Antimicrobial Cocktails to Control Bacterial and Fungal Contamination in Chlamydomonas Reinhardtii Cultures

Abstract

Chlamydomonas reinhardtii is a unicellular green alga widely used for research in photosynthesis, cell cycle regulation, ciliary biogenesis, and other physiological processes. Sterile cultures are needed for these studies, but contamination from bacteria and fungi occurs frequently. Although the One-shot Solution cocktail consisting of carbendazim, ampicillin, and cefotaxime has been developed for removing these contaminants from algal cultures, it is not always effective. Here we report two new antimicrobial cocktails for treating mixed bacterial and fungal contamination of Chlamydomonas cultures. A combination of the bactericide nalidixic acid with one of two fungicides, azoxystrobin or tebuconazole, was more effective than the One-shot Solution cocktail. In some of our tests, we find that alternating use of our new cocktails with One-shot Solution is needed to remove obstinate contaminants.

Rapid and high efficiency transformation of Chlamydomonas reinhardtii by square-wave electroporation

“…C. reinhardtii wild-type strain 21gr (CC-1690, wild-type, mt+), 6145C (CC-1691, wild-type, mt−), CC-125 (137c, wild-type, mt+), CC-124 (137c, wild-type, mt−) were originally provided from Chlamydomonas Resource Center, University of Minnesota, U.S.A., and kept in our laboratory. Cells were cultured in Tris-acetate-phosphate (TAP) plate at 23 ± 0.5°C under 14/10 h light/dark cycles with light intensity of 8000 Lx [25,27].…”

Design of a nanobiosystem with remote photothermal gene silencing in Chlamydomonas reinhardtii to increase lipid accumulation and production

“…We initially grew C. reinhardtii under specific conditions to determine its photoheterotrophic growth kinetics. C. reinhardtii is a single-cell green alga; because of its remarkable adaptability to different environments and short generation time, it is considered a model organism for various studies, such as those related to lipid metabolism [23]. The sequencing of its whole genome and the availability of several molecular research tools allowing genetic studies make this organism an attractive model system to study at the molecular level [24].…”