Bacterial wilt biocontrol bacterium ANTI-8098A and its culturing medium, culturing method and biocontrol uses

Abstract

The invention provides a microbe-wilt bacterium preventing ANTI-8098A and the culture medium, culture method and biology application thereof. In particular a wilt bacterium preventing strain ANTI-8098A is provided, which is preserved in the Deutsche Sammlung von Mikroorganismen und Zellkulturen(DSMZ), with the preservation number being DSM ID01-100, the classification designation of the microbe being Bacillus cereus. The wilt bacterium preventing microbe collecting method, screen method, ferment production method and the culture medium thereof are provided, the supernatant containing the wilt bacterium preventing microbe or its ferment liquid, the biological pesticide and biological bactericide germicide containing its metabolic product are also provided. The prevention and cure effect of the wilt bacterium preventing microbe and the biological pesticide and biological bactericide germicide on the wilt bacteria of various kinds of plants in the field control experiment is over 80%, in addition, the quality and the output of the treated fruits are increased.

Description

technical field

[0001] The invention relates to a microorganism and its screening, cultivation method and culture medium, in particular to a bacterial wilt biocontrol bacteria and its culture medium, cultivation method and biocontrol application. Background technique

[0002] Bacterial wilt is a devastating soil-borne disease caused by Ralstonia solanacearum, which can harm more than 300 species of plants in 44 families and is widely distributed in tropical, subtropical and temperate regions. , the hosts with severe damage and wide distribution mainly include potato, tomato, tobacco, eggplant, pepper, peanut, ginger, banana, ganru, hemp and so on. In recent years, it has been reported successively in woody plants such as olive, mulberry, casuarina, cassava, custard apple, Australian pine, eucalyptus, broad bean, soybean, cowpea, lupine, pumpkin, Chinese cabbage, radish, fennel, cotton, clover, and comfrey. The bacterial wilt that occurs on herbaceous plants such as strawber...

Examples

Embodiment 1

[0152] The experiment started with the collection of bacterial wilt biocontrol strains for screening. Soil samples were collected from tomato, pepper, tobacco, and ginger in nine cities in Fujian Province, including Fuzhou, Putian, Sanming, Longyan, Ningde, Nanping, Xiamen, Zhangzhou, and Quanzhou. , peanut and other Solanaceae crop root soil, with 87 samples.

[0153] The collection method is as follows (respectively make 87 soil samples carry out the following operations):

[0154] 1. Provide one of the above 87 kinds of soil samples,

[0155] 2. Mix the soil with nine parts of sterile water (by weight) in a glass vessel,

[0156] 3. Shake the mixture for 20 minutes and let it stand for 20-30 seconds to obtain 10 -1 Diluent,

[0157] 4. Put a 10 -1 Diluent mixed with nine parts of sterile water (weight ratio) to obtain 10 -2 Dilution, and so on, serially diluted to make 10 -4 Diluent, 10 -5 Diluent, 10 -6 Diluent,

[0158] 5. Put 10 -4 Diluent, 10 -5 Diluent, 10 ...

Embodiment 2

[0164] The more than 12,100 strains isolated in Example 1 were screened as follows.

[0165] (1) Provide a strain for screening,

[0166] (2) Spotting the bacterial strains for screening on the culture medium in the petri dish,

[0167] (3) Place the petri dish in a 30°C incubator and cultivate it for 48 hours to form the first bacterial group,

[0168] (4) provide a kind of R. solanacearum indicator bacteria,

[0169] (5) The concentration of the indicator bacteria is 10 7 cfu / ml suspension,

[0170] (6) put 10 7 The cfu / ml suspension was sprayed in the first bacterial colony, and cultivated in the incubator for 48 hours,

[0171] (7) Observe the size of the bacteriostatic zone around the colony. Those with a bacteriostatic zone greater than 2mm are strains with antagonistic effect on R. solanacearum, and name the strains.

[0172] More than 12,100 bacterial strains were collected, and 10 strains of Pseudomonas solanacearum were selected as indicator strains (these bact...

