The small, annual grass Brachypodium distachyon (L.) Beauv., a close relative of wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.), is a powerful model system for cereals and bioenergy grasses. Genome-wide association studies (GWAS) of natural variation can elucidate the genetic basis of complex traits but have been so far limited in B. distachyon by the lack of large numbers of well-characterized and sufficiently diverse accessions. Here, we report on genotyping-by-sequencing (GBS) of 84 B. distachyon, seven B. hybridum, and three B. stacei accessions with diverse geographic origins including Albania, Armenia, Georgia, Italy, Spain, and Turkey. Over 90,000 high-quality single-nucleotide polymorphisms (SNPs) distributed across the Bd21 reference genome were identified. Our results confirm the hybrid nature of the B. hybridum genome, which appears as a mosaic of B. distachyon-like and B. stacei-like sequences. Analysis of more than 50,000 SNPs for the B. distachyon accessions revealed three distinct, genetically defined populations. Surprisingly, these genomic profiles are associated with differences in flowering time rather than with broad geographic origin. High levels of differentiation in loci associated with floral development support the differences in flowering phenology between B. distachyon populations. Genome-wide association studies combining genotypic and phenotypic data also suggest the presence of one or more photoperiodism, circadian clock, and vernalization genes in loci associated with flowering time variation within B. distachyon populations. Our characterization elucidates genes underlying population differences, expands the germplasm resources available for Brachypodium, and illustrates the feasibility and limitations of GWAS in this model grass. Core Ideas• Genotyping diverse Brachypodium accessions expands research tools for grasses.• The B. hybridum genome is a mosaic of B. distachyonand B. stacei-like sequences.• Three distinct, genetically defined populations of B. distachyon were identified.• Flowering time, more than geography, distinguishes B. distachyon populations.• Results support the feasibility of genome-wide association studies in a model grass.

Isoetes is a widely distributed lycophyte genus of at least 200 species occurring in diverse habitats. The species can be difficult to identify because Isoetes, with its apparent simplicity of form and conserved morphology, provides few diagnostic features to reliably distinguish its species. The last worldwide monograph, published nearly a century ago, listed 77 taxa. The first step in producing a flora or monograph of all known species of a genus is to compile a list of the acceptable species names. The list presented here is a compilation of 192 accepted names representing taxa from regions around the world: chromosome numbers were assigned to 101 of them, with polyploidy settled on 46.7%. Distribution mapping of the accepted species indicates that South America is the center of diversity for Isoetes and species diversity is the highest in temperate regions. Many of the species on this list are rare and have limited ranges. The list of taxa can be used to initiate floristic studies and conservation efforts in keeping with the target goals of the Global Strategy for Plant Conservation.

In this study we focused on four taxa of the genus Calendula (C. maritima, C. suffruticosa subsp. fulgida, C. arvensis, and the hybrid between the first two ones), collected in Mediterranean area (Sicily). Six extracts for each species were obtained using solvents with increasing polarity (hexane, ethanol 80%, acetone 70%, and water) and through extraction by supercritical fluids (SFE). It has been observed that the solvent with the highest extraction efficiency was ethanol 80% for all species. However, SFE extracts showed high antioxidant activity comparable to the ethanol 80% extract (polyphenol, DPPH, and reducing power method). These findings were confirmed by in vitro analysis (MTT assay) where it was observed that the tested concentration (24 µg/mL), obtained from ethanol 80% and SFE extracts, showed a protective effect comparable to that induced by a synthetic antioxidant. Extraction with SFE ensured a great selectivity by avoiding the use of toxic organic solvents and thus consisted of a promising technique for sustainable production of Calendula extracts.

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Our understanding of the richness and uniqueness of the flora growing on gypsum substrates in Italy has grown significantly since the 19th century and, even today, new plant species are still being discovered. However, the plants and plant communities, growing on gypsum substrates in Italy, are still a relatively unknown subject.The main aim of this paper was to elaborate a checklist of the Italian gypsophilous flora, to increase knowledge about this peculiar flora and for which conservation efforts need to be addressed.Through a structured group communication process of experts (application of the Delphi technique), a remarkable number of experienced Italian botanists have joined together to select focal plant species linked to gypsum substrates. From the results obtained, 31 plant species behave as absolute or preferent taxa (gypsophytes and gypsoclines) and form the ‘core’ Italian gypsophilous flora. The most abundant life forms were chamaephytes and hemicryptophytes, belonging to Poaceae and Brassicaceae; as for chorotypes, the most represented are Mediterranean and narrow endemics. By improving on previously available information about the flora with a clear preference for gypsum in Italy, this undertaking represents an important contribution to the knowledge of a habitat which is today considered a priority for conservation.

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Hybridization and competition between the endangered sea marigold (Calendula maritima, Asteraceae) and a more common congener ARTICLE in PLANT BIOSYSTEMS · FEBRUARY 2015 Impact Factor: 1.91 · AbstractOccurrences of hybridization between the rare, endangered Sicilian endemic, Calendula maritima, and other congeners in the wild have been suspected by several authors, but never demonstrated. In San Cusumano (Trapani), one of the only three Sicilian mainland populations, C. maritima occurs in close proximity to the more common congener C. suffruticosa subsp. fulgida, and individuals morphologically intermediate between the two taxa have been observed. In order to determine whether hybridization is occurring at this site, and, if so, begin to assess the impact it could have on C. maritima, two independent tests of hybridity were conducted by studying (1) the pollen morphology and viability and (2) sequence variation in one chloroplast marker (atpI-atpH) and in the nuclear ribosomal internal transcribed spacer region (ITS). Both palynological and molecular data provided evidence of hybridization between C. maritima and C. suffruticosa subsp. fulgida, and suggest that although fertility of hybrids appeared to be reduced, they are likely capable of back-crossing with parents. Thus, hybridization may be putting C. maritima at risk of extinction via introgression. Anthropogenic habitat disturbance and expansion of C. suffruticosa subsp. fulgida into remaining populations of C. maritima could pose a serious threat to its successful conservation.