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Bacillus subtilis capable of degrading organophosphorus and preventing diseases

A technology of Bacillus subtilis and organophosphorus, which is applied in the field of agricultural microorganisms, can solve the problems of single function of Bacillus, and achieve the effects of difficult drug resistance, broad antibacterial spectrum and good inhibitory effect.

Active Publication Date: 2018-05-18
齐永新
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The functions of the above-mentioned known bacillus are relatively single, or they only have the function of dissolving phosphorus, or only have the function of preventing and controlling diseases; secondly, with the change of environmental conditions or the evolution of pathogenic bacteria, the function of bacillus may also evolve and change accordingly Therefore, continuous isolation and screening of Bacillus with high-efficiency phosphorus-dissolving function and broad antibacterial spectrum from the soil will help to adjust the contradiction between supply and demand of soil phosphorus, improve the effect of plant disease prevention, reduce production costs, and protect the ecological environment. It is of great significance to promote the sustainable development of agriculture

Method used

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  • Bacillus subtilis capable of degrading organophosphorus and preventing diseases
  • Bacillus subtilis capable of degrading organophosphorus and preventing diseases
  • Bacillus subtilis capable of degrading organophosphorus and preventing diseases

Examples

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Effect test

Embodiment 1

[0047] Example 1 Screening and isolation process and classification identification of bacterial strain WKPHY-54 of the present invention

[0048] (1) Collection, isolation and screening process of WKPHY-54 strain

[0049] In September 2015, Baoding Microcontrol Biotechnology Co., Ltd. collected 5 soil samples, each 200g, at five points in the Phosphate Mine Area of ​​Fanshan, Hebei Province, Zhulu County, Hebei Province. Weigh 1.0g of air-dried soil samples and add them to the Erlenmeyer flask with sterilized glass beads, then add 99mL of sterile water, let it stand for 20min, fully shake it on the shaker at 30°C and 180r / min for 30min, and then proceed according to the 10-fold dilution method Gradient dilution, respectively take 10 -3 , 10 -4 , 10 -5 100 μL of the diluted solution of the above-mentioned solution was applied on the organophosphate solution medium, and each concentration was repeated three times. 7d. The strains with phosphorus-solubilizing function were s...

Embodiment 2

[0058] Embodiment 2 The present invention contains the preparation of the microbial bacterial agent of WKPHY-54 bacterial strain

[0059] Follow the steps below:

[0060] (1) Strain activation: Activate the Bacillus subtilis strain WKPHY-54 (preservation number: CGMCCNo.14951) stored at -80°C on LB plate medium (30°C), pick a single colony and culture it on the LB slant Cultivate on the base for 12 hours at 30°C to obtain activated strains; the composition and weight ratio of LB plate medium or LB slant medium are: tryptone 10g, yeast extract 5g, sodium chloride 5g, agar powder 15g, water 1000mL;

[0061] (2) Preparation of seed liquid: 100 mL of LB liquid culture medium (its composition and weight ratio: tryptone 10 g, yeast extract 5 g, sodium chloride 5 g, water 1000 mL) was placed in a 250 mL conical flask, and high pressure damp heat Sterilize, after the temperature drops to room temperature, insert an inoculation loop into each bottle of the activated strain in step (1...

Embodiment 3

[0065] Example 3 Qualitative Determination Test of the Bacterial Strain WKPHY-54 of the Present Invention for Degrading Organophosphorus Ability

[0066] Proceed as follows:

[0067] With sterilized toothpick the WKPHY-54 bacterial strain that embodiment 2 step (1) is activated is spot-inoculated on solution organophosphate plate culture medium (its composition and its weight ratio are: glucose 10.0g, (NH 4 ) 2 SO 4 0.2g, MgSO 4 ·7H 2 O 0.5g, KCl 0.1g, MgCl 2 ·6H 2 O 5.0g, calcium phytate 2.0g, agar 20.0g, distilled water 1000mL, pH: 7.0-8.0), placed in a constant temperature incubator at 30 ℃ for 72 hours, measure the diameter of the transparent circle and the diameter of the colony.

[0068]As a result, it was found that the WKPHY-54 bacterial strain of the present invention produced a transparent circle with a diameter of 13.4 mm on the organophosphorus plate medium containing calcium phytate, indicating that the bacterial strain WKPHY-54 of the present invention can...

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Abstract

The invention discloses a Bacillus subtilis strain WKPHY-54, collected under CGMCC No. 14951, and also discloses a microbial agent containing the strain. The strain WKPHY-54 can degrade organophosphorus and prevent diseases, utilization rate of phosphate fertilizer or organophosphorus in soil can be increased, crop growth can be promoted, fertilizer input can be decreased, and production cost canbe decreased; in addition, the strain WKPHY-54 can effectively prevent plant diseases such as eggplant verticillium wilt, averagely up to 70.0% and above; further, the strain WKPHY-54 has a wide antibacterial spectrum, and has good inhibitory effect for eggplant verticillium, cucumber verticillium, Fusarium vasinfectum, potato Phyllachora and the like; more additionally, the strain WKPHY-5 is highly specific to the plant diseases such as eggplant verticillium wilt, rarely causes resistance and has long-lasting effect.

Description

technical field [0001] The invention belongs to the field of agricultural microorganisms, and in particular relates to a bacillus subtilis with dual functions of degrading organic phosphorus and preventing diseases, and also relates to a microbial agent containing the bacillus subtilis, and their application in degrading organic phosphorus and preventing and controlling crop diseases . Background technique [0002] Phosphorus is one of the mineral elements necessary to maintain the normal growth of plants, and is an important component of nucleic acids, various enzymes, coenzymes, and ATP in plants. The phosphorus needed for plant growth and development mainly comes from the supply of fertilizers and soil. The phosphorus fertilizer applied to the soil is easy to form insoluble phosphate, which is difficult to be absorbed and utilized by crops. Therefore, the key to improving the phosphorus nutrition of plants is to transform and utilize the insoluble phosphorus in the soil....

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P3/00C09K17/14C12R1/125C09K101/00
CPCA01N63/00C09K17/14C09K2101/00C12N1/20C12N1/205C12R2001/125
Inventor 李博超马毅楠孔令佳
Owner 齐永新
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