Phosphorus-solubilizing and growth-promoting bacillus tropicalis SG15, biological agent and application of bacillus tropicalis SG15 and biological agent in potato seed dressing agent
A technology of bacillus and biological inoculum, which is applied in the field of agricultural biology to achieve the effects of remarkable resistance to heavy metal Cd stress, promoting growth and increasing yield
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Embodiment 1
[0033] Screening, isolation and purification of strains in Example 1
[0034] The red soil rhizosphere soil from the farm of South China Agricultural University was used as the screening soil sample, and 10.0 g of the soil sample was accurately weighed and placed in a 250 mL conical flask (with 5-7 glass beads) filled with 90 mL of sterile water. -1 , shake the soil sample for 30min to fully disperse the soil sample, let it stand for 20-30s, take 5mL of the supernatant in a test tube, and heat it at 90°C for 10min. Take 1 mL of the supernatant after the water bath and add 25 mL of phosphorus-dissolving medium (glucose 10.0 g, tricalcium phosphate 5.0 g, MgCl 2 ·6H 2 O 5.0 g, MgSO 4 ·7H 2 O 0.25g, KCl 0.2g, (NH 4 ) 2 SO 40.1g, distilled water 1000mL, pH 7.0) in a 100mL conical flask, placed in a shaker for enrichment culture, 180r·min -1 , 30℃. Taking 5 d as an enrichment culture cycle, after each sampling, 1 mL of culture solution was taken into a new phosphorus-dissol...
Embodiment 2
[0036] Example 2 Characterization
[0037] (1) Morphological characteristics of colonies
[0038] The colonies formed by SG15 after culturing on nutrient agar medium for 24 hours were irregular in shape, white, moist and smooth on the surface, not sticky and easy to provoke.
[0039] (2) Growth characteristics
[0040] Pick a single colony on the plate and incubate it in LB liquid medium at 180 rpm and a temperature of 37°C for 12 hours, then insert it into fresh LB liquid medium according to 1% of the inoculum, and cultivate for 12 hours. , the number of viable bacteria in the fermentation broth of the strain was 3.2 × 10 9 cfu·mL -1 .
[0042] Pick the colonies formed after culturing on the nutrient agar medium for 48 hours and place them on the glass slides, draw 20 μL of sterile water and mix them with the strains on the glass slides, dry naturally, fix them by flame heating, add dropwise 5% malachite green staining solution, and heat , ma...
Embodiment 3
[0049] The dissolution effect test of embodiment 3SG15 to different insoluble phosphates
[0050] The SG15 cells in glycerol were picked and streaked on LB solid medium plates, and cultured at 37°C for 12h. Pick a single SG15 colony on the plate and put it into a test tube containing LB liquid medium, and culture at 37°C, 180rpm, and shaker for 12h. The culture medium was transferred to fresh LB liquid medium and shaken to OD600=0.4 in a shaker at 37°C and 180 rpm. The bacterial solution was inoculated into 20 mL of liquid phosphorus-dissolving medium with calcium phosphate, iron phosphate and aluminum phosphate as the only phosphorus source according to the amount of 1% medium, and cultured at 30° C., 180 rpm, and shaker for 10 days. On the 3rd, 5th, 7th, and 10th days, 2 mL of culture solution was taken and centrifuged at 8000 rpm for 5 min. The supernatant was taken, and the water-soluble phosphorus content was determined by the molybdenum-antimony antibody method. The am...
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