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

Metabolite profiling and antioxidative activity of Sage (Salvia fruticosa Mill.) under the influence of genotype and harvesting period

Two cultivated accessions of Salvia fruticosa Mill. were investigated and evaluated for their essential oil, phenolic composition and antioxidant activity, during different harvesting time. The essential oil and its major compound 1.8 cineole, presented their higher yields during the early summer harvesting. The advanced analytical LC–MS/MS method applied in this work led to the identification of thirty five compounds with rosmarinic acid, the diterpene artefact carnosol and several flavones and flavonols as the main phenolic constituents, the concentration of which varied largely from spring to autumn. The antioxidant activity of respective methanolic extracts was determined using the Ferric Reducing Ability of Plasma (FRAP), 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assays, as a quality control tool. High positive correlations were observed between FRAP and ABTS antioxidant activities and total phenolic/flavonoid content, and particular phenolic constituent

Sweet cherry fruit cracking: follow-up testing methods and cultivar-metabolic screening

Background Rain-induced fruit cracking is a major physiological problem in most sweet cherry cultivars. For an in vivo cracking assay, the ‘Christensen method’ (cracking evaluation following fruit immersion in water) is commonly used; however, this test does not adequately simulate environmental conditions. Herein, we have designed and evaluated a cracking protocol, named ‘Waterfall method’, in which fruits are continuously wetted under controlled conditions. Results The application of this method alone, or in combination with ‘Christensen method, was shown to be a reliable approach to characterize sweet cherry cracking behavior. Seventeen cherry cultivars were tested for their cracking behavior using both protocols, and primary as well as secondary metabolites identification was performed in skin tissue using a combined GC–MS and UPLC-MS/MS platform. Significant variations of some of the detected metabolites were discovered and important cracking index–metabolite correlations were identified. Conclusions We have established an alternative/complementary method of cherry cracking characterization alongside to Christiansen assay

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

Emergence of sequence type 11 Klebsiella pneumoniae coproducing NDM-1 and VIM-1 metallo-β-lactamases in a Greek hospital

Sequence type 11 Klebsiella pneumoniae, coproducing NDM-1 and VIM-1 metallo-β-lactamases, were isolated in a Greek hospital. blaNDM-1 was part of a Tn125 derivative, located on an ~90-kb plasmid similar to the NDM-1-encoding plasmid pB-3002cz. blaVIM-1 was located in an In-e541-like integron, carried on a multireplicon (IncA/C and IncR) plasmid of ~180 kb. © 201

Characterisation of a ST100 Staphylococcus epidermidis producing an LnuB nucleotidyltransferase: Evidence for interspecies spread of an lnuB-carrying transposon

[No abstract available

Antimicrobial Agent Susceptibility and Typing of Staphylococcal Isolates from Subclinical Mastitis in Ewes

Objective was to study susceptibility to antimicrobial agents of 142 staphylococcal isolates from subclinical mastitis in ewes. In total, 41.5% of these were resistant and 5.6% multidrug resistant. More coagulase-negative staphylococci (47.0%) were resistant than Staphylococcus aureus (18.5%) isolates. Resistance was greater to penicillin (22.5%), tetracycline, or ampicillin (18.3%). More biofilm-forming (20.6%) isolates were resistant to tetracycline than nonbiofilm-forming (0.0%) ones. Presence of tetK was associated with presence of icaA in the same strains. Further, 76.6% of resistant isolates versus 57.7% of susceptible ones were recovered immediately postpartum and 23.4% of resistant isolates versus 9.9% of susceptible ones were recovered in farms that practiced routine administration of antimicrobial agents at the end of a lactation period. Most S. aureus (59.3%) were classified in ST133 and most Staphylococcus epidermidis were classified in ST100, ST142, or ST152 (19.0% each). There was no association of sequence types with resistance. Whole genome sequencing showed that, in a Staphylococcus lentus strain, the ermB gene was part of transposon Tn917 integrated into the chromosome; also, a small plasmid was observed in an ermC-carrying Staphylococcus hominis strain and, finally, in an S. aureus and an S. epidermidis strains, small tetK-carrying plasmids (pSau-2716Lar, pSau-3893Lar) of 4.439 kb were found. © Copyright 2019, Mary Ann Liebert, Inc., publishers 2019