In Vitro Multiplication and NMR Fingerprinting of Rare Veronica caucasica M. Bieb.

Micropropagation of rare Veronica caucasica M. Bieb. was achieved by successful in vitro cultivation of mono-nodal segments on MS medium supplemented with 1.0 mg L-1 6-benzylaminopurine (BA) and then transferring the regenerated plants on hormone free basal MS medium for root development. In vitro multiplicated plants were successively acclimated in a growth chamber and a greenhouse with 92% survival. The number of plastid pigments and the total phenolics content in in vitro cultivated and ex vitro adapted plants were unchanged, and no accumulation of reactive oxygen species (ROS) was detected by staining with 3-3'-diaminobenzidine (DAB) and 2',7'-dichlorofluorescein diacetate (DCF-DA). Nuclear Magnetic Resonance (NMR) fingerprinting allowed for the identification of the major alterations in metabolome of V. caucasica plants during the process of ex situ conservation. Iridoid glucosides such as verproside, aucubin and catalpol were characteristic for in vitro cultivated plants, while in ex vitro acclimated plants phenolic acid-protocatechuic acid and caffeic acid appeared dominant. The successful initiation of in vitro and ex vitro cultures is an alternative biotechnological approach for the preservation of V. caucasica and would allow for further studies of the biosynthetic potential of the species and the selection of lines with a high content of pharmaceutically valuable molecules and nutraceuticals.

Chemical Profiling and Assessment of Antineurodegenerative and Antioxidant Properties of Veronica teucrium L. and Veronica jacquinii Baumg.

Neuroprotective potential of V. teucrium and V. jacquinii methanol extracts was analyzed. Chemical analysis of investigated extracts showed the presence of phenolic acid derivatives, flavonoids and one secoiridoid. The detected flavonoids derived from flavones (luteolin and isoscutellarein in V. jacquinii; apigenin, isoscutellarein and luteolin in V. teucrium) and flavonol (quercetin in V. jacquinii). Acteoside was the dominant compound in V. jacquinii, while plantamajoside and isoscutellarein 7-O-(6‴-O-acetyl)-ß-allosyl (1‴→2‴)-ß-glucoside were the major phenolics in V. teucrium. Additionally, the antineurodegenerative activity was tested at concentrations of 25, 50, and 100 µg/ml using acetylcholinesterase (AChE) and tyrosinase (TYR) assays. The inhibition of both enzymes was achieved with the investigated extracts, ranging from 22.78 to 35.40% for AChE and from 9.57 to 16.38% for TYR. There was no statistical difference between the activities of the analyzed extracts. Our data indicate that V. teucrium and V. jacquinii may have beneficial effects against Alzheimer's and Parkinson's disease.

Examination of secondary metabolites and antioxidant capacity of Anthyllis vulneraria, Fuchsia sp., Galium mollugo and Veronica beccabunga.

Anthyllis vulneraria L., Fuchsia sp., Galium mollugo L., and Veronica beccabunga L. were selected to analyse the phenolic content and the antioxidant activity by ferric ion reducing antioxidant power (FRAP) and trolox equivalent antioxidant capacity (TEAC) assays. The highest polyphenol, tannin, and flavonoid contents were measured in Fuchsia species (7.40 ± 0.8, 5.62 ± 0.7 and 0.72 ± 0.1 g/100 g dry weight), while the lowest values were detected in Anthyllis vulneraria (0.68 ± 0.02, 0.17 ± 0.03 and 0.45 ± 0.01 g/100 g dry weight) and Galium mollugo (1.77 ± 0.05, 0.49 ± 0.04 and 0.16 ± 0.06 g/100 g dry weight). The leaf extract of Fuchsia sp. had the highest, while the herb of A. vulneraria had the lowest antioxidant effect measured by both methods, which is probably related to total polyphenol, tannin, and flavonoid contents.

Monitoring of selenium in macrophytes - the case of Slovenia.

This paper examines macrophytes from various locations in Slovenian streams for selenium (Se) content in an attempt to discover if Se contamination is present and if Se uptake varies between sampling sites. For this purpose, macrophytes and water from ten locations in the Notranjska and Central regions (Slovenia) with different land use in the catchment were sampled. To assess the environmental conditions of the streams the Riparian, Channel, and Environment (RCE) inventory was applied, which revealed that investigated stretches of streams fall into RCE classes III, IV and V. The concentration of Se in water at all locations was less than 1µgSeL(-1). The Se content in macrophytes differed between sampling sites, with the highest content of Se in samples from Zerovniscica stream and the lowest in samples from Lipsenjscica stream. The content of Se was the highest in moss samples (3038ngSeg(-1) DM) and in the amphibious species Veronica anagallis-aquatica (1507ngSeg(-1) DM).

