Biodiversity, Ecology and Distribution of Mediterranean Charophytes in Southern Italy.

Charophytes are amongst the most endangered primary producers in freshwater and coastal ecosystems. In spite of the extensive research on the group and its ecological and conservational relevance, scarce information is available on Mediterranean environments, especially rivers and small water reservoirs, where charophytes face challenging summer droughts and changes in hydrological regimes, as well as pervasive anthropogenic pressures. This research aimed, through repeated field observations, detailed analyses of population traits, and extensive characterization of the colonized environments, to foster an understanding of the distribution, biodiversity, and ecology of charophytes in an area of exceptional environmental value and that is still uninvestigated in relation to its charophyte flora, the southern Campania region (Italy). Overall, 17 populations were discovered, belonging to 4 taxa of the Chara genus: C. globularis, C. gymnophylla, C. vulgaris, and C. vulgaris var. papillata, reduced to 12 populations and to the first 3 taxa by the end of the study. The species occupied different ecological niches and colonized environments such as rivers and small ponds, with environment-dependent morphotypes. The occurrence of few taxa with a wide distribution, often forming ephemeral populations, suggests ongoing constraints on charophyte biodiversity in the area, favoring opportunistic species that are able to benefit from temporary refugia.

A multidisciplinary approach to the comparison of three contrasting treatments on both lampenflora community and underlying rock surface.

Removing lampenflora, phototrophic organisms developing on rock surfaces in tourist cavities due to the artificial lighting, is a challenge for sustainable and appropriate long-term management of caves. Photosynthetic-based biofilms usually cause rock biodeterioration and an ecological imbalance in cave ecosystems. In this work, a detailed investigation of the effects of the 3 most commonly used lampenflora cleaning operations (NaClO, H2O2 and UVC) was carried out in Pertosa-Auletta Cave (Italy). The application of NaClO showed good disinfection capability over extended periods of time without causing any appreciable rock deterioration. The H2O2 treatment showed to be corrosive for the rock surfaces covered with vermiculation deposits. The chemical alteration of organic and inorganic compounds by H2O2 did not remove biomass, favoring biofilm recovery after three months of treatment. Both NaClO and H2O2 treatments were effective at removing photoautotrophs, although the bacterial phyla Proteobacteria and Bacteroidetes as well as Apicomplexa and Cercozoa among the Eukaryotes, were found to be resistant to these treatments. The UVC treatments did not show any noticeable effect on the biofilms.

Underground Ecosystem Conservation Through High-resolution Air Monitoring.

In cave ecosystems tourists represent moving sources of discontinuous disturbances, able to induce transient system responses whose knowledge is crucial in defining appropriate conservation measures. Here we propose an approach to evaluate the amplitude and scales of cave alterations based on high-resolution air monitoring, through the use of purposely developed low-cost monitoring stations and a consistent analytical framework for information retrieval based on time series analysis. In particular, monitoring stations adopt a modular structure based on physical computing platforms acquiring data through several sensors, with means of preventing humidity damages and guaranteeing their continuous operation. Data are then analyzed using wavelet periodograms and cross-periodograms to extract the scales of tourism-induced alterations. The approach has been exemplified in the Pertosa-Auletta Cave, one of the most important underground environments in Southern Italy, highlighting the development of monitoring stations and the information obtainable with the proposed analytical workflow. Here, 2 monitoring stations acquiring data for 1 year at 1' sampling time on temperature, relative humidity, CO2, VOCs, and particulate matter were deployed in trails subjected to different levels of tourism. In terms of Pertosa-Auletta Cave air dynamics, the approach allowed estimating the temporal and spatial scales of tourism-induced alterations in the order of minutes and meters, respectively, with parameter-dependent variations. On more general terms, the approach proved reliable and effective, with its modularity and low-cost fostering its straightforward adoption in other underground ecosystems, where it can support the development of tailored management strategies.

Microbial Community Characterizing Vermiculations from Karst Caves and Its Role in Their Formation.

