Targeting mutations to the plastidial psbA gene of Chlamydomonas reinhardtii without direct positive selection.

Targeting mutations to specific genomic loci is invaluable for assessing in vivo the effect of these changes on the biological role of the gene in study. Here, we attempted to introduce a mutation that was previously implicated in an increased heat stability of the mesophilic cyanobacterium Synechocystis sp. PCC6803 via homologous recombination to the psbA gene of Chlamydomonas reinhardtii. For that, we established a strategy for targeted mutagenesis that was derived from the efficient genome-wide homologous-recombination-based methodology that was used to target individual genes of Saccharomyces cerevisiae. While the isolated mutants did not show any benefit under elevated temperature conditions, the new strategy proved to be efficient for C. reinhardtii even in the absence of direct positive selection.

Temperature dependence of photosynthesis and thylakoid lipid composition in the red snow alga Chlamydomonas cf. nivalis (Chlorophyceae).

Here, we report an effect of short acclimation to a wide span of temperatures on photosynthetic electron transfer, lipid and fatty acid composition in the snow alga Chlamydomonas cf. nivalis. The growth and oxygen evolution capacity were low at 2 °C yet progressively enhanced at 10 °C and were significantly higher at temperatures from 5 to 15 °C in comparison with the mesophilic control Chlamydomonas reinhardtii. In search of the molecular mechanisms responsible for the adaptation of photosynthesis to low temperatures, we have found unprecedented high rates of QA to QB electron transfer. The thermodynamics of the process revealed the existence of an increased structural flexibility that we explain with the amino acid changes in the D1 protein combined with the physico-chemical characteristics of the thylakoid membrane composed of > 80% negatively charged phosphatidylglycerol.