The cumulative impact of temperature and nitrogen availability on the potential nitrogen fixation and extracellular polymeric substances secretion by Dolichospermum.

Nitrogen-fixing cyanobacteria not only cause severe blooms but also play an important role in the nitrogen input processes of lakes. The production of extracellular polymeric substances (EPS) and the ability to fix nitrogen from the atmosphere provide nitrogen-fixing cyanobacteria with a competitive advantage over other organisms. Temperature and nitrogen availability are key environmental factors in regulating the growth of cyanobacteria. In this study, Dolichospermum (formerly known as Anabaena) was cultivated at three different temperatures (10 °C, 20 °C, and 30 °C) to examine the impact of temperature and nitrogen availability on nitrogen fixation capacity and the release of EPS. Initially, confocal laser scanning microscopy (CLSM) and the quantification of heterocysts at different temperatures revealed that lower temperatures (10 °C) hindered the differentiation of heterocysts under nitrogen-deprived conditions. Additionally, while heterocysts inhibited the photosynthetic activity of Dolichospermum, the secretion of EPS was notably affected by nitrogen limitation, particularly at 30 °C. Finally, real-time quantitative polymerase chain reaction (qPCR) was used to measure the expression of nitrogen-utilizing genes (ntcA and nifH) and EPS synthesis-related genes (wzb and wzc). The results indicated that under nitrogen-deprived conditions, the expression of each gene was upregulated, and there was a significant correlation between the upregulation of nitrogen-utilizing and EPS synthesis genes (P < 0.05). Our findings suggested that Dolichospermum responded to temperature variation by affecting the formation of heterocysts, impacting its potential nitrogen fixation capacity. Furthermore, the quantity of EPS released was more influenced by nitrogen availability than temperature. This research enhances our comprehension of interconnections between nitrogen deprivation and EPS production under the different temperatures.

How does Microcystis aeruginosa respond to elevated temperature?

Cyanobacteria and their toxins widely exist in freshwater ecosystems. Microcystis aeruginosa is among dominant bloom-forming cyanobacteria. Water temperature is a key factor influencing the life cycle of M. aeruginosa. We simulated elevated temperature (4-35 °C) experiment and cultured M. aeruginosa during the overwintering, recruitment and rapid growth phases. The results showed that M. aeruginosa recovered growth after overwintering at 4-8 °C and recruited at 16 °C. The total extracellular polymeric substance (TEPS) concentration increased rapidly at 15 °C. The actual quantum yield of photosystem II (Fv'/Fm') peaked at 20 °C during the rapid growth phase, and the optimum temperature of M. aeruginosa growth was 20-25 °C. Additionally, TEPS and microcystins (MCs) secretion peaked at 20-25 °C. The cell density accumulated rapidly from 26 °C to 35 °C. Furthermore, enzymes of RuBisCO and FBA related to photosynthetic activity were confirmed to contribute to the metabolism, as well as mcyB gene was affected by elevated temperature. Our results provide insights of the physiological effects and metabolic activity during annual cycle of M. aeruginosa. And it is predicted that global warming may promote the earlier recruitment of M. aeruginosa, extend the optimum growth period, enhance the toxicity, and finally intensify M. aeruginosa blooms.

Effects of extracellular polymeric substances on the aggregation of Aphanizomenon flos-aquae under increasing temperature.

Aphanizomenon flos-aquae (A. flos-aquae) blooms are serious environmental and ecological problems. Extracellular polymeric substances (EPSs) are among the most important indicators for the growth and aggregation of A. flos-aquae. In this study, the secretion of the EPS matrix under temperature rise (7-37°C) was investigated and the role of this matrix in A. flos-aquae aggregation was quantified. First, when the temperature increased, the aggregation ratio increased from 41.85 to 91.04%. Meanwhile, we found that when soluble EPSs (S-EPSs), loosely bound EPSs (LB-EPSs), and tightly bound EPSs (TB-EPSs) were removed successively, the aggregation ratio decreased from 69.29 to 67.45%, 61.47%, and 41.14%, respectively. Second, the content of polysaccharides in the EPS matrix was higher than the content of proteins under temperature change. The polysaccharide in TB-EPSs was closely related to the aggregation ability of A. flos-aquae (P < 0.01). Third, PARAFAC analysis detected two humic-like substances and one protein-like substance in EPSs. Furthermore, Fourier transforms infrared spectroscopy (FTIR) showed that with increasing temperature, the polysaccharide-related functional groups increased, and the absolute value of the zeta potential decreased. In conclusion, these results indicated that a large number of polysaccharides in TB-EPSs were secreted under increasing temperature, and the polysaccharide-related functional groups increased correspondingly, which reduced the electrostatic repulsion between algal cells, leading to the destruction of the stability of the dispersion system, and then the occurrence of aggregation. This helps us to understand the process of filamentous cyanobacterial aggregation in lakes.