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Bacillus megatherium T482, microbial agent and preparation method of microbial agent

A technology of Bacillus megaterium and T482, applied in the field of microbial inoculants, 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, optimization of fermentation conditions, and interaction with potassium solubilizing bacteria, but the nitrogen fixation of Bacillus megaterium is rare.

Method used

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  • Bacillus megatherium T482, microbial agent and preparation method of microbial agent
  • Bacillus megatherium T482, microbial agent and preparation method of microbial agent
  • Bacillus megatherium T482, microbial agent and preparation method of microbial agent

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Effect test

Embodiment 1

[0084] A type of Bacillus megaterium T482 in this embodiment, said Bacillus megaterium T482 is preserved in China Center for Type Culture Collection, and the preservation number is CCTCCM 2015754. The Bacillus megaterium T482 has the DNA sequence of No. 1 in the sequence table.

[0085] The nitrogenase activity of the Bacillus megaterium T482 was 1100 nmol / (mL·h), and the secretion amount of indole acetic acid during the growth and metabolism process was 250 mg / L.

[0086] A method for preparing a bacterial agent of bacillus megaterium T482, comprising the following steps:

[0087] (1) Separation and screening

[0088]Select soil samples containing nitrogen-fixing bacteria colonies of Bacillus megaterium T482, and obtain nitrogen-fixing bacteria colonies through isolation and screening culture medium;

[0089] (2) Purification and preservation

[0090] Purify the isolated and screened nitrogen-fixing bacteria colony on the purification preservation medium, culture and isola...

Embodiment 2

[0101] 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:

[0102] (S1) Gram staining: perform Gram staining on the purified single colony, and screen to obtain positive bacteria;

[0103] (S2) Spore staining: the positive bacteria are stained with spores, and a single colony of Gram-positive bacteria containing spores is obtained by screening.

[0104] Specifically, the Gram staining method is:

[0105] (1) Smear: In a sterile operating table, take a 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.

[0106] (2) Initial dyeing: Add 2-5 drops of ammonium oxalate crystal violet dye solution, dye for 1 min, pou...

Embodiment 3

[0115] The difference between this embodiment and Embodiment 1 or 2 is that in the step (3) of the solid fermentation culture of the present embodiment, the above-mentioned Bacillus megaterium T482 single colony is selected and cultivated with a solid medium, and the pH of the fermentation environment is 7.1, the inoculum size is 0.5%, the culture temperature is 30°C, and the culture time is 60 hours.

[0116] Said step (4) bacterial agent preparation: the product after the solid fermentation culture is dried and crushed, the detection result of the number of viable bacteria shows that the range of the number of viable bacteria is 300 million / g, that is, the bacterial agent of Bacillus megaterium T482 is obtained .

[0117] In the step (1), the separation and screening medium is made from raw materials of the following quality: CaCO 3 1.0g, MgSO 4 ·7H 2 O 0.6g, K 2 HPO 4 1.0g, NaCl 0.1g, FeSO 4 ·7H 2 O 0.001g, NaMO 4 2H 2 O 0.05g, sucrose 5g, agar 18g, distilled wat...

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Abstract

The invention relates to the technical field of microbial agents, in particular to bacillus megatherium T482, a microbial agent and a preparation method of the microbial agent. The bacillus megatherium T482 is collected in China Center for Type Culture Collection, and the collection number is CCTCCM 2015754. 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 bacillus megatherium T482. The microbial agent has the advantages that the bacillus megatherium T482 has higher nitrogenase activity and IAA (indole acetic acid) secreting capability, the nitrogenase activity is 1000-1200nmol / (mL h), and the IAA secreting amount is 200-300mg / L in the growth and metabolism process; the microbial agent is a new microbial agent, and the application prospect in agricultural production is good.

Description

technical field [0001] The invention relates to the technical field of microbial bacterial agents, in particular to a bacillus megaterium T482, its bacterial agent and a preparation method thereof. Background technique [0002] The effective supply of chemical fertilizers is one of the basic guarantees for the sustainable development of agriculture. However, with the large-scale use of chemical fertilizers, especially nitrogen fertilizers, while meeting the needs of high-yield crops, the rate of return of chemical fertilizers is diminishing, soil physical properties deteriorate, and water resources are polluted. Such negative effects cannot be ignored. Because the application of traditional chemical fertilizers is accompanied by various disadvantages, it is very necessary to develop a new type of fertilizer that can reduce its dosage and meet the requirements of sustainable agricultural development. At present, as a new agricultural measure, the role of microbial fertilizer...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01P21/00C12R1/11
CPCC12N1/20C12N1/205C12R2001/11
Inventor 王亚君杨国平林敏孙旭生杨盼盼尹坤张学贤沈世华陈三凤谭志远燕永亮
Owner DONGGUAN BAODE BIOLOGICAL ENG
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