Production of β-Cyclocitral and Its Precursor β-Carotene in Microcystis aeruginosa: Variation at Population and Single-Cell Levels

Abstract

Bloom-forming cyanobacteria produce and release odorous compounds and pose threats to the biodiversity of aquatic ecosystem and to the drinking water supply. In this study, the concentrations of β-cyclocitral in different bacterial growth phases were investigated using GC-MS to determine the growth stage of Microcystis aeruginosa at high risk for β-cyclocitral production. Moreover, the synchronicity of the production of β-cyclocitral and its precursor β-carotene at both population and single-cell levels was assessed. The results indicated that β-cyclocitral was the main odorous compound produced by M. aeruginosa cells. The intracellular concentration of β-cyclocitral (C_β-cc) as well as its cellular quota (Q_β-cc) increased synchronously in the log phase, along with the increase of cell density. However, they reached the maximum values of 415 μg/L and 10.7 fg/cell in the late stationary phase and early stationary phase, respectively. The early stage of the stationary phase is more important for β-cyclocitral monitoring, and the sharp increase in Q_β-cc is valuable for anticipating the subsequent increase in C_β-cc. The molar concentrations of β-cyclocitral and β-carotene showed a linear relationship, with an R² value of 0.92, suggesting that the production of β-cyclocitral was linearly dependent on that of β-carotene, especially during the log phase. However, the increase in Q_β-cc was slower than that in β-carotene during the stationary phase, suggesting that β-cyclocitral production turned to be carotene oxygenase-limited when the growth rate decreased. These results demonstrate that variations of β-cyclocitral production on a single-cell level during different bacterial growth phases should be given serious consideration when monitoring and controlling the production of odorous compounds by M. aeruginosa blooms.

Keywords: Microcystis aeruginosa; cellular quota; cyanobacteria; growth phase; β-carotene; β-cyclocitral.

Figure 1

Cell density, percentage of membrane-damaged cells (P_md) (a), and photosynthetic activity (b) of Microcystis aeruginosa cells during incubation.

Figure 2

Concentration of β-cyclocitral produced by Microcystis aeruginosa cells during incubation: (a) total and extracellular concentrations of β-cyclocitral, (b) intracellular concentrations of β-cyclocitral.

Figure 2

Concentration of β-cyclocitral produced by Microcystis aeruginosa cells during incubation: (a) total and extracellular concentrations of β-cyclocitral, (b) intracellular concentrations of β-cyclocitral.

Figure 3

Cellular production quota (Q_β-cc) of Microcystis aeruginosa cells during the incubation.

Figure 4

Variation of β-cyclocitral along with β-carotene in Microcystis aeruginosa cells during incubation. (a) Variations of cell density and intracellular concentrations of β-cyclocitral and β-carotene during 14 days, (b) relationship between intracellular β-cyclocitral and β-carotene, (c) variations of quotas of β-cyclocitral (Q_β-cc) and β-carotene (Q_β-Car) and their molar ratios.

Figure 4

Variation of β-cyclocitral along with β-carotene in Microcystis aeruginosa cells during incubation. (a) Variations of cell density and intracellular concentrations of β-cyclocitral and β-carotene during 14 days, (b) relationship between intracellular β-cyclocitral and β-carotene, (c) variations of quotas of β-cyclocitral (Q_β-cc) and β-carotene (Q_β-Car) and their molar ratios.