Integrative Taxonomy of Armeria Taxa (Plumbaginaceae) Endemic to Sardinia and Corsica.

Sardinia and Corsica are two Mediterranean islands where the genus Armeria is represented by 11 taxa, 10 out of which are endemic. An integrative approach, using molecular phylogeny, karyology, and seed and plant morphometry was used to resolve the complex taxonomy and systematics in this group. We found that several taxa are no longer supported by newly produced data. Accordingly, we describe a new taxonomic hypothesis that only considers five species: Armeria leucocephala and A. soleirolii, endemic to Corsica, and A. morisii, A. sardoa, and A. sulcitana, endemic to Sardinia.

Genotyping-by-sequencing provides new genetic and taxonomic insights in the critical group of Centaurea tenorei.

Centaurea L. is one of the most widespread, differentiated, and critical genera of Asteraceae in the Euro-Mediterranean area, with more than 100 currently recognized species inhabiting the region. The controversial C. tenorei group, narrowly endemic to the Peninsula of Sorrento (Campania region, southern Italy), includes three weakly differentiated microspecies: C. tenorei Guss. ex Lacaita, C. montaltensis (Fiori) Peruzzi and C. lacaitae Peruzzi. However, their taxonomic distinctiveness and relationships with close or sympatric species are still unclear. In particular, the existence in several localities of individuals with intermediate morphology suggests inadequate taxonomic assessment within the group or hybridization and introgression with other species. In this study we aimed at defining population structure in this complex. With this objective, we sampled the three currently accepted species from their loci classici (i.e., the localities in which the taxa were originally described) and from other localities throughout the range, including populations of difficult identification occurring where the ranges of different taxa overlap. We employed a panel of SNPs obtained via genotyping-by-sequencing for investigations on genetic structure, admixture and ploidy inference, the latter also compared with chromosome counts. Our results showed that Centaurea tenorei s.l. is consistently tetraploid, contradicting the current taxonomy that was also based on ploidy level. Population structure analyses indicated the presence of four to seven clusters, most of which with clear evidence of admixture. Furthermore, contrarily to what previously supposed, we demonstrated a remarkable contribution of C. deusta, more that of C. cineraria in the genetic make-up of C. tenorei. However, we found a population of C. cineraria outside its ecological range, probably driven by climate change, which could be responsible in the future of further hybridization phenomena.

A Morphometric Analysis of the Santolina chamaecyparissus Complex (Asteraceae).

The genus Santolina (Asteraceae, Anthemideae) includes 26 species of aromatic evergreen shrubs endemic to the western Mediterranean Basin. Santolina is widely used as ornamental plant, in xerigardening, and in ethnobotany. The Santolina chamaecyparissus complex, including about half of the known species diversity, has been properly investigated on systematic and taxonomic grounds only recently, and a complete morphometric study is still missing. Here we provide a morphometric characterization and comparison of all the 14 species of this complex, using both univariate and multivariate analyses. Our results suggest that species of this complex can be distinguished using combinations of quantitative and qualitative character-states, mostly related to the leaf morphology. The analysis of S. villosa, a tetraploid/hexaploid Spanish endemic, showed that the two cytotypes cannot be safely identified based on morphology. Coupling this evidence with available phylogenetic information, we conclude that there is no reason to split the two cytotypes of S. villosa in two distinct taxa. An identification key for all the species of the complex is presented.

Variability and Nativeness in the Mediterranean Taxa: Divergence and Phylogeography of Genista etnensis (Fabaceae) Inferred from Nuclear and Plastid Data.