The microbiota associated with vermiculations from karst caves is largely unknown. Vermiculations are enigmatic deposits forming worm-like patterns on cave walls all over the world. They represent a precious focus for geomicrobiological studies aimed at exploring both the microbial life of these ecosystems and the vermiculation genesis. This study comprises the first approach on the microbial communities thriving in Pertosa-Auletta Cave (southern Italy) vermiculations by next-generation sequencing. The most abundant phylum in vermiculations was Proteobacteria, followed by Acidobacteria > Actinobacteria > Nitrospirae > Firmicutes > Planctomycetes > Chloroflexi > Gemmatimonadetes > Bacteroidetes > Latescibacteria. Numerous less-represented taxonomic groups (< 1%), as well as unclassified ones, were also detected. From an ecological point of view, all the groups co-participate in the biogeochemical cycles in these underground environments, mediating oxidation-reduction reactions, promoting host rock dissolution and secondary mineral precipitation, and enriching the matrix in organic matter. Confocal laser scanning microscopy and field emission scanning electron microscopy brought evidence of a strong interaction between the biotic community and the abiotic matrix, supporting the role of microbial communities in the formation process of vermiculations.

Acute effects of PAH contamination on microbial community of different forest soils.

Polycyclic aromatic hydrocarbons (PAHs) are hazardous organic compounds with mutagenic, genotoxic and carcinogenic properties. Although PAHs in soil can cause toxicity to microorganisms, the microbial community is able to degrade these compounds. For this reason, it is important to study acute and short-term effects of PAH contamination on soil microbial community, also to shed light on its possible exploitation in soil restoration. The effects of acute PAH contamination on the structure and metabolic activity of microbial communities in three forest (beech, holm oak, black pine) soils were studied. The soils were spiked with phenanthrene, pyrene or benzo[a]pyrene and incubated in experimental mesocosms, under controlled conditions. Enzymatic activities (laccase, total peroxidase and hydrolase), as well as microbial biomass and community structure (through phospholipid fatty acid and ergosterol analyses), were evaluated in the three soil systems 4 days after contamination and compared to no-spiked soils. In soil under holm oak, there was a stimulation of Gram+ bacteria after contamination with all the 3 PAHs, whereas in soil under pine, pyrene and phenanthrene additions mainly stimulated fungi and actinomycetes.

Multivariate spatial analysis for the identification of criticalities and of the subtended causes in river ecosystems.

In statistics, the identification of environmental criticalities, one of the primary goals of environmental monitoring and management, translates into the detection of spatial outliers. Detected in relation to purposely defined sets of indicators, both global and local outliers are pivotal in the identification not only of the severity and spread of criticalities, but also of their nature and causes. The present research exemplifies a procedural framework to identify environmental criticalities, using two different approaches for the detection of spatial outliers in river ecosystems related to several sets of parameters (organic C, inorganic C, Ca, Co, Cr, Fe, K, Mg, Mn, N, Na, P, S, Si, V, Zn, Cl-, F-, NO3-, SO42-, chlorophyll a, chlorophyll b, pheophytin a, pheophytin b, total carotenoids, pH, and electrical conductivity), including emerging contaminants. To this end, indicator sets diagnostic for specific criticalities, derived from an empirical dataset of water quality parameters, were employed, using detection techniques based on geographically weighted principal component analysis and a modified pairwise Mahalanobis distance-based algorithm. Clear and accurate criticality scenarios were derived, highlighting both the strengths and the limitations of the proposed approach, especially in relation to the classic threshold-based methods.

Low copper availability limits Helicobacter infection in mice.

Copper is an essential element for all living organisms; however, it becomes toxic at high concentrations due to its ability to participate in many redox reactions. This vital micronutrient balance plays an important role in the battle between host and pathogen, due to its use by the host to intoxicate pathogens. In this study, we explore the effects of copper deprivation on Helicobacter infection in mice using the copper chelator tetrathiomolybdate. Our results reveal that Helicobacter infection significantly reduces copper concentration in mice stomachs without affecting its circulating levels. Moreover, in copper-deprived mice, bacteria hardly colonize the epithelium and mice show less gastric damage in comparison with the infected ones. However, when the copper chelator is administered after infection, the condition of the mouse stomachs declines. This could be explained by the lower copper availability in tetrathiomolybdate-treated mice, which would reduce macrophages' action against the pathogen. In this scenario, we observe that the protective factor trefoil factor 1 is induced upon copper-deprived conditions, probably contributing to the inefficacy of infection, whereas, when the chelator is administered after infection, the gene is already silenced by bacteria and cannot be restored. In conclusion, our data suggest that Helicobacter takes advantage of gastric copper reducing its availability for the host and that copper levels have an impact on the outcome of infection.

Role of different microorganisms in remediating PAH-contaminated soils treated with compost or fungi.

