Bacillus megaterium D122, Bacillus megaterium D122 agent, and Bacillus megaterium D122 agent preparation method

A technology of Bacillus megaterium and inoculum, which is applied in the field of Bacillus megaterium D122 and the preparation of inoculants and inoculants, can solve the problem of rare research on Bacillus megaterium inoculants protection agents, low effective viable bacteria, and short validity period And other issues

Active Publication Date: 2016-11-30
DONGGUAN BAODE BIOLOGICAL ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the products produced by my country's microbial fertilizer industry show many problems, such as low effective number of viable bacteria, high rate of miscellaneous bacteria, short validity period, etc., which seriously affect the enthusiasm of farmers to use and market development
Domestic research on Bacillus megaterium in microbial fertilizers mainly focuses on strain selection, phosphorus-solubilizing effect, optimization of fermentation conditions, and interaction with potassium-solubilizing bacteria.

Method used

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  • Bacillus megaterium D122, Bacillus megaterium D122 agent, and Bacillus megaterium D122 agent preparation method
  • Bacillus megaterium D122, Bacillus megaterium D122 agent, and Bacillus megaterium D122 agent preparation method
  • Bacillus megaterium D122, Bacillus megaterium D122 agent, and Bacillus megaterium D122 agent preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] A kind of Bacillus megaterium D122 in this embodiment, said Bacillus megaterium D122 is preserved in China Center for Type Culture Collection, and the preservation number is CCTCCM 2015751. The Bacillus megaterium D122 has the DNA sequence of No. 1 in the sequence table.

[0082] The nitrogenase activity of the Bacillus megaterium D122 is 500 nmol / (mL·h), and it can produce indole acetic acid during growth and metabolism, and the secretion amount of indole acetic acid is 100 mg / L.

[0083] A method for preparing a bacterial agent of bacillus megaterium D122, comprising the following steps:

[0084] (1) Separation and screening

[0085] Select soil samples containing nitrogen-fixing bacteria colonies of Bacillus megaterium D122, and obtain nitrogen-fixing bacteria colonies through isolation and screening medium culture;

[0086] (2) Purification and preservation

[0087] Purify the isolated and screened nitrogen-fixing bacteria colony on the purification preservation ...

Embodiment 2

[0097] The difference between this embodiment and Embodiment 1 is that the following steps are also included between the step (2) and step (3) of this embodiment:

[0098] (S1) Gram staining: Perform Gram staining on the purified single colony, and screen to obtain positive bacteria;

[0099] (S2) Spore staining: The positive bacteria were stained with spores, and a single colony of Gram-positive bacteria containing spores was screened.

[0100] Specifically, the Gram staining method is:

[0101] (1) Smear: In a sterile operating table, take a piece of glass slide and bake it slightly above the flame lamp to remove impurities on the slide. Drop a drop of sterile water in the center of the slide, pick a single colony in the water drop, and spread evenly with a burnt inoculation loop. Pass the sample slide back and forth 3 times over the fire lamp to fix the cells.

[0102] (2) Initial dyeing: Add 2-5 drops of ammonium oxalate crystal violet dye solution, dye for 1 minute, po...

Embodiment 3

[0111] The difference between this example and Example 1 or 2 is that the nitrogenase activity of the Bacillus megaterium D122 in this example is 520 nmol / (mL h), which can produce indole acetic acid during growth and metabolism , the secretion of indole acetic acid is 120mg / L.

[0112] The initial bacterial count of the Bacillus megaterium D122 bacterial agent was determined to be 250 million / mL.

[0113] The mass concentration of sodium chloride in the step (4) is 6.5%, and the mass concentration of sodium acetate is 1.5%.

[0114] In the step (1), the separation and screening medium is made of raw materials of the following quality: CaCO 3 1.2g, MgSO 4 ·7H 2 O 0.8g, K 2 HPO 4 1.5g, NaCl 0.2g, FeSO 4 ·7H 2 O 0.002g, NaMO 4 2H 2 O 0.06g, sucrose 6g, agar 18g, distilled water 1000ml, the pH of separation and screening medium is 7.2.

[0115] The rest of the content of this embodiment is the same as that of Embodiment 1 or 2, and will not be repeated here.

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Abstract

The invention relates to the technical field of microbial agents, specifically to Bacillus megaterium D122, a Bacillus megaterium D122 agent, and a Bacillus megaterium D122 agent preparation method. According to the present invention, the Bacillus megaterium D122 is preserved in China Center for Type Culture Collection and has the preservation number of CCTCCM 2015751; the preparation method comprises: (1) separating and screening; (2) purifying and preserving; (3) culturing with a culture medium; and (4) D122 agent preparation to obtain the Bacillus megaterium D122 agent; and the Bacillus megaterium D122 has functions of nitrogenase activity and auxin indoleacetic acid secretion.

Description

technical field [0001] The invention relates to the technical field of microbial bacterial agents, in particular to a bacillus megaterium D122, its bacterial agent and a preparation method thereof. Background technique [0002] Agricultural production has long relied on chemical fertilizers, which not only wastes a lot of non-renewable resources, but also leads to problems such as soil quality deterioration, agricultural product quality decline, and environmental pollution, which greatly affects human health and survival. [0003] Bacillus megaterium ( Bacillus megaterium ) is a bacterium that decomposes phosphorus and promotes potassium and nitrogen fixation. It can degrade phosphorus and potassium in the soil that cannot be used by plants, improve soil fertility, and at the same time fix nitrogen in the air to promote crop production. However, the products produced by my country's microbial fertilizer industry show many problems, such as low effective number of viable bac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/11
CPCC12N1/20C12N1/205C12R2001/11
Inventor 张学贤林敏杨国平孙旭生王亚君杨盼盼尹坤沈世华谭志远燕永亮
Owner DONGGUAN BAODE BIOLOGICAL ENG
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