Genista etnensis is a remarkable and well-known tree endemic to Sicily, Sardinia, and Corsica (Mediterranean Basin). Nevertheless, its morphological variability and its native status throughout its range need to be further investigated. In this study, we aim to clarify some aspects of this infraspecific variability by molecular means. Sequences of one nuclear and five plastid markers were analyzed under maximum parsimony by using TCS software. Plastid data were also time-calibrated under a Bayesian Inference framework. Plastid data revealed strong isolation between the populations from the Cyrno-Sardinian biogeographical province, which are also the most diverse and presumably the most archaic, and those from Sicily and Southern Italy (in this latter area, the species is naturalized). The calibration analysis indicates that the last common ancestor between G. etnensis and its sister group G. fasselata dates back to the middle Pliocene or slightly later, when sclerophyllous Mediterranean vegetation spread, whereas G. etnensis itself might have originated in the middle Pleistocene. The current, rather unusual distribution of G. etnensis could be explained by long-range seed dispersal from the western part of the range or by anthropogenic introduction into Sicily, with extinctions of transported haplotypes in the region of origin. Interestingly, the Vesuvius population, introduced from Sicily in recent times and locally naturalized, shows private genotypes, and was richer in both genotypes and haplotypes than the Sicilian ones.

Relationships within Mcneillia Indicate a Complex Evolutionary History and Reveal a New Species of Minuartiella (Caryophyllaceae, Alsinoideae).

The genus Mcneillia has been recently segregated from Minuartia L. based on molecular results, also supported by morphology. However, to date, a comprehensive study on the phylogenetic relationships within this genus is lacking. In this paper, we provide a multigene phylogeny of all the species and subspecies of Mcneillia employing two nuclear and six chloroplast markers. We documented extensive gene flow between taxa, sometimes separated at specific rank. In addition, Mcneillia as currently circumscribed, is not monophyletic. In fact, Mcneillia graminifolia subsp. brachypetala, strictly endemic to Greece, truly belongs to Minuartiella, a genus otherwise limited to South-West Asia. Moreover, even after removal of this taxon, our results do not support the monophyly of the taxa included in M. graminifolia s.l., the most variable and widespread species of the genus. Further controversial subspecies of Mcneillia graminifolia, i.e., subsp. hungarica and subsp. rosanoi, are shown to deserve taxonomic recognition as separate species, whereas Mc. moraldoi is not distinct at specific rank. In addition, Mc. saxifraga subsp. tmolea is here regarded as a further distinct species. A consistent taxonomic treatment is therefore proposed with six new combinations and nomenclatural notes, providing the necessary typifications.

Characterisation of Environmental Biofilms Colonising Wall Paintings of the Fornelle Cave in the Archaeological Site of Cales.

Caves present unique habitats for the development of microbial communities due to their peculiar environmental conditions. In caves decorated with frescoes, the characterization of microbial biofilm is important to better preserve and safeguard such artworks. This study aims to investigate the microbial communities present in the Fornelle Cave (Calvi Risorta, Caserta, Italy) and their correlation with environmental parameters. The cave walls and the wall paintings have been altered by environmental conditions and microbial activity. We first used light microscopy and scanning electron microscopy (SEM) and X-ray diffraction to characterise the biofilm structure and the mineral composition of substrata, respectively. Then, using both culture-dependent (Sanger sequencing) and culture-independent (automated ribosomal intergenic spacer analysis, ARISA) molecular methods, we demonstrated that the taxonomic composition of biofilms was different across the three substrata analysed and, in some cases, positively correlated with some environmental parameters. We identified 47 taxa in the biofilm samples, specifically 8 bacterial, 18 cyanobacterial, 14 algal and 7 fungal taxa. Fungi showed the highest number of ARISA types on the tuff rock, while autotrophic organisms (cyanobacteria and algae) on the frescoes exposed to light. This study confirms that caves constitute a biodiversity-rich environment for microbial taxa and that, in the presence of wall paintings, taxonomic characterization is particularly important for conservation and restoration purposes.

Nomenclatural Synopsis of Cirsium Sect. Eriolepis (Asteraceae) in Italy.

The names of the Italian taxa in Cirsium sect. Eriolepis are discussed. The accepted names are: Cirsium echinatum, C. eriophorum subsp. eriophorum, C. eriophorum subsp. spathulatum, C. ferox, C. italicum, C. lacaitae, C. lobelii, C. morisianum, C. scabrum, C. tenoreanum, C. vallis-demonii subsp. vallis-demonii, C. vallis-demonii subsp. calabrum comb. nov., and C. vulgare (= C. crinitum, C. sylvaticum). Four accepted names are typified by specimens preserved at FI (one lectotype), G (one lectotype and one neotype), P (one lectotype), and by illustrations (two lectotypes). Several other heterotypic synonyms of taxa described from Italy are discussed and six of them are typified. A new combination and status are proposed: C. vallis-demonii subsp. calabrum, based on C. eriophorum var. vallis-demonii f. calabrum.

