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BaciIllus polymyxa KN-03 and culturing method and use thereof

A technology of Bacillus polymyxa and culture, applied in the direction of botany equipment and methods, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of low bacterial production efficiency, poor spore ability, high cost, etc., and achieve increased production The effect is obvious, the production cost is low, and the fermentation cycle is short

Active Publication Date: 2015-05-13
WUHAN KERNEL BIO-TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The Bacillus polymyxa disclosed in CN102851250A can only effectively prevent and treat tobacco black shank
[0003] In addition, the existing Bacillus polymyxa has poor spore-producing ability, resulting in low bacterial production efficiency, long cycle and high cost, thus limiting its application

Method used

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  • BaciIllus polymyxa KN-03 and culturing method and use thereof
  • BaciIllus polymyxa KN-03 and culturing method and use thereof
  • BaciIllus polymyxa KN-03 and culturing method and use thereof

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Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses baciIllus polymyxa KN-03 capable of simultaneously preventing and treating various crop diseases, and the preservation number is CCTCC NO:M2012077. The invention also discloses a culturing method and use of the baciIllus polymyxa. The baciIllus polymyxa can effectively simultaneously prevent and treat crop bacterial wilt, fusarium wilt, verticillium wilt, sclerotinia disease, take-all disease, early blight, gray mold and anthracnose, and has strong sporulation ability, the fermentation liquid spore number can reach 30 billion CFU / ml, at the same time, the baciIllus polymyxa KN-03 also has obvious growth promoting effect, and can significantly increase crop yield.

Description

technical field [0001] The invention relates to a bacillus polymyxa and its cultivation method and application. Background technique [0002] Plant bacterial wilt and fusarium wilt are known as plant "cancers". They are the most harmful, the heaviest, and the most widely distributed soil-borne diseases worldwide. and fungal soil-borne diseases, and there is no microbial fungicide that can simultaneously control bacterial and fungal soil-borne diseases such as bacterial wilt and fusarium wilt and leaf diseases. For example: the Bacillus polymyxa disclosed in CN101712941A can only be used for preventing and treating plant bacterial wilt. The Bacillus polymyxa disclosed in CN102851243A can be used to prepare broad-spectrum biocontrol agents for inhibiting pathogenic fungi and pathogenic bacteria. The pathogenic fungi are apple ringworm, cotton Verticillium dahliae, peach brown rot, wheat sheath blight, Lilium base rot fungus and cabbage wilt fungus; the pathogenic bacteria ar...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P3/00C12R1/12
CPCA01N63/00C12N1/20C12N1/205C12R2001/12
Inventor 周莉余娜邹维李青程治国刘华梅唐纬坤杨克华王晓辉姚金伍胡虓孙刚忠刘荷梅向东万俊
Owner WUHAN KERNEL BIO-TECH CO LTD
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