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Single spore isolation method of grape powdery mildew

A technology for the separation of powdery mildew bacteria and single spores, which is applied in the methods of using spores, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of growth, unsuitable single spore separation of grape powdery mildew bacteria, etc., and achieves a high success rate. Effect

Inactive Publication Date: 2013-09-25
云南劳味劳味农业科技有限公司
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Problems solved by technology

However, because grape powdery mildew is an obligate parasite, it needs to obtain the necessary nutrients from living host tissues and cannot grow on synthetic media, the above method is not suitable for the isolation of single spores of grape powdery mildew

Method used

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  • Single spore isolation method of grape powdery mildew
  • Single spore isolation method of grape powdery mildew

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Embodiment 1

[0019] Embodiment 1, the single spore isolation of cabernet sauvignon variety grape powdery mildew

[0020] In the Bolongbao winery in Fangshan District, Beijing, the leaves suspected of grape powdery mildew were collected, and the pathogenic bacteria of this disease were isolated from single spores. The specific steps are as follows:

[0021] 1. Collect 10 powdery mildew leaves of Cabernet Sauvignon varieties, and place them in sealed bags numbered 1-10;

[0022] 2. Take two disposable petri dishes with a diameter of 90mm (each petri dish consists of a round bottom and a lid) and stack them together, and use a 5mm diameter punch to culture the bottom and bottom of the upper petri dish. Punch holes at the same position on the lid of the dish, and add 20mL of sterilized ultrapure water to the lower Petri dish to obtain a double-layer Petri dish device ( figure 1 , the punching position is near the edge of the bottom of the upper Petri dish and the lid of the lower Petri dish);...

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Abstract

The invention provides a single spore isolation method of grape powdery mildew. The method comprises the following steps: firstly, inducing a collected powdery mildew sample to regenerate conidia; transferring the newly generated single or single series of conidia to a sterile grape powdery mildew sensitive grape leaves treated by 1% sodium hypochlorite by an acupuncture pin; marking an inoculating point by a marking pen; cultivating for 10-14 days in a cultivation box periodically with light in 12 hours at 22 DEG C; obtaining bacterial colonies at the inoculating point, namely the pure culture of the isolated grape powdery mildew. The method is simple, convenient and accessible, and can obtain the pure culture of single spore of grape powdery mildew with higher success rate, so that the problem that conventional fungus isolation method is not suitable for single spore isolation of grape powdery mildew is solved.

Description

technical field [0001] The invention relates to a method for isolating single spores of biological pathogens, in particular to a method for isolating single spores of obligate parasitic grape powdery mildew bacteria. Background technique [0002] Grape Powdery Mildew (Grape Powdery Mildew) is a worldwide fungal disease, its pathogen is grape powdery mildew Erysipher necator Schw.[syn. Grapevine powdery mildew (Erysiphe necator): a fascinating system for the study of the biology, ecology and epidemiology of an obligate biotroph. Molecular Plant Pathology, 2012,13(1):1-16.). This disease is distributed in all grape growing countries and regions in the world (Reisch, B.I., Pratt C.1996.Grapes, p.297–369.In:J.Janick and J.N.Moore (eds.).Fruit breeding.Vine and small fruit crops.vol.2.Wiley,New York.Gadoury D M.,Seem R C.,Ficke A.,et al.Ontogenic Resistance to Powdery Mildew in Grape Berries.Phytopathology,2003,93(5):547-555 .), seriously affecting the grape yield, tree vigor ...

Claims

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Application Information

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IPC IPC(8): C12N3/00C12R1/645
Inventor 张玮燕继晔李兴红刘梅周莹严红
Owner 云南劳味劳味农业科技有限公司
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