UV-C Irradiation as a Tool to Reduce Biofilm Growth on Pompeii Wall Paintings.

This study focuses on the experimentation of a method based on the use of UV-C irradiation to eliminate the biofilms present in a tomb located in the necropolis of Porta Nocera, in Pompeii. For this study, the autotrophic component of the biofilm was isolated in the laboratory, while, contemporarily, the characterization of the composition of the pigments of the frescoes took place on original fragments, which had already detached from the tomb and were examined in situ. These preliminary analyses were necessary for the recreation of test samples in the laboratory, which closely matched the original surfaces. Artificial biofilms were used for experimental exposure to UV-C radiation. The exposure to UV-C radiation was carried out at different distances for a fixed time interval. The effectiveness of the biocidal action was assessed by employing optical microscopy techniques, through a careful visual assessment of the area occupied by the biofilm on the different test samples, using a photographic survey, as well as by means of colorimetric measurements using spectrometric techniques. In order to obtain an additional parameter to evaluate the death rate of microorganism cultures exposed to the UV-C radiation, the concentrations of the photosynthetic pigments were also measured by spectrophotometry. Results showed that biofilms were completely eradicated by radiation, and no change in pigment color was observed.

Use of high-strength electromagnetic radiation to remove phototrophic biofilms from terracotta artifacts.

A novel technique, effective in eliminating biodeteriogens from biofilms encrusting terracotta artifacts, is presented here. This method is based on the use of high-strength electromagnetic radiation (EMR) in the radiofrequency band. Shards of terracotta from historical pots at the Botanical Garden of Naples, Italy, were used. The shards, after sterilization, were inoculated with several phototrophic microorganisms previously isolated from whole terracotta pots. The newly formed biofilms were exposed to EMR amplitude modulated by a train of rectangular pulses with Tr = 200 ns repetition time and 10% duty cycle. The exposure protocol consisted of three applications of 2 h each, every other day. Denaturing gradient gel electrophoresis analyses conducted on the newly formed biofilms showed that, after the first exposure to EMR, all species in the biofilms but one were still alive. The second exposure resulted in the disappearance of 9 out of 13 species that were initially present on the samples. After the third exposure, all species disappeared. Superficial layers of terracotta from the exposed samples, transferred to a culture medium at 24 °C for 72 h, did not show any re-growing of organisms. Petrographic analyses of the sampleswere carried out before and after the treatments; they showed that exposure to EMR did not cause structural alterations in the treated substrates. Moreover, the amplitude of the EMR that samples were exposed to was not high enough to cause any significant increase in the temperature of the substrates; that is, no thermal effect, which is the most relevant effect when matter or organisms containing water are exposed to EMR, was observed. Finally, the field strength of the EMR showed to be non-invasive for the artifacts and non-dangerous for operators and the environment as compared to other techniques adopted in the field of conservation of cultural heritage.

Conservation and genetic characterisation of common bean landraces from Cilento region (southern Italy): high differentiation in spite of low genetic diversity.

Since its introduction from Central-South America to Italy almost 500 years ago, the common bean (Phaseolus vulgaris L.) was largely cultivated across the peninsula in hundreds of different landraces. However, globalisation and technological modernisation of agricultural practices in the last decades promoted the cultivation of few varieties at the expense of traditional and local agro-ecotypes, which have been confined to local markets or have completely disappeared. The aim of this study was to evaluate the genetic diversity and differentiation in 12 common bean landraces once largely cultivated in the Cilento region (Campania region, southern Italy), and now the object of a recovery program to save them from extinction. The analysis conducted using 13 nuclear microsatellite loci in 140 individuals revealed a high degree of homozygosity within each landrace and a strong genetic differentiation that was reflected in the success in assigning individuals to the source landrace. On the contrary, internal transcribed spacers 1 and 2, analysed in one individual per landrace, were highly similar among common bean landraces but allowed the identification of a cowpea variety (Vigna unguiculata Walp.), a crop largely cultivated in the Old World before the arrival of common bean from Americas. In conclusion, our study highlighted that conservation of landraces is important not only for the cultural and socio-economic value that they have for local communities, but also because the time and conditions in which they have been selected have led to that genetic distinctiveness that is at the basis of many potential agronomical applications and dietary benefits.

