Asperula calabra (Rubiaceae) and allied taxa in southern Apennines,Italy

The present work is aimed at studying several Asperula (Rubiaceae) sect. Cynanchicae populations in southern Apennines, Italy, with particular reference to those referred to A. calabra, by employing biometrical methods on macromorphological data. Among other historical misapplications, A. cynanchica subsp. cynanchica seems to be very rare or missing in southern Italy. The enigmatic Asperula calabra, confirmed to be limited to a single mountain population in Calabria, appears to be strictly related to A. aristata subsp. scabra, so we suggest to treat it as a further subspecies of A. aristata. This latter species is otherwise distributed throughout southern Italy with subsp. scabra and subsp. aristata, characterised by slight morphological differences, which are correlated to altitudinal ranges.     

Seed germination in the Sicilian subspecies of Dianthus rupicola Biv. (Caryophyllaceae)

Abstract

Five populations belonging to three subspecies of Dianthus rupicola Biv. (D. rupicola subsp. rupicola, D. rupicola subsp. aeolicus, and D. rupicola subsp. lopadusanus) and growing in different geographical areas of Sicily were tested for seed germination at various temperatures. All populations showed high germination rates with an optimum temperature between 15°C and 25°C. Efficient seed germination might contribute significantly to the preservation of these subspecies which are currently exposed to several environmental threats.     

Prenylated flavonoids and total extracts from Morus nigra L. root bark inhibit in vitro growth of plant pathogenic fungi

Total extracts and kuwanon G from Morus nigra root bark showed antifungal activity against several phytopathogenic fungi, with minimal inhibitory concentration (MIC₅₀) ranging from 32 to 128 μg/ml and from 16 to 64 μg/ml, respectively. Acetonic extracts inhibited 60% B. cinerea biofilm formation at concentration of 128 μg/ml.     

Molecular identification of root fungal associates in Orchis pauciflora Tenore

Abstract

The terrestrial orchid, Orchis pauciflora Ten., growing in poor grassland and garrigue of Central Mediterranean region, is local and rare and has been included in the red lists of several Italian regions. We investigated the diversity of fungal associates in O. pauciflora adult plants collected in two protected areas of Tuscany (Central Italy). Genomic DNA was extracted from mycorrhizal roots of 12 orchid plants and the fungal ITS were amplified and sequenced. Several fungal associates, belonging to different taxa of basidiomycetes (Tulasnellaceae) and ascomycetes such as Leptodontidium, Exophiala and Phialophora species, were recovered. The trophic role of these fungi and their impact on O. pauciflora growth and conservation are discussed.     

Submediterranean dry grasslands along the Tyrrhenian sector of central Italy: Synecology,syndynamics and syntaxonomy

Abstract

Semi-natural dry grasslands are habitats of high conservation concern. These ecosystems have not been extensively explored in the Tyrrhenian sector of the Italian peninsula, particularly in the Submediterranean climatic region. In order to address this issue and to define the synecology, syndynamics and syntaxonomy of calcareous grasslands in this area, we considered 127 phytosociological relevés. Our sampling was performed in the Lazio region according to a stratified sampling scheme based on homogeneous land units, defined by means of an ecological land classification process. We analyzed the vegetation data using multivariate methods. Two new associations, whose typical aspects occur in the mesotemperate phytoclimatic belt, were identified:Erysimo pseudorhaetici-Dasypyretum villosi (Taenianthero-Aegilopion) and Scorpiuro muricati-Brometum erecti (Phleo-Bromion). Phytoclimatic belts within the Submediterranean region cause a significant degree of differentiation within Scorpiuro-Brometum, which was described at the subassociation and variant levels. The overall relevance of therophytes represents the most important feature distinguishing Scorpiuro-Brometum from the published Bromus erectus associations. At a finer scale, the presence of the two physiognomically different grassland communities is related to different soil types. All these communities are dynamically linked to Quercus virgiliana and Q. pubescens woods, and contribute to the coenological differentiation of the vegetation series related to such woodlands.     

Chemical composition of the essential oil of Centaurea grinensis Reuter and Centaurea apiculata Ledeb: Growing wild in Croatia and Bulgaria,respectively and PCA analysis of subgenus Lopholoma (Cass.) Dobrocz

In the present study, the chemical composition of the essential oils from aerial parts of Centaurea grinensis Reuter (K) collected at ?uta Lova, near Senj (Croatia), and aerial parts (L1) and flowers (L2) of C. apiculata Ledeb. collected near Rebro village, near Breznik town, Znepole Region (Bulgaria), both belonging to subgenus Lopholoma (Cass.) Dobrocz, was evaluated by GC and GC-MS. The main components of K were 4-vinyl guaiacol (21.5%), hexadecanoic acid (16.2%), acetophenone (12.5%). Caryophyllene oxide (15.8%) together with sphathulenol (14.5%) and humulene epoxide II (9.4%) were recognized as the main constituent of both the aerial part (L1) and flowers (L2) of C. apiculata. Furthermore, a complete review on the composition of the essential oils of all the Centaurea taxa belonging to subgenus Lopholoma studied so far has been inserted and cluster analysis (PCA) was carried out.     

