Pinarosa Avato

Metabolites in cell suspension cultures, calli, and in vitro regenerated organs of Hypericum perforatum cv. Topas

Abstract Methanolic extracts from cell suspension cultures, calli, and in vitro regenerated shoots and roots of Hypericum perforatum cv. Topas have been evaluated for their ability to produce active metabolites (hypericins, hyperforins and flavonoids). Biosynthesis of hypericins is connected with the formation of secretory structures (black globules) in regenerated vegetative buds. A further degree of leaf development is necessary to stimulate the production of hyperforins and flavonoids. Xanthones are the main metabolic products in suspension cells, undifferentiated calli and roots regenerated from plantlets or formed by callus. No xanthones are detected in the aerial parts of regenerated plantlets accumulating hypericins, hyperforins and flavonoids.

Phytochemical analysis of a herbal tea from Artemisia annua L.

Strategies to control diffusion of malaria needs to account for the increase of resistance of the parasite to the conventional antimalarial drugs. It has been proposed that a traditional aqueous preparation from Artemisia annua, with a low content of the active compound, artemisinin, may reduce the risk of resistance of the protozoa and be relatively more effective in the treatment of the disease. The solubility properties of the molecule have been the matter of concern about the therapeutic usefulness of herbal teas from A. annua. The present study aimed at analysing the chemical profile of a tea infusion from A. annua. Tea from A. annua was prepared through infusion of the plant aerial parts in water for 1, 24 and 48 h. Content of artemisinin was determined by HPLC-ELSD. Overall chemical characterization of the extracts was carried out by a combination of metabolomic techniques. The artemisinin content varied only slightly in the three different extracts (about 0.12%). A series of mono-caffeoyl- and mono-feruloyl-quinic acids, di-caffeoyl- and di-feruloyl-quinic acids was identified as main components of the tea infusion, together with some flavonoids. Reconstitution of the same extracts in less polar or apolar solvents resulted in a different composition with no phenolics and a much lower concentration of artemisinin.

A comparison of headspace solid-phase microextraction and classic hydrodistillation for the identification of volatile constituents from Thapsia spp. provides insights into guaianolide biosynthesis in Apiaceae.

INTRODUCTION Thapsia spp. (Apiaceae) are the major natural source of polyoxygenated guaianolide sesquiterpene lactones known as thapsigargins, which induce apoptosis in mammalian cells via a high affinity inhibition of the sarco/endoplasmic reticulum Ca(2+) ATPase. The mechanism of biosynthesis of thapsigargins has not been elucidated, and probable biochemical precursors such as hydrocarbon or oxygenated sesquiterpenes have not been identified in previous phytochemical analyses of essential oils from this genus. OBJECTIVE To investigate the utility of solid phase micro-extraction (SPME), when compared with classical essential oil distillates, for identifying potential precursors of guaianolide sesquiterpene lactones from Thapsia garganica L. and Thapsia villosa L. type II. METHODOLOGY A systematic description of the volatile components of roots, flowers, stems and fruits of T. villosa and of root, flower and fruits of T. garganica was constructed via GC-MS analyses of SPME-adsorbed compounds and of essential oils obtained through hydrodistillation of the same tissues. RESULTS The sesquiterpenoids δ-cadinene, α- and δ-guaiene, elemol and guaiols were found to be major volatile constituents of the roots of T. garganica and T. villosa trapped using SPME. In contrast, these sesquiterpenoids were not detected or were at negligible levels in essential oils, where sesquiterpenoids are potentially converted to azulenes during hydrodistillation. CONCLUSION The new data reported in this study demonstrates that SPME is a valuable tool for the identification of volatile sesquiterpenes when compared with analysis of essential oils, and we postulate that guaiene is the likely precursor of guaianolide sesquiterpenes from Thapsia.