Embodiment 3

[0186] The fermentation production method of the bacterial wilt biocontrol bacterium, the bacterial strain used is the bacterial wilt biocontrol bacterium (Bacillus cereus ANTI-8098A DSM ID 01-1000) obtained by isolation and screening, and the specific steps are as follows:

[0187] (1) provide a kind of bacterial wilt biological control bacterial classification and some glasswares that the first culture medium is housed,

[0188] (2) Inoculate the bacterial strain on the first culture medium, and obtain the bacterial strain A by activating the culture at a temperature of 30° C. for 18 hours,

[0189] (3) Mix strain A with sterile water to obtain a suspension,

[0190] (4) provide a kind of seed jar and second culture medium,

[0191] (5) inoculate the suspension containing strain A into the seed bottle equipped with the second culture medium by 5% inoculation amount, and ferment for 12 hours to obtain the seed liquid,

[0192] (6) providing a fermenter and a third culture m...

technical field

[0001] The invention relates to a microorganism and its screening, cultivation method and culture medium, in particular to a bacterial wilt biocontrol bacteria and its culture medium, cultivation method and biocontrol application. Background technique

[0002] Bacterial wilt is a devastating soil-borne disease caused by Ralstonia solanacearum, which can harm more than 300 species of plants in 44 families and is widely distributed in tropical, subtropical and temperate regions. , the hosts with severe damage and wide distribution mainly include potato, tomato, tobacco, eggplant, pepper, peanut, ginger, banana, ganru, hemp and so on. In recent years, it has been reported successively in woody plants such as olive, mulberry, casuarina, cassava, custard apple, Australian pine, eucalyptus, broad bean, soybean, cowpea, lupine, pumpkin, Chinese cabbage, radish, fennel, cotton, clover, and comfrey. The bacterial wilt that occurs on herbaceous plants such as strawber...

Examples

Embodiment 1

[0152] The experiment started with the collection of bacterial wilt biocontrol strains for screening. Soil samples were collected from tomato, pepper, tobacco, and ginger in nine cities in Fujian Province, including Fuzhou, Putian, Sanming, Longyan, Ningde, Nanping, Xiamen, Zhangzhou, and Quanzhou. , peanut and other Solanaceae crop root soil, with 87 samples.

[0153] The collection method is as follows (respectively make 87 soil samples carry out the following operations):

[0154] 1. Provide one of the above 87 kinds of soil samples,

[0155] 2. Mix the soil with nine parts of sterile water (by weight) in a glass vessel,

[0156] 3. Shake the mixture for 20 minutes and let it stand for 20-30 seconds to obtain 10 -1 Diluent,

[0157] 4. Put a 10 -1 Diluent mixed with nine parts of sterile water (weight ratio) to obtain 10 -2 Dilution, and so on, serially diluted to make 10 -4 Diluent, 10 -5 Diluent, 10 -6 Diluent,

[0158] 5. Put 10 -4 Diluent, 10 -5 Diluent, 10 ...

Embodiment 2

[0164] The more than 12,100 strains isolated in Example 1 were screened as follows.

[0165] (1) Provide a strain for screening,

[0166] (2) Spotting the bacterial strains for screening on the culture medium in the petri dish,

[0167] (3) Place the petri dish in a 30°C incubator and cultivate it for 48 hours to form the first bacterial group,

[0168] (4) provide a kind of R. solanacearum indicator bacteria,

[0169] (5) The concentration of the indicator bacteria is 10 7 cfu / ml suspension,

[0170] (6) put 10 7 The cfu / ml suspension was sprayed in the first bacterial colony, and cultivated in the incubator for 48 hours,

[0171] (7) Observe the size of the bacteriostatic zone around the colony. Those with a bacteriostatic zone greater than 2mm are strains with antagonistic effect on R. solanacearum, and name the strains.

[0172] More than 12,100 bacterial strains were collected, and 10 strains of Pseudomonas solanacearum were selected as indicator strains (these bact...

Embodiment 3

[0186] The fermentation production method of the bacterial wilt biocontrol bacterium, the bacterial strain used is the bacterial wilt biocontrol bacterium (Bacillus cereus ANTI-8098A DSM ID 01-1000) obtained by isolation and screening, and the specific steps are as follows:

[0187] (1) provide a kind of bacterial wilt biological control bacterial classification and some glasswares that the first culture medium is housed,

[0188] (2) Inoculate the bacterial strain on the first culture medium, and obtain the bacterial strain A by activating the culture at a temperature of 30° C. for 18 hours,

[0189] (3) Mix strain A with sterile water to obtain a suspension,

[0190] (4) provide a kind of seed jar and second culture medium,

[0191] (5) inoculate the suspension containing strain A into the seed bottle equipped with the second culture medium by 5% inoculation amount, and ferment for 12 hours to obtain the seed liquid,

[0192] (6) providing a fermenter and a third culture m...

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