Effects of arginine on Polytomella parva growth, PII protein levels and lipid body formation.

MAIN CONCLUSION: L-Arginine supports growth and resulted in increased PII signaling protein levels and lipid droplet accumulation in the colorless green alga Polytomella parva. Polytomella parva, a model system for nonphotosynthetic green algae, utilizes ammonium and several carbon sources, including ethanol and acetate. We previously reported that P. parva accumulates high amounts of arginine with the key enzyme of the ornithine/arginine biosynthesis pathway, N-acetyl-L-glutamate kinase, exhibiting high activity. Here we demonstrate that L-arginine can be used by this alga as a nitrogen source. Externally supplied arginine directly influenced the levels of PII signaling protein and formation of triacylglycerol (TAG)-filled lipid bodies (LBs). Our results suggest that the nitrogen source, but not nitrogen starvation, may be critical for the accumulation of LBs in a PII-independent manner in P. parva.

Regulation by light of ammonium transport systems in Chlamydomonas reinhardtii.

The unicellular green alga Chlamydomonas reinhardtii is able to take up methylammonium/ammonium from the medium at different stages of its sexual life cycle. Vegetative cells and pre-gametes mostly used a low-affinity system (LATS) component, but gametes obtained after light treatment of N-deprived pre-gametes expressed both LATS and high-affinity system (HATS) components for the uptake of methylammonium/ammonium. The activity of the LATS component was stimulated by light in only 5 min in a process independent of protein synthesis. By using the lrg6 mutant that produces sexually competent gametes in the dark, light effects on ammonium transport and gamete differentiation have been separately analysed. We have found light regulation of four Amt1 genes: Amt1; 1, Amt1; 2, Amt1; 4 and Amt1; 5. Whereas light-dependent expression of Amt1; 1, Amt1; 2 and Amt1; 4 was independent of gametogenesis, and that of Amt1; 5 was activated in the lrg6 mutant, suggesting a connection between this transporter and the subsequent events taking place during gametogenesis.

Chemotaxis to ammonium/methylammonium in Chlamydomonas reinhardtii: the role of transport systems for ammonium/methylammonium.

Ammonium is the preferred nitrogen source and chemoattractant for the unicellular green alga Chlamydomonas reinhardtii. Here we describe that chemotaxis to methylammonium, a non-metabolizable analogue to ammonium, that occurs only in vegetative cells and pregametes but not in gametes. A new methylammonium-resistant mutant, hat1 that was generated by insertional mutagenesis, has been isolated and found to be affected at multiple loci. At the physiological level, hat1 showed altered Km and Vmax for high-affinity ammonium and methylammonium transport. Transcript levels of the ammonium transporter genes CrAmt1.(1-8) were similar to the wild type except for a lower expression level of CrAmt1.5, 6 and 8. Treatment with the potassium-channel inhibitor tetraethylammonium (TEA) blocked chemotaxis to ammonium/methylammonium and [14C]-methylammonium uptake. Our results suggest that the hat1 mutant could be affected in genes encoding regulatory elements of AMT1 transporter activities and that chemotaxis to ammonium/methylammonium is mediated by ammonium transporters sensitive to the potassium-channel inhibitor TEA.

Phototropin plays a crucial role in controlling changes in chemotaxis during the initial phase of the sexual life cycle in Chlamydomonas.

During sexual differentiation, Chlamydomonas reinhardtii changes its chemotactic behavior in response to ammonium. Just like gamete formation, the change in chemotaxis mode is controlled by the sequential action of two environmental cues, removal of ammonium or nitrate from the medium and light. Thus, vegetative cells and mating incompetent pre-gametes, the latter being generated by nitrogen starvation in the dark, exhibit chemotaxis towards ammonium. Irradiation of pre-gametes results in a loss of chemotaxis and the gaining of mating competence. Incubation of these gametes in the dark resulted in their regaining chemotactic activity; re-illumination again resulted in its loss. Blue light was shown to be most effective in switching-off chemotaxis. RNA-interference strains with reduced levels of the blue-light receptor phototropin showed an attenuated inactivation of chemotaxis that could be partially compensated by the application of higher fluence rates, suggesting that these light responses are mediated by phototropin. The sharing of photoreceptor and signal transduction components as well as similar temporal patterns observed for changes in chemotaxis towards ammonium and gametic differentiation suggest an integration of the signaling pathways that control these two responses.

Chemotactic behavior of Chlamydomonas reinhardtii is altered during gametogenesis.

Chemotactic behavior of Chlamydomonas reinhardtii is altered during the sexual life cycle. Unlike vegetative cells and noncompetent pregametes, mature gametes did not show chemotaxis to ammonium. Loss of chemotaxis to ammonium in mating-competent cells is controlled by gamete-specific genes that are common for both mating-type gametes. Change of chemotaxis mode requires the sequential action of the two environmental signals: removal of ammonium from the medium and light. The mutants lrg1, lrg3, and lrg4 affected in the light-dependent step of sexual differentiation exhibited the loss of chemotaxis to ammonium in the absence of light. These data indicate that there are common components in the signaling pathways that control change of chemotactic behavior and forming of mating competence in gametes.