Microbial degradation is the main responsible for polycyclic aromatic hydrocarbons (PAHs) removal from contaminated soils, and the understanding of this process is pivotal to define effective bioremediation approaches. To evaluate the contribution of several microbial groups in soil anthracene and benzo[a]pyrene degradation, the analysis of phospholipid fatty acid (PLFA) profiles and machine learning techniques were employed. To this end, PLFAs and PAH concentrations were analysed, along 274 days of incubation in mesocosms, in soils artificially contaminated with anthracene and benzo[a]pyrene, subjected to different treatments: untreated soil and soils treated with biowaste compost or fungal consortium. Random forest models, figuring anthracene or benzo[a]pyrene concentrations as dependent variables and PLFAs as predictors, were then built to evaluate the contribution of each variable in PAH degradation. PLFA profiles varied substantially among soil treatments and along time, with the increase of Actinomycetes in soils added with fungi and other Gram+ bacteria in compost amended soils. The former, together with fungi, are primarily responsible for anthracene and benzo[a]pyrene degradation in both treated soils, a process in which also metanotrophs and other Gram+ and Gram- bacteria participate. In untreated soil, the cooperation of a multitude of different microorganisms was, instead, responsible for PAH removal, a process with lower efficiency in respect to treated soils.

Geomicrobiology of a seawater-influenced active sulfuric acid cave.

Fetida Cave is an active sulfuric acid cave influenced by seawater, showing abundant microbial communities that organize themselves under three main different morphologies: water filaments, vermiculations and moonmilk deposits. These biofilms/deposits have different cave distribution, pH, macro- and microelement and mineralogical composition, carbon and nitrogen content. In particular, water filaments and vermiculations had circumneutral and slightly acidic pH, respectively, both had abundant organic carbon and high microbial diversity. They were rich in macro- and microelements, deriving from mineral dissolution, and, in the case of water filaments, from seawater composition. Vermiculations had different color, partly associated with their mineralogy, and unusual minerals probably due to trapping capacities. Moonmilk was composed of gypsum, poor in organic matter, had an extremely low pH (0-1) and low microbial diversity. Based on 16S rRNA gene analysis, the microbial composition of the biofilms/deposits included autotrophic taxa associated with sulfur and nitrogen cycles and biomineralization processes. In particular, water filaments communities were characterized by bacterial taxa involved in sulfur oxidation and reduction in aquatic, aphotic, microaerophilic/anoxic environments (Campylobacterales, Thiotrichales, Arenicellales, Desulfobacterales, Desulforomonadales) and in chemolithotrophy in marine habitats (Oceanospirillales, Chromatiales). Their biodiversity was linked to the morphology of the water filaments and their collection site. Microbial communities within vermiculations were partly related to their color and showed high abundance of unclassified Betaproteobacteria and sulfur-oxidizing Hydrogenophilales (including Sulfuriferula), and Acidiferrobacterales (including Sulfurifustis), sulfur-reducing Desulfurellales, and ammonia-oxidizing Planctomycetes and Nitrospirae. The microbial community associated with gypsum moonmilk showed the strong dominance (>60%) of the archaeal genus Thermoplasma and lower abundance of chemolithotrophic Acidithiobacillus, metal-oxidizing Metallibacterium, Sulfobacillus, and Acidibacillus. This study describes the geomicrobiology of water filaments, vermiculations and gypsum moonmilk from Fetida Cave, providing insights into the microbial taxa that characterize each morphology and contribute to biogeochemical cycles and speleogenesis of this peculiar seawater-influenced sulfuric acid cave.

Nutrients and non-essential elements in edible crops following long-term mineral and compost fertilization of a Mediterranean agricultural soil.

The effects of long-term soil fertilizations on nutrient and non-essential element concentrations in edible parts of three crops important in human diet were investigated repeating four treatments (biowaste compost, biowaste compost plus mineral nitrogen, mineral NPK, unfertilized control) for seven consecutive years (2007-2014). Fruits of Solanum lycopersicum cv San Marzano collected in 2011 and 2012, bulbs of Allium cepa cv Bianca di Pompei collected in 2012 and 2013, and bulbs of Foeniculum vulgare cv Orbit collected in 2014 were analyzed. Wide variations in element concentrations were observed along time and among species, with Ca, K, Mg, and Na higher in fennel bulbs and Cd, Cr, Mn, Ni, Pb, and Zn higher in tomato fruits, where Cd reached concentrations up to ninefold higher than the permitted values (EU Regulation n. 488/2014). Despite the enrichments in soil total Cu and available Cd, Fe, K, Mn, and Zn concentrations due to long-term fertilization with biowaste compost (alone or with mineral fertilizers), plants showed lower micronutrient and non-essential element concentrations in respect to those on unfertilized soils. Considering the potential toxicity for human beings of these mobile and persistent elements, the obtained findings reassure on the safe use of biowaste compost in agriculture. Overall, this study suggests the use of compost as the most advisable fertilization practice and highlights the need of multiple crops analysis in evaluating the effects of long-term soil fertilization on their chemical composition.

