Chemical fingerprinting of single glandular trichomes of Cannabis sativa by Coherent anti-Stokes Raman scattering (CARS) microscopy
| Autor: | Erik Freier, Paul Ebersbach, Oliver Kayser, Felix Stehle |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0106 biological sciences
0301 basic medicine Coherent anti-stokes Raman scattering Hyperspectral imaging Acyclic Monoterpenes Secondary Metabolism Plant Science Biology Alkenes Cannabis sativa Spectrum Analysis Raman 01 natural sciences Terpene 03 medical and health sciences symbols.namesake chemistry.chemical_compound Imaging Three-Dimensional CBDA lcsh:Botany Oils Volatile Dronabinol Cannabis THCA Plants Medicinal Cannabinoids Terpenes Secondary metabolites Trichomes Unmixing Fluorescence Trichome lcsh:QK1-989 030104 developmental biology chemistry Mapping Myrcene symbols Microscopy Electron Scanning Monoterpenes Biological system Two-photon fluorescence Chemical fingerprinting Raman scattering 010606 plant biology & botany Research Article |
| Zdroj: | BMC Plant Biology BMC Plant Biology, Vol 18, Iss 1, Pp 1-12 (2018) |
| ISSN: | 1471-2229 |
| Popis: | Background Cannabis possesses a rich spectrum of phytochemicals i.e. cannabinoids, terpenes and phenolic compounds of industrial and medicinal interests. Most of these high-value plant products are synthesised in the disk cells and stored in the secretory cavity in glandular trichomes. Conventional trichome analysis was so far based on optical microscopy, electron microscopy or extraction based methods that are either limited to spatial or chemical information. Here we combine both information to obtain the spatial distribution of distinct secondary metabolites on a single-trichome level by applying Coherent anti-Stokes Raman scattering (CARS), a microspectroscopic technique, to trichomes derived from sepals of a drug- and a fibre-type. Results Hyperspectral CARS imaging in combination with a nonlinear unmixing method allows to identify and localise Δ9-tetrahydrocannabinolic acid (THCA) in the secretory cavity of drug-type trichomes and cannabidiolic acid (CBDA)/myrcene in the secretory cavity of fibre-type trichomes, thus enabling an easy discrimination between high-THCA and high-CBDA producers. A unique spectral fingerprint is found in the disk cells of drug-type trichomes, which is most similar to cannabigerolic acid (CBGA) and is not found in fibre-type trichomes. Furthermore, we differentiate between different cell types by a combination of CARS with simultaneously acquired two-photon fluorescence (TPF) of chlorophyll a from chloroplasts and organic fluorescence mainly arising from cell walls enabling 3D visualisation of the essential oil distribution and cellular structures. Conclusion Here we demonstrate a label-free and non-destructive method to analyse the distribution of secondary metabolites and distinguish between different cell and chemo-types with high spatial resolution on a single trichome. The record of chemical fingerprints of single trichomes offers the possibility to optimise growth conditions as well as guarantee a direct process control for industrially cultivated medicinal Cannabis plants. Moreover, this method is not limited to Cannabis related issues but can be widely implemented for optimising and monitoring all kinds of natural or biotechnological production processes with simultaneous spatial and chemical information. Electronic supplementary material The online version of this article (10.1186/s12870-018-1481-4) contains supplementary material, which is available to authorized users. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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