Effect of Hydrogenase Deficiency on Accumulation of Phosphorus-Rich Inclusions in Chlamydomonas reinhardtii.

Photogeneration of hydrogen in microalgae is thought to be among the mechanisms increasing their resilience to stresses including those caused by nutrient deprivation by re-routing the flow of electrons and reducing power in the cell. Metabolism of phosphorus (P), an essential nutrient, and its reserve forms such as polyphosphate (PolyP), is affected by and plays a role in the responses to diverse stresses, too. However, the potential interplay of the capability of photogeneration of hydrogen and turnover of phosphorus-rich inclusions in stressed microalgae cells so far escaped the attention of researchers. Here, we present a quantitative ultrastructural view of the turnover of P-rich inclusions in the model microalga Chlamydomonas reinhardtii strains, the parent strain CC-425 and its hydEF-1 mutant lacking hydrogenase activity as a function of sulfur and oxygen availability in the medium. In addition to the electron microscopy cell image analysis of the studied strains, we followed the elemental composition of the inclusions in different (sub) compartments of the cells obtained with energy dispersive X-ray spectroscopy. The stress caused by sulfur deprivation and subsequent transition of the microalgae culture to anaerobic conditions declined the size of phosphorus-containing inclusions but increased their number in the parent strain. Overall, the accumulation of the phosphorus-rich inclusions in hydEF-1 mutant was much lower than in the fully functional parent strain regardless of the cultivation conditions. We believe that impaired hydrogenase activity and correspondingly reduced sink of electrons and reducing power in the mutant strain indirectly affects the turnover of P and its reserves in the cell. These effects were manifested by the changes in the abundance, morphology, and elemental composition of the P-containing inclusions. We hypothesized that the sulfur-deprivation stress increased the initiation of the biosynthesis of PolyP chains, but their elongation and hence the formation of large PolyP-containing inclusions was hindered by anaerobiosis. [ABSTRACT FROM AUTHOR]