Biomonitoring of nutrient and toxic element concentrations in the Sarno River through aquatic plants.

The Sarno River is considered the most polluted river in Europe and one of the ten most polluted rivers in the world. So far, its quality has been usually evaluated by water and sediment analyses of either inorganic or organic pollutants. However, a biomonitoring approach would be of paramount importance in the evaluation of river quality, since it integrates pollutant temporal fluctuations, as in the case of discontinuous inputs from urban, industrial and agricultural activities. To this end, a passive biomonitoring study of the Sarno River was carried out, using two native aquatic plants accumulators of inorganic pollutants. The spring area was monitored analysing the roots of the semi-submerged Apium nodiflorum, whereas the whole river course was monitored analysing the shoots of the submerged Potamogeton pectinatus. The information on the four macronutrient (Ca, K, Mg, P), the six micronutrient (Cu, Fe, Mn, Na, Ni, Zn) and the four toxic element (Cd, Cr, Pb, V) concentrations were separately combined in the Nemerow Pollution Index. Results evidenced a severe pollution degree of the Sarno River, attributable to toxic elements > micronutrients > macronutrients. In particular, the spring area showed high K concentrations, as well as high concentrations of several micronutrients and toxic elements. A generalized Zn contamination and a progressive macronutrient (above all Ca and P), micronutrient (above all Ni, Cu and Fe) and toxic element (above all Cr and Pb) accumulation toward the mouth was related to pollution from agricultural and urban activities. Industrial sources, especially tanneries along the Solofrana tributary, accounted for high Mn concentrations, whereas the volcanic origin of the substrate accounted for a generalized V contamination.

Antibiotic effects on seed germination and root development of tomato (Solanum lycopersicum L.).

Antibiotics are emerging pollutants released into the environment through wastewater and manure or effluents from livestock plants. Compared to the wide literature on the effects of antibiotics on the development of drug-resistant bacteria and on the adverse effects on animals and human beings, the effects on plants are less investigated. Here we evaluated the effects of four antibiotics (cloramphenicol: CAP, spiramycin: SPR, spectinomycin: SPT, vancomycin: VAN) belonging to different chemical groups, on seed germination and root development of tomato (Solanum lycopersicum L. cv. San Marzano). Specifically, seed germination and root elongation kinetics, as well as the number of mithotic figures in root apical meristem, were studied in relation to different concentrations of each antibiotic (0, 0.1, 1, 10, 100, 1000mgL-1) for 10 and 7 days, respectively. Results showed that seed germination was not affected, but root development (root elongation kinetics and cell division) was impaired at concentrations from 10mgL-1 (SPT) and 100mgL-1 (CAP) to 1000mgL-1 (SPR and VAN).

Anthracene and benzo(a)pyrene degradation in soil is favoured by compost amendment: Perspectives for a bioremediation approach.

In order to validate the use of compost in soil PAH bioremediation, the degradation of anthracene and benzo(a)pyrene was monitored in soils artificially contaminated and incubated in mesocosms under controlled conditions. The dynamics observed in compost amended soil were compared to those observed in soil added with a fungal consortium and untreated soil. At the same time, three microbial enzyme activities usually involved in PAH degradation (laccase, o-diphenol oxidase and peroxidase activities) were monitored. Both PAHs decreased along the time in the three mesocosms, with anthracene, with lower molecular weight, degrading with a higher rate and reaching lower residual values than benzo(a)pyrene. Although at the end of incubation, the residual values of investigated PAHs are similar in the three mesocosm types, PAH dynamics showed a higher degradation rate in the early stage in mesocosms added with the fungal mycelium and amended with compost. Among the three enzyme activities, only peroxidase showed higher values in treated than untreated mesocosms. Considering the ameliorating effects of compost on degraded soils, its use can be suggested in PAH bioremediation.

Heavy metal and polycyclic aromatic hydrocarbon concentrations in Quercus ilex L. leaves fit an a priori subdivision in site typologies based on human management.

