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Klebsiella pneumoniae, bacterium agent of klebsiella pneumoniae and preparation method and application

A technology of Klebsiella pneumonia and Klebsiella pneumoniae, which is applied in the field of microorganisms, can solve the problems of no Klebsiella pneumoniae and other problems, and achieve the effects of good removal effect, convenient use, and low production cost

Inactive Publication Date: 2017-05-31
YUNNAN ACAD OF TOBACCO AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, a number of strains with thiram degradation ability have been isolated, such as Pseudomonas, Mycobacterium, Bacillus, etc., but there is no report on the use of Klebsiella pneumoniae in thiram degradation

Method used

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  • Klebsiella pneumoniae, bacterium agent of klebsiella pneumoniae and preparation method and application
  • Klebsiella pneumoniae, bacterium agent of klebsiella pneumoniae and preparation method and application
  • Klebsiella pneumoniae, bacterium agent of klebsiella pneumoniae and preparation method and application

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preparation example Construction

[0034] According to the present invention, the preparation method of Klebsiella pneumoniae as an active ingredient is characterized in that it includes the steps of strain activation, seed cultivation, production cultivation and preparation preparation, specifically comprising:

[0035] Strain activation: the preserved species of the FM84 bacterial strain is cultivated according to the plate culture method until a single colony occurs; seed culture: the single colony is inserted into a seed culture bottle and cultivated with the FM84 bacterial strain seed medium until the Logarithmic growth phase; production cultivation: the culture solution obtained by step seed cultivation is inserted into the fermenter, and cultivated to the logarithmic growth phase with the FM84 strain production medium, and the insertion amount of the culture solution is the production medium of the FM84 strain 4-8% of the total volume; preferred formulations for seed culture and production media contain: ...

Embodiment 1

[0039] Screening and Identification of Strains

[0040] Leaf samples were collected from tobacco grown in the field, and used as a bacterial source for natural enrichment culture. Glucose 2g, tryptone 4g, beef extract 4g, MgSO 4 Add 0.8g into 400mL of distilled water, sterilize the prepared culture solution at 121°C for 20 minutes, cultivate on this medium for three days, then prepare inorganic salt base medium, add thiram, and prepare a solution Finally, add distilled water to make it constant to 500mL. The amount of thiram added above should make the final concentration of thiram in the prepared culture solution be 100mg / mL, and then prepare the prepared culture solution at a temperature of 121°C Sterilize for 20 minutes to obtain the required culture solution. After culturing on this medium for three days, a strain was isolated by dilution coating and named FM84, which was deposited in the "General Microbiology Center of China Committee for the Collection of Microbial Cult...

Embodiment 2

[0043] Preparation of bacteria

[0044] The preserved species of the FM84 strain was inoculated into the LB solid plate culture method and cultivated for 24 hours until a single colony appeared; 2 PO 4 3g / L, MgSO 4 •7H 2 O 0.1g / L, glucose 5 g / L culture medium in the Erlenmeyer flask, cultivate to the logarithmic growth phase to obtain the seed liquid; the culture liquid that the seed culture obtains is connected in the fermenter, the inoculation ratio is 5%, 28 ℃, 200rpm, the air flow rate is 0.8vvm, cultivated for 48h to obtain the production culture solution. Centrifuge the fermented broth obtained from the production culture (6000rpm, 10min), suspend the precipitate with an aqueous solution containing 0.5% Tween-20 and 0.9% sodium chloride, and subpackage to obtain the thiram pesticide residue-degrading bacterial agent.

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Abstract

The invention discloses Klebsiella pneumoniae, a bacterium agent of the Klebsiella pneumoniae and a preparation method and application. The Klebsiella pneumoniae FM84 is a thiram degradation bacterium; the preservation unit is China General Microbiological Culture Collection Center (CGMCC); the preservation date is December 15, 2015; the preservation number is CGMCC N0. 11663; the Klebsiella pneumoniae belongs to bacteria, proteobacteria, gamma-proteobacteria, enterobacteriales, enterobacteriaceae and Klebsiella. The bacterium agent with the Klebsiella pneumoniae as an active ingredient is a bacterium agent prepared by activating the Klebsiella pneumoniae FM84 through strains, culturing seeds, producing and culturing and preparing with a preparation. The preparation method comprises an activation culture step and a fermentation culture step; the invention provides application of the bacterium agent to preparation of agricultural plants capable of degrading thiram. The bacterium agent comprising the strains has a simple preparation process, is low in cost and convenient to use, and has very good application prospect.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to a Klebsiella pneumoniae and its bacterial agent, preparation method and application. Background technique [0002] Thiram is a commonly used pesticide in the prevention and control of plant fungal diseases. Its chemical name is tetramethylthioperoxydicarbonic acid diamide, and it is often called thiuram, saiosan, etc. English name: tetramethylthiuram disulfide; Thiram. White or off-white, with special smell, crystalline powder. Density slightly greater than water. Insoluble in water, insoluble in dilute lye, gasoline, soluble in ethanol, benzene, chloroform, carbon disulfide, etc. Thiram and mancozeb are often used as two active ingredients to make a compound fungicide, which is a low-toxicity fungicide and has an irritating effect on the skin and mucous membranes. It can treat fungal diseases such as blight, and is widely used as a plant fungicide. It has good c...

Claims

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Application Information

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IPC IPC(8): C12N1/20A62D3/02A62D101/04C12R1/22
CPCA62D3/02A62D2101/04C12N1/20C12N1/205C12R2001/22
Inventor 夏振远雷丽萍吴玉萍莫笑晗师君丽马雁军
Owner YUNNAN ACAD OF TOBACCO AGRI SCI
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