Mo primarily improves the nitrogen fixation to the plant and increases its antioxidant potential, which we observed in our study. This also might be the reason for the enhanced antidiabetic potential of Canavalia species assessed by GI, α-amylase inhibition activity, and α-glucosidase inhibition activity. Terpenoids and flavonoids of Canavalia gladiata are reported to play a role in lowering glucose levels and possessing antioxidant potential . The Mo-mediated enhanced concentrations of terpenoids and flavonoids might have led to the increased antioxidant and antidiabetic activities of Canavalia sprouts in our study. For sample preparation, we used an ETA 0067 grinder with grinding stones, VIPO mini grinder, followed by homogenization by Vibrom S2 , and a cryogenic grinder accompanied by liquid nitrogen. Supercritical fluid extraction using SE-1 extractor and a steam distillatory apparatus according to CSN 58 0110 and CSN 6571 were successively applied for extraction and subsequent determination of the total content of Canavalia oils.

Approximately 500 mg of each sample was transferred into an extraction column for SFE extraction. The extraction was performed at 40 MPa for 60 min, and extractor and restrictor temperatures were 80 and 120 ◦C, respectively. The extract was further trapped into a hexane layer inside a trapping vessel.The determination of phenolics and their precursor metabolites was carried out using an ultra-performance liquid chromatography system coupled with a quadrupole mass spectrometer provided with an ESI source according to Wang et al.’s method . Phenolic and flavonoid levels were identified by comparing the standard mixture of different phenols and flavonoids to the relative retention time. The concentration of each compound was calculated using the peak area of the corresponding standard. In addition, deoxy-d-arabino 201 heptulosonate-7-phosphate synthase activity was analyzed according to Wang et al.. This enzyme catalyzes the reactions in cinnamic and shikimic acid pathways that are involved in phenylpropanoid biogenesis and therefore in the biosynthesis of coumarins.The assay mixture contained 0.1 mM erythrose-4-phosphate, 0.2 mM phosphoenolpyruvate, and 0.1 mM MnSO4/0.1 mM CoCl2.

In total, the reaction was initiated by enzyme addition and terminated by 25% trichloroacetic acid addition. The activity of DAHPS was measured at 549 nm. Regarding PAL, it was extracted in 1 mL of 200 mM sodium borate buffer and then evaluated by measuring the production of trans-cinnamic acid at 290 nm. For centuries, Cannabis sativa L. has been widely used around the world for various applications . These days, interest has been focused on medicinal and recreational facets, furthering commercial expansion. With Canada recently adopting the more globally appreciated view of cannabis, there exists an ever evolving, multi-billion-dollar industry focused on vegetative propagation. Despite the reliance on clonal propagation, there is a continual need to germinate seeds to select new elite genotypes, perform pheno-hunting, as well as supporting breeding programs. To select new elite genotypes, plants are started from seed. During the vegetative phase of growth, a cutting is taken and maintained as a vegetative plant while the seedling is grown to maturity. Once the elite genotypes are selected, the cutting is then used as a source to propagate the clonal line. Maintaining the large population of cuttings during the phenotyping exercise represents a significant cost to producers and leaves the cutting derived mother plants exposed to insects and diseases.

To address the issues of insect, disease, and viral infections in mother plants, many producers use plant tissue culture to ensure that they are starting with clean material. Forthis process, nodal segments are disinfected and established in culture, a time consuming and relatively expensive endeavor. A potential alternative to the traditional approach is to first establish the seed in tissue culture. Once the seedlings are established and multiplied, micropropagated clones can be transferred into the growth facility and cultivated to maturity to identify elite genotypes. After selecting the elite genotypes, the in vitro parent material would be available for clonal propagation. This approach would greatly reduce the amount of space required for selecting new cultivars and provide a ready source of clean planting material once elite genotypes are identified. However, this approach requires an effective in vitro seed germination protocol with high germination speed and frequency. An efficient in vitro seed germination system would also support downstream biotechnologies in which seedling-derived tissues are preferred .We previously reported the effect of different types and strengths of media in addition to carbohydrate types and levels as primarily important factors contributing to in vitro cannabis seed germination indices and morphological seedling traits . Our results demonstrated that maximum germination percentage was achieved with 0.43 strength mMS medium and 2.3% sucrose . While the germination rate was over 80%, this was after 40 days of culture. Typically, the cannabis seed germinates within several days in the greenhouse/growth room, suggesting that something during the disinfection process was interfering with subsequent germination.