Species-specific secondary metabolites from Primula veris subsp. veris obtained In Vitro adventitious root cultures: an alternative for sustainable production

Primula veris subsp. veris L. is a perennial herbaceous and medicinal plant species the roots and flowers of which are a source of valuable pharmaceutical raw materials. The plant tissues are used to produce expectorant and diuretic drugs due to their high content of triterpene saponins and phenolic glycosides. Underground roots of P. veris can be obtained only through a destructive process during the plant’s harvesting. In the present study, an in vitro adventitious root production protocol was developed as an alternative way of production, focused on four species-specific secondary metabolites. Root explants were cultured in Murashing & Skoog liquid medium supplemented with 5.4 µM α-naphthaleneacetic acid, 0.5 µM kinetin, L-proline 100 mg/L, and 30 g/L sucrose, in the dark and under agitation. The effect of temperature (10, 15 and 22 ◦C) on biomass production was investigated. The content of two flavonoid compounds (primeverin and primulaverin), and two main triterpene saponins (primulic acid I and II) were determined after 60 days of culture and compared with 1.5-year-old soil-grown plants. The accumulated content (mg/g DW) of bioactive compounds of in vitro adventitious roots cultured under 22 ◦C was significantly higher than the other two temperatures of the study, being 9.71 mg/g DW in primulaverin, 0.09 mg/g DW in primeverin, 6.09 mg/g DW in primulic acid I, and 0.51 mg/g DW in primulic acid II. Compared to the soil-grown roots (10.23 mg/g DW primulaverin, 0.28 mg/g DW primeverin, 17.01 mg/g DW primulic acid I, 0.09 mg/g DW primulic acid II), the in vitro grown roots at 22 ◦C exhibited a 5.67-fold higher content in primulic acid II. However, primulic acid I and primeverin content were approximately three-fold higher in soil-grown roots, while primulaverin content were at similar levels for both in vitro at 22 ◦C and soil-grown roots. From our results, tissue culture of P. veris subsp. veris could serve not only for propagation but also for production of species-specific secondary metabolites such as primulic acid II through adventitious root cultures. This would therefore limit the uncontrolled collection of this plant from its natural environment and provide natural products free from pesticides in a sustainable wa

Foliar calcium effects on quality and primary and secondary metabolites of white-fleshed ‘Lemonato’ peaches

‘Lemonato’ is a Greek peach melting-flesh white-flesh cultivar with high nutritional value highly appreciated by the consumers. This study aimed to evaluate the effect of pre-harvest foliar calcium application on fruit quality, primary metabolite profile, antioxidant activity, total phenolic content, and phenolic profile of the ‘Lemonato’ peach, clone ‘Stamatis’. The experiment was conducted for two years, 2019 and 2020, in two commercial orchards at Kato Lehonia and Agios Vlasios regions, central Greece, where the ‘Lemonato’ clone ‘Stamatis’ is traditionally cultivated. The treatments were organic calcium (Ca), calcium–silicate in nanoparticles (Ca–Si), and calcium chloride (CaCl2). Foliar application of the different Ca formulations, commonly used as a horticultural practice, were not effective at improving the fruit quality characteristics in this clone, which is characterized by fruit softening during ripening. The study revealed the sugars and organic acid composition and phenolic profile of the ‘Lemonato’ peach, clone ‘Stamatis’. Peach fruit quality, primary metabolites, and phenolic compounds of the two orchards showed a different response to organic Ca and Ca–Si, indicating that genetic or environmental factors may also be involved. A higher concentration of organic Ca and CaCl2 increased the peach fruit phenolic compounds content and the total antioxidant activity, improving the fruit nutritional qualit

Chemical and Biological Characterization of the Anticancer Potency of \u3ci\u3eSalvia fruticosa\u3c/i\u3e in a Model of Human Malignant Melanoma

Malignant melanoma is one of the most aggressive types of skin cancer with an increasing incidence worldwide. Thus, the development of innovative therapeutic approaches is of great importance. Salvia fruticosa (SF) is known for its anticancer properties and in this context, we aimed to investigate its potential anti-melanoma activity in an in vitro model of human malignant melanoma. Cytotoxicity was assessed through a colorimetric-based sulforhodamine-B (SRB) assay in primary malignant melanoma (A375), non-malignant melanoma epidermoid carcinoma (A431) and non-tumorigenic melanocyte neighbouring keratinocyte (HaCaT) cells. Among eight (8) different fractions of S. fruticosa extracts (SF1-SF8) tested, SF3 was found to possess significant cytotoxic activity against A375 cells, while A431 and HaCaT cells remained relatively resistant or exerted no cytotoxicity, respectively. In addition, the total phenolic (Folin–Ciocalteu assay) and total flavonoid content of SF extracts was estimated, whereas the antioxidant capacity was measured via the inhibition of tert-butyl hydroperoxide-induced lipid peroxidation and protein oxidation levels. Finally, apoptotic cell death was assessed by utilizing a commercially available kit for the activation of caspases - 3, - 8 and - 9. In conclusion, the anti-melanoma properties of SF3 involve the induction of both extrinsic and intrinsic apoptotic pathway(s), as evidenced by the increased activity levels of caspases - 8, and - 9, respectively

Identification and characterization of genetic structures coding for carbapenemases in enterobacteria from Central Greece

The dissemination of carbapenemases among different species of Enterobacteriaceae was investigated in the University Hospital of Larissa, Central Greece. The presence of the isoform (Tn4401a) of the transponson carrying bla(KPC-2) and 5 divergent bla(VIM)-carrying class I integrons, including a novel structure, suggests interspecies transfer of these mobile elements and underscores their ongoing evolution. (C) 2015 Elsevier Inc. All rights reserved

Characterization of a novel lsa(E)- and lnu(B)-carrying structure located in the chromosome of a Staphylococcus aureus sequence type 398 strain

[No abstract available

Characterization of a Novel lsa (E)- and lnu (B)-Carrying Structure Located in the Chromosome of a Staphylococcus aureus Sequence Type 398 Strain : Letter to the editor

Microbial Biotechnolog

Slime-producing staphylococci as causal agents of subclinical mastitis in sheep

Hitherto, research work in slime production from staphylococcal strains of mastitis origin has focused in laboratory properties of these organisms. Objective of present work was to study subclinical mastitis in sheep, caused specifically by slime-producing staphylococci: to investigate its frequency and to identify potential factors playing a role therein. Slime production was evaluated in 708 staphylococcal isolates recovered from cases of subclinical mastitis in a field study in 2198 ewes performed in an extensive countrywide field investigation across Greece. Isolates were studied by means of microbiological and molecular methods. Of these strains, 262 were characterised as slime-producing, 227 as weak slime-producing and 219 as non slime-producing. Most frequently detected genes were eno and icaB; Staphylococcus aureus possessed more genes than coagulase-negative strains; greater number of genes was detected in slime-producing than in weak slime-producing or non-slime-producing strains. Subclinical mastitis caused specifically by slime-producing staphylococci was detected in 337 ewes: prevalence in population sampled was 0.153. A multivariable mixed-effects model revealed that milking mode (highest prevalence in hand-milked flocks) and flock management system (highest prevalence in semi-intensive flocks) were the two factors associated with increased prevalence of mastitis in flocks. The results confirmed the significance of slime producing staphylococcal strains of importance in the aetiology of subclinical mastitis of sheep. Hand-milking was identified as the most important factor predisposing to that infection. © 2018 Elsevier B.V