Cell Suspensions of Cannabis sativa (var. Futura): Effect of Elicitation on Metabolite Content and Antioxidant Activity

Abstract

Cannabis sativa L. is one of the most-studied species for its phytochemistry due to the abundance of secondary metabolites, including cannabinoids, terpenes and phenolic compounds. In the last decade, fiber-type hemp varieties have received interest for the production of many specialized secondary metabolites derived from the phenylpropanoid pathway. The interest in these molecules is due to their antioxidant activity. Since secondary metabolite synthesis occurs at a very low level in plants, the aim of this study was to develop a strategy to increase the production of such compounds and to elucidate the biochemical pathways involved. Therefore, cell suspensions of industrial hemp (C. sativa L. var. Futura) were produced, and an advantageous elicitation strategy (methyl jasmonate, MeJA) in combination with precursor feeding (tyrosine, Tyr) was developed. The activity and expression of phenylalanine ammonia-lyase (PAL) and tyrosine aminotransferase (TAT) increased upon treatment. Through 1H-NMR analyses, some aromatic compounds were identified, including, for the first time, 4-hydroxyphenylpyruvate (4-HPP) in addition to tyrosol. The 4-day MeJA+Tyr elicited samples showed a 51% increase in the in vitro assay (2,2-diphenyl-1-picrylhydrazyl, DPPH) radical scavenging activity relative to the control and a 80% increase in the cellular antioxidant activity estimated on an ex vivo model of human erythrocytes. Our results outline the active metabolic pathways and the antioxidant properties of hemp cell extracts under the effect of specific elicitors.

Advances and Perspectives in Tissue Culture and Genetic Engineering of Cannabis

“…However, cell suspension cultures may still be promising for producing other secondary metabolites such as terpenes, polyphenols, lignans, and alkaloids [ 7 , 10 ]. For instance, Gabotti et al [ 55 ] reported that the activity and expression of tyrosine aminotransferase (TAT) and phenylalanine ammonia-lyase (PAL) increased in cannabis cell suspension cultures using a methyl jasmonate elicitor in combination with tyrosine precursor. Some aromatic compounds such as 4-hydroxyphenylpyruvate (4-HPP) were also identified.…”

Elicitation-Based Method for Increasing the Production of Antioxidant and Bactericidal Phenolic Compounds in Dionaea muscipula J. Ellis Tissue

“…Similar findings have been shown by Ansari et al [46], where transformed hairy root culture of Ligularia fischeri were characterized by increased synthesis of phenolic compounds and higher radical scavenging activity, than control plants. Moreover, Cannabis sativa cell suspension culture treated with jasmonates and some precursors of phenylpropanoid pathway accumulated more phenolic derivatives and had increased radical scavenging activity against DPPH [47].…”

Effects of Abiotic Elicitors on Expression and Accumulation of Three Candidate Benzophenanthridine Alkaloids in Cultured Greater Celandine Cells

“…Plant cell culture, on the other hand, is introduced as a potential alternative to up-scale production of SMs in a shorter time and less space than traditional manners [ 7 ]. Moreover, a higher level of SMs production of different medicinal plants through cell culture has already been reported [ 8 , 9 ], although the production level of SMs through cell culture is still considerably lower than the intact plant in some cases. Particular substances called “elicitors” are applied as stress agents to boost the SM production in cultured plant cells [ 10 ].…”