Abstract: | With the aim of explaining the variations in microcystin (MC) concentrations during cyanobacterial blooms, we studied several Microcystis aeruginosa populations blooming in different freshwater ecosystems located in the same geographical area. As assessed by real-time PCR, it appeared that the potentially MC-producing cells (mcyB+) were predominant (70 to 100%) in all of these M. aeruginosa populations, with the exception of one population in which non-MC-producing cells always dominated. Apart from the population in the Grangent Reservoir, we found that the proportions of potentially MC-producing and non-MC-producing cells varied little over time, which was consistent with the fact that according to a previous study of the same populations, the intergenic transcribed spacer (ITS) genotype composition did not change (38). In the Grangent Reservoir, the MC-RR variant was the dominant microcystin variant throughout the bloom season, despite changes in the ITS composition and in the proportions of mcyB+ cells. Finally, the variations in total MC concentrations (0.3 to 15 μg liter−1) and in the MC cellular quotas (0.01 to 3.4 pg cell−1) were high both between and within sites, and no correlation was found between the MC concentrations and the proportion of mcyB+ cells. All of these findings demonstrate that very different results can be found for the proportions of potentially MC-producing and non-MC-producing cells and MC concentrations, even in M. aeruginosa populations living in more or less connected ecosystems, demonstrating the importance of the effect of very local environmental conditions on these parameters and also the difficulty of predicting the potential toxicity of Microcystis blooms.Microcystins (MCs) are the most common cyanotoxins and have been involved in several animal and human poisoning episodes (8). These hepatotoxic cyclic heptapeptides are synthesized by a multifunctional enzyme complex (10, 40), and the discovery of the gene cluster encoding this complex has made it possible in recent years to develop molecular tools for studying the relative proportions of MC-producing and non-MC-producing cells in natural cyanobacterial populations. Potentially MC-producing and non-MC-producing cells can coexist in these populations, but the factors and processes governing the dynamics of these subpopulations remain unclear.Some recent papers on the Microcystis genus have shown that the proportions of potentially MC-producing cells can differ considerably from lake to lake. For example, in Lake Wannsee, Germany, this proportion was between 0 and 40% (28), as it was in Lake Oneida, United States (18), and in Lake Mikata, Japan (48). In contrast, large variations over time (6 to 93%) of potentially MC-producing cells were found in the Grangent Reservoir, France (4). Major variations (30 to 80%) were also found in a natural French population of Planktothrix agardhii (3), and the variations in the proportions of potentially MC-producing cells reflected those of the MC concentrations. However, only 54% of the variation in MC concentrations could be explained by changes in the proportion of MC-producing cells, suggesting that a considerable part of the MC concentrations was also due to variations in MC cell quotas. These findings suggest that the toxic risks during cyanobacterial proliferations are due to variations in both the proportion of MC-producing cells and the production of MC by the toxic cells.Numerous papers have already investigated the impact of various biotic and abiotic environmental factors on microcystin production by toxic cells. These studies demonstrate that MC production can be influenced by temperature (35), light (46), nutrients such as nitrogen and phosphorus (12, 32), pH (39), iron (42), xenobiotics (17, 34, 45), and predators (22, 23, 47). Despite inconsistent results, the production of microcystins by the cells does seems to be linked to their growth rate (11, 31, 33), which is itself affected by environmental conditions. On the other hand, several studies of variations in the proportions of MC-producing cells have demonstrated the potential influence of nutrient concentrations (9, 48) and light and temperature (5), and two papers (3, 5) have suggested that there is a negative correlation between the proportions of MC-producing cells and the abundance of cyanobacterial cells. These findings are consistent with the data of Kardinaal and Visser (26), showing that in Dutch lakes there is a negative relationship between the densities of cyanobacterial cells and the mean MC concentration in the cells.In an overall attempt to explain the variations of toxicity during cyanobacterial blooms, we studied the spatiotemporal variations in MC concentrations and in the proportions of MC-producing and non-MC-producing cells in several Microcystis aeruginosa populations blooming in different freshwater ecosystems located in the same geographical area. The point of this study was to analyze these variations in terms of the characteristics of these ecosystems and the population dynamics of the M. aeruginosa populations. In addition, these data were compared to the variations in the intergenic transcribed spacer (ITS) composition of the same populations recently reported by Sabart et al. (38). The proportion of potentially MC-producing cells was estimated by a real-time quantitative PCR approach, the change in threshold cycle (ΔCT) method recently developed by Briand et al. for Planktothrix (3) and Microcystis (4) and targeting the mcyB (mcyA for Planktothrix) and phycocyanin (PC) genes. |