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
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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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