Ruta chalepensis L. In Vitro Cultures as a Source of Bioactive Furanocoumarins and Furoquinoline Alkaloids

SIMPLE SUMMARY: Of late, the interest in alternative sources of bioactive compounds to field crops has been increasing due to decreasing crop acreages and climate change. Plant biotechnology offers the possibility of cultivating valuable medicinal plant species using in vitro methods. In vitro cultures are characterized by a significant increase in biomass, and one of their primary advantages is that they are independent of climate and soil conditions. This study aimed to investigate the ability of Ruta chalepensis bioreactor cultures to produce the following valuable bioactive substances: linear furanocoumarins and furoquinoline alkaloids. Bioreactor cultures of R. chalepensis may be used as a good raw material for the production of important bioactive compounds from the groups of coumarins and alkaloids. ABSTRACT: Recently, due to the decreasing areas of cultivation and climate change, the use of biotechnological methods to obtain biomass, which is a source of valuable bioactive metabolites, is becoming more and more interesting. In this study, Ruta chalepensis in vitro cultures were investigated in RITA(®) temporary immersion bioreactors. Biomass growth and the production of secondary metabolites in 4- and 5-week growth cycles on three variants of the Linsmaier and Skoog (LS) medium (naphthyl-1-acetic acid/6-benzylaminopurine (NAA/BAP): 0.5/1.0, 0.1/0.1, and 1.0/1.0 mg/L) were analyzed. Using high-performance liquid chromatography of methanolic extracts of biomass, the presence of linear furanocoumarins (bergapten, isoimperatorin, isopimpinellin, psoralen, and xanthotoxin) and furoquinoline alkaloids (γ-fagarine, 7-isopentenyloxy-γ-fagarine, and skimmianine) was confirmed. The highest content of linear furanocoumarins (1170 mg/100 g DW (dry weight)) was observed in the LS medium variant containing 0.5/1.0 mg/L NAA/BAP (4-week growth cycle). The highest content of furoquinoline alkaloids (449 mg/100 g DW) was observed in the LS medium variant containing 0.1/0.1 mg/L NAA/BAP (5-week growth cycle). Hence, R. chalepensis bioreactor cultures may be used as a biotechnological source of linear furanocoumarins (xanthotoxin and bergapten) and furoquinoline alkaloids (skimmianine and γ-fagarine).