RESUMO
Cannabis sativa L. is a plant with a wide range of potential medicinal applications. In recent years, polyploidy has gained attention as a potential strategy for rapidly improving C. sativa, which, unlike other modern crops, has not yet benefitted from this established biotechnological application. Currently, no reports on high THCA and CBDA drug-type polyploid cultivars have been published. Moreover, it still needs to be clarified if different cultivars react similarly to polyploidization. For these reasons, we set out to evaluate and compare the phenotype and chemotype of three high Δ9-tetrahydrocannabinolic acid (THCA) and one high cannabidiolic acid (CBDA) drug-type cultivars in their diploid, triploid and tetraploid state through agronomic and metabolomic approaches. Our observations on plant morphology revealed a significant increase in plant height and leaf size with increasing ploidy levels in a cultivar-dependent manner. In contrast, cannabinoids were negatively affected by polyploidization, with the concentration of total cannabinoids, THCA, CBDA and cannabigerolic acid (CBGA) decreasing significantly in higher ploidy levels across all four cultivars. Headspace analysis of volatiles revealed that total volatile content decreased in triploids. On the other hand, tetraploids reacted differently depending on the cultivars. Two THCA dominant cultivars showed an increase in concentrations, while in the other two cultivars, concentrations decreased. Additionally, several rare compounds not present in diploids appeared in higher ploidy levels. Moreover, in one high THCA cultivar, a couple of elite tetraploid genotypes for cannabinoid and volatile production were identified, highlighting the role of cultivar and genotypic variability as an important factor in Cannabis sativa L. polyploids. Overall, our observations on plant morphology align with the giga phenotype observed in polyploids of other plant species. The decrease in cannabinoids and volatiles production in triploids have relevant implications regarding their commercial use. On the other hand, this study found that tetraploidization is a suitable approach to improve Cannabis sativa L. medicinal potential, although the response is cultivar and genotype-dependent. This work lays the ground for further improving, evaluating and harnessing Cannabis sativa L. chemical diversity by the breeding, biotechnological and pharmaceutical sectors.
RESUMO
Citrus black spot (CBS) is a disease caused by the fungus Phyllosticta citricarpa that affects citrus plants, causing fruit blemish and premature drop that result in severe economic losses in commercial citrus orchards. However, CBS symptoms and effects may vary depending on the citrus species: Citrus limon (lemon) is susceptible and highly affected by the disease, while no CBS-related damage has ever been observed for Citrus latifolia (Tahiti lime), implying that it must be resistant to the disease. The difference in the response to this disease provided the opportunity to gain insight into the metabolites responsible for the resistance by comparison of the metabolomic profiles of these two citrus species. Metabolic variations of C. limon and C. latifolia inoculated with P. citricarpa were analyzed using various metabolomic-based platforms including 1H NMR for overall metabolic profiling, and LC-MS and HPTLC for targeted analysis. The 1H NMR spectra of the samples demonstrated that certain phenolics were strongly induced after pathogenic inoculation, especially in the resistant species. The induced phenolics were identified from C. latifolia by further 1H NMR, LCMS and HPTLC analysis yielding six prenylated and methoxy coumarins, i.e., 5,7-dimethoxycoumarin, 5-geranyloxy-7-methoxycoumarin, 7-geranyloxycoumarin, 8-methoxypsoralen, 5,8-dimethoxypsoralen and 5-geranyloxypsoralen. These isolated coumarins and a coumarin-rich fraction were tested against the fungal pathogen, P. citricarpa, to evaluate their activity. None of the individual coumarins exhibited a significant inhibition, while the coumarin fraction exhibited a strong antifungal activity suggesting a synergistic interaction of its components. To obtain further insight into the roles of these compounds in the plant defense, the possible mechanisms of the individual coumarins were tested using an in-silico model, the PASS Online Tool; the analysis showed that each coumarin appeared to have a unique defense mechanism, even with very slight variations in the chemical structures. The results could provide evidence of the existence of a complex plant defense mechanism consisting in a multitude of synergistic interactions between compounds.
