The physical state of osmoregulatory solutes in unicellular algae. A natural-abundance carbon-13 nuclear-magnetic-resonance relaxation study.

Natural-abundance 13C n.m.r. spin-lattice relaxation-time measurements have been carried out on intact cells of the unicellular blue--green alga Synechococcus sp. and the unicellular green alga Dunaliella salina, with the aim of characterizing the environments of the organic osmoregulatory solutes in these salt-tolerant organisms. In Synechococcus sp., all of the major organic osmoregulatory solute, 2-O-alpha-D-glucopyranosylglycerol, is visible in spectra of intact cells. Its rotational motion in the cell is slower by a factor of approx. 2.4 than in aqueous solution, but the molecule is still freely mobile and therefore able to contribute to the osmotic balance. In D. salina, only about 60% of the osmoregulatory solute glycerol is visible in spectra of intact cells. The rotational mobility of this observable fraction is approximately half that found in aqueous solution, but the data also indicate that there is a significant concentration of some paramagnetic species in D. salina which contributes to the overall spin-lattice relaxation of the glycerol carbon atoms. The non-observable fraction, which must correspond to glycerol molecules that have very broad 13C resonances and that are in slow exchange with bulk glycerol, has not been properly characterized as yet, but may represent glycerol in the chloroplast. The implications of these findings in relation to the physical state of the cytoplasm and the mechanism of osmoregulation in these cells are discussed.

Carbon-13 nuclear magnetic resonance study of osmoregulation in a blue-green alga.

The process of osmoregulation in a unicellular blue-green alga, Synechococcus sp., has been studied by natural-abundance carbon-13 nuclear magnetic resonance spectroscopy of intact cells and cell extracts. 2-O-alpha-D-Glucopyranosylglycerol was identified as the major organic osmoregulatory solute. This demonstrates the presence of a major osmoregulatory solute in a blue-green alga and is also an example of an osmoregulatory role for glucosylglycerol.

The salt relations of Dunaliella. Further observations on glycerol production and its regulation.

Dunaliella tertiolecta (marine) and D. viridis (halophilic) were each trained by serial transfer to grow at salt concentrations previously regarded as the other's domain. D. viridis then had a salt optimum at 1.0-1.5 M sodium chloride whereas that for D. tertiolecta was less than 0-2 M. Nevertheless D. tertiolecta grew faster than the halophil at all salt concentrations up to 3.5 M, the highest at which they were compared. Both species accumulate glycerol, which is necessary for growth at elevated salinities and which responds in its content to water activity (aw) rather than specifically to salt concentration. Variation in glycerol content is a metabolic process which occurs in the dark from accumulated starch as well as photosynthetically. Regulation of glycerol content by aw does not require protein synthesis. The NADP-specific glycerol dehydrogenase of each of the algae is likely to be directly involved in the regulation of glycerol content. Kinetic studies, together with those described in an earlier publication, show that the enzyme has regulatory properties and that both glycerol and dihydroxyacetone act as effectors as well as reactants. A mechanism of the reaction is tentatively proposed.

Role of silicon in diatom metabolism. VIII. Cyclic AMP and cyclic GMP in synchronized cultures of Cylindrotheca fusiformis.

Levels of the cyclic nucleotides, cAMP and cGMP, were determined in four species of pennate diatoms; changes in their levels and ratios were monitored in silicon-starved and light-dark synchronized cultures of Cylindrotheca fusiformis. Content of both cAMP and cGMP changed during the cell cycles: when silicate was added to starved cultures, cAMP, cGMP and DNA levels rose rapidly; cAMP and cGMP declined befor DNA synthesis was complete and continued to fall during the events leading to cell separation. In unstarved synchronies, net synthesis of DNA continued until cell separation; 1 h before cell separation cAMP levels fell while those of cGMP rose. The results support of the proposal that cAMP and cGMP may play a part in the process of cell division in the diatom, possible involving silicon.