Bacillus pumilus N103-1 and application thereof
A technology of Bacillus pumilus and biocontrol fungicides, applied in the direction of application, bacteria, fungicides, etc., to achieve the effects of reducing residues, human and animal safety, and enriching biocontrol resources
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Embodiment 1
[0023] Inhibition of three pathogenic bacteria by Bacillus pumilus N103-1
[0024] A. The pathogenic bacteria tested
[0025] Pathogenic bacteria include Phytophthora amylovora, Phytophthora oryzae bacterial spot and Ralstonia solanacearum, provided by the Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences. Use LB culture solution (peptone 10.0g, yeast powder 5.0g, NaCl 10g, distilled water 1000mL) at 28°C, 150rpm shaking culture for 2 days, dilute to 10 with sterile water 8 cfu / mL, spare.
[0026] B. Test method:
[0027]The Bacillus pumilus N103-1 screened by Jiangsu Academy of Agricultural Sciences was activated on LB medium, then transferred into 50mL LB medium, cultured with shaking at 28°C and 150rpm for 2 days, and diluted with sterile water to 5×10 8 cfu / mL, spare.
[0028] Take 5 μL of Bacillus pumilus N103-1 bacteria solution, spot it on the center of LB plate, incubate at 28°C for 48 hours, spray with pathogenic bacteria diluent, incubate ov...
Embodiment 2
[0033] Inhibitory Effects of Bacillus pumilus N103-1 on Four Kinds of Phytopathogenic Fungi
[0034] A. For testing pathogenic fungi:
[0035] Pathogenic fungi include Strawberry Anthracnose, Tomato Fusarium, Rice Sheath Blight and Cabbage Black Spot, all provided by the Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences. Use PDA medium (200g potato, 20g glucose, 20g agar and 1000mL distilled water, pH 5.6-6.6) to culture at 28°C for 5d, set aside.
[0036] B. Test method:
[0037] The preparation of Bacillus pumilus N103-1 bacterial solution is the same as in Example 1.
[0038] Place the pathogenic fungus bacterium cake (6mm in diameter) on one side of the blank PDA plate, 30mm away from the edge of the plate. On the other side of the plate, 30 mm from the edge of the plate, 5 μL of Bacillus pumilus N103-1 bacterial solution was spotted. Cultivate at a constant temperature of 28°C, set up clean water as a control, and investigate the growth diameter ...
Embodiment 3
[0046] Metabolic secretions of Bacillus pumilus N103-1-production of antibacterial related substances
[0047] Test materials: protease detection medium: skimmed milk powder 100g, agar 20g, dilute to 1000mL. Cellulase detection medium: peptone 10g, yeast powder 10g, sodium carboxymethylcellulose 10g, NaCl 5g, KH 2 PO 4 1g, 20g agar, dilute to 1000mL, pH 7.0. Siderophilic detection medium: CAS 60.5mg, 10mL ferric iron solution (1mmol L -1 FeCl 3 ·6H 2 O), HDTMA 72.9mg, agar 20g, set to 1000mL, pH7.0.
[0048] The preparation of Bacillus pumilus N103-1 bacterial solution is the same as in Example 1. Spot 5 μL of Bacillus pumilus N103-1 bacterial solution in the center of each test medium plate, and repeat 3 times for each treatment. Incubate at a constant temperature of 28°C for 3-7 days and observe.
[0049] Protease detection: observe whether there is a transparent circle. If a transparent circle is produced, it indicates that the antagonistic bacteria can secrete pr...
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