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Bacillus methylotrophicus, bactericide and preparation method and application of bactericide

A methylotrophic and bacillus technology, applied in the field of microorganisms, can solve the problems such as potato early blight of methylotrophic bacillus, and achieve the effects of short fermentation production period, prolonging the preservation period and inhibiting the growth of pathogenic bacteria.

Active Publication Date: 2016-06-15
山东京青农业科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Through literature search, there is no report on the use of methylotrophic bacillus to prepare biological agents and apply them to control potato early blight in field

Method used

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  • Bacillus methylotrophicus, bactericide and preparation method and application of bactericide
  • Bacillus methylotrophicus, bactericide and preparation method and application of bactericide
  • Bacillus methylotrophicus, bactericide and preparation method and application of bactericide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Isolation and screening of methylotrophic bacillus JQ-018 strain:

[0030] 1. Separation

[0031] (1) Soil collection: 100g of soil samples were collected from the potato field in Daquanquan Village, Zhangbei County, Hebei Province, from the surface layer to 10cm. Five samples were collected in parallel at one location, and mixed through a 40-mesh grading sieve. Take 100g of sieved soil for separation.

[0032] (2) Medium preparation: prepared separately

[0033] NA medium: peptone 10g / L, beef extract 3.0g / L, sodium chloride 5g / L, agar 18g / LpH=7.0-7.2;

[0034] And PDA medium: potato 200g, glucose 20g, agar powder 20g, distilled water 1000mL;

[0035] Make media plates.

[0036] (3) Accurately weigh 1 g of soil and place it in a 250 ml Erlenmeyer flask, add 99 ml of sterile water, and shake on a shaker at 200 rpm at 37°C for 30 min. Make a soil suspension and dilute to 10 by a 10-fold gradient -8 . Single colonies were isolated in NA medium and PDA med...

Embodiment 2

[0046] Embodiment two: the identification of methylotrophic bacillus JQ-018 bacterial strain:

[0047] 1. Microbiological characteristics: JQ-018 bacteria are short rod-shaped, with perinatal flagella, can produce spores, and Gram staining is positive. After the strain was cultured on PDA medium at 34°C for 12 hours, the colony was round, translucent and white, with a smooth surface and a raised center, no pigment and stickiness. Colonies are small or do not grow on NA medium. Bacterium was formed in liquid culture, and spores were produced after 24 hours, brownish yellow, turbid, and precipitated. The strain exhibited aerobic growth.

[0048] 2. Physiological and biochemical experiments:

[0049] Table 3 Physiological and biochemical characteristics of methylotrophic Bacillus JQ-018

[0050] Test items

characteristics

Test items

characteristics

Test items

characteristics

D-Fructose

+

Salicin

-

Mannitol

+

Malt...

Embodiment 3

[0054] Embodiment three: the preparation of methylotrophic bacillus liquid inoculum

[0055] (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.

[0056] (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), at a speed of 250 rpm, at a temperature of 36° C., and cultivate it on a shaking table for 20 h, as a grade seed liquid.

[0057] (3) Secondary seed liquid preparation: insert the primary seed liquid prepared by step (2) into the seed culture medium according to the inoculum size of 3% (volume percentage), the ventilation rate is 1.0vvm, the stirring speed is 2...

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Abstract

The invention relates to bacillus methylotrophicus, bactericide and a preparation method and application of the bactericide.The bacillus methylotrophicus is bacillus methylotrophicus JQ-018 and is preserved in China General Microbiological Culture Collection Center on 7th December, 2015, and the preservation register number is CGMCC No:11838.The bactericide can be prepared through seed activation, seed culture and fermentation culture.The invention further provides the preparation method of the bactericide of the bacillus methylotrophicus and the application to preparation of an agent for preventing and controlling potato early blight.The bactericide has high bioactivity and good environment compatibility, is free of poisons to people and livestock and convenient to use, and can adjust plant microorganism ecological florae.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a methylotrophic bacillus, a bacterial agent, a method for preparing the bacterial agent and an application of the bacterial agent. Background technique [0002] Potato early blight is a serious disease of potatoes, which occurs in all areas where potatoes are grown, seriously affecting the yield of potatoes. Potato early blight is mainly caused by Alternaria solani, which mainly occurs on leaves and can also infect tubers. Spores produced by diseased spots on potato leaves can be dispersed by wind, rain, insects, etc., and infect surrounding healthy plants. When the leaf surface is wet, the spores that land on the leaves germinate and invade through the stomata and wounds. After a few days, new lesions are formed, and spores are produced on the lesions to spread. During a growing season, multiple infestations can occur repeatedly, resulting in disease in the entire fiel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01P3/00C12R1/07
CPCA01N63/00C12N1/20C12N1/205C12R2001/07
Inventor 史盼盼李元鑫贾振宇郭婧
Owner 山东京青农业科技有限公司
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