Raman spectroscopy as a potential method for the detection of extremely halophilic archaea embedded in halite in terrestrial and possibly extraterrestrial samples

Abstract

Evidence for the widespread occurrence of extraterrestrial halite, particularly on Mars, has led to speculations on the possibility of halophilic microbial forms of life; these ideas have been strengthened by reports of viable haloarchaea from sediments of geological age (millions of years). Raman spectroscopy, being a sensitive detection method for future astrobiological investigations onsite, has been used in the current study for the detection of nine different extremely halophilic archaeal strains which had been embedded in laboratory-made halite crystals in order to simulate evaporitic conditions. The cells accumulated preferentially in tiny fluid inclusions, in simulation of the precipitation of salt in natural brines. FT-Raman spectroscopy using laser excitation at 1064 nm and dispersive micro Raman spectroscopy at 514.5 nm were applied. The spectra showed prominent peaks at 1507, 1152 and 1002 cm −1 which are attributed to haloarchaeal C 50 carotenoid compounds (mainly bacterioruberins). Their intensity varied from strain to strain at 1064-nm laser excitation. Other distinguishable features were peaks due to peptide bonds (amide I, amide III) and to nucleic acids. No evidence for fatty acids was detected, consistent with their general absence in all archaea.These results contribute to a growing database on Raman spectra of terrestrial microorganisms from hypersaline environments and highlight the influence of the different macromolecular composition of diverse strains on these spectra.

Raman Spectroscopy of Microbial Pigments

“…Two laser excitation wavelengths were used: nonresonant 1,064 nm and resonant 514.5 nm. The main peaks collected (1,507, 1,152, and 1,002 cm Ϫ1 ) could be mainly assigned to bacterioruberin (95).…”

The Miniaturized Raman System and Detection of Traces of Life in Halite from the Atacama Desert: Some Considerations for the Search for Life Signatures on Mars

“…It has already been demonstrated that benchtop instrumentation is able to detect biomolecules associated with extremophilic colonization of rocks (Russell et al, 1998;Edwards, 2000a, 2000b;Edwards et al, 2005;Villar et al, 2005;Marshall et al, 2006;Fendrihan et al, 2009), which provide model terrestrial organisms as analogues for those that may have been present on Mars. A specially constructed miniaturized Raman system, however, is an essential requirement for implementation of these in situ robotic measurements on Mars.…”

Miniaturized Raman instrumentation detects carotenoids in Mars-analogue rocks from the Mojave and Atacama deserts

“…In another case of salinixanthin from S. ruber Raman spectra indicate that there are few structural differences [52]. Other studies also demonstrated in laboratory conditions that benchtop instrumentation is able to detect biomolecules associated with the extremophilic endolithic colonization of rocks [38,[53][54][55][56][57][58][59][60].…”