Concentrations of four heavy metals (HMs) (Cd, Cr, Fe, Pb) and four polycyclic aromatic hydrocarbons (PAHs) (fluoranthene, phenanthrene, chrysene, benzo[a]pyrene) in Quercus ilex L. leaves collected at the Campania Region (Southern Italy) in previous air biomonitoring studies were employed to (1) test the correspondence with an a priori site subdivision (remote, periurban, and urban) and (2) evaluate long temporal trends of HM (approximately 20 years) and PAH (approximately 10 years) air contaminations. Overall, Q. ilex leaf HM and PAH concentrations resulted along the gradient: remote < periurban < urban sites, reflecting the a priori subdivision based on human management. Over a long time, although a clear decrease of leaf Pb, chrysene, fluoranthene, and phenanthrene concentrations occurred at the urban sites, a high contamination level persists.

Nutrient and toxic element soil concentrations during repeated mineral and compost fertilization treatments in a Mediterranean agricultural soil.

Agricultural soils of semi-arid Mediterranean areas are often subjected to depletion of their chemical, physical, and biological properties. In this context, organic fertilization, in addition to providing nutrients for a longer time in respect to mineral fertilization, improves many other characteristics related to soil fertility. Moreover, the combined use of organic and mineral fertilizers may promote a more sustainable crop production. However, a concern on the long-term use of organic fertilizers arises in relation to the possible accumulation of toxic elements in soil and their transfer to human beings. For this reason, a long-term study on nutrient and toxic element total concentrations and availabilities during fertilization treatments was carried out. In particular, mineral NPK fertilized soils, soils amended with biowaste compost, soils amended with biowaste compost plus mineral nitrogen, and unfertilized soils were analyzed for 11 chemical elements. The results highlighted that temporal variations in total and bioavailable concentrations of both nutrients and toxic elements, occurring also in unfertilized soils, are wider than those related to fertilization treatments. Anyway, soil amendments with biowaste compost, alone or in combination with mineral fertilizers, reduce Cu bioavailability but improve K, Fe, Mn, and Zn availabilities, excluding at the same time a long-term accumulation in soil. Total and bioavailable toxic element concentrations (apart from available Cd) do not vary in relation to fertilization treatments.

Usefulness of different vascular plant species for passive biomonitoring of Mediterranean rivers.

Choosing native vascular plants as nutrient and toxic element accumulators for passive biomonitoring of urban river quality is not an easy task in Mediterranean rivers, due to the particular climate determining high variations in river hydrology. To identify potential biomonitors for this area, the roots of seven species (Angelica sylvestris, Apium nodiflorum, Tradescantia fluminensis, Nasturtium officinale, Persicaria lapathifolia, Arctium lappa, Typha latifolia), growing in seven sites along the River Irno (Southern Italy), were collected in July 2010 and analyzed regarding their capability to accumulate Cd, Cr, Cu, Fe, K, Mg, Mn, Na, Ni, Pb, V, and Zn through atomic absorption spectrometry. Notwithstanding the expected different accumulation degree among the species, they highlighted similar spatial contamination gradients, and all of them appeared suitable, alone or in combination, for river passive biomonitoring. A. nodiflorum, in particular, appeared the best biomonitor for the River Irno, where severe anthropogenic impacts were detected: high Cu and Cd contamination from vine cultivation in the upper stretch, and Pb, Zn, and Mn contamination in the medium stretch from airborne dusts coming from a cast iron foundry.

Polymer functionalized nanocomposites for metals removal from water and wastewater: An overview.

Pollution by metal and metalloid ions is one of the most widespread environmental concerns. They are non-biodegradable, and, generally, present high water solubility facilitating their environmental mobilisation interacting with abiotic and biotic components such as adsorption onto natural colloids or even accumulation by living organisms, thus, threatening human health and ecosystems. Therefore, there is a high demand for effective removal treatments of heavy metals, making the application of adsorption materials such as polymer-functionalized nanocomposites (PFNCs), increasingly attractive. PFNCs retain the inherent remarkable surface properties of nanoparticles, while the polymeric support materials provide high stability and processability. These nanoparticle-matrix materials are of great interest for metals and metalloids removal thanks to the functional groups of the polymeric matrixes that provide specific bindings to target pollutants. This review discusses PFNCs synthesis, characterization and performance in adsorption processes as well as the potential environmental risks and perspectives.

Soil compost amendment enhances tomato (Solanum lycopersicum L.) quality.

