The high-altitude pre-Andean region of the Atacama Desert is characterized by its stark volcanic rock formations and unique hydrothermal gypsum outcrops (gypcrete) that it hosts. This study delves into the biomolecular composition of the endolithic phototrophic microbes that thrive within these gypcretes. Using advanced Raman spectroscopy techniques, including Raman imaging (complemented by microscopic and 3D microscopic observations), herein we unveil new insights into the adaptive strategies of these gypsum-inhabiting algae. Our Raman imaging results provide a detailed chemical map of carotenoids associated with microbial colonization. This map reveals a significant gradient in pigment content, highlighting a critical survival mechanism for algae and cyanobacteria in this polyextreme environment. Intriguingly, we detected signals for carotenoids not only in the algae-colonized layer, but also deeper within the gypsum matrix - indicating pigment migration following cell disruption. In addition, we conducted an in-depth analysis of individual algal cells from the Trebouxiaceae family, noting their color variations from green to orange, plus describing the spectral differences in detail. This investigation identified in-vivo pigments (carotenoids, chlorophyll) and lipids at the cellular level, offering a comprehensive view of the molecular adaptations enabling life in one of the Earth's most extreme habitats.

Departamento Biología Vegetal Ecología y Ciencias de la Tierra and IACYS Universidad de Extremadura Plasencia 10600 Spain

Global Change Research Institute of the Czech Academy of Sciences Bělidla 986 4a Brno 603 00 Czech Republic

Museo Nacional de Ciencias Naturales CSIC c Serrano 115 dpdo Madrid 28006 Spain

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