Carvacrol: From Ancient Flavoring to Neuromodulatory Agent

Oregano and thyme essential oils are used for therapeutic, aromatic and gastronomic purposes due to their richness in active substances, like carvacrol; however, the effects of the latter on the central nervous system have been poorly investigated. The aim of our study was to define the effects of carvacrol on brain neurochemistry and behavioural outcome in rats. Biogenic amine content in the prefrontal cortex and hippocampus after chronic or acute oral carvacrol administration was measured. Animals were assessed by a forced swimming test. Carvacrol, administered for seven consecutive days (12.5 mg/kg p.o.), was able to increase dopamine and serotonin levels in the prefrontal cortex and hippocampus. When single doses were used (150 and 450 mg/kg p.o.), dopamine content was increased in the prefrontal cortex at both dose levels. On the contrary, a significant dopamine reduction in hippocampus of animals treated with 450 mg/kg of carvacrol was found. Acute carvacrol administration only significantly reduced serotonin content in either the prefrontal cortex or in the hippocampus at the highest dose. Moreover, acute carvacrol was ineffective in producing changes in the forced swimming test. Our data suggest that carvacrol is a brain-active molecule that clearly influences neuronal activity through modulation of neurotransmitters. If regularly ingested in low concentrations, it might determine feelings of well-being and could possibly have positive reinforcer effects.

Control of plant parasitic nematodes with active saponins and biomass from Medicago sativa

Medicago sativa L., alfalfa, is the most known plant species within the Medicago genus. The plant has been extensively studied for its content of saponins, mainly consisting of triterpene glycosides of medicagenic acid, possessing several biological properties including a biocidal activity on different soil microorganisms. Phytoparasitic nematodes are responsible for heavy economic damages to numerous agricultural crops and, due to their large distribution, they are among the most difficult crop pests to control. Attention on environmental safety and human and animal health has led to the progressive dismission of many synthetic formulations for the control of those pests and to the search of alternative strategies, including the use of natural metabolites from plants. Saponins from M. sativa may be good candidates for natural nematicide formulations, as in our in vitro studies the saponin mixtures from M. sativa tissues have been found effective in vitro against the virus-vector nematode Xiphinema index, the root-knot nematode Meloidogyne incognita and the potato cyst parasite, Globodera rostochiensis. A structure–activity relationship among saponins and related prosapogenins and sapogenin, respectively, has also been analyzed. The nematicidal efficacy differed among the three assayed nematode species, G. rostochiensis being the most susceptible to the active compounds from alfalfa. The in vitro results were also confirmed by experiments in potting mixes infested by M. incognita or G. rostochiensis and amended with dry top and root material from M. sativa, and in field trials on M. incognita and carrot cyst nematode Heterodera carotae with M. sativa pelleted meal. All amendments reduced root and soil population densities of target nematode species compared to non-treated and chemical controls, with a general improvement of plant growth and yield performances.

Biosynthesis of saponins in the genus Medicago

AbstractsSaponins from Medicago species are glycosidic compounds with an aglycone moiety formed through the enzymatic cyclization of 2,3-oxidosqualene by the β-amyrin cyclase. All the saponins from Medicago genus possess the triterpenic pentacyclic nucleus belonging to the class of β-amyrin. The so formed β-amyrin skeleton can be further modified by oxidative reactions, mediated by cytochromes belonging to the class of cytochrome P450, to give different saponin compounds, characterized by the presence of hydroxyl or carboxyl groups located in specific positions of the triterpenic skeleton. Based on the position and the oxidation degree of the substituents, it is possible to distinguish two groups of saponins (sapogenins) in Medicago spp: (1) sapogenins possessing an OH group on C-24 (soyasapogenols A, B and E) without any substituent at the C-28 atom, and (2) sapogenins possessing the COOH group at C-28 that are associated with different oxidation degrees (zero, OH, CHO, COOH) at C-23. These results seem to indicate that the oxidation at C-24 and the presence of the COOH group at C-28 are mutually exclusive. The subdivision in the aglycone moiety is reflected also in the sugar moiety, operated by glycosyltranferases, as the saponins of the two groups differ for the position and the nature of the sugar chains. Based on these findings, new considerations on the biosynthesis of saponins in the genus Medicago can be drawn and a biosynthetic scheme is proposed.

New triterpenic saponins from the aerial parts of Medicago arabica (L.) huds.

