Untargeted Metabolomic Analyses Reveal Chemical Complexity of Dioecious Cannabis Flowers
Matthew T. Welling
A , Myrna A. Deseo A B , Antony Bacic A B and Monika S. Doblin A B C
A La Trobe Institute for Agriculture and Food, Department of Animal, Plant and Soil Sciences, School of Life Sciences, AgriBio Building, La Trobe University, Bundoora, Vic. 3086, Australia.
B Australian Research Council Research Hub for Medicinal Agriculture, AgriBio Building, La Trobe University, Bundoora, Vic. 3086, Australia.
C Corresponding author. Email: m.doblin@latrobe.edu.au
Australian Journal of Chemistry 74(6) 463-479 https://doi.org/10.1071/CH21033
Submitted: 30 January 2021 Accepted: 26 April 2021 Published: 21 May 2021
Journal Compilation © CSIRO 2021 Open Access CC BY-NC-ND
Abstract
Cannabis is a mostly dioecious multi-use flowering plant genus. Sexual dimorphism is an important characteristic in Cannabis-based commercial production systems, which has consequences for fibre, seed, and the yield of secondary metabolites, such as phytocannabinoid and terpenes for therapeutic uses. Beyond the obvious morphological differences between male and female plants, metabolic variation among dioecious flowers is largely undefined. Here, we report a pilot metabolomic study comparing staminate (male) and pistillate (female) unisexual flowers. Enrichment of the α-linolenic acid pathway and consensus evaluation of the jasmonic acid (JA) related compound 12-oxo-phytodienoicacid (OPDA) among differentially abundant metabolites suggests that oxylipin signalling is associated with secondary metabolism and sex expression in female flowers. Several putative phytocannabinoid-like compounds were observed to be upregulated in female flowers, but full identification was not possible due to the limitation of available databases. Targeted analysis of 14 phytocannabinoids using certified reference standards (cannabidiolic acid (CBDA), cannabidiol (CBD), Δ9-tetrahydrocannabinolic acid A (Δ9-THCAA), Δ9-tetrahydrocannabinol (Δ9-THC), cannabichromenic acid (CBCA), cannabichromene (CBC), cannabigerolic acid (CBGA), cannabigerol (CBG), cannabinolic acid (CBNA), cannabinol (CBN), cannabidivarinic acid (CBDVA), cannabidivarin (CBDV), tetrahydrocannabivarinic acid (THCVA), and tetrahydrocannabivarin (THCV)) showed a higher total phytocannabinoid content in female flowers compared with the male flowers, as expected. In summary, the development of a phytocannabinoid-specific accurate-mass MSn fragmentation spectral library and gene pool representative metabolome has the potential to improve small molecule compound annotation and accelerate understanding of metabolic variation underlying phenotypic diversity in Cannabis.
Keywords: liquid chromatography, mass spectrometry, phytochemistry, metabolism, Cannabis, cannabinoids, untargeted metabolomics, flowering.
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