Dunaliella tertiolecta, Butcher, 1959

Sorochan Armstrong, Michael D., Ren, O., Campos, e Arredondo, Bannon, Catherine C., Mata, A. Paulina de la, Case, Rebecca J. & Harynuk, James J., 2022, Global metabolome analysis of Dunaliella tertiolecta, Phaeobacter italicus R 11 Co-cultures using thermal desorption - Comprehensive two-dimensional gas chromatography - Time-of-flight mass spectrometry (TD-GC × GC-TOFMS), Phytochemistry (113052) 195, pp. 1-8 : 5

4.1. Preparation of D. tertiolecta , P. italicus R11, and co-cultured samples

4.1.1. Growth and maintenance of algal and bacterial strains

The Dunaliella tertiolecta Butcher (Dunaliellaceae) CCMP 1320 strain was obtained from the Provasoli-Guillard National Centre for Marine Algae and Microbiota (NCMA). The chlorophyte was maintained in L1-Si media made with artificial seawater (35 g /L of Instant Ocean, Blacksburg, VA, USA), at 18 ◦ C with a diurnal incubator cycle (12:12 h dark-light cycle). Samples from the cultures of microalgae were examined microscopically to rule out bacterial contamination before experimental use. These samples were also inoculated onto marine agar plates, and incubated at 28 ◦ C for 3 days to identify any colony forming units (CFUs). (18.7 g of Difco Marine Broth 2216 with 9 g NaCl and 15 g Difco agar in 1 L). Experimental use of the algal cultures proceeded once a cell concentration of 104 cells/mL was reached.

Samples of Phaeobacter italicus R 11 (Vandecandelaere) Wirth & Whitman ( Rhodobacteraceae ) were acquired from Botany Bay, Australia ( Case et al., 2011) and identified during a parallel study by sequencing its entire genome ( Fernandes et al., 2011). The bacterial cultures were maintained at 28 ◦ C on the aforementioned marine agar plates, then transferred to 5 mL 50% dilute marine broth media (2216 Marine Broth, Difco) where it was grown until reaching a stationary phase for 24 h before the experiments. Cell concentration for stationary phase P. italicus R11 was similar to the cell concentration of the algae cultures, at 10 4 cells/mL.