Bacillus subtilis n24 and its application
A technology of bacillus subtilis and bacteria agent, applied in the field of bacillus subtilis N24 and its application
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Embodiment 1
[0041] Example 1 Isolation, screening and identification of strain N24
[0042] 1. Isolation and screening of strain N24
[0043] In 2019, soil was collected from the pepper rhizosphere of the experimental and demonstration base of Genlituo Biotechnology Co., Ltd., Wei County, Xingtai City, Hebei Province. Shaker at 200rpm for 30min, add 1ml to 9ml sterile saline, and then dilute 10 2 , 10 3 , 10 4 Take 100 μl of the above soil suspensions and spread them evenly on LB medium (yeast powder 5g / L, peptone 10g / L, NaCl 5g / L, agar 15g / L), and culture them in an incubator at 28°C for 48h. Sterile toothpicks were used to pick out bacterial colonies of different shapes, streak and purify them, and save them for later use. A total of 892 bacterial strains were isolated.
[0044] The biocontrol effect of the isolated strains was detected by the confrontation culture method. Firstly, Botrytis cinerea was activated on a PDA plate, and a 5mm hole punch was used to make a pathogenic bac...
Embodiment 2
[0055] Embodiment 2 Phosphorus-dissolving function detection
[0056] Strain N24 was first activated on LB medium. The activated strain was inoculated into the phosphorus-dissolving medium shown in Appendix A of NY / T 1847-2010 (glucose 10.00 g, (NH 4 ) 2 SO 4 0.50g, MnSO 4 ·7H 2 O0.3g, NaCl 0.3g, KCl0.30g, FeSO 4 ·4H 2 O0.036g, MnSO 4 ·4H 2 O 0.03g, distilled water 1000mL, pH 7.0. Calcium phytate was selected as the phosphorus source, and the addition amount was 2 g; the solid medium was added with 1.5% agar powder in proportion). Detect whether it has the function of dissolving phosphorus. After testing, N24 has the ability to dissolve phosphorus, and its dissolving circle diameter (D) reaches 11.49±0.31mm ( figure 2 ).
[0057] After confirming that the strain has the function of dissolving phosphorus, the ability of dissolving phosphorus of the strain is quantitatively determined, and the content of available phosphorus in the fermentation supernatant is determi...
Embodiment 3
[0059] Example 3 Determination of protease production ability
[0060] First, the strain N24 was activated on LB medium, and the activated strain was inoculated on the medium for protease detection (tryptone 5.00g, yeast extract 3.00g, glucose 1.00g, agar 15.00g, distilled water 1000mL, pH 7.0, Autoclave sterilization at 121°C for 30min. When the sterilized detection medium is cooled to about 50°C, add skim milk at a ratio of 10% to the medium, mix well, pour it into a petri dish, and wait for it to cool for later use), incubate at 28°C 48h, observe whether there is a dissolution circle, the results show that N24 has a good ability to dissolve proteins, high protease production, its dissolution circle diameter (D) reaches 25.72±0.79mm, colony diameter (d) 17.67±0.53mm, the amount of enzyme produced (D / d) is 1.46±0.31( image 3 ).
[0061] Protease activity detection: Inoculate N24 strain with 5% inoculum into liquid detection medium (tryptone 5.00g, yeast extract 3.00g, gluc...
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