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Dynamic protrusions mediate crawling motility in Asgard archaea
Cultivation of Ca. L. ossiferum and M. peptidophilum Ca. L. ossiferum enrichment cultures were grown protected from light in lokiarchaeal media in sealed, pressurized serum bottles in an 80:20% N2:CO2 atmosphere (0.3 bars) at 20 °C without shaking, as described previously7. The growth medium composition (per litre) was as follows: 20.7 g NaCl, 5 g MgCl2·6H2O, 2.7 g NaHCO3, 1.36 g CaCl2·2H2O, 0.54 g NH4Cl, 0.14 g KH2PO4, 0.03 g Na2S·9H2O, 0.03 g cysteine·HCl, 0.5 ml of acid trace element solution, 0.5 ml of alkaline trace element solution, 1 ml Se/W solution, 0.1% casein hydrolysate (w/v). The acid trace element solution contained (per litre): 1.49 g FeCl2·4H2O, 0.062 g H3BO3, 0.068 g ZnCl2, 0.017 g CuCl2·H2O, 0.099 g MnCl2·4H2O, 0.119 g CoCl2·6H2O, 0.024 g NiCl2·6H2O and 4.106 ml of 37% HCl. The alkaline trace element solution contained (per litre): 0.017 g Na2SeO3, 0.033 g Na2WO4, 0.021 g Na2MoO4 and 0.4 g NaOH. The pH was adjusted to 7.5, and the medium was supplemented with ampicillin, kanamycin and streptomycin (200 μg ml−1 each). The cultures were continuously passaged into serum flasks containing fresh medium—typically using an early stationary-phase culture as inoculum (1:10 dilution). Growth of L. ossiferum cultures was monitored by...
