, covering the leaf discs with the agar blocks with monosporangia in an inverted manner under a microscope to facilitate infection, cultivating the agar blocks for 4-7 days, and forming colonies on the leaf discs. The diameter of the hole puncher is 2cm, the mass percentage concentration of the water agar slabs is 3%, and the colonies are pure cultures of monosporangia of the grape downy mildew pathogens.

Description

technical field [0001] The invention relates to a method for rapid and large-scale propagation and cultivation of pathogenic microorganisms in vitro, in particular to a method for rapid and large-scale propagation of grape downy mildew pathogens in vitro. Background technique [0002] Grape downy mildew (Downy mildew) is caused by grape downy mildew [Plasmopara viticola (Berk.dtCurtis) Berl.Et de Toni]. belongs to. The pathogen is a diploid biotrophic obligate parasitic fungus (Wong et al., 2001), which is one of the most serious and widespread diseases (Song Rungang et al., 1998). It is an ancient worldwide fungus Diseases, distributed in all grape-growing countries and regions in the world (Wong, F.P., Burr, H.N, and Wilcox, W.F.2001.Heterothallism inPlasmopara viticola.Plant Pathol.50:427-432), seriously affect the yield and quality of grapes , In serious years, the production can be reduced by 70%-80%. [0003] Peronospora vine is an obligate parasite, which cannot be...

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Problems solved by technology

[0003] Peronospora vine is an obligate parasite, which cannot be cultivated on synthetic medium, and can only obtain nutrients from living host tissues for growth. The incubation period of the disease is generally 4-12 days (Wang Zhongyue, 2009), mainly affected by host resistance. And the impact of environmental conditions, and it is not easy to multiply the original bacteria in a large number under indoor culture conditions, and the cultivation is difficult, which once became an obstacle restricting the research of this pathogen

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