Luigi Minuto

Molecular phylogeny of the Caryophyllaceae (Caryophyllales) inferred from chloroplast matK and nuclear rDNA ITS sequences

Caryophyllaceae is a principally holarctic family including around 2200 species often classified into the three subfamilies Alsinoideae, Caryophylloideae, and Paronychioideae. Complex and possibly homoplasious morphological characters within the family make taxa difficult to delimit and diagnose. To explore part of the morphological evolution within the family, we investigated the phylogeny of the Caryophyllaceae by means of analyzing plastid and nuclear sequence data with parsimony and Bayesian methods. We describe a mode of tracing a stable phylogenetic signal in ITS sequences, and a significant common signal is shared with the plastid data. Parsimony and Bayesian analyses yield some differences in tree resolution. None of the subfamilies appear monophyletic, but the monophyly of the Caryophylloideae is not contradicted. Alsinoideae are paraphyletic, with Arenaria subg. Eremogone and Minuartia subg. Spergella more closely related to the Caryophylloideae. There is strong support for the inclusion of Spergula-Spergularia in an Alsinoideae-Caryophylloideae clade. Putative synapomorphies for these groupings are twice as many stamens as number of sepals and a caryophyllad-type of embryogeny. Paronychioideae form a basal grade, where tribe Corrigioleae are sister to the rest of the family. Free styles and capsules with simple teeth are possibly plesiomorphic for the family.

Seed morphology in Moehringia L. and its taxonomic significance in comparative studies within the Caryophyllaceae

Seeds of 30 species of Moehringia and 12 representatives of the Caryophyllaceae were examined with the Scanning Electron Microscope. High diversity was found in seed coat micromorphology, and a number of novel morphological features have been observed. Comparison with other representatives of the Caryophyllaceae has provided new insight into the potential taxonomic value of discrete morphological characters. Though many characters appear informative within Moehringia, they are affected by a high level of homoplasy when considered in a wider phylogenetic context. Smooth seed is the common condition in Moehringia, except for Eastern Balkan and Iberian species that maintain a secondary ornamentation of the testa cells resembling the plesiomorphic Caryophyllaceae-type. Five types of strophioles were recognized, in accordance with previous literature. Among these, the strophiole of the Iberian species appears very distinctive and a proper strophiole could not be verified in M. glochidisperma and M. fontqueri.

Ecogeographic and genetic evaluation of endemic species in the Maritime Alps: the case of Moehringia lebrunii and M. sedoides (Caryophyllaceae)

Abstract The Maritime Alps are one of the ‘hot spots’ in the Mediterranean basin. This study investigated two endemic plants, Moehringia lebrunii and Moehringia sedoides (Caryophyllaceae) in order to increase knowledge of the vegetation of this region, and to investigate possible conservation strategies. Ecogeographic surveys and molecular analyses were undertaken. Gene diversity, the Shannon index and GST were calculated within and among populations of the two species based on ISSR data. The populations of M. lebrunii had a density ranging between 0.04 and 0.86 individual/m2 and a rather low inner genetic variability value. According to IUCN Red List Criteria, the current status of M. lebrunii is Endangered [EN B2ab(ii, iv)]. M. sedoides is an endemic of the SW Alps (not exclusive of the Maritime Alps), and is very abundant within the core of the range. Its range of occurrence is smaller than previously reported; nevertheless, the species is not under threat. This taxon showed a population density ranging between 0.03 and 0.58 individual/m2. Genetic variability values revealed a high variation among the species. Only peripheral populations seemed to suffer from their segregated position. Thus, M. sedoides is to be considered Critically Endangered [CR B1ab(i, ii, iii, iv) + 2ab(i, ii, iii, iv)] for Italy according to Regional Guidelines.

Evaluation of biodiversity and conservation strategies in Pancratium maritimum L. for the NorthernTyrrhenian Sea

Abstract.Pancratium maritimum L. is an Amaryllidaceous species whose presence is severely endangered in its original range, the sandy coasts of the Mediterranean sea. A molecular analysis has been performed to evaluate the genetic distance among populations coming from different locations, in order to define the best repopulating strategy. The plant genome, analysed by AFLP markers, was found to be extremely homogeneous and conserved, evoking vegetative or autogamous reproductive habits. Seeds from two different locations showed a good germination capability in greenhouse tests, indicating the potential presence of an efficient sexual reproduction. The combination of molecular data and germination tests would support the hypothesis of an autogamous reproduction for this species.

Polyploid evolution and Pleistocene glacial cycles: A case study from the alpine primrose Primula marginata (Primulaceae)

BackgroundRecent studies highlighted the role of Pleistocene climatic cycles in polyploid speciation and of southern Alpine refugia as reservoirs of diversity during glacial maxima. The polyploid Primula marginata, endemic to the southwestern Alps, includes both hexaploid and dodecaploid cytotypes that show no ecological or morphological differences. We used flow cytometry to determine variation and geographic distribution of cytotypes within and between populations and analyses of chloroplast (cp) and nuclear ribosomal (nr) DNA sequences from the Internal Transcribed Spacer (ITS) region to infer the evolutionary history of the two cytotypes and the auto- vs. allopolyploid origin of dodecaploid populations.ResultsWe did not detect any intermediate cytotypes or variation of ploidy levels within populations. Hexaploids occur in the western and dodecaploids in the eastern part of the distributional range, respectively. The cpDNA and nrDNA topologies are in conflict, for the former supports shared ancestry between P. marginata and P. latifolia, while the latter implies common origins between at least some ITS clones of P. marginata and P. allionii.ConclusionsOur results suggest an initial episode of chloroplast capture involving ancestral lineages of P. latifolia and P. marginata, followed by polyploidization between P. marginata-like and P. allionii-like lineages in a southern refugium of the Maritime Alps. The higher proportion of ITS polymorphisms in dodecaploid than in hexaploid accessions of P. marginata and higher total nucleotide diversity of ITS clones in dodecaploid vs. hexaploid individuals sequences are congruent with the allopolyploid hypothesis of dodecaploid origin.

