A kind of alkalophilic bacillus strain wp-1 and its application
A technology of Bacillus alkalophilus, wp-1, which is applied in the field of agricultural microorganisms and can solve problems such as poor plant growth and soil salinization
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Embodiment 1
[0048] 1.1 Isolation and purification of strains
[0049] The soil was collected from the saline-alkali land of the 146th regiment of the Eighth Division of Xinjiang, put into a clean sampling bag, marked, and stored in a 4°C refrigerator for later use. Weigh 10g of the collected soil sample and place it in a 250mL Erlenmeyer flask of 90mL sterilized water. Mix thoroughly in the test tube, then use a pipette to draw 1mL from this test tube and add it to another test tube containing 9mL sterile water, mix well, and so on to make 10 -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 After the solutions with different dilutions, draw 0.1 mL each and spread them evenly on the bacterial culture medium (LB) plate, invert them in a 30°C incubator for 2-3 days, pick a single colony and connect it to the LB slant, and wait for it to grow out. Store in a 4°C refrigerator after the bacterial lawn.
[0050] LB medium: 5g of yeast extract, 10g of tryptone, 10g of sodium chloride, 1000mL of ...
Embodiment 2
[0055] 2.1 Study on growth characteristics of strain wp-1
[0056] 2.1.1 Effect of different pH on the growth of strain wp-1
[0057]The medium is LB-based medium, and the pH is adjusted to 6, 7, 8, 9, 10, 11, and sterilized at 121°C for 20 minutes when preparing the medium. Cultivate the strain in LB culture medium overnight, inoculate it with 1.0% inoculum amount in LB culture medium with different pH, culture with shaking at 150r / min, 30°C for 24h, and use 722 spectrophotometer to detect the optical density (D 600 nm), the results are shown in Table 1, and the optimum growth pH is 10.
[0058] Table 1 Effects of different pH on the growth of strain wp-1
[0059]
[0060] 2.1.2 Effects of different temperatures on the growth of strain wp-1
[0061] Cultivate the strain in LB culture medium overnight, inoculate the strain in LB culture medium (1% NaCl, pH 8) with 1.0% inoculum, respectively place them in a temperature gradient of 24-36°C, shake at 150r / min for 24h, and...
Embodiment 3
[0069] 3.1 Quantitative determination of phosphorus-dissolving ability of strain wp-1
[0070] Montina organophosphate medium (1L): glucose 10.0g, (NH4) 2 SO 4 0.5 g, MgSO 4 ·7H 2 O 0.3g, NaCl 0.3g, KCl 0.3g, FeSO 4 ·7H 2 O 0.03g, MnSO 4 ·H 2 O 0.03g, lecithin 0.2g, CaCO 3 1.0 g, yeast powder 0.5 g, agar 20.0 g; distilled water 1000 mL; liquid medium without agar, sterilized at 121°C for 20 minutes.
[0071] PKO inorganic phosphorus medium (1L): glucose 10.0g, MgSO 4 ·7H 2 O 0.3g, (NH 4 ) 2 SO 4 0.5g, NaCl0.3g, Ca 3 (PO 4 ) 22.0g, KCl 0.3g, MnSO 4 ·H 2 O 0.03g, FeSO 4 ·7H 2 O 0.036g, agar 20g, distilled water 1000mL; liquid medium without agar, sterilized at 121°C for 20 minutes.
[0072] 2μg·mL -1 Gradiently dilute the phosphorus standard solution to the phosphorus content of 0.00, 0.04, 0.08, 0.24, 0.40, 0.80, 1.20μg·mL -1 Make a standard curve. Add molybdenum antimony anti-color developing agent at room temperature, develop color for 20min, measure its...
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