A kind of isolation method of lawn dollar spot bacteria

Abstract

The invention discloses a method for separating Sclerotinia homoeocarpa. The method comprises the following steps: collection of disease samples: cutting off a diseased and healthy junction part of a plant having a typical dollar spot symptom and a soil sample as the disease samples; moisturized culture of the disease samples; pathogenic fungi separation: using a transfer needle to pick up filament-shaped spawn growing on the surfaces of the disease samples, putting the filament-shaped spawn on an APDA culture medium, and carrying out culture for 2 days under a dark condition at 22 to 25 DEG C; pathogenic fungi purification: after aerial hyphae grow out, cutting tips of the aerial hyphae on the edge of abacterial colony, putting the tips onto a new APDA culture medium, and carrying out culture for 3 to 5 daysunder a dark condition at 22 to 25 DEG C, so as to obtain target pathogenic bacteria. As moisturized culture pretreatment is conducted on the disease samples, and the acidized PDA culture medium is used, the problem of various non-target fungus contamination in a Sclerotinia homoeocarpa separation process can be solved; the operation is simple; the whole process is free of strict disinfection and sterilization operation. Compared with the traditional Sclerotinia homoeocarpa separation method, the separation efficiency and the success rate of the method disclosed by the invention are improved obviously.

Description

technical field

[0001] The invention belongs to the field of plant pathogenic microorganisms, and relates to a method for isolating turf coin spot bacteria. Background technique

[0002] Sclerotinia homoeocarpa is an important plant pathogenic fungus, which can infect a variety of cool-season and warm-season turfgrasses with a variety of hosts. Since it was first discovered and reported in the United States in 1932, it has now spread widely all over the world. Preliminary survey results show that turf dollar spot has occurred in more than 20 provinces and autonomous regions in my country. Dollar spot disease mainly harms the blades of turfgrass, causing the turfgrass to turn yellow and even die, seriously affecting the landscape beauty of the lawn.

[0003] Dollar spot disease mainly has two peaks in early summer and early autumn. When the soil temperature is higher than 18°C ​​and the humidity is high at night, dollar spot disease begins to develop. At the early stage of ...

Examples

Embodiment 1

[0032] 1. Experimental method

[0033] Adopt the inventive method to separate the turf coin spot bacterium, comprise the steps:

[0034] (1) Disease sample collection: Use a soil sampler to cut out the plants and soil samples at the junction of disease and health with typical dollar spot symptoms with a diameter of 4 cm and a height of 6-8 cm as disease samples, put them in an envelope, transport them at room temperature, and bring them back to the laboratory Then transferred to 4 ℃ refrigerator for storage;

[0035] (2) Moisturizing treatment of diseased samples: Take out the diseased samples preserved in step (1), place them in flower pots or trays that have been sterilized (dried at 80°C for 2 hours), and spray sterile water to make the humidity not lower than 80%, after sealing with parafilm, place the diseased samples in a light incubator for moisturizing culture for 2-3 days, and the culture conditions are 22-25°C light for 16h / 18-20°C dark for 8h alternately;

[0036]...

Embodiment 2

[0051] 1. Experimental method

[0052] Isolation of turfgrass dollar spot bacterium by traditional method comprises the following steps:

[0053] (1) Disease sample collection: collect disease samples with typical symptoms of dollar spot in the field, and transfer them to the laboratory for isolation of pathogenic bacteria within 24 hours;

[0054] (2) Disease sample pretreatment: select leaf tissues with typical disease symptoms, use 0.5% sodium hypochlorite to disinfect them for 60 seconds, and then rinse them several times with aseptic;

[0055] (3) Isolation of pathogenic bacteria: after the disinfection treatment in step (2), the clean leaves were placed on the APDA medium, and cultured in the dark at 25°C for 2 days;

[0056] (4) Purification of pathogenic bacteria: after the mycelium grows out, cut the tip of mycelium at the edge of the colony and transfer it to a new APDA medium for 3-5 days at 25°C in dark conditions to obtain purified pathogenic bacteria;

[0057] ...

technical field

[0001] The invention belongs to the field of plant pathogenic microorganisms, and relates to a method for isolating turf coin spot bacteria. Background technique

[0002] Sclerotinia homoeocarpa is an important plant pathogenic fungus, which can infect a variety of cool-season and warm-season turfgrasses with a variety of hosts. Since it was first discovered and reported in the United States in 1932, it has now spread widely all over the world. Preliminary survey results show that turf dollar spot has occurred in more than 20 provinces and autonomous regions in my country. Dollar spot disease mainly harms the blades of turfgrass, causing the turfgrass to turn yellow and even die, seriously affecting the landscape beauty of the lawn.

[0003] Dollar spot disease mainly has two peaks in early summer and early autumn. When the soil temperature is higher than 18°C ​​and the humidity is high at night, dollar spot disease begins to develop. At the early stage of ...

Examples

Embodiment 1

[0032] 1. Experimental method

[0033] Adopt the inventive method to separate the turf coin spot bacterium, comprise the steps:

[0034] (1) Disease sample collection: Use a soil sampler to cut out the plants and soil samples at the junction of disease and health with typical dollar spot symptoms with a diameter of 4 cm and a height of 6-8 cm as disease samples, put them in an envelope, transport them at room temperature, and bring them back to the laboratory Then transferred to 4 ℃ refrigerator for storage;

[0035] (2) Moisturizing treatment of diseased samples: Take out the diseased samples preserved in step (1), place them in flower pots or trays that have been sterilized (dried at 80°C for 2 hours), and spray sterile water to make the humidity not lower than 80%, after sealing with parafilm, place the diseased samples in a light incubator for moisturizing culture for 2-3 days, and the culture conditions are 22-25°C light for 16h / 18-20°C dark for 8h alternately;

[0036]...

Embodiment 2

[0051] 1. Experimental method

[0052] Isolation of turfgrass dollar spot bacterium by traditional method comprises the following steps:

[0053] (1) Disease sample collection: collect disease samples with typical symptoms of dollar spot in the field, and transfer them to the laboratory for isolation of pathogenic bacteria within 24 hours;

[0054] (2) Disease sample pretreatment: select leaf tissues with typical disease symptoms, use 0.5% sodium hypochlorite to disinfect them for 60 seconds, and then rinse them several times with aseptic;

[0055] (3) Isolation of pathogenic bacteria: after the disinfection treatment in step (2), the clean leaves were placed on the APDA medium, and cultured in the dark at 25°C for 2 days;

[0056] (4) Purification of pathogenic bacteria: after the mycelium grows out, cut the tip of mycelium at the edge of the colony and transfer it to a new APDA medium for 3-5 days at 25°C in dark conditions to obtain purified pathogenic bacteria;

[0057] ...

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