Ecomorphology of the giant bear-dogs Amphicyon and Ischyrocyon

Giant bear-dogs of the genera Amphicyon and Ischyrocyon (Carnivora, Amphicyonidae, Amphicyoninae) were the largest carnivorans in North America during middle and late Miocene (17.5–8.8 Mya) with a dental and skeletal morphology that combined features found in living Ursidae, Canidae, and Felidae. This study tests previously proposed models of diet and hunting behaviour of these extinct carnivorans. Relative grinding area (RGA) of lower molars and wear pattern on upper molars suggest that bear-dogs were carnivorous. Amphicyon and Ischyrocyon possessed skeletal features of both ambush (short distal limb segments) and pursuit (caudally bent olecranon process of ulna) living predators. Therefore, bear-dogs probably pursued their prey (mediportal ungulates) for a longer distance but at a slower speed than do living ambush predators. Upon catching up to its prey a bear-dog probably seized it with powerfully muscled forelimbs and killed it by tearing into its ribcage or neck with canines set in a narrow rostrum.     

Phytochemical analysis and antibacterial activity towards methicillin-resistant Staphylococcus aureus of leaf extracts from Argania spinosa (L.) Skeels

Argania spinosa (L.) Skeels is an endemic Moroccan species belonging to Sapotaceae family. In this work, lipophilic and aqueous extracts were obtained from leaves and subjected to a chemical profiling by MS and LC-MS/MS. Pentacyclic terpenoids were identified and quantified in the lipophilic fraction, while phenolic compounds (mainly belonging to flavonols and flavan-3-ols) were identified in the aqueous fraction. The antibacterial activities of fractions were evaluated in vitro against both reference Gram-positive and -negative bacterial strains and clinical isolates of methicillin-sensitive and methicillin-resistant Staphylococcus aureus (MSSA and MRSA); in addition, the compounds quantified as main components in each extract were assayed against reference strains. A relevant antibacterial activity was observed against reference MSSA and MRSA strains of S. aureus: for the lipophilic fraction, MIC₅₀ values obtained were 177.8 and 170.6 μg/mL for the former and the latter, respectively, while for the aqueous fraction were 215.5 and 233.3 μg/mL. These inhibitory activities could be mainly ascribed to ursolic and oleanolic acids, among pentacyclic terpenoids, and to quercetin concerning phenolic compounds. A remarkable antibacterial activity was also observed against clinical isolates, thus argan leaves can be considered of interest in the chemotherapy of human infections.     

Morphological analysis,chorology and ecology of Juniperus navicularis Gand. in the Iberian Peninsula

Juniperus navicularis is an endemic taxon of Iberian Peninsula that develops on paleodunes of coastal ecosystems and whose distribution range is not well defined. In this paper, we present the results of the morphological analysis in populations attributed to J. navicularis of southern Spain. In order to assess the similarity between Portuguese and Spanish populations, the Mann–Whitney U test was performed, the same test was also applied between this taxon and Juniperus oxycedrus subsp. oxycedrus. In order to distinguish this species from other taxa of the Juniperus oxycedrus group (subsp. oxycedrus, subsp. badia, subsp. macrocarpa) in the southwestern Mediterranean region, a principal component analysis has been carried out. New data about the chorology and ecology of Spanish populations considered as a priority habitat in the Habitat Directive as well as number of individuals and conservation status are provided.     

Biomass and nutrient dynamics in Mediterranean seasonal dimorphic shrubs: Strategies to face environmental constraints

Abstract

To analyze biomass and nutrient dynamics of Mediterranean seasonal dimorphic shrubs, aboveground biomass (AGB) and belowground biomass (BGB), litterfall (LF), aboveground net annual primary production (ANPP), and nutrient allocation and turnover were estimated in Cistus salviifolius and C. ladanifer, during two years, in southern Portugal. AGB, BGB, LF, and ANPP of C. salviifolius are within the range reported for other seasonal dimorphic plants, while those of C. ladanifer are closer to evergreen sclerophylls. Leaf renewal was higher for C. salviifolius than for C. ladanifer, especially for winter leaves, while root-to-shoot ratio and accumulation of live wood was greater in the latter. The concentrations of N, K, Ca, and Mg were lower in leaves of C. ladanifer than in those of C. salviifolius, suggesting less nutritive requirements for the former. Moreover, leaf nutrient translocation was higher in C. ladanifer than in C. salviifolius. Therefore, C. ladanifer shrubs seem to be more efficient with respect to carbon uptake, even during severe drought, while those of C. salviifolius act as weaker sinks more susceptible to the negative effects of drought. Additionally, C. ladanifer seems to have a competitive advantage in more arid conditions, as a great proportion of its annual nutrient requirement may derive from internal sources.     

