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The brown seaweed Ectocarpus siliculosus is an emerging model species distributed worldwide in temperate coastal ecosystems. Over 1500 strains of E. siliculosus are available in culture from a broad range of geographic locations and ecological niches. To elucidate the molecular mechanisms underlying its capacity to cope with different environmental and biotic stressors, genomic and transcriptomic studies are necessary; this requires the co-isolation of genomic DNA and total RNA. In brown algae, extraction of nucleic acids is hindered by high concentrations of secondary metabolites that co-precipitate with nucleic acids. Here, we propose a reliable, rapid and cost-effective procedure for the co-isolation of high-quality nucleic acids using small quantities of biomass (25-, 50- and 100 mg) from strains of E. siliculosus (RHO12; LIA4A; EC524 and REP10-11) isolated from sites with different environmental conditions. The procedure employs a high pH extraction buffer (pH 9.5) which contains 100 mM Tris-HCl and 150 mM NaCl, with the addition of 5 mM DTT and 1% sarkosyl to ensure maximum solubility of nucleic acids, effective inhibition of nuclease activity and removal of interfering contaminants (e.g. polysaccharides, polyphenols). The use of sodium acetate together with isopropanol shortened precipitation time and enhanced the yields of DNA/RNA. A phenol:chlorophorm:isoamyl alcohol step was subsequently used to purify the nucleic acids. The present protocol produces high yields of nucleic acids from only 25 mg of fresh algal biomass (0.195 and 0.284 µg mg(-1) fresh weigh of RNA and DNA, respectively) and the high quality of the extracted nucleic acids was confirmed through spectrophotometric and electrophoretic analyses. The isolated RNA can be used directly in downstream applications such as RT-PCR and the genomic DNA was suitable for PCR, producing reliable restriction enzyme digestion patterns. Co-isolation of DNA/RNA from different strains indicates that this method is likely to have wider applications for intra- and inter-specific studies on other brown algae.

The Antarctic Peninsula is one of the regions to be most affected by increase in sea surface temperatures (SSTs) mediated by Global Climate Change; indeed, most negative predictions imply an up to 6 °C increment by the end of the XXI century. Temperature is one of the most important factors mediating diversity and distribution of macroalgae, although there is still no consensus as to the likely effects of higher SSTs, especially for polar seaweeds. Some available information suggests that potential strategies to withstand future increases in SSTs will be founded upon the glutathione-ascorbate cycle and the induction of chaperone-functioning heat shock proteins (HSPs); however, their eventual role, even for general stress responses, is unclear. The intertidal green, brown and red macroalgae species Monostroma hariotii, Adenocystis utricularis and Pyropia endiviifolia, respectively, from King George Island, Antarctic Peninsula, were exposed to 2 °C (control) and 8 °C (climate change scenario) for up to 5 days (d). Photosynthetic activity (αETR and ETRmax, and EkETR), photoinhibition (Fv/Fm) and photoprotection processes (αNPQ, NPQmax, and EkNPQ) provided no evidence of negative ecophysiological effects. There were moderate increases in H2O2 production and levels of lipid peroxidation with temperature, results supported by stable levels of total glutathione and ascorbate pools, with mostly higher levels of reduced ascorbate and glutathione than oxidized forms in all species. Transcripts of P. endiviifolia indicated a general upregulation of all antioxidant enzymes and HSPs genes studied under warmer temperature, although with different levels of activation with time. This pioneering investigation exploring different levels of biological organization, suggested that Antarctic intertidal macroalgae may be able to withstand future rise in SSTs, probably slightly altering their latitudinal distribution and/or range of thermal tolerance, by exhibiting robust glutathione-ascorbate production and recycling, as well as the induction of associated antioxidant enzymatic machinery and the syntheses of HSPs.