Amounts Of Individual Compounds Research Articles

Endogenous production of specific flavonoids and verbascoside in agar and agitated microshoot cultures of Scutellaria lateriflora L. and biotransformation potential

Methanolic extracts of microshoots from agar cultures and of microshoots and media from agitated cultures of Scutellaria lateriflora grown on identical five variants of the Murashige-Skoog (MS) and Linsmaier-Skoog (LS) media with different 6-benzyladenine (BA) and 1-naphthaleneacetic acid (NAA) concentrations (0.5–3.0 mg/l) were analyzed for flavonoids (27 compounds), phenylpropanoid glycosides (2 compounds) and phenolic acids (19 compounds) using the DAD-HPLC method. The amounts of individual compounds and total amounts of the above mentioned metabolite groups in the biomass from both types of the tested cultures were dependent on the composition of basal media and concentrations of BA and NAA in the media variants. In media extracts, no metabolites were confirmed. Higher total amounts of flavonoids were obtained in agar cultures (max. 722.04 and 2989.55 mg/100 g DW on MS and LS media variants, respectively), which were 1.1 and 1.7 times higher, respectively, than in agitated cultures. In both types of cultures, glycosidic conjugates: baicalin and wogonoside were the main flavonoids (max. amounts: 513.93 and 1838.18 mg/100 g DW, and 305.92 and 700.85 mg/100 g DW in agar cultures on MS and LS variants, respectively). High amounts of verbascoside in agar cultures were also confirmed (max. 384.99 and 543.17 mg/100 g DW on MS and LS media variants, respectively). The cells grown in agitated cultures converted exogenous precursors, i.e. hydroquinone (HQ) and 4-hydroxybenzoic acid (4-HBAc) into arbutin (β-d-glucoside of HQ). The highest amount of this product reached 5.63 and 1.45 g/100 g DW after the addition of these two precursors, respectively. This is the first large-scale report documenting in vitro biosynthetic potential of Scutellaria lateriflora microshoots cultivated in two tested culture.

Selective responses of Trissolcus basalis and Telenomus podisi to chemical footprints of preferred hosts

AbstractChemical trails of the hosts (footprints) are important cues for the host searching behaviour of egg parasitoids of the family Scelionidae. The present study aims to determine the influence of the footprints of three neotropical stink bugs (Euchistus heros, Dichelops melacanthus and Nezara viridula) on the foraging behaviour of two parasitoids, Trissolcus basalis and Telenomus podisi, as well as a possible selective response to fooprints of their preferred hosts. Accordingly, Tr. basalis and Te. podisi females are allowed to forage on open arenas where E. heros, D. melacanthus or N. viridula had walked or on open arenas that had been treated with samples of an extract from each stink bug's footprints. Hexane extracts of stink bug footprints are obtained from solvent‐washed Petri dishes where insects were allowed to walk for 24 h, and these extracts were analyzed by gas chromatography‐flame ionization detection and gas chromatography‐mass spectrometry. Each parasitoid responds selectively to the footprints of their preferred host (Tr. basalis to footprints of N. viridula and Te. podisi to footprints of E. heros). Twenty‐six compounds comprising of C18 to C35 (saturated and unsaturated) and methyl branched hydrocarbons were identified in extracts of E. heros, D. melacanthus and N. viridula, respectively. There are significant differences in the total amount of the compounds identified in the footprint stink bug's extracts and also a difference in the amounts of individual compounds. In addition, the behavioural assays showed that footprints of stink bugs are stimuli that are used by egg parasitoids to search, discriminate and selectively locate their preferred host.

The importance of applied light quality on the production of lignans and phenolic acids in Schisandra chinensis (Turcz.) Baill. cultures in vitro

Shoot-differentiating callus cultures of Schisandra chinensis were cultivated under different spectral properties of monochromatic light (far-red, red, blue, UV-A), and in darkness and under white light (control conditions). The applied lighting conditions influenced biomass growth and the accumulation of secondary metabolites—dibenzocyclooctadiene lignans (fourteen compounds) and phenolic acids (seven compounds). Total amounts of the estimated metabolites increased 1.71 and 1.98 times, respectively, depending on light quality. Blue light was found to be the most effective lighting to the production of both groups of compounds. Their total amounts reached the maximum values of 376.41 mg/100 g DW and 46.57 mg/100 g DW, and were correspondingly 1.31 and 1.37 times greater than under white light. The amounts of individual compounds from the tested groups increased from 1.51 to 3.38 times (lignans), and from 1.74 to 2.72 times (phenolic acids), depending on the lighting conditions. The main bioactive compounds obtained in high amounts were: schisandrin (67.70 mg/100 g DW), deoxyschisandrin (55.19 mg/100 g DW), gomisin A (36.97 mg/100 g DW), and also chlorogenic acid (15.33 mg/100 g DW) and protocatechuic acid (13.11 mg/100 g DW). This is the first report providing evidence of the importance of light quality on the production of dibenzocyclooctadiene lignans and phenolic acids in the biomass of S. chinensis cultivated in vitro.

