Pollinator specificity has traditionally been considered the main reproductive isolation mechanism in orchids. Among Mediterranean orchids, however, many species attract and deceive pollinators by mimicking nectar-rewarding plants. To test the extent to which deceptive orchid species share pollinators, we collected and identified hemipollinaria-carrying insects, and used ribosomal sequences to identify the orchid species from which hemipollinaria were removed. We found that social and solitary bees, and also flies, carried hemipollinaria belonging to nine orchid species with different degrees of specialization. In particular, Anacamptis morio , Dactylorhiza romana and Orchis mascula used a large set of pollinator species, whereas others such as Orchis quadripunctata seemed to be pollinated by one pollinator species only. Out of the insects with hemipollinaria, 19% were found to carry hemipollinaria from more than one orchid species, indicating that sympatric food-deceptive orchids can share pollinators. This sharing was apparent even among orchid sister-species, thus revealing an effective overlap in pollinator sets among closely related species. These results suggest varying degrees of pollinator specificity in these orchids, and indicate that pollinator specificity cannot always act as the main isolation mechanism in food-deceptive temperate orchids.

Drinking tea constitutes a tradition which is deeply rooted in the culture of several countries. Moreover, in recent years, tea consumption is growing all over the world. Improper herbal tea storage (long periods, humid environments) represents a relevant health hazard for consumers because of the growth of bacteria and molds. This study analyzed 32 samples of commercially available black and green teas – purchased from southern Italy markets and online-shops – and the monitoring of microbiological quality of the tea bag content was performed. Evaluations were conducted with the aim of characterizing pathogens indicated by the European and American guidelines (total bacterial count, fungi and Escherichia coli) and on the research of Pseudomonas spp. and Clostridium perfringens. The presence of ochratoxin A in tea matrix-leaves and infusions was further assessed, using a validated and accredited HPLC-FLD method. Microbial loads, for over 80% samples, ranged from 1.0 × 102 to 2.8 × 105 CFU/g tea: most of identified microorganisms were classified as Bacillaceae. The utilization of rapid detection and identification methods (PCR and sequencing), allowed the characterization of strains of Pseudomonas psychrotolerans, Staphylococcus warneri, Pantoea gaviniae and the isolation of one strain of Clostridium perfringens, whose ability to produce toxins can result in harmful outcomes for consumers. Fungi were isolated from 70% samples: the most prevalent molds were Aspergillus niger strains, followed by Aspergillus tubingensis. Ochratoxin A was detected in 22 of 32 tea solid samples investigated: concentrations resulted over the indicated limits for food products for 50% samples. Results obtained demonstrated the need to develop targeted regulations for the safety of herbal teas.

Background and AimsCyperus 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.

The phylogenetics and biogeography of Pancratium (Amaryllidaceae) were investigated, with a focus on the Mediterranean and adjacent areas, with the aim of contributing new information towards a better understanding of the evolutionary history of the genus and the taxonomic placement of P. linosae and P. hirtum. To address these questions, we sequenced four plastid DNA markers: the ndhF and rbcL genes, the trnL^(UAA)–trnF^(GAA) intergenic spacer and the trnL^(UAA) intron, analysing them using parsimony, likelihood and Bayesian approaches. The results show that the relationships among the majority of the species are resolved; however, the relationships of one of the major clades of the genus are unresolved compared with the others. The phylogenetic and the dispersal–vicariance analyses show that Pancratium appears as a well-structured group with interesting patterns of speciation. Notably, P. arabicum and P. linosae fall within the P. maritimum complex. In addition, P. hirtum is identical, in terms of plastid DNA sequences, to the P. trianthum accessions. The results provide new insights and help to formulate new working hypotheses for evolutionary biology of the genus.

Petagnaea gussonei (Apiaceae) is an endangered species endemic to the Nebrodi mountains (north-eastern Sicily). Although an increasing number of studies have been performed on this species, its reproductive biology remains poorly understood. The aim of this study was to investigate in detail the structure of the flower and the fruit of Petagnaea, and the possible implications for its breeding system and seed dispersal mechanism. Results from fieldwork, light microscopy and scanning electron microscopy suggest (1) the presence of protandrous hermaphrodite flowers; (2) geitonogamy, autogamy, and allogamy as breeding system mechanisms of P. gussonei, even if asexual reproduction is preferred by the plant; and (3) epizoochory and hydrochory as possible modalities of fruit dispersal.