Proliferations of cyanobacteria have detrimental effects on ecosystem functioning, and on the global freshwater food chain. Many studies have focused on the “in situ” dynamics of bloom-forming cyanobacteria, including Cylindrospermopsis raciborskii and Planktothrix agardhii. Few have used experimental assays to explore the fast-growing ability of naturally co-occurring species. Here we investigated the growth of these species when exposed separately (i.e., in monocultures) to a range of light and nutrient conditions, plus their interactive performances in mixed cultures in a short-time experiment (6 days). The use of microplates made it possible to carry out multiple measurements of in-vivo fluorescence (IVF), and to monitor species-dependent biovolumes. No allelopathic effect was significantly observed for any target species, while significantly lower growth rates were obtained in mixed cultures, which may reflect other interference interactions between the species. We showed that Planktothrix grew faster with low light intensity and high nutrient concentrations, and was drastically inhibited by nitrogen deprivation, in contrast to Cylindrospermopsis. However, Cylindrospermopsis outgrew Planktothrix at high NH4+ concentrations, suggesting that this species may be a serious competitor for the native species in many water systems.