New seed morphological features in Moehringia L. (Caryophyllaceae) and their taxonomic and ecological significance

Abstract Seeds of 27 species of Moehringia and four related representatives of Arenaria (old M. sect. Pseudomoehringia) were examined by scanning electron microscopy. Novel morphological features were observed in addition to those found in a previous similar study. The new information thus obtained provided new insight into the taxonomic value of discrete morphological characters considered in a recent phylogenetic context. Main Moehringia sections are undoubtedly defined by different types of strophioles, and by the corresponding testa cell features. This study also shows the different ontogenesis of hilar appendages previously attributed to Iberian taxa, now belonging to Arenaria. In addition to phylogenetic differences, ecology seems to play a role in the differentiation of seed morphological features with clear convergences among plants living on cliffs.

Floral features and reproductive ecology in Thymelaea hirsuta (L.) Endl.

Abstract.Thymelaea hirsuta is a circum-Mediterranean subdioecious shrub used to model the evolutionary pathway from heterodichogamy to dioecy. Since discordant data on the pollination mechanism of this plant have been reported, in this study we have tried to clarify the complex of features and events involved in the breeding system of this species at the Northern edge of its range (Tyrrhenian coast, Italy and France). Flower and pollen features related to pollination mechanism were analyzed, and the seasonal presence of insects visiting the plant was monitored. Despite the presence of small pollen grains, normally associated with wind pollination, the Crotonoid-pattern of exine, the large amount of pollenkitt and the presence of nectar are typical entomophilous adaptations. Among insects visiting the plant, thrips (Thysanoptera) might have a role in pollination owing to their large number and for the constant presence of T. hirsuta pollen on their body. In contrast, no evidence of anemophilous transport of pollen was obtained, clashing with reports of a wind pollination system for Egyptian populations. Based on our observations wind seems not to play a key role in the pollination process of this species in the Northern Mediterranean region, while insects might play a major role.

Allopatric divergence and secondary contacts in Euphorbia spinosa L: Influence of climatic changes on the split of the species

Euphorbia spinosa, a perennial xerophilous shrub naturally distributed across the Italian peninsula, was selected for examination of the role of the Ligurian Alps and Apennines in glacial survival. The Italian Peninsula is considered to be one of the principal glacial refugia in Europe, but few plant population genetic and phylogeography studies have been undertaken in this region. The combined analysis of chloroplast and nuclear loci (ITS, cpSSR and ISSR) enabled us to detect extensive DNA variation and proved to be a very powerful tool for the reconstruction of the phylogeography. Molecular data support the hypothesis of a long-term separation of the Northwestern (Maritime Alps, Sardinia, Corsica, Northern Apennines) and Southeastern (Southern Apennines and Balkan area) lineages in glacial refugia. The existence of allopatrically fragmented lineages is most probably the result of isolation in different glacial refugia, possibly due to the Last Glacial Maximum cooling and the topographic complexity of the Italian peninsula. The most plausible hypothesis assumes the formation of two migration paths during more recent periods: the first one starting with southward migration and the second one moving northwards. The Central Apennines should be considered the confluence of migration routes radiating from separate refugia according to this hypothesis.

Pollination ecology in the narrow endemic winter-flowering Primula allionii (Primulaceae).

Reduction of pollen flow can affect plant abundance and population viability and cause selection on plant mating system and floral traits. Little is known on the effect of this phenomenon in species naturally restricted to small and isolated habitats, that may have developed strategies to cope with long-term isolation and small population size. We investigated the pollination ecology of the endemic distylous winter-flowering P. allionii to verify the possible limitation of female fitness due to reduced pollinator visits. We recorded a higher production of pollen grains in long-styled morph, and a higher seed set in short-styled morph. The high intra-morph variability of sexual organ position may explain the hybridization phenomena allowing and easier intra-morph pollination. The fruit set is constant, although its winter-flowering period might decrease pollen transfer. Nevertheless, the lower competition for pollinators with neighbouring plants and the long-lasting anthesis may offset its reproductive success. Even if our results show no evidence of imminent threats, changes in plant–pollinator interactions might increase inbreeding, resulting in an increased extinction risk.

Molecular diversity in Ligurian local races of common bean (Phaseolus vulgaris L.)

Abstract Eight varieties of Ligurian common beans (Phaseolus vulgaris L.) were analysed using molecular approaches. Results were compared with two commercial cultivars (‘Cannellino’ and ‘Borlotto’). Data suggest that all Ligurian bean varieties have a low genetic variability and are very close to the commercial varieties. In particular, the three ‘Bianco’ varieties showed a molecular affinity, probably due to their common genomic origin.