Culture of a high-chlorophyll-producing and halotolerant Chlorella vulgaris

In order to increase the value of freshwater algae as raw ingredients for health foods and feed for seawater-based farmed fish, we sought to breed high-chlorophyll halotolerant Chlorella with the objective of generating strains with both high chlorophyll concentrations (≥ 5%) and halotolerance (up to 1% NaCl). We used the Chlorella vulgaris K strain in our research institute culture collection and induced mutations with UV irradiation and acriflavine which is known to effect mutations of mitochondrial DNA that are associated with chlorophyll production. Screenings were conducted on seawater-based "For Chlorella spp." (FC) agar medium, and dark-green-colored colonies were visually selected by macroscopic inspection. We obtained a high-chlorophyll halotolerant strain (designated C. vulgaris M-207A7) that had a chlorophyll concentration of 6.7% (d.m.), a level at least three-fold higher than that of K strain. This isolate also exhibited a greater survival rate in seawater that of K strain.