## 2 Results
### Phytohormonal Effects on Cannabinoid Metabolism  
Phytohormonal effects on disc cell cannabinoidoid content were measured and compared to the CTR_24h values (Figure 6). DIECA application resulted in a significant 87% decrease in CBGA, and while not significant (\(p\) = 0.058), THCA trended to decrease by 29%. ETH also caused a notable trend (\(p\) = 0.058) by increasing CBGA by 43%. CBCA and total measured cannabinoid content did not change for any of the treatments. Figure 5: Comparison of differentially abundant proteins (DAPs) in cannabis (_Camabis sativa_ L.) glandular trichome disc cells when treated _in vitro_ (\(n\) = 4) with either kinetin (KIN), jasmonic acid (JA), or salicylic acid (SA). Venn diagrams represent intersecting increasing (**A**) and decreasing (**B**) differentially DAPs between KIN, JA, and SA. Key differentially abundant proteins affected by at least two of the phytohormones included those related to plastitial function (**C–E**), cytosolic acetyl-CoA synthesis (**F**), and fatty acid metabolism (**G–I**). Boxplots represent median, interquartile range, maximum, minimum, and outliers (closed circles). Significance (denoted by \(\alpha\)) determined by two-sample Student’s \(t\)-test (\(p\)-value \(<\) 0.05) by comparing phytohormone treatments (\(n\) = 4) to the control (CTR_24h). Abbreviations: WTF1, WHAT’s THIS FACTOR 1; cpSecA, chloroplastic protein translocase subunit SECA1; TIC214, translocon at the inner envelope membrane of chloroplasts 214; ACLA-1, ATP citrate lyase A-1; CER10, very-long-chain enoyl-CoA reductase; CER2, eceriferum 2; LACS9, chloroplastic long chain acyl-CoA synthetase 9. Listed in titles are protein names, closest arabidopsis homolog, and UniProt accession number. SWATH-MS allowed the relative abundance levels of seven key cannabinoid biosynthesis protein (Figure 7) to be calculated: tetraketide synthase (TKS), olivetolic acid cyclase (OAC), aromatic preyltransferasees1 (CsPT1), cannabinoidic acid synthase (CBDAS), tetrahydroocannabinolic acid synthase (THCAS), geranylgeranyl pyrophosphate synthase 1 GPPLsu (GPPS.lsu), and geranylgeranyl pyrophosphate reductase GPPSsu (GPPS.ssu). Neither GA, ETH, nor ABA had any effect on the abundance of these seven proteins. Both BR and DIECA increased TKS abundance by 100% and 88%, respectively. DIECA, KIN, SA, and JA decreased OAC abundance by 9%, 80%, 77%, and 64%, respectively. BR, KIN, and JA decreased CBDAS abundance by 15%, 23%, and 22%, respectively. SA increased THCAS abundance by 17%. KIN and SA increased GPPS.ssu abundance by 34% and 31%, respectively. None of the treatments had any effect on the abundance of CsPT1 or GPPS.lsu. Figure 6: CBGA (**A**), THCA (**B**), CBCA (**C**), and total measured cannabinoid (**D**) content [mg g\({}^{-1}\) (DW)] of isolated cannabis (_Camabis sativa_ L) glandular trichome disc cells when treated _in vitro_ with phytohormes (\(n=4\)). Boxplots represent median, interquartile range, maximum and minimum, and outliers (closed circles). Significance (denoted by _a_) was determined by two-sample Student’s _t_-test (_p_-value \(<0.05\)) by comparing phytohormone treatments to the control (CTR_24h). Abbreviations: CBGA, cannabinoid acid; THCA, tetrahydroocannabinolic acid; CBCA, cannabinoid acid; ABA, abscisic acid; BR, brassinolide; DIECA, diethylditicarbamate; ETH, ethephon; GA, gibberell acid; KIN, kinetin; JA, jasmonic acid; SA, salicylic acid. Figure 7: Abundance of key proteins involved in cannabinoid biosynthesis (**A**–**E**) and geranyl pyrophosphate synthesis (**F**,**G**) in cannabis (_Cannabis sativa_ L.) glandular trichome disc cells when treated _in vitro_ with phytohormes (_n_ = 4). Boxplots represent median, interquartile range, maximum, minimum, and outliers (closed circles). Significance (denoted by _a_) was determined by two-sample Student’s _t_-test (_p_-value < 0.05) by comparing phytohorme treatments to the control (CTR_24h).