Effect of carbon source and elicitors on biomass production and accumulation of friedelin and epifriedelanol in hairy roots of hemp (Cannabis sativa L.)

Abstract

Friedelin and epifriedelanol are pentacyclic triterpenoids that preferentially accumulate in the roots of hemp (Cannabis sativa L.) and are valued for their antidiabetic, hypolipidemic, antioxidant, liver protective, anti-ulcer, anti-in ammatory, antimicrobial, anticancer, and antisenescence properties. The aim of the present study was to investigate the in uence of media, carbon sources, and elicitation on the production of C. sativa hairy root biomass and these metabolites. The MS liquid medium promoted the highest fresh weight (9.45 ± 0.00 g/100 mL ask) biomass production in hairy root cultures after 28 days.The highest levels of epifriedelanol (3.79-fold) and friedelin (3.25-fold) were found at the end of the exponential phase. The presence of 3% sucrose provided the highest accumulation of epifriedelanol (0.930 ± 0.013 mg/g DW) and friedelin (0.574 ± 0.024 mg/g DW) in the roots. The effects of methyl jasmonate (MJ) and salicylic acid (SA) on the enhancement of friedelin and epifriedelanol in C. sativa hairy root cultures were investigated. Between the two elicitors, SA showed the highest production of epifriedelanol (up to 5.018 ± 0.35 mg/g DW) and friedelin up to 1.56 ± 0.34 mg/g DW in 28-day-old stationary phase hairy roots. These represented 5.22-and 2.88-fold increase over the control (0.96 ± 0.01 mg/g DW and 0.54 ± 0.03 mg/g DW) after 96 h of treatment, respectively. The maximum accumulations of epifriedelanol (3.59 ± 0.12 mg/g DW) and friedelin (1.31 ± 0.01 mg/g DW) were observed in the treatment with MJ (100 µM) after 24 h of exposure and were 3.73-and 2.44-fold higher than the control, respectively. These ndings suggest that elicitation is an effective technique for enhancing the yields of these valuable bioactive pentacyclic triterpenoids in C. sativa hairy root cultures in a relatively short period of time.

Elicitation boosts withaferin A and withanolide A production in Withania somnifera (L.) Dunal hairy root cultures

“…As stated in prior research, the time of elicitation, exposure period, and type and concentration of the elicitor play a pivotal role in the pattern of targeted/speci c metabolite accumulation in hairy root cultures (Maciel et al 2022;Mahendran and Vimolmangkang 2024). According to our growth curve and metabolite accumulation results, the 25th day of hairy root growth at the end of the exponential stage was identi ed as the most effective day for elicitation.…”

“…PCR ampli cation was conducted using a PCR (Eppendorf, Hamburg, Germany). The ampli cation conditions and primer pair, consisting of forward primer 5'-CTCCTGACATCAAACTCGTC-3', reverse primer: 5'-TGCTTCGAGTTATGGGTACA-3', were used to amplify a ∼582-bp fragment of the rolC gene, as previously described in our earlier reports (Mahendran and Vimolmangkang, 2024).For the PCR procedure, a nontransformed root was utilized as a negative control, while a positive control containing R. rhizogenes ATCC 43057 was included for comparison.…”