The reinvestigation of saponin composition from Medicago arabica from Italy allowed the detection of nineteen (1-19) saponins. All of them were purified by reverse-phase chromatography and their structures elucidated by spectroscopic and spectrometric (1D and 2D NMR; ESI-MS/MS) and chemical methods. Fourteen were known saponins, previously found in other plants including other Medicago species. They have been identified as glycosides of oleanolic acid, 2beta,3beta-dihydroxyolean-12-en-28-oic acid, hederagenin, bayogenin and soyasapogenol B. Five saponins, identified as 3-O-[-alpha-L-arabinopyranosyl(1-->2)-beta-D-glucuronopyranosyl]-30-O-beta-D-glucopyranosyl 2beta,3beta,30-trihydroxyolean-12-en-28-oic acid (1), 3-O-[alpha-L-arabinopyranosyl(1-->2)-beta-D-glucuronopyranosyl]-30-O-[beta-D-glucopyranosyl] 3beta,30-dihydroxyolean-12-en-28-oic acid (2), 3-O-[beta-D-glucuronopyranosyl]-30-O-[alpha-L-arabinopyranosyl(1-->2)-beta-d-glucopyranosyl] 2beta,3beta,30-trihydroxyolean-12-en-28-oic acid (3), 3-O-[beta-D-glucuronopyranosyl]-30-O-[alpha-L-arabinopyranosyl(1-->2)-beta-D-glucopyranosyl] 3beta,30-dihydroxyolean-12-en-28-oic acid (4) and 3-O-[beta-D-glucuronopyranosyl]-30-O-[beta-D-glucopyranosyl] 2beta,3beta,30-trihydroxyolean-12-en-28-oic acid (5), are reported here as new natural compounds. These new saponins, possessing a hydroxy group at the 30-methyl position of the triterpenic skeleton, have never been previously found in the genus Medicago.

Acetylenes and terpenoids of Bellis perennis

Abstract The essential oils of leaves and flowers from Bellis perennis , the common daisy, have been investigated. Polyacetylenes were one of the major chemical classes (18–21%) mainly consisting of two new C 10 acetylenic compounds which were identified, by spectroscopic and chemical means, as methyl deca-4,6-diynoate (2,8-tetrahydromatricaria ester) and deca-4,6-diynoic acid.

Nematicidal potential of Brassicaceae

Brassicaceae Burnett (syn. Cruciferae A. L. de Jussieu) include many important economic plants used as edibile or ornamental. They are commonly known as the “mustard” plant family due to the sharp, potent flavour of their main metabolites, the glucosinolates (GLSs) which contain sulfur. Glucosinolates coexist in vivo with glycosylated thioglucosidases, myrosinase(s), responsible of their hydrolysis with the production of bioactive cognate isothiocyanates (ITC). GLSs and ITCs function as defence bioactive metabolites against plant pathogens, insects and herbivores. The present review paper focus on GLSs role as bionematicides. The current knowledge on the efficacy of these phytochemicals against the most common phytoparasitic nematodes affecting crops of agriculture importance such as tomato, potato and grapevine is reported. Data from our ongoing research on the in vitro biocidal activity of glucosinolate extracts, and their main components, against the virus-vector nematode Xiphinema index Thorne & Allen and the carrot cyst nematode Heterodera carotae Jones are also described.

Determination of Major Constituents in St. John's Wort Under Different Extraction Conditions

Different extraction procedures have been evaluated to quantify the major constituents (hypericins, hyperforins and flavonoids) of H. perforatum drug samples. The total amount of hypericins has been determined by UV and HPLC. Analysis of hyperforins and flavonoids has been performed by HPLC. Sonication with MeOH resulted the best extraction method to obtain a homogenous drug sample, representative of all the major metabolites and with less degradation products. The procedure was standardized with a commercial drug sample and then applied to quantify the active metabolites in H. perforatum plants collected in various localities in South Italy. Two of the drug samples resulted particularly rich in hypericin, namely, Altamura, 0.21% and Casamassima, 0.27%. Analysis of the flavonoid content confirmed the presence, in Italy, of a chemotype of H. perforatum without rutin.