Molecular characterization of a seed-fern ovule (Pennsylvanian, Sydney Coalfield, Canada) by FTIR, ¹³C NMR, and Py-GC-MS

Abstract

Despite the abundance of detached seed-fern ovules (Euramerican and Cathaysian floral provinces, Pennsylvanian - Permian) as compression / impression, their palaeobiochemistry remains unknown. The Late Pennsylvanian strata of the Sydney Coalfield, Canada, have yielded numerous ovulate trigonocarpalean compressions, 6-8cm long, with preserved cuticles assigned to Trigonocarpus grandis (Lesquereux) Cleal et Zodrow. The macerated cuticles are analyzed by Fourier Transform Infrared spectroscopy (FTIR), Carbon-13 Nuclear Magnetic Resonance (¹³C NMR), and Pyrolysis Gas Chromatography-mass spectrometry (Py-GC-MS). FTIR and ¹³C NMR data reveal a predominantly aliphatic structure including C-H groups (with or without hetero-substitution such as in alkyl and aryl alcohols, ethers, esters, and ketones). Larger CH₂/CH₃ ratios (above 20) suggest the presence of long and straight aliphatic side chains linked to the main macromolecular structure. Other groups present include C-O (in phenols and phenoxy structures, and aryl and alkyl ethers and alcohols), C=O carbonyl groups (in carboxylic acids and conjugated and highly conjugated structures such as ketones), C=C (in aromatic carbon structures with and without bridging to other carbon groups), and O-H (in aliphatic or aromatic alcohols). In addition, ¹³C NMR shows, compared to other cuticles in related seed ferns, a high amount of acetal groups as they occur in di- and polysaccharides. In agreement with FTIR and NMR results, Py-GC-MS data show the presence of highly aliphatic molecules (alkanes/alkenes) with carbons between C₄ and C₈ (including a complex mixture of mono and di unsaturated hydrocarbon isomers up to C₈). Other compounds detected include benzene and toluene. However, the most striking chemical feature of the pyrolysates markers is the presence of 5 isomers of C₅H₈ including isoprene (0.5 - 1.0 % abundance). The latter could be derived from some tocopherol precursor (vitamin E-like compound). If confirmed, this is the earliest chemical evidence for the presence of isoprenoids in vascular plants.