Identification of five wood species of the Meliaceae family on the basis of their wood anatomical and chemical characteristics

Swietenia, Cedrela, Khaya et Toona sont des genres d'arbres tropicaux à bois précieux. Une identification précise du bois est nécessaire pour contrôler l'exploitation forestière illégale et le commerce connexe ainsi qu'à des fins de certification dans l'industrie des produits forestiers et du bois. Swietenia mahagoni, Swietenia macrophylla et Cedrela odorata ont été inscrites à l'Annexe II de la Convention sur le commerce international des espèces de faune et de flore menacées d'extinction (CITES) et le commerce de ces espèces a été restreint par l'imposition de documentation spécifique et de permis pour l'importation et l'exportation de marchandises. En conséquence, certaines espèces (par exemple, Khaya ivorensis et Toona ciliata) ont été introduites comme substituts de ces espèces et leur identification est maintenant un problème difficile pour l'application de la réglementation liée à la CITES à travers le monde. Ainsi, le but de l'étude est d'identifier et de différencier des espèces étroitement apparentées de la famille des Meliaceae, à savoir Swietenia mahagoni, Swietenia macrophylla, Cedrela odorata, Khaya ivorensis et Toona ciliata sur la base d'analyses anatomiques, de vision et d'intelligence artificielles et de caractérisation chimique du bois. Dans cette étude, trois approches ont été utilisées, la structure du bois par microscopie optique, le système de vision artificielle et le profilage du métabolome par chromatographie en phase gazeuse bidimensionnelle combinée à la spectrométrie de masse à temps de vol (GC*GC-TOFMS). Au total, nous avons analysé 50 échantillons de bois des espèces susmentionnées provenant des xylothèques du Service canadien des forêts et du Centre de recherche sur les matériaux renouvelables de l'Université Laval. Des variations considérables ont été observées dans les caractéristiques anatomiques du bois de ces espèces, mais ces variations n'étaient pas suffisantes pour les différencier au niveau de l'espèce. De même, le système de vision artificielle (Xylotron) est simple et efficace pour l'identification du bois, mais cet outil est encore en développement et les bases de données de référence sont encore insuffisantes pour pouvoir différencier efficacement les espèces étudiées. En revanche, la technique de GC*GC-TOFMS combinée à des statistiques non supervisées (Analyse en Composantes Principales) a montré des signatures chimiques distinctes à partir d'échantillons de bois des cinq espèces, ce qui a permis une identification rapide au niveau de l'espèce. Ainsi, GC*GC-TOFMS fournit une méthode rapide d'identification du bois qui peut être facilement appliquée à de petits échantillons de bois de duramen et peut avoir des applications potentielles dans un contexte d'application de la loi et d'analyse/identification judiciaire des bois illégaux, et dans diverses recherches liées à la foresterie, aux sciences du bois et à l'écophysiologie des arbres.Swietenia, Cedrela, Khaya and Toona are tree genera providing valuable tropical timber. Accurate identification of timber is necessary to control illegal logging and related trade as well as for certification purposes in the forest and wood products industry. Swietenia mahagoni, Swietenia macrophylla and Cedrela odorata have been included in Appendix II of Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES) and trade in these species has been restricted by the imposition of specific documentation and permits for importation and exportation of goods. As a result of these essential restrictions to protect species survival, other closely related non-CITES species including Khaya ivorensis and Toona ciliata have been introduced as substitutes for these CITES- regulated species and their accurate identification represents a challenge for the application of CITES regulation worldwide. Thus, the aim of the study is to evaluate identification methods to identify and differentiate closely related species of the Meliaceae family namely Swietenia mahagoni, Swietenia macrophylla, Cedrela odorata, Khaya ivorensis and Toona ciliata based on anatomical, machine-vision and chemical analysis of the wood. In this study, three approaches were used, analysis of the wood structure by optical microscopy, machine-vision system and metabolome profiling by two-dimensional gas chromatography combined with time-of-flight mass spectrometry (GC*GC-TOFMS). In total, 50 wood samples of the above-mentioned species from the Xylaria of the Canadian Forest Service and the Centre de recherche sur les matériaux renouvelables, Université Laval were analyzed. Considerable variations were observed in the anatomical characteristics of the wood of these species, but these variations were not sufficient to differentiate them at the species level. Similarly, machine-vision system (Xylotron) is simple and effective in wood identification, but this tool is still in development and require the construction of sufficient reference databases to cover a greater number of wood species. In contrast, GC*GC-TOFMS combined with unsupervised statistics (Principal Component Analysis) showed distinct chemical signatures from wood samples of five species, which allowed rapid identification to the species level. Thus, GC*GC-TOFMS provides a rapid method for wood identification that can be easily applied to small samples of heartwood and forensics and have potential applications in a context of law enforcement analysis/identification of illegal wood, and in related to forestry, wood science and tree ecophysiology

COMPARING GC×GC-TOFMS-BASED METABOLOMIC PROFILING AND WOOD ANATOMY FOR FORENSIC IDENTIFICATION OF FIVE MELIACEAE (MAHOGANY) SPECIES

Illegal logging and associated trade have increased worldwide. Such environmental crimes represent a major threat to forest ecosystems and society, causing distortions in market prices, economic instability, ecological deterioration, and poverty. To prevent illegal imports of forest products, there is a need to develop wood identification methods for identifying tree species regulated by the Convention on International Trade in Species of Wild Fauna andFlora in Trade (CITES) and other look-alike species. In this exploratory study, we applied metabolomic profiling of five species (Swietenia mahagoni, Swietenia macrophylla, Cedrela odorata, Khaya ivorensis, and Toona ciliata) using two-dimensional gas chromatog- raphy combined with time-of-flight mass spectrometry (GC3GC-TOFMS). We also performed qualitative, quantitative (based on the measurement of vessel area, tangential vessel lumina diameter,vessel element length, ray height, and ray width), and machine-vision aided (XyloTron) wood anatomy on a subsample of wood specimens to explore thepotential and limits of each approach. Fifty dried xylaria wood specimens were ground, extracted with methanol, and subsequently analyzed by GC3GC-TOFMS. In this study, the four genera could easily be identified using qualitative wood anatomy and chemical profiling. At the spe- cies level, Swietenia macrophylla and Swietenia mahagoni specimens were found to share many major metabolites and could only be differentiated after feature selection guided by cluster resolution (FS-CR) and visualization using Principal Component Analysis (PCA). Expectedly, specimens from the two Swiete- nia spp. could not be distinguished based on qualitative wood anatomy. However, significant differences in quantitative anatomical features were obtained for these two species. Excluding T. ciliata that was not included in the reference database of end grain images at the time of testing (2021), the XyloTron could successfully identify the majority of the specimens to the right genus and 50% of the specimens to the right species. The machine-vision tool was particularly successful at identifying Cedrela odorata samples, where all samples were correctly identified. Despite the limited number of specimens available for thisstudy, our preliminary results indicate that GC3GC-TOFMS-based metabolomic profiles could be used as comple- mentary method to differentiate CITES-regulated wood specimens at the genus and species levels.