Raman Spectroscopy: A Novel Approach for Studying Hydrothermal Microbial Communities
The survival strategies of extremophiles rely on the production of protective biomolecules to thrive in stressed terrestrial conditions; however, it is still unclear as to what dictates or influences the production of these select taxonomic markers. Resonance Raman (RR) and Fourier-transform Infrared (FTIR) are complementary techniques that can provide a complete and unique vibrational ‘fingerprint’ of microbial taxonomy. While pigments offer distinctive diagnostic RR signatures owing to resonance enhancement associated with their conjugated double bond systems, FTIR provides taxonomic information regarding cellular biomarkers (fatty acids, proteins, lipids, and carbohydrates) that may define microbial response to stress. Spectroscopic data of microorganisms from YNP will undergo chemometric analyses to elucidate the physical relationships that exist between biological classification and the chemical traits necessary for survival under extreme conditions. The project goals of this work are:
1.DESCRIMINATING TAXONOMIC ORIGIN WITHIN YNP MICROBIAL COMMUNITIES USING PIGMENT AND CELLULAR BIOMARKERS. This project aims to discern any interdependence between the cellular structure and the choice of pigment production for survival. Chemometric methods will be used to correlate any pattern recognition that may exist between the structural moieties present in the YNP spectral data (RR, FTIR) with microbial phylogeny and/or environmental conditions. Separation techniques will be coupled to reveal information pertaining to the qualitative and spatial distribution of these biomolecules under varying conditions of pH and temperature.
2.CORRELATING THE CELLULAR GENETIC MATERIAL OF CYANIDIALES (RED ALGAE) WITH THE GEOLOGICAL RECORD. This project will evaluate how the cellular framework of cyanidiales will differ based on geological origin. FTIR data generated from cyanidiales specimens isolated from YNP, Japan, Iceland, Antarctica, and Spain will undergo chemometric treatment to determine how genetic differences and/or similarities may reflect the geological record.
3.MONITORING ULTRAVIOLET (UV) STRESS RESPONSE IN CYANIDIALES MEROLAE USING VIBRATIONAL SPECTROSCOPY. This project intends to explain the phenomena responsible for the seasonal color variations observed in cyanidiales mats from YNP. Spectral monitoring of cyanidiales merolae isolates cultured under varying conditions of UV exposure will be used to determine microbial propagation (growth curves), cell viability, and chlorophyll production. Significantly, vibrational data will be generated during different kinetic growth phases (lag, exponential, stationary, death) of the pure cultures and compared with cyanidiales specimens collecture under controlled UV exposure in the field (Lemonade Creek and Dragon Springs, YNP) on seasonal basis to improve our understanding of the microbial evolution of these organisms.
This research combines microbiological and physical methods to unravel the chemical characteristics necessary for biological adaptation in diverse environments. Not only will this work demonstrate a novel and innovative application of vibrational spectroscopy, but also provides unique opportunities for students to learn key concepts and methodologies endemic to the areas of chemistry, biology, and geology. Developed for the undergraduate level with a focused confluence of research and education, students will interact with complementary disciplines to learn specialized techniques and gain exposure to the applications of modern biological and chemical methods.