Isolation of cyanobacteria from soil

media preparation

[[Cyanobacteria-Medium-BG11]] [[Cyanobacteria-Medium-Soil-enriched]] * Note:

isolation procedure:

• dry soil for 1 week at 25C in dark
• After drying, soak dried soil with sterile nanopure H2O, allow to sit for 1 day.
• After soaking, add nanopure H2O to cover soil, place in shaker at 150 rpm, leave on shaker for 2 hrs
• Make 3 - 10-fold dilutions, then spread 100ul of each dilution onto AGAR plates.
• Place under continuous light at 22-30C.
• Incubate for up to 1 month.
• Evaluate and note growth daily.

• After incubation period, streak colonies for isolation.

• Identification of pure cultures can be achieved by Cyanobacterial specific primers CYA-359F (5′-GGGGAATYTTCCGCAATGGG-3′) and CYA-781R (5′-GACTACWGGGGTATCTAATCCCW-3′) @Nubel1997 @Lee2017 ‌

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Citations

• Katoh et al 2012. Isolation and purification of an axenic diazotrophic drought-tolerant cyanobacterium, Nostoc commune, from natural cyanobacterial crusts and its utilization for field research on soils polluted with radioisotopes. Biochimica et Biophysica Acta
• Marks et al 2017. Application of a microalgal slurry to soil stimulates heterotrophic activity and promotes bacterial growth. Science of the Total Environment
• Lee et al 2017. Higher production of C-phycocyanin by nitrogen-free (diazotrophic) cultivation of Nostoc sp. NK and simplified extraction by dark-cold shock. Bioresource Technology
• Rippka et al 1979. Generic assignments, strain histories and properties of pure cultures of cyanobacteria. J. Gen. Microbiol.
• Nubel et al 1997. PCR primers to amplify 16S rRNA genes from cyanobacteria. Applied Environ Microbial.
• Rippka 1988. Isolation and Purification of Cyanobacteria. Methods in Enzymology.
• Allen MM, Stanier RY (1968) Growth and division of some unicellular blue-green algae. J Gen Microbiol 51(2):199–202
• Ferris MJ, Hirsch CF (1991) Method for isolation and purification of cyanobacteria. Appl Environ Microbiol 57(5):1448–1452