Bacillus velezensis strain and fungicide and application thereof
A technology of Bacillus Velez and Bacillus, applied in the field of microorganisms, can solve the problems of chemical prevention and control of environmental pollution and unsafety
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Embodiment 1
[0027] Example 1 Isolation and Identification of Bacillus Veles strain Bv01
[0028] Bacillus Velez strain Bv01 was obtained from soil, and soil samples were collected from Dayin Village, Raoyang County, Hengshui City, Hebei Province.
[0029] (1) Separation: Take a Erlenmeyer flask, put 15 glass beads into it, add 90 ml of distilled water, and sterilize at 121°C for 30 min. Weigh 10 g of soil sample into the above-mentioned sterilized Erlenmeyer flask, put it into a shaker at 150 r / min and shake for 15 min to obtain a 10-fold diluted soil suspension. Take 4 sterilized test tubes and add 9 ml sterilized distilled water to each test tube. Take 1 ml of the 10-fold diluted soil suspension in the above-mentioned test tube filled with 9 ml of distilled water to obtain a dilution of 10 2 times the soil dilution solution, and so on, until it is diluted to 10 5 times the soil dilution. will be diluted 10 3 times, 10 4 times and 10 5 Put the diluted soil solution at 80°C in a wa...
Embodiment 2
[0032] Example 2 Determination of the inhibitory effect of Bacillus veleisi strain Bv01 on phytopathogenic fungi
[0033] Preparation of PDA medium: Weigh 200 g of potatoes, wash, peel and chop them, add 800 ml of distilled water to boil for half an hour, filter with gauze, then add 20 g of glucose and 20 g of agar, dilute to 1000 ml with distilled water, 121 °C for 30 minutes.
[0034] Strain activation: Inoculate Curvularia geniculateum, Pythium ultima, Sclerotinia sclerotium, Alternaria solani, Fusarium graminearum, and Fusarium oxysporum on PDA medium, and cultivate at 28°C until the colony diameter is about 6 cm spare.
[0035] Confrontation culture: use a puncher with a diameter of 5 mm to punch the bacteria cake at the edge of the above-mentioned activated colony, and inoculate it in the center of the PDA plate. Then, take a new PDA plate, spread 200 ul of the suspension of Bacillus veleisi Bv01 (1 billion / ml) evenly on it, let it dry, and use a puncher with a diamete...
Embodiment 3
[0039] Example 3 Determination of antibacterial activity of metabolites of Bacillus veleisi strain Bv01
[0040] Preparation of metabolites: Streak-inoculate the Bacillus veleisi strain Bv01 on the slant medium, culture at 28°C for 48 hours, take a ring with an inoculation loop and inoculate it in 50 ml liquid LB medium, at 28°C, 180 r / After 24 h of min culture, 15 ml was transferred to 150 ml LB medium, and cultured in a shaker at 28 °C and 180 r / min for 72 h to obtain the fermentation broth of Bacillus veleisi strain Bv01. After centrifugation at 6000 r / min for 10 min, and then filtered with a bacterial filter, the metabolites of Bacillus Velez strain Bv01 can be obtained.
[0041] Preparation of PDA plates containing metabolites: 20 ml of the above fermentation products were added to 80 ml of melted PDA medium to prepare PDA plates containing 20% metabolites of Bacillus veleisi strain Bv01 for use. Take the PDA plate added with the same amount of LB liquid medium as the...
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