Identification of Ruta graveolens L. Metabolites Accumulated in the Presence of Abiotic Elicitors

Abstract

The study aimed to elucidate the effects of benzothiadiazole (BTH) and saccharin on the biosynthesis of simple coumarins, linear furanocoumarins, dihydrofuranocoumarins, and furoquinolone alkaloids in shoots of R. graVeolens cultivated in vitro. The biosynthesized metabolites were analyzed and identified by GC-MS and by comparison of Kovats indices. Eight coumarin metabolites were identified: bergapten, chalepin, isopimpinelin, pinnarin, psoralen, rutacultin, rutamarin, and xanthotoxin, and also four alkaloids: dictamnine, γ-fagarine, skimmianine, and kokusaginine. Each of the tested BTH concentrations induced a significant production of furanocoumarins and furoquinolone alkaloids. The use of saccharin also increased the production of bergapten, isopimpinelin, pinnarin, psoralen, and xanthotoxin several times. IntroductionThe in vitro culture of R. graVeolens is a useful biotechnological source of biologically active linear furanocoumarins and furoquinolone alkaloids. Linear furanocoumarins, particularly xanthotoxin, bergapten, and isopimpinelin, have been applied in the treatment of skin diseases characterized by excessive cell proliferation (e.g., psoriasis, mycosis fungoides) or in pigmentation disorders (e.g., vitiligo) (1) and also in neurology (the symptomatic treatment of demyelinating diseases, particularly multiple sclerosis) (2). In addition, alkaloids like dictamnine and methoxydictamnine from R. graVeolens tissues are wellknown antimicrobial factors (3). Apart from their antibiotic effect, furoquinolone alkaloids appear to have a spasmolytic effect (4). The biological effects of furanocoumarins and furoquinolone alkaloids makes them attractive for pharmaceutical uses, hence the considerable interest shown in their availability and sources. However, the commercial production of secondary metabolites is usually limited by their low yield. Elicitation has therefore become an extensively used tool for enhancing secondary metabolites: it is an integral part of any large-scale process for secondary metabolite production.The mechanisms of action of biotic and abiotic elicitors are thought to be different; they are complex, and many hypotheses have been set up in this regard. Moreover, since little is known about the biosynthetic pathways of secondary metabolites, the effect of an elicitor on a plant cell or tissue culture is not easily predictable; the majority of elicitation approaches are therefore empirical (5). The search for elicitors is of the utmost importance in order to obtain the most effective biotechnological system for producing the required metabolites. The objective of our study was to elucidate the effects of saccharin, as well as benzothiadiazole (BTH, benzo(1,2,3)-thiadiazole-7-carbothionic acid S-methyl ester) on the biosynthesis of simple coumarins, linear furanocoumarins, dihydrofuranocoumarins, simple coumarins, and furoquinolone alkaloids in shoots of R. graVeolens cultivated in vitro.Benzothiadiazole is the main component of the BION preparation (6, 7). Chemically, it is a functional...

Bioprocess Optimization of Furanocoumarin Elicitation by Medium Renewal and Re-elicitation: A Perfusion-Based Approach

“…al. [9][10][11] recently reported enhanced FC production in response to various biotic and abiotic elicitors in R. graveolens shoot cultures. However, they did not investigate the effect of elicitation on product release and biomass.…”

Rutamarin, an active constituent from Ruta angustifolia Pers., induced apoptotic cell death in the HT29 colon adenocarcinoma cell line

“…Eight compounds [ Figure 2 ], namely graveoline (1), kokusaginine (2), bergapten (3), arborinine (4), chalepin (5), moskachan D (6), chalepensin (7), and rutamarin (8), were identified through their mass spectral and NMR data, which were consistent with published data. [ 9 10 11 ] However, only the mass spectral and NMR data for rutamarin are provided in Appendixes section. Psoralen (9) and moskachan B (10) were identified through their mass spectral data, which were consistent with published data.…”

“…Psoralen (9) and moskachan B (10) were identified through their mass spectral data, which were consistent with published data. [ 9 10 ] Gas chromatography-mass spectrometry (GC-MS) analysis was performed using an Agilent Technologies 6980 N gas chromatograph equipped with a 5979 Mass Selective Detector (70eV direct inlet). The column used here was an HP-5 ms capillary column (5% phenylmethylsiloxane) (30.0 m × 25 mm × 25 μm), which was initially set at 100°C, then increased to 300°C at 5°C/min, and then held for 10 min using helium as the carrier gas at a flow rate of 1 ml/min.…”

Avaliação de furanocumarinas como inibidores da fotossíntese através de ensaios de fluorescência da clorofila a

“…12,14 Os parâmetros funcionais calculados por este ensaio, chamado de test-JIP, são específicos (centro de reação-RC) e fenomenológicos (seção transversal-CS), medindo o acúmulo de energia absorvida (TR) e conversão de energia de excitação para a cadeia transportadora de elétrons (ET). 8 Dando continuidade aos estudos sobre a interação de produtos naturais com os sítios da fotossíntese, 6,7,15,16 o potencial herbicida das furanocumarinas psoraleno, bergapteno, chalepensina e chalepina foram avaliados dentro da cadeia transportadora de elétrons.…”