Metabolomics, proteomics, and transcriptomics of Cannabis sativa L. trichomes
Creators: name: Happyana, Nizar
Publisher: name: Technische Universität Dortmund
Year: 2014
Subjects: subject: Cannabis
subject: Cannabinoid
subject: Trichomes
subject: Metabolomics
subject: Proteomics
subject: Transcriptomics
subject: 660
subject: Hanf
subject: Sekundärmetabolit
subject: Pflanzenhaar
subject: Proteomanalyse
subject: Transkriptomanalyse

description: Cannabis sativa L. trichomes are the main site for synthesizing and storing cannabinoids and other secondary metabolites on this plant. Metabolomics, proteomics, and transcriptomics were applied in this research as analytical approaches in order to study secondary metabolites, proteins, and genes that are synthesized in the trichomes. The function of specific trichomes and their individual parts in the cannabinoid biosynthesis was investigated as well. 1H NMR-based metabolomics has been successfully applied for monitoring the production of metabolites, especially cannabinoids in the Cannabis trichomes during the last weeks of flowering period. Proteomics analysis of the Cannabis trichomes revealed that many enzymes corresponding in the biosynthesis of secondary metabolites, including cannabinoids, flavonoids, and terpenoids were successfully recorded. This finding supported the function of Cannabis trichomes as the main site of secondary metabolite production. Although there is no flavonoid reported from the trichomes, however the identification of enzymes related to its biosynthesis indicated that this compound might be present in this organ. Interestingly, identification of enzymes involved in the biosynthesis of cannabinoids, terpenoids, and flavonoids in the proteomic work has been also confirmed by the detection of their putative transcripts in the cDNA library of Cannabis trichomes. Analysis of cannabinoids in the laser-microdissected trichomes of Cannabis showed that these compounds were detected not only in the head of capitate-stalked trichomes but also in its stem part. This finding suggest that cannabinoid biosynthesis is not only limited to the expected head cells, but also the stems of Cannabis capitate-stalked trichomes might play a role in the cannabinoids biosynthesis.
description: Cannabis sativa L. Trichome sind der hauptsächliche Ort der Cannabinoid-Biosynthese. Im Rahmen dieser Arbeiten wurden Techniken angewandt und verbessert, um das Metabolom, Transkriptom und Proteom der biosynthetisierenden Zellen der sezernierenden Trichome zu erforschen. Die Biosyntheleistung der drei Trichomtypen wurde mit Hilfe von LC-MS, Laser Dissection Mikroskopie und 1H-NMR Metabolomics untersucht, um die Biosynthese über 8 Wochen qualitativ und quantitativ zu erfassen. Weitere Analysen des Proteom zeigten auf, das Genexpression und Funktion biosynthetischer Proteine für Cannabinoide, Flavonoide und Terpene des ätherischen Öls ist stark abhängig von Alter und Blütenbildung ist. Analyse und Annotierung der Gene ist durch die eigens erstellte cDNA Bank ermöglicht worden. Die Ergebnisse zeigen, das eine Biosynthese in den sezernierenden Kopfzellen sehr stark aber auch in den Stielzellen zu finden ist. Auf Grund der Proteomuntersuchungen im ätherischen Öl ist eine Biotransformation im nicht wässrigen Milieu ausgeschlossen worden. Mit Hilfe von 1H-NMR Metabolomics und statistischer PCA-Analyse konnten Strategien entwickelt werden, um Zuchtlinien von Cannabis sativa zu identifizieren und auf Basis des metabolomischen Profil voneinander zu unterscheiden.