Singlet oxygen-induced signalling depends on the metabolic status of the Chlamydomonas reinhardtii cell

Using a mutant screen, we identified trehalose 6-phosphate phosphatase 1 (TSPP1) as a functional enzyme dephosphorylating trehalose 6-phosphate (Tre6P) to trehalose in Chlamydomonas reinhardtii . The tspp1 knock-out results in reprogramming of the cell metabolism via altered transcriptome. As a secondary effect, tspp1 also shows impairment in 1 O 2 -induced chloroplast retrograde signalling. From transcriptomic analysis and metabolite profiling, we conclude that accumulation or deficiency of certain metabolites directly affect 1 O 2 -signalling. 1 O 2 -inducible GLUTATHIONE PEROXIDASE 5 ( GPX5 ) gene expression is suppressed by increased content of fumarate and 2-oxoglutarate, intermediates in the tricarboxylic acid cycle (TCA cycle) in mitochondria and dicarboxylate metabolism in the cytosol, but also myo-inositol, involved in inositol phosphate metabolism and phosphatidylinositol signalling system. Application of another TCA cycle intermediate, aconitate, recovers 1 O 2 -signalling and GPX5 expression in otherwise aconitate-deficient tspp1 . Genes encoding known essential components of chloroplast-to-nucleus 1 O 2 -signalling, PSBP2, MBS, and SAK1, show decreased transcript levels in tspp1 , which also can be rescued by exogenous application of aconitate. We demonstrate that chloroplast retrograde signalling involving 1 O 2 depends on mitochondrial and cytosolic processes and that the metabolic status of the cell determines the response to 1 O 2 .