Anti‐quorum sensing activity of Psidium guajava L. flavonoids against Chromobacterium violaceum and Pseudomonas aeruginosa PAO1

Psidium guajava L., which has been used traditionally as a medicinal plant, was explored for anti‐quorum sensing (QS) activity. The anti‐QS activity of the flavonoid (FL) fraction of P. guajava leaves was determined using a biosensor bioassay with Chromobacterium violaceum CV026. Detailed investigation of the effects of the FL‐fraction on QS‐regulated violacein production in C. violaceum ATCC12472 and pyocyanin production, proteolytic, elastolytic activities, swarming motility and biofilm formation in Pseudomonas aeruginosa PAO1 was performed using standard methods. Possible mechanisms of QS‐inhibition were studied by assessing violacein production in response to N‐acyl homoserine lactone (AHL) synthesis in the presence of the FL‐fraction in C. violaceum ATCC31532 and by evaluating the induction of violacein in the mutant C. violaceum CV026 by AHL extracted from the culture supernatants of C. violaceum 31532. Active compounds in the FL‐fraction were identified by liquid chromatography–mass spectrometry (LC–MS). Inhibition of violacein production by the FL‐fraction in a C. violaceum CV026 biosensor bioassay indicated possible anti‐QS activity. The FL‐fraction showed concentration‐dependent decreases in violacein production in C. violaceum 12472 and inhibited pyocyanin production, proteolytic and elastolytic activities, swarming motility and biofilm formation in P. aeruginosa PAO1. Interestingly, the FL‐fraction did not inhibit AHL synthesis; AHL extracted from cultures of C. violaceum 31532 grown in the presence of the FL‐fraction induced violacein in the mutant C. violaceum CV026. LC–MS analysis revealed the presence of quercetin and quercetin‐3‐O‐arabinoside in the FL‐fraction. Both quercetin and quercetin‐3‐O‐arabinoside inhibited violacein production in C. violaceum 12472, at 50 and 100 μg/mL, respectively. Results of this study provide scope for further research to exploit these active molecules as anti‐QS agents.     

Micropropagation of Aristolochia rotunda L.

An original protocol of micropropagation of Aristolochia rotunda L., an important herb for the survival of an endangered butterfly was developed. The cytokinin 6-benzylaminopurine affected the production of new shoots but had no effect on shoot length. The addition of indole-3-butyric acid (IBA) had a negative effect on shoot length. Best results in terms of rooting percentage and root length were achieved with 1.5 μM IBA. The protocol can be employed to enhance the number of A. rotunda in the environment.     

Diurnal variations in physiology and behaviour of the noble crayfish Astacus astacus and the signal crayfish Pacifastacus leniusculus

Heart rate, locomotor activity, and oxygen consumption were recorded simultaneously and continuously in seven individuals of the noble crayfish Astacus astacus (Linneus 1758) and seven individuals of the signal crayfish Pacifastacus leniusculus (Dana 1852). The recordings were made in the laboratory over 7 days at 15°C under a 12?:?12?h dark?:?light regime. Circadian rhythms in heart rate, locomotor activity and oxygen consumption were found both in A. astacus and P. leniusculus. Increased heart rate, locomotor activity, and oxygen consumption levels during night time in both A. astacus and P. leniusculus illustrated expression of nocturnal behaviour. No differences in oxygen consumption levels were observed between A. astacus and P. leniusculus. Also, no significant difference between heart rate levels or heart rate variances was found in A. astacus and P. leniusculus at night. During day, however, heart rate levels, heart rate variances and locomotor activity were higher in P. leniusculus than in A. astacus. The higher activity level in P. leniusculus than in A. astacus during daytime indicates that P. leniusculus is less strictly nocturnal than is A. astacus.     

A new Centaurea L. (Asteraceae) species from Turkey

A new Centaurea L. (Asteraceae) species from Turkey is described and illustrated. Centaurea mersinensis Uysal and Hamzao?lu exists on calcareous slopes in Pinus brutia forests of Ayd?nc?k (Mersin) in southern Anatolia. It belongs to C. sect. Phalolepis (Cass.) DC., and taxonomically its closest relative is C. lycaonica. Diagnostic morphological characters from very similar taxa are provided, and a key is provided that includes related species of sect. Phalolepis from Turkey. The geographical distribution of the new species and relatives of the same section are mapped. The chromosome number of C. mersinensis, 2n = 18, counted in root tips, is also reported and illustrated.     

