Bacillus megatherium T317, microbial agent and preparation method of microbial agent

A technology of Bacillus megaterium and bacterial agent, applied in the field of microbial bacterial agent, can solve the problem of rare nitrogen fixation of Bacillus megaterium

Active Publication Date: 2016-10-12
DONGGUAN BAODE BIOLOGICAL ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, domestic research on Bacillus megaterium in microbial fertilizers mainly focuses on strain selection, phosphorus solubilizing effect, ...

Method used

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  • Bacillus megatherium T317, microbial agent and preparation method of microbial agent
  • Bacillus megatherium T317, microbial agent and preparation method of microbial agent
  • Bacillus megatherium T317, microbial agent and preparation method of microbial agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0107] One kind of Bacillus megaterium T317 in this example, the Bacillus megaterium T317 is deposited in the China Center for Type Culture Collection, and the deposit number is CCTCCM 2015753. The Bacillus megaterium T317 has the DNA sequence of Sequence Listing No. 1.

[0108] The nitrogenase activity of the Bacillus megaterium T317 was 800 nmol / (mL·h), and the secretion amount of indoleacetic acid in the process of growth and metabolism was 312.48 mg / L.

[0109] A method for preparing a bacterial agent of Bacillus megaterium T317, comprising the following steps:

[0110] (1) Separation and screening

[0111] The soil samples containing the nitrogen-fixing bacteria colonies of Bacillus megaterium T317 were selected, and the nitrogen-fixing bacteria colonies were obtained by separating and selecting the culture medium;

[0112] (2) Purification and preservation

[0113] The nitrogen-fixing bacteria colonies obtained by separation and screening are purified on the purificat...

Embodiment 2

[0122] The difference between this embodiment and Embodiment 1 is that the steps (2) and (3) of this embodiment further include the following steps:

[0123] (S1) Gram staining: Gram staining is performed on the purified single colony, and positive bacteria are obtained by screening;

[0124] (S2) Spore staining: the positive bacteria are stained with spores, and a single colony of Gram-positive bacteria containing spores is obtained by screening.

[0125] Specifically, the Gram staining method is:

[0126] (1) Smear: In a sterile operating table, take a glass slide and bake it slightly above the flame lamp to remove impurities on the glass slide. Drop a drop of sterile water in the center of the slide, pick a single colony into the water drop, and spread it evenly with a cauterized inoculation loop. Pass the sample slide 3 times back and forth over the fire light to fix the cells.

[0127] (2) Initial dyeing: Add 2-5 drops of ammonium oxalate crystal violet dye solution dr...

Embodiment 3

[0136] The difference between this example and Example 1 or 2 is that the nitrogenase activity of the Bacillus megaterium T317 in this example is 830 nmol / (mL·h), and the secretion amount of indoleacetic acid in the process of growth and metabolism is 830 nmol / (mL·h). is 340mg / L.

[0137] Step (3) Solid Fermentation Culture

[0138] The above-mentioned Bacillus megaterium T317 was selected and cultured on a solid medium. The pH of the fermentation environment was 7.2, the inoculum amount was 0.6%, the culture temperature was 32°C, and the culture time was 58 hours.

[0139] In the solid fermentation culture of step (3), the medium is composed of the following raw materials by weight: bran 70%, soybean meal 24%, NaCl 4.3%, CaCO 3 1.6%, MnSO 4 ·H 2 O 0.04%, MgSO 4 ·7H 2 O 0.06%, the pH of the medium is 7.2.

[0140] In the described step (1), the separation and screening medium is made from the raw materials of the following quality: CaCO 3 1.2g, MgSO 4 ·7H 2 O 0.8g, ...

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Abstract

The invention relates to the technical field of microbial agents, in particular to bacillus megatherium T317, a microbial agent and a preparation method of the microbial agent. The bacillus megatherium T317 is collected in China Center for Type Culture Collection, and the collection number is CCTCCM 2015753. The preparation method of the microbial agent comprises the following steps of (1) separating and sieving; (2) purifying and storing; (3) solid fermenting and culturing; (4) preparation of microbial agent, so as to obtain the microbial agent of the bacillus megatherium T317. The microbial agent has the advantages that the bacillus megatherium T317 is used as an original strain, and the nitrogenase activity and the auxin secreting capability are measured; the bacillus megatherium T317 has higher nitrogenase activity and IAA (indole acetic acid) secreting capability, the nitrogenase activity of bacillus megatherium T317 is 800-900nmol/(mL h), and the IAA secreting amount is 300-400mg/L in the growth and metabolism process.

Description

technical field [0001] The invention relates to the technical field of microbial inoculants, in particular to a Bacillus megaterium T317, inoculum and a preparation method thereof. Background technique [0002] The long-term over-reliance on chemical fertilizers and pesticides in agricultural production has resulted in the decline of farmland soil quality and fertility, the deterioration of crop quality, and the increasingly serious environmental pollution such as food and groundwater. With the rise and development of ecological agriculture and green food production, coupled with the lack of available phosphorus, potassium and other nutrients in most soils in my country, microbial fertilizers, as an important fertilizer source for the development of biotechnology and agricultural production, have gradually attracted people's attention. focus on. [0003] Bacillus megaterium (Bacillus megaterium) is an aerobic spore-producing Gram-positive bacterium with a research history of...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P21/00C12R1/11
CPCA01N63/00C12N1/20C12N1/205C12R2001/11
Inventor 沈世华杨国平林敏孙旭生王亚君尹坤杨盼盼张学贤陈三凤谭志远燕永亮
Owner DONGGUAN BAODE BIOLOGICAL ENG
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