Diatoms are a hugely diverse microalgal class, which possesses unique biological features and complex metabolic pathways and may activate sophisticated mechanisms to respond to environmental changes. Abiotic stress factors may limit growth rate of diatoms, but may also trigger intracellular signaling pathways that cause cells to undergo programmed cell death (PCD). Here we investigate the gene expression of different target genes related to cell death, namely programmed cell death 4 (PDCD4), tumor susceptibility gene 101 (TSG101), developmental and cell death (DCD) domain, death specific protein (DSP) and metacaspase (MC), using RT-qPCR in the cosmopolitan coastal centric diatom species Skeletonema marinoi, which contributes significantly to phytoplankton blooms in temperate waters. To this end, we undertook a detailed study of the best reference genes to analyze gene expression in S. marinoi under different experimental conditions (i.e. in different growth phases or under silica starvation). Results showed that DSP gene expression had a clear and constant increase along the S. marinoi growth curve reaching its maximum during the senescent phase. On the contrary, PDCD4, DCD, TSG101 and MC did not show any significant variation. These findings indicate that the DSP gene is a possible PCD marker induced by aging in this diatom species. In contrast, levels of DSP transcripts induced by silica starvation were relatively low compared to those induced by cell aging suggesting differential activation and/or regulation of the PCD machinery in response to different stressful conditions. Our study also expands the list of reference genes available for the diatom S. marinoi for normalization of RT-qPCR data of cells cultivated under different growth phases or under silica starvation.
Keywords: Death specific protein; Diatom; Gene expression; Nutrient starvation; Programmed cell death; Reference gene.
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