BACKGROUND: Tomato (Solanum lycopersicum L.) is one of the most important crops in the world and represents a key crop in southern Italy. With the aim to evaluate the nutritional characteristics of tomato fruits in relation to NPK and compost fertilisation, the concentrations of the main nutrients, toxic elements, primary metabolites and total phenols were determined in two varieties (Lido and San Marzano). Each variety was cultivated in a different experimental field, subjected to different agronomic techniques. RESULTS: Concentrations of toxic elements (Cd and Pb) were below the limits indicated by the EU Regulation (2011) in all the fruits analysed. Moreover, fruits obtained from San Marzano plants grown on organic amended soils showed a better overall quality than those obtained on mineral fertilised soil, being characterised by lower N (attributed to lower nitrate and nitrite concentrations), lower Cd, and higher soluble sugar concentrations. Higher concentrations of soluble sugars in fruits from organic amended soils were also observed in the Lido variety. CONCLUSIONS: The agricultural use of quality compost represents an effective strategy to obtain high quality products in an economically and environmentally sustainable way. © 2016 Society of Chemical Industry.

Cadmium accumulation in leaves of leafy vegetables.

Leafy vegetables have a relatively high potential for Cd uptake and translocation, and are thus considered Cd accumulators. For this reason, leaves and roots of lettuce (Lactuca sativa L.) and endive (Cichorium endivia L.) plants, grown on different agricultural soils in Campania region (southern Italy), subjected to different fertilisation treatments (unfertilisation, compost amendment and mineral fertilisation), were analysed for Cd concentrations. Moreover, to clarify if the highest concentrations found are linked to older and inedible or to younger and edible leaves, external and internal endive leaves were separately analysed. All the leafy vegetables analysed showed on average 2-fold higher Cd concentrations in leaves than in roots. Leaf Cd concentrations in both lettuce and endive plants significantly differed among fertilisation treatments, with values highest in the plants grown on mineral fertilised soils. Apart from the soil fertilisation treatments, however, Cd leaf concentrations were often higher (up to 4-fold) than the threshold deduced by the EU 420/2011 Regulation, although the plants grew on unpolluted soils. Anyway, external leaves of endive plants showed significantly higher concentrations than internal leaves (in some cases the values were 3-fold higher), partly reassuring on the consumption of the younger leaves. Moreover, this study points out two major drawbacks in the Italian and European regulatory frameworks: (1) metal concentration (as total and/or available fraction) limits in agricultural soils are lacking; (2) metal concentration thresholds (currently existing only for Cd and Pb in crops) reported in the EU 420/2011 Regulation, expressed on the fresh weight basis rather than on the dry weight basis, appear not suitable.

Evolution, ecology and systematics of Soldanella (Primulaceae) in the southern Apennines (Italy).

BACKGROUND: The populations of Soldanella (Primulaceae) of the southern Apennines (Italy) are unique within the genus for their distribution and ecology. Their highly fragmented distribution range, with three main metapopulations on some of the highest mountains (Gelbison, Sila and Aspromonte massifs) of the area, poses intriguing questions about their evolutionary history and biogeography, and about the possibility of local endemisms. AIMS AND METHODS: In order to clarify the phylogeny and biogeography of the three metapopulations of Soldanella in the southern Apennines, attributed to S. calabrella to date, and to identify possible local endemisms, a comparative approach based on the study of molecular, morphological and ecological characteristics of the populations was employed. Specifically, one nuclear (total ITS) and two plastid (rbcL and trnL) markers were used for the phylogenetic analyses, performed through both maximum likelihood and Bayesian techniques. Among the morphological features, the glandular hair and leaf biometric traits were analysed, and the environment in which the populations grew was characterised for altitude, forest canopy composition and soil pH, C, N and organic matter. RESULTS AND CONCLUSIONS: Our findings demonstrate that the lineage of Soldanella of southern Italy diverged from the Carpathians lineage during the Middle Pleistocene, and underwent an evolutionary radiation during the Late Pleistocene. The populations of the Sila and Aspromonte massifs diverged from the populations of the Gelbison massif around 380000 years ago and are probably undergoing a progressive differentiation due to their isolation. The populations on the Gelbison massif, moreover, have different morphological features from those of the Sila and Aspromonte massifs and a different ecological niche. The molecular, morphological and ecological data clearly demonstrate that the metapopulation of Soldanella on the Gelbison massif belongs to a new taxonomic unit at the species level, which we name Soldanella sacra A. & L. Bellino from the name of the massif on which it was discovered, the "Holy Mountain".