Watson, Jonathon S.; Sephton, Mark A.; Sephton, Sarah V.; Self, Stephen; Fraser, Wesley T.; Lomax, Barry H.; Gilmour, Iain; Wellman, Charles H. and Beerling, David J.
Rapid determination of spore chemistry using thermochemolysis gas chromatography-mass spectrometry and micro-Fourier transform infrared spectroscopy.
Photochemical and Photobiological Sciences, 6 pp. 689–694.
Spore chemistry is at the centre of investigations aimed at producing a proxy record of harmful ultraviolet radiation (UV-B) through time. A biochemical proxy is essential owing to an absence of long-term (century or more) instrumental records. Spore cell material contains UV-B absorbing compounds that appear to be synthesised in variable amounts dependent on the ambient UV-B flux. To facilitate these investigations we have developed a rapid method for detecting variations in spore chemistry using combined thermochemolysis gas chromatography-mass spectrometry and micro-Fourier transform infrared spectroscopy. Our method was tested using spores obtained from five populations of the tropical lycopsid Lycopodium cernuum growing across an altitudinal gradient (650–1981 m a.s.l.) in S.E. Asia with the assumption that they experienced a range of UV-B radiation doses. Thermochemolysis and subsequent pyrolysis liberated UV-B pigments (ferulic and para-coumaric acid) from the spores. All of the aromatic compounds liberated from spores by thermochemolysis and pyrolysis were active in UV-B protection. The various functional groups associated with UV-B protecting pigments were rapidly detected by micro-FTIR and included the aromatic CC absorption band which was exclusive to the pigments. We show increases in micro-FTIR aromatic absorption (1510 cm–1) with altitude that may reflect a chemical response to higher UV-B flux. Our results indicate that rapid chemical analyses of historical spore samples could provide a record ideally suited to investigations of a proxy for stratospheric O3 layer variability and UV-B flux over historical (century to millennia) timescales.
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