Subtropical freshwater phytoplankton show a greater response to increased temperature than to increased pCO2.

Global increases in atmospheric CO2 and temperatures will impact aquatic systems, with freshwater habitats being affected. Some studies suggest that these conditions will promote cyanobacterial dominance. There is a need for a clearer picture of how algal species and strains within species will respond to higher temperatures and CO2, especially in combination. This study examined two chlorophytes (Monoraphidium and Staurastrum), and two strains of the cyanobacterium Raphidiopsis raciborskii (straight S07 and coiled C03), to determine how the combination of higher temperature and CO2 levels will affect their growth and maximum cell concentrations. Continuous cultures were used to compare the steady state cell concentrations at 28 °C and 30 °C, and CO2 partial pressures (pCO2), 400 and 750 ppm for all cultures, and in addition 1000 ppm at 28 °C for R. raciborskii strains. This study showed that, for all species, water temperature had a greater effect than higher pCO2 on cell concentrations. There were clear differences in response between the chlorophyte species, with Monoraphidium preferring 28 °C and Staurastrum preferring 30 °C. There were also differences in response of the R. raciborskii strains to increasing temperature and pCO2, with S07 having a greater increase in cell concentration. Genome analysis of R. raciborskii showed that the straight strain has five additional carbon acquisition genes (ß-CA, chpY, cmpB, cmpD and NdhD4), indicative of increased carbon metabolism. These differences in the strains' response to elevated pCO2 will lead to changes in the species population structure and distribution in the water column. This study shows that it is important to examine the effects of both pCO2 and temperature, and to consider strain variation, to understand how species composition of natural systems may change under future climate conditions.

Differential expression of phosphorus acquisition genes in response to phosphorus stress in two Raphidiopsis raciborskii strains.

The cyanobacterium Raphidiopsis raciborskii is a nuisance in freshwater ecosystems. Strains vary in their physiological responses to environmental drivers, thus a greater understanding of the magnitude of strain variation is required to characterize the species. In this study, two strains of R. raciborskii isolated from a tropical Australian water reservoir were grown with and without phosphorus (P) to determine any relative response to P stress. The strains had the same growth rates and under P free conditions, cells grew at the same rate as P replete conditions until day 9 when cell growth ceased. There was no difference in the alkaline phosphatase activity per cell for the P replete and P free conditions, but the level of activity per cell was greater in CS-505 than CS-506. P acquisition genes were identified from the sequenced genomes; both strains contained the same genes, but with differences in copy number of phoA (7 and 6), phnK (3 and 1) and phnH (2 and 1) between CS-505 and CS-506 (respectively). The expression of P acquisition genes under P stress was measured throughout the experiment and shown to vary in magnitude and timing across strains, and in P replete versus P free cultures. In strain CS-505, upregulation of the pstS1 and phoA genes occurred late in the growth phase and into senescence. These genes are involved in phosphate uptake and use of various forms of organic P. For strain CS-506, there was upregulation of the phosphate uptake gene, pit, and organic P utilization genes, phoA, phoU, phnD and phnK, commencing late in the growth phase. Our study shows that despite the fact that these two strains were isolated from the same waterbody, they differed markedly in their gene expression response to P free conditions. This capacity of R. raciborskii to vary in strain responses to P conditions gives the organism flexibility in responding to environmental change, particularly P stress conditions.

Nitrogen fixation by the diazotroph Cylindrospermopsis raciborskii (Cyanophyceae).

Nitrogen fixation has been proposed as a mechanism that allows the diazotrophic cyanobacterium, Cylindrospermopsis raciborskii, to bloom in nitrogen-limited freshwater systems. However, it is unclear whether dinitrogen fixation (N2 fixation) can supplement available dissolved inorganic nitrogen (DIN) for growth, or only provides minimum nitrogen (N) for cell maintenance under DIN deplete conditions. Additionally, the rate at which cells can switch between DIN use and N2 fixation is unknown. This study investigated N2 fixation under a range of nitrate concentrations. Cultures were grown with pretreatments of nitrate replete (single dose 941 µmol NO3- · L-1 ) and N-free conditions and then either received a single dose of 941 µmol NO3- · L-1 (N941), 118 µmol NO3- · L-1 (N118) or 0 N. Heterocysts appeared from days 3 to 5 when treatments of high NO3- were transferred to N free media (N941:N0), and from day 5 in N941 transferred to N118 treatments. Conversely, transferring cells from N0 to N941 resulted in heterocysts being discarded from day 3 and day 5 for N0:N118. Heterocyst appearance correlated with a detectable rate of N2 fixation and up-regulation of nifH gene expression, the discard of heterocysts occurred after sequential reduction of nifH expression and N2 fixation. Nitrate uptake rates were not affected by pretreatment, suggesting no regulation or saturation of this uptake pathway. These data demonstrate that for C. raciborskii, N2 fixation is regulated by the production or discard of heterocysts. In conclusion, this study has shown that N2 fixation only provides enough N to support relatively low growth under N-limited conditions, and does not supplement available nitrate to increase growth rates.

Intraspecific variation in growth, morphology and toxin quotas for the cyanobacterium, Cylindrospermopsis raciborskii.

Cylindrospermopsis raciborskii is a bloom forming cyanobacterium with complex population dynamics and toxicity. In January of 2013 a single sample was collected from surface waters in Lake Wivenhoe, Australia, and twenty-four individual trichomes were isolated. Each isolate exhibited differences in growth rate, toxin cell quota and morphology, in the absence of phylogenetic heterogeneity. This study demonstrates substantial intraspecific isolate variation within a small sample and this has implications for understanding the population dynamics of this species.