α-cycloPNA: 1-Aminocylopentane-1-carboxylic Acid-Derived Peptide Nucleic Acid

Abstract

All four diastereoisomers of 3-thymine-1-(^tbutoxycarbonyl)aminocyclopentane- 1-carboxylic acid have been synthesised from (S)-dimethyl malate and thymine monomer 12 has been incorporated into an α-cycloPNA oligomer.     

A field experiment on the use of Pistacia lentiscus L. and Scrophularia canina L. subsp. bicolor (Sibth. et Sm.) Greuter for the phytoremediation of abandoned mining areas

Abstract

A two-year study has been conducted in an abandoned Pb/Zn mining site, with the aim of investigating the feasibility of phytoremediation using two native Mediterranean plants (Pistacia lentiscus and Scrophularia bicolor) and of assessing the performance of amendments able to reduce the toxic effects of heavy metals. The amendments used were compost, chemical fertilizer, and zeolites, used singly or in combination. Depending on the amendments applied, the two species showed different mortality rates in the different plots, but all produced an increase in P. lentiscus survival, while S. bicolor survival improved only when amended with zeolite or zeolite + fertilizer. Scrophularia bicolor proved to be a more efficient accumulator than P. lentiscus, especially for Pb uptake. Pistacia lentiscus accumulated metals mostly in the roots. The effect of amendments was to generally reduce the bioavailable metal fraction, especially lead, in the plots amended with compost. Pistacia lentiscus proved to be the most suitable species for phytostabilization and environmental restoration, both for its resistance to metals and high phytomass production. The experiments demonstrate that the use of compost not only encourages this kind of revegetation in degraded areas, but is also an economical option that uses a by-product of solid municipal waste treatment.     

Genetic and morphological variability analysis revealed a complex network in South-Eastern Sicilian Helichrysum occurrences

The Helichrysum occurrences spread over Hyblaean area (Sicily) are characterized by an unusual richness of morphotype variants which ancestral origin is still unclear. Morphological and genetic variability of seven populations attributable to six taxonomic entities (Helichrysum archimedeum, H. stoechas subsp. conglobatum, H. stoechas subsp. barrelieri, H. hyblaeum, H. pendulum and H. nebrodense) was examined, in order to highlight their relationships within the Helichrysum sect. Stoechadina. Morphological analysis showed a clear isolation of populations referred to H. pendulum, H. nebrodense and H. stoechas subsp. conglobatum. Whereas, genetic structure profiling, with 21 ISSRs, displayed a strong genetic relationship within H. stoechas and H. archimedeum accessions, where the latter exhibited evident genetic similarity with H. hyblaeum. Analysis of rbcL cpDNA fragment allowed to identify a shared haplotype between H. stoechas and H. rupestre species, while, through the construction of haplotype network based on trnL-F/rpl32 region, the sampled accessions were properly differentiated as confirmed by the cpDNA consensus network. These results confirmed the high affinity of H. archimedeum with other H. stoechas populations and the closer relationship of H. hyblaeum to H. stoechas respect to H. rupestre.     

Effect of Biomass Pre-Treatment and Solvent Extraction on β-Carotene and Lycopene Recovery from Blakeslea trispora Cells

Abstract

The production of carotenoids from Blakeslea trispora cells in a synthetic medium has been reported, with the main products being β-carotene, lycopene, and γ-carotene. The effect of biomass pretreatment and solvent extraction on their selective recovery is reported here. Eight solvents of class II and III of the International Conference of Harmonization: ethanol, methanol, acetone, 2-propanol, pentane, hexane, ethyl acetate, and ethyl ether, and HPLC analysis were used for the evaluation of their selectivities towards the three main carotenoids with regard to different biomass pre-treatment. The average C_max values (maximum concentration of caronoids in a specific solvent) were estimated to 16 mg/L with the five out of eight solvents investigated, whereas methanol, pentane, and hexane gave lower values of 10, 11, and 9 mg/L, respectively. The highest carotenoid yield was obtained in the case of wet biomass, where 44–56% is recovered with one solvent and three extractions and the rest is recovered only after subsequent treatment with acetone; thus, four extractions of 2.5 h are needed. Two extractions of 54 min are enough to recover carotenoids from dehydrated biomass, with the disadvantage of a high degree of degradation. Our results showed that, for maximum carotenoid recovery, ethyl ether, 2-propanol, and ethanol could be successfully used with biomass without prior treatment, whereas fractions enriched in β-carotene or lycopene can be obtained by extraction with the proper solvent, thus avoiding degradation due to time-consuming processes.