Bacillus cereus for activating soil nutrient and application of bacillus cereus
A technology of Bacillus cereus and soil nutrients, applied in the direction of application, soil conditioning materials, bacteria, etc., to achieve the effects of increasing diversity, promoting sustainable use of soil, and saving mineral resources
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Embodiment 1
[0029] The isolation, purification and identification of embodiment 1 bacillus cereus NC-1
[0030] (1) Separation
[0031] Prepare insoluble inorganic phosphorus medium: glucose 10.0g, ammonium sulfate 0.5g, sodium chloride 0.3g, potassium chloride 0.3g, ferrous sulfate heptahydrate 0.03g, manganese sulfate tetrahydrate 0.03g, yeast extract 0.4g, phosphoric acid Calcium 5g, distilled water 1L, pH value 7.0~7.5.
[0032] The soil collected from the paddy field of South China Agricultural University in Guangzhou City, Guangdong Province was used as the screening soil sample, and 10.0g of the soil sample to be tested was accurately weighed by the flat plate coating method, and put into a 250mL triangular flask filled with 90mL sterile water (add glass beads), shake the shaker for 30 minutes to fully disperse the microorganisms, let it stand for 20~30s, and take the supernatant for 10-fold decreasing dilution, using 10 3 ~10 5 Dilution: Use a pipette gun to pipette 0.1mL of ...
Embodiment 2
[0036] Example 2 Characterization
[0037] (1) Morphological characteristics of bacteria
[0038] a-8 bacteria are Gram-positive bacteria with spores, and the bacterium does not expand after the formation of spores. The bacteria are rod-shaped, non-capsulated, motile, and aerobic. Gram staining effect see figure 1 , the cells were stained blue-purple, and the spores were not colored. Spore staining effect see figure 2 , the thallus is yellow-brown, and the spores are light blue.
[0039] (2) Colony Morphological Characteristics
[0040] The colonies formed on the nutrient agar medium are round or nearly round, soft, non-pigmented, and slightly glossy white colonies.
[0041] (3) Molecular biological characteristics
[0042]Bacterial total DNA was extracted using the Mobio PowerSoil DNA Isolation Kit (Bio-rad). Using bacterial 16S rDNA universal primers F63 (CAG GCC TAA CAC ATG CAA GTC, SEQ ID NO: 2) and R1087 (CTC GTTGCG GGA CTT ACC CC, SEQ ID NO: 3), routine PCR a...
Embodiment 3
[0043] Example 3 Effect experiment of NC-1 bacteria producing water-soluble phosphorus under insoluble inorganic phosphorus medium culture
[0044] Put 50mL of sterilized insoluble inorganic phosphorus medium (concentration of calcium phosphate is 5.0g / L) into the conical flask, add a-3, a-4, a -5, a-6, a-7, a-8 (that is, NC-1 bacteria) and a-9 bacteria (inoculum size 5 rings), after making the bacteria solution, culture it on a shaking table at 28°C and 70rpm for 7 days. Centrifuge the cultured bacterial solution at 10,000rpm and 4°C for 15min, take the supernatant and use the molybdenum antimony anti-colorimetric method to measure the content of water-soluble phosphorus (the unit is mg / L, which means the mg of water-soluble phosphorus contained in each liter of sample) number), the culture medium without any bacteria was set as the control, and each treatment was repeated 3 times. The test results are shown in Table 1 and image 3 . The results showed that the water-sol...
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