A molecular survey concerning the origin of Cyperus esculentus (Cyperaceae, Poales): two sides of the same coin (weed vs. crop).

BACKGROUND AND AIMS: Cyperus esculentus is widespread in tropical and temperate zones and is also present in cooler regions. It is used as a crop plant, but it also occurs in the wild and as a weed. As a consequence of its ecological plasticity, C. esculentus has remarkable variability, with several morphotypes. Four wild-type varieties are presently recognized, in addition to the cultivated form. This study investigates the phylogenetic position and biogeography of C. esculentus with the objective of contributing new data to increase the understanding of its evolutionary history. METHODS: Genealogical relationships among genotypes were inferred by using plastid DNA haplotype and nuclear ribosomal (nr) DNA ribotype sequences for 70 specimens either collected in the field or obtained from herbaria. Statistical dispersal-vicariance (S-DIVA) and Bayesian binary method (BBM) analyses were used to reconstruct the possible ancestral ranges of C. esculentus. In order to determine the age of C. esculentus, a time-measured phylogenetic analysis was performed. KEY RESULTS: Considerable variation between the chosen nuclear and plastid markers was detected (27 ribotypes vs. six haplotypes). No geographical structure was displayed among the haplotypes, but information on the dispersal pattern may be deduced. Two types of ribotypes were detected in nrDNA, with an evident geographical segregation into an Old World group and a polymorphic New World group. Both S-DIVA and BBM analyses suggested a biogeographical history in which dispersal from the African region has been crucial in shaping the current distribution pattern of C. esculentus. The most recent common ancestor between C. esculentus races has an age of 5.1 million years (95 % highest posterior density 2.5-10.2). CONCLUSIONS: The molecular analysis provides novel insights into the evolutionary history of C. esculentus. The results have various taxonomic and phylogenetic implications, including a hypothesis on the origin and phylogeography of this species, which probably originated in the late Cenozoic in Africa, and reached the Americas repeatedly, independently of Columbian exchanges.

Rotational thromboelastometry analysis and management of life-threatening haemorrhage in isolated craniofacial injury.

Massive haemorrhage from facial fractures is rare but the associated mortality rate is high. Here, we describe a case in which thromboelastometry [rotational thromboelastometry (ROTEM)]-guided administration of prothrombin complex concentrate and fibrinogen concentrate was effective in correcting coagulopathy in a 68-year-old man with serious craniofacial trauma and massive haemorrhage. The patient, a cyclist who collided with a car, was transferred to the emergency department of our hospital with signs of shock and significant bleeding from multiple fractures and soft tissue injuries to the face. Blood gas analysis and standard laboratory tests revealed the presence of anaemia and acidosis, and our massive haemorrhage protocol was initiated. E-FAST and total-body computed tomography scans excluded the possibility of bleeding from other sites. All efforts were directed towards stopping bleeding from craniofacial lesions, but the surgeon experienced difficulty in maintaining haemostasis. ROTEM analysis revealed severe coagulopathy and was indispensable in guiding transfusion: 2 g tranexamic acid, followed by 1000 IU prothrombin complex concentrate, 5 g fibrinogen and 2 U platelet concentrate. Two hours later, ROTEM analysis showed that coagulopathy had been corrected, and haemostasis was confirmed by cessation of bleeding. This report highlights the potential for using ROTEM to guide treatment with fibrinogen and prothrombin complex concentrates in the presence of profuse multifocal bleeding and severe coagulopathy.