Method for high throughput screening of microorganisms for preventing plant soil-borne fungal diseases

Abstract

The invention discloses a method for high throughput screening of microorganisms for preventing plant soil-borne fungal diseases, and belongs to the technical field of agricultural microorganisms. Themethod includes preparing the diseased oil with a high-concentration wheat root rot pathogenic bacteria culture and the soil, sowing germinated wheat seeds into the soil containing pathogenic bacteria to simulate the natural growth conditions and environment of plants and the pathogenic bacteria in the soil to the maximum extent. The method for screening for a pathogen-plant pathogenesis system of antagonistic microorganisms of the wheat bipolaris sp, the dense planting of the wheat reduces the space occupied by the screening, and the symptoms of the wheat root rot appear early, which reducesthe screening time, so that the high-throughput screening can be achieved. The screening efficiency of the microorganisms with better inhibition of pathogenic bacteria obtained in the early screeningis greatly improved, and a large number of microorganisms that cannot exert control effects in practical applications can be removed, so that the pace of microorganism screening, microorganism pesticide development and microorganism fertilizer research and development can be accelerated.

Description

technical field

[0001] The invention relates to a method for high-throughput screening of microorganisms used for preventing and controlling plant soil-borne fungal diseases, and belongs to the technical field of agricultural microorganisms. Background technique

[0002] Soil-borne fungal diseases caused by phytopathogenic fungi account for about 70-80% of all plant diseases, and their symptoms are various and can appear in various parts of plants and in various stages of growth. Biological control has been recognized all over the world as an environmentally friendly, pollution-free, green, friendly, and durable method of disease control that can replace traditional chemical pesticides. How to obtain microbial resources that have good biocontrol effects and can achieve large-scale industrial production It is the core technology of scientific research institutes and companies.

[0003] According to the types of system carriers used, the current conventional screening methods...

Examples

Embodiment 1

[0033] Embodiment 1: the cultivation of wheat root rot bacteria

[0034] The Umbilophilus tritici used in the present invention was purchased from the China Agricultural Microorganism Culture Collection and Management Center, and the strain number is ACCC 36529. This strain can grow on PDA medium. Under the environment of 25~30℃, the hyphae can cover the whole surface of the medium within 2-3 days, and then begin to grow aerial hyphae, and produce brown or brown branches in 4-6 days. Sporulation. Conidia are oblong or fusiform, with 3-9 septa. Umbilical point broad, base truncated ( figure 1 ).

Embodiment 2

[0035] Embodiment 2: the preparation of wheat root rot fungus diseased soil

[0036]Preparation of culture medium: Soak the wheat seeds overnight, filter the water the next day, put the wheat seeds that have absorbed enough water into the triangular flask, the amount of seeds in each bottle (by volume, v / v) 30-40%, intermittently Sterilize twice.

[0037] Cultivation of diseased soil: Cut the spore-forming Helminthospora tritici together with the culture medium into small pieces, inoculate them into a triangular flask containing sterilized wheat, mix well, and culture in the dark at 28°C for 5~ 8 days until the mycelium of the pathogenic fungus is covered with wheat seeds; light culture at 28°C for 2 weeks, until Helminthosporium tritici produces black spores on the wheat seeds; dry with natural ventilation for 1-2 days, and use a pulverizer to slightly pulverize it to obtain Containing the diseased soil of umbilical sp. tritici, the concentration of umbilical sp. spores in t...

Embodiment 3

[0038] Embodiment 3: Determination of the use ratio of wheat root rot fungus diseased soil

[0039] In the soil with nutrient soil and sandy soil = 1:1, the diseased soil containing wheat root rot bacteria produced in Example 2 was added according to different volume ratios, and the growth of wheat was observed. When the wheat seedlings show slow emergence, short growth, and lesions on the leaves, it is considered as a symptom of the disease. Select the ratio of symptoms appearing under the minimum addition amount to repeat. The results showed that when the addition ratio of wheat root rot fungus diseased soil was greater than 10% (v / v), compared with the blank control, the emergence rate was seriously delayed or no emergence, the growth was seriously damaged or basically could not grow; When the addition ratio of rot fungus soil is 2.5% (v / v), compared with the blank control, there is no obvious phenomenon of slow growth and short plant; when the addition volume is 5% to 10%...

technical field

[0001] The invention relates to a method for high-throughput screening of microorganisms used for preventing and controlling plant soil-borne fungal diseases, and belongs to the technical field of agricultural microorganisms. Background technique

[0002] Soil-borne fungal diseases caused by phytopathogenic fungi account for about 70-80% of all plant diseases, and their symptoms are various and can appear in various parts of plants and in various stages of growth. Biological control has been recognized all over the world as an environmentally friendly, pollution-free, green, friendly, and durable method of disease control that can replace traditional chemical pesticides. How to obtain microbial resources that have good biocontrol effects and can achieve large-scale industrial production It is the core technology of scientific research institutes and companies.

[0003] According to the types of system carriers used, the current conventional screening methods...

Examples

Embodiment 1

[0033] Embodiment 1: the cultivation of wheat root rot bacteria

[0034] The Umbilophilus tritici used in the present invention was purchased from the China Agricultural Microorganism Culture Collection and Management Center, and the strain number is ACCC 36529. This strain can grow on PDA medium. Under the environment of 25~30℃, the hyphae can cover the whole surface of the medium within 2-3 days, and then begin to grow aerial hyphae, and produce brown or brown branches in 4-6 days. Sporulation. Conidia are oblong or fusiform, with 3-9 septa. Umbilical point broad, base truncated ( figure 1 ).

Embodiment 2

[0035] Embodiment 2: the preparation of wheat root rot fungus diseased soil

[0036]Preparation of culture medium: Soak the wheat seeds overnight, filter the water the next day, put the wheat seeds that have absorbed enough water into the triangular flask, the amount of seeds in each bottle (by volume, v / v) 30-40%, intermittently Sterilize twice.

[0037] Cultivation of diseased soil: Cut the spore-forming Helminthospora tritici together with the culture medium into small pieces, inoculate them into a triangular flask containing sterilized wheat, mix well, and culture in the dark at 28°C for 5~ 8 days until the mycelium of the pathogenic fungus is covered with wheat seeds; light culture at 28°C for 2 weeks, until Helminthosporium tritici produces black spores on the wheat seeds; dry with natural ventilation for 1-2 days, and use a pulverizer to slightly pulverize it to obtain Containing the diseased soil of umbilical sp. tritici, the concentration of umbilical sp. spores in t...

Embodiment 3

[0038] Embodiment 3: Determination of the use ratio of wheat root rot fungus diseased soil

[0039] In the soil with nutrient soil and sandy soil = 1:1, the diseased soil containing wheat root rot bacteria produced in Example 2 was added according to different volume ratios, and the growth of wheat was observed. When the wheat seedlings show slow emergence, short growth, and lesions on the leaves, it is considered as a symptom of the disease. Select the ratio of symptoms appearing under the minimum addition amount to repeat. The results showed that when the addition ratio of wheat root rot fungus diseased soil was greater than 10% (v / v), compared with the blank control, the emergence rate was seriously delayed or no emergence, the growth was seriously damaged or basically could not grow; When the addition ratio of rot fungus soil is 2.5% (v / v), compared with the blank control, there is no obvious phenomenon of slow growth and short plant; when the addition volume is 5% to 10%...

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