A method for in-situ aseptic enrichment and cultivation of Synechococcus
A technology of enrichment culture and Synechococcus, applied in the field of aseptic culture of marine picocyanobacteria, can solve the problems of unsatisfactory effect, cell damage of Synechococcus, long experimental period, etc.
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Embodiment 1
[0028] 1) Cultivation of algae strains: Algae strains were grown in SNAX medium and cultured in a light incubator at 25 °C, with a light intensity of 2000 lx and a light-to-dark ratio of 12 h: 12 h.
[0029] 2) Antibiotic preparation: weigh Kanamycin Monosulfate (Km), Ampicillin (Amp), Cefotaxime Sodium (CTX), Streptomycin Solution (STR), Gentamic Sulfate Gentamicin Sulfate (Gm) and Neomycin Sulphate (Neomycin Sulphate, N) were used to prepare an antibiotic solution with a mass concentration of 10 g / L in sterilized ultrapure water.
[0030]3) Antibacterial effect of antibiotics: Take 100 μL of the above-mentioned Synechococcus culture solution cultured to the logarithmic growth phase and spread it on the 2216E solid medium; soak a filter paper piece with a diameter of 5.5 mm in the above antibiotic solutions, Paste 4 pieces of filter paper on each plate (one plate only uses one antibiotic); incubate at 25°C for 3 days, and measure the size of the inhibition zone. The measurem...
Embodiment 2
[0036] Utilize the conclusion of above-mentioned embodiment 1 to further verify:
[0037] 1) Cultivation of algae strains: Algae strains were grown in SNAX medium at 25 °C, in a light incubator with a light intensity of 2000 lx and a light-to-dark ratio of 12 h: 12 h until the logarithmic growth phase.
[0038] 2) Antibiotic preparation: STR, Gm, and N were prepared with sterilized ultrapure water to prepare antibiotic solutions with a mass concentration of 50 g / L.
[0039] 3) Sterile enrichment culture: Culture the above-mentioned Synechococcus liquid culture solution cultured to the logarithmic growth phase at 25 °C in the dark for 30 h, so that the Synechococcus is in the stagnant stage of division; after the dark culture, take For three enriched Synechococcus liquids, add an antibiotic solution to each algae liquid at the dose of 1 mL antibiotic solution per 100 mL, and continue to culture in the dark for 12 h; after the dark culture, use sterilized natural seawater The c...
Embodiment 3
[0044] 1) Cultivation of algae strains: Algae strains were grown in SNAX medium at 25 °C, in a light incubator with a light intensity of 2000 lx and a light-to-dark ratio of 12 h: 12 h until the logarithmic growth phase.
[0045] 2) Antibiotic preparation: Dissolve STR, Gm, and N in sterilized ultrapure water to prepare antibiotic solutions with mass concentrations of 50 g / L, 50 g / L, and 50 g / L, respectively, and the volume ratio is 1:1:1 Mix as mixed antibiotic working solution.
[0046] 3) Sterile enrichment culture: culture the above-mentioned Synechococcus liquid culture solution cultured to the logarithmic growth phase at 25 °C in the dark for 30 h, so that the Synechococcus is in the stagnant stage of division; after the dark culture, every 100 Add 3 mL of antibiotic working solution to the Synechococcus liquid, and continue to culture in the dark for 12 h; after the dark culture, wash the culture solution with sterilized natural seawater for 4 times, and then resuspend ...
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