Bacillus polymyxa kn-03 and its culture method and application
A technology of Bacillus polymyxa and culture medium, applied in the directions of botanical equipment and methods, microorganism-based methods, biochemical equipment and methods, etc. The effect of increasing production is obvious, the production cost is low, and the fermentation period is short.
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Embodiment 1
[0030] The screening of embodiment 1 bacillus polymyxa KN-03
[0031] 1. Heat treatment screening of strains:
[0032] Weigh 1.0 g of soil sample (soil sample from Shandong tomato planting demonstration base) and put it into a small conical flask, add 9 mL of sterilized PDA culture solution (glucose 20 g, peeled potatoes 200 g (boil with tap water for 30 min), agar powder 15 g, Dilute to 1L, pH7.0). After culturing on a constant temperature shaker at a temperature of 30° C. and 220 rpm for 3 days, they were placed in a water bath at 80° C. for 30 minutes. After fully shaking, let it stand still, and take the supernatant for concentration gradient dilution. Use a pipette gun to draw 100ul of the diluted solution and evenly spread it on the PDA nutrient agar medium (the same composition as before, add 1.5% agar powder), culture in a temperature-controlled incubator at 30°C for 3-5 days, and then pick a single colony for microscopic examination , Select the strains with the mo...
Embodiment 2
[0049] The cultivation of embodiment 2 bacillus polymyxa KN-03
[0050] (1) Strain activation:
[0051] Prepare a slant medium containing the following ingredients: glucose 20g, peeled potatoes 200g (boiled with tap water for 30min), agar powder 15g, dilute to 1L, pH7.0;
[0052] A ring of Bacillus polymyxa KN-03 was picked on the slant medium, and the inoculated slant was placed in a temperature-controlled incubator at 30° C. for 72 hours.
[0053] (2) Seed cultivation:
[0054] Prepare a seed medium containing the following ingredients: sucrose 5-15g / l, yeast extract 5-20g / l, rice flour 8-16g / l, corn flour 5-20g / l, bran 10-30g / l , calcium chloride 0.1-1g / l, manganese chloride 0.09-1g / l, magnesium sulfate 3-10g / l, pH7.2-7.5;
[0055] Get step (1) activated Bacillus polymyxa KN-03 loop and inoculate in described seed culture medium (fill 100ml liquid seed culture medium in 500ml Erlenmeyer flask), place the inoculated seed culture solution at temperature 30 Cultivate for 1...
Embodiment 3
[0060] Indoor antibacterial activity test of embodiment 3 bacillus polymyxa KN-03
[0061] 1. Determination of the antibacterial activity of Bacillus polymyxa KN-03 against Ralstonia solanacearum of tomato, Fusarium wilt of pepper, Verticillium dahliae of rice, Phytophthora maize take-all and Sclerotinia sclerotiorum
[0062] The test steps are as follows:
[0063] 1) Test agent: Bacillus polymyxa KN-03 powder.
[0064] 2) Drug preparation: Dilute the Bacillus polymyxa KN-03 powder directly with sterile water to an appropriate concentration, and set a blank control without drug.
[0065] 3) Determination by plate culture method: take 0.1 ml of the diluted and mixed medicament and evenly spread it on a sterilized PDA medium plate. Use an inoculation needle to streak and smear R. solanacearum on the PDA plate containing the drug, and set up a blank control. The cultivated pathogenic bacteria (tomato solanacear, capsicum wilt, rice verticillium wilt, corn total rot and rapesee...
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