Wettable Bacillus methylotrophicus powder and preparation method and application thereof
A methyl nutritional type, wettable powder technology, applied in agriculture and biological fields, can solve problems such as unfavorable potato production and food, ecological safety, easy to be infected by other pathogenic bacteria, insufficient attention, etc., to improve crop quality, No toxic side effects, the effect of stimulating plant growth
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Embodiment 1
[0036] Example 1: Acquisition, Identification and Preservation of JQ-018 Strains
[0037] 1. Obtaining JQ-018 strain
[0038] 1. The methylotrophic bacillus (JQ-018 bacterial strain) of the present invention is isolated from the potato root soil in Daquanquan Village, Zhangbei County, Hebei Province. The specific separation method is as follows:
[0039] (1) Soil collection: Collect 100g soil samples from the soil surface to 10cm, collect 5 samples in parallel at one location, and mix them through a 40-mesh grading sieve. Take 100g for separation.
[0040] (2) Medium configuration: NA medium and PDA medium were respectively prepared to make a medium plate.
[0041] (3) Accurately weigh 1g of the soil sample and place it in a 250ml Erlenmeyer flask, add 99ml of sterile water, and shake on a shaking table at 200rpm at 30-37°C for 30min. Make a soil suspension and dilute to 10 by a 10-fold gradient -8 . Single colonies were isolated in NA medium and PDA medium respectively,...
Embodiment 2
[0064] Embodiment 2: Determination of antagonistic effect of JQ-018 bacterial strain fermentation broth
[0065] Solid medium: NA medium: peptone 10g / L, beef extract 3.0g / L, sodium chloride 5g / L, agar 18g / LpH=7.0-7.2.
[0066] Gao's No. 1 medium: soluble starch 20g, KNO 3 1g, NaCl0.5g, K 2 HPO 4 0.5g, MgSO 4 0.5g, FeSO 4 0.01g agar powder 16g, distilled water 1000ml, pH7.4, when preparing, first use a small amount of cold water to make starch into a paste, pour into boiling water, heat on the fire, add other ingredients while stirring, after melting, make up When the water content reaches 1000ml, then use 100ml Erlenmeyer flasks to evenly distribute into 13 bottles, and sterilize at 121°C for 30 minutes.
[0067] PDA medium: Weigh 200g of potatoes, wash, peel and chop them, add 1000ml of water to boil for half an hour, filter with gauze, add 20g of glucose and 16g of agar, filter with hot gauze after fully dissolving, bottle them, each bottle is about 75ml, After sterili...
Embodiment 3
[0085] Embodiment 3: the preparation and application of methylotrophic bacillus JQ-018 wettable powder
[0086] One, the preparation of bacterial agent:
[0087] (1) Strain activation: the cryopreserved methylotrophic Bacillus JQ-018 strain was inoculated on LB solid medium, and cultured at 36°C for 36 hours; a single colony was picked and transferred to LB slant medium, and incubated at 36°C. Cultivate at ℃ for 20 h, wash the bacterial cells on the surface of the culture medium with sterile water as the inoculum.
[0088] (2) Preparation of first-level seed solution: add the inoculum prepared in step (1) to the LB seed medium according to the inoculum amount of 10% (volume percentage), the rotation speed is 220rpm, the temperature is 36°C, and the shaking table is cultivated for 20h, as a grade seed liquid.
[0089] (3) Secondary seed liquid preparation: insert the primary seed liquid prepared in step (2) into the seed medium according to the inoculum size of 3% (volume per...
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