Co-pigmentation and flavonoid glycosyltransferases in blue Veronica persica flowers.

Glycosylation is one of the key modification steps for plants to produce a broad spectrum of flavonoids with various structures and colors. A survey of flavonoids in the blue flowers of Veronica persica Poiret (Lamiales, Scrophulariaceae), which is native of Eurasia and now widespread worldwide, led to the identification of highly glycosylated flavonoids, namely delphinidin 3-O-(2-O-(6-O-p-coumaroyl-glucosyl)-6-O-p-coumaroyl-glucoside)-5-O-glucoside (1) and apigenin 7-O-(2-O-glucuronosyl)-glucuronide (2), as two of its main flavonoids. Interestingly, the latter flavone glucuronide (2) caused a bathochromic shift on the anthocyanin (1) toward a blue hue in a dose-dependent manner, showing an intermolecular co-pigment effect. In order to understand the molecular basis for the biosynthesis of this glucuronide, we isolated a cDNA encoding a UDP-dependent glycosyltransferase (UGT88D8), based on the structural similarity to flavonoid 7-O-glucuronosyltransferases (F7GAT) from Lamiales plants. Enzyme assays showed that the recombinant UGT88D8 protein catalyzes the 7-O-glucuronosylation of apigenin and its related flavonoids with preference to UDP-glucuronic acid as a sugar donor. Furthermore, we identified and functionally characterized a cDNA encoding another UGT, UGT94F1, as the anthocyanin 3-O-glucoside-2''-O-glucosyltransferase (A3Glc2''GlcT), according to the structural similarity to sugar-sugar glycosyltransferases classified to the cluster IV of flavonoid UGTs. Preferential expression of UGT88D8 and UGT94F1 genes in the petals supports the idea that these UGTs play an important role in the biosynthesis of key flavonoids responsible for the development of the blue color of V. persica flowers.

Red (anthocyanic) leaf margins do not correspond to increased phenolic content in New Zealand Veronica spp.

BACKGROUND AND AIMS: Red or purple coloration of leaf margins is common in angiosperms, and is found in approx. 25 % of New Zealand Veronica species. However, the functional significance of margin coloration is unknown. We hypothesized that anthocyanins in leaf margins correspond with increased phenolic content in leaf margins and/or the leaf entire, signalling low palatability or leaf quality to edge-feeding insects. METHODS: Five species of Veronica with red leaf margins, and six species without, were examined in a common garden. Phenolic content in leaf margins and interior lamina regions of juvenile and fully expanded leaves was quantified using the Folin-Ciocalteu assay. Proportions of leaf margins eaten and average lengths of continuous bites were used as a proxy for palatability. KEY RESULTS: Phenolic content was consistently higher in leaf margins compared with leaf interiors in all species; however, neither leaf margins nor more interior tissues differed significantly in phenolic content with respects to margin colour. Mean phenolic content was inversely correlated with the mean length of continuous bites, suggesting effective deterrence of grazing. However, there was no difference in herbivore consumption of red and green margins, and the plant species with the longest continuous grazing patterns were both red-margined. CONCLUSIONS: Red margin coloration was not an accurate indicator of total phenolic content in leaf margins or interior lamina tissue in New Zealand Veronica. Red coloration was also ineffective in deterring herbivory on the leaf margin, though studies controlling for variations in leaf structure and biochemistry (e.g. intra-specific studies) are needed before more precise conclusions can be drawn. It is also recommended that future studies focus on the relationship between anthocyanin and specific defence compounds (rather than general phenolic pools), and evaluate possible alternative functions of red margins in leaves (e.g. antioxidants, osmotic adjustment).

Physiological and biochemical stress responses in grassland species are influenced by both early-season ozone exposure and interspecific competition.

The effects of two-year early season ozone exposure on physiological and biochemical stress response were investigated in model plant communities. Achillea millefolium and Veronica chamaedrys target plants were grown in monocultures and in mixed cultures with Poa pratensis (phytometer) and exposed in open-top chambers over two years for five weeks to charcoal-filtered (CF) air plus 25 nl l(-1) O3 (control) and non-filtered (NF) air plus 50 nl l(-1) O3. Significant O3 effects were detected in different physiological and biochemical parameters, evidencing interspecific differences in metabolic stress responses and a strong influence of the competition factor. O3 induced strong oxidative effects in Achillea irrespective to the different growth modality. Veronica showed less O3-induced effects in monoculture than when grown in competition with the phytometer. Poa exhibited a different behaviour against O3 depending on the species in competition, showing an overall higher sensitivity to O3 when in mixture with Achillea.