Algae. 1989; 4(2): 183-189.
A Simple Method of Seaweed Axenic Culture
Masakazu Tatewaki
Institute of Algological Research, Faculty of Science, Hokkaido University
The axenic culture is an essential of modern biological research, including tissue culture, protoplast isolation, cell fusion and gene transfer. However, seaweed axenic cultures are still believed to be extremely difficult to establish and various studies using axenic strains of seaweeds have been limited in comparison with those of unicellular microscopic algae. Here, a simple method obtaining seaweed axenic cultures combining the action of antibiotics with mechanical washing of reproductive cells (spores) is reported for various species, especially red seaweeds which do not have any motile stages in their life cycle. Fresh spores after washing serially with capillary pipettes are first introduced into a screw-cap test tube containing 10 ml of PES medium to establish unialgal cultures. Then 3-4 drops of antibiotic mixture are added to each unialgal culture with a Pasteur pipette, lthough periods of antibiotic treatment vary with the different species. The antibiotic mixture contains 100,000 units penicillin G potassium, 250 mg streptomycin sulfate,25,000 units polymyxin B sulfate, and 1 mg chloramphenicol in 10 ml of distilled water. With this method, axenic cultures are obtained with high percentage of success for various species examined.
Keywords : Antibiotic mixture, axenic culture, mechanical washing, seaweed spore-washing medium