Accumulation of biologically active phenolic acids in agitated shoot cultures of three Hypericum perforatum cultivars: ‘Elixir’, ‘Helos’ and ‘Topas’

Agitated shoot cultures of three Hypericum perforatum cultivars: ‘Elixir’, ‘Helos’ and ‘Topas’ were established and maintained on Linsmaier and Skoog (LS) and Murashige and Skoog (MS) media containing varying concentrations (0.1–3.0 mg l−1) of α-naphthaleneacetic acid (NAA) and 6-benzyladenine (BA). In methanolic extracts of the biomass, the amount s of free phenolic acids and cinnamic acid were determined by HPLC. Six of the seventeen compounds analyzed were detected in all the extracts: 3,4-dihydroxyphenylacetic acid, p-coumaric acid, protocatechuic acid, syringic acid, and two depsides—chlorogenic and neochlorogenic acids. The amounts of individual compounds and the total amount of phenolic acids depended on the media variants used and the increases ranged from 1.2- to 15.0-fold and from 1.2- to 1.9-fold, respectively. The maximum total amounts in the biomass of ‘Elixir’, ‘Helos’ and ‘Topas’ were 212, 222, and 191 mg 100 g−1 DW, respectively. The main metabolites in all the cultivars were neochlorogenic acid (max. 118.81 mg 100 g−1 DW) and 3,4-dihydroxyphenylacetic acid (max. 129.29 mg 100 g−1 DW). Three metabolites that also accumulated in high amounts were protocatechuic acid (max. 21.43 mg 100 g−1 DW), syringic acid (max. 23.63 mg 100 g−1 DW) and chlorogenic acid (max. 18.79 mg 100 g−1 DW). The richest s ource of the estimated compounds and hence a potential biotechnological platform were the shoots of ‘Helos’ cultivated on LS and MS media containing low concentrations of NAA and BA (0.1–1.0 mg l−1).

Production of bioactive phenolic acids and furanocoumarins in in vitro cultures of Ruta graveolens L. and Ruta graveolens ssp. divaricata (Tenore) Gams. under different light conditions

Extracts from the biomass of Ruta graveolens and Ruta graveolens ssp. divaricata cultured in vitro under different light conditions (far-red, red and blue light, UV-A irradiation, in darkness and white light) were tested for the amounts of free phenolic acids and cinnamic acid (twelve compounds) as well as furanocoumarins and umbelliferone (seven compounds) using HPLC methods. Total amounts of the investigated groups of compounds in the cultures of both plants increased from 2.6 to 6.7 times, depending on light quality, and the maximum values reached were 106.50 and 1,276.74 mg 100 g−1 DW (in R. graveolens), and 106.97 and 262.54 mg 100 g−1 DW (in the subspecies), respectively. Both white light and blue light were equally beneficial for the total production of phenolic acids in cultures of both plants, whereas the total production of furanocoumarins was clearly better stimulated by blue light in R. graveolens and by darkness in the subspecies (i.e. the amounts were respectively 1.44 and 1.7 times higher than in the biomass cultivated under white light). The amounts of individual compounds in both plant cultures increased from about 2.2 to 26.3 times depending on light quality. The following bioactive compounds were obtained in quantities which are of interest from a practical perspective: in R. graveolens culture—protocatechuic acid (45 mg 100 g−1 DW), isopimpinellin (about 500 mg 100 g−1 DW) and bergapten (about 270 mg 100 g−1 DW), and in the subspecies culture: p-coumaric acid (70 mg 100 g−1 DW) and isopimpinellin (about 210 mg 100 g−1 DW).

Variation of phenolic acids from herb and roots of Salsola kali L.

Qualitative and quantitative analysis of phenolic acids present as free and liberated from their conjunction with sugars and alcohols after hydrolysis in <em>Salsola kali</em> L was done, together with comparison of their content in roots and herb. Analysis was conducted by 2D-TLC and HPLC methods. Eleven phenolic acids were identified such as: protocatechuic, caffeic, gentisic, p-coumaric, p-hydroxybenzoic, p-hydroxyphenylacetic, syryngic, vanillic, ferulic, α- and ß-resorcylic. Differences in composition of fraction of free phenolic acids and those liberated after hydrolysis were noticed. The amount of individual compounds were in the range between 0.10±0.04 mg/100 g of dry weight for p-OH-benzoic acid and 6.40±1.75 mg/100 g of dry weight for ferulic acid.

Sites of production and occurrence of volatiles in Douglas-fir beetle,Dendroctonus pseudotsugae hopkins

Volatiles were found to be distributed throughout adult Douglasfir beetles,Dendroctonus pseudotsugae Hopkins, suggesting that the hindgut may not be the sole site of production. The volatile content of individual beetles increased dramatically with feeding; qualitative changes reflected the different bark habitats of newly emerged and established beetles. All detected volatiles were also found in beetle frass. Myrcene was the predominant monoterpene of emergent beetles and α-pinene of fed beetles. Linalool and bornyl acetate occurred in significant amounts after feeding, and diacetone alcohol and sulcatol were also detected for the first time. The presence of sulcatol explains the reported cross-attraction withGnathotrichus spp.cis-Verbenol was also found to occur in addition totrans-verbenol. The volatile content of male beetles was similar to that of females but amounts of individual compounds were less. Acetic acid was found in both emerged and fed beetles and in lesser amounts in frass, suggesting that microbial metabolism could be a potential source of volatile production.