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CrocusseriesScardicihas only two members,C. scardicusandC. pelistericusdistributed at higher elevations in the Balkan Peninsula. In the course of our research, we discovered two new species growing on alpine meadows and pastures in the serpentine massifs in Albania and limestones in North Macedonia, respectively. The morphology and phylogenetic analyses (two nuclear single-copy markers, nrITS, two chloroplast markers) place the new species within seriesScardici. Both new species are closely related toC. scardicus, but morphologically and ecologically clearly differentiated. We also measured genome sizes for all seriesScardicispecies and report here the largest genomes found up to now in crocuses. The impact of different habitat types and isolation on separate mountain ranges in this alpine group of plants is discussed, too.

Crocus asymmetricus (Iridaceae) is described as a new species endemic to the southern part of the Anatolian diagonal in Turkey. It is phylogenetically related to C. vitellinus and morphologically to C. antalyensis, but differs from these species in showing an asymmetric corm and a single point of root emergence. Both C. antalyensis and C. asymmetricus are illustrated and compared in this paper. A Bayesian phylogenetic tree of the nuclear rDNA ITS region confirms the affiliation of C. asymmetricus to C. ser. Flavi, and its close relationship to C. vitellinus. A new identification key to the species of C. ser. Flavi occurring in Turkey is also presented.

The Italian endemic Crocusbiflorus usually shows white or lilac flowers with three-to-five striking violet longitudinal stripes on the outer tepals, but unstriped plants were recorded in the past. These plants were originally described as C.annulatussubvar.estriatus, and subsequently recombined as a variety of C.biflorus. The rediscovery of such plants in Toscana gave us the opportunity to clarify their systematic relationships, so that we typified the name, and performed karyological and ITS analyses. These plants share the same chromosome number (2n = 2x = 8) and ITS sequence with C.biflorus s. str.

The most important center of speciation in the genus Gagea is thought to be in Central Asia. Here, we focus on species diversity in southeastern Kazakhstan (around Almaty, Ili-Alatau range of the Western Tian-Shan mountains). We studied an elevational transect, reaching from lowland steppes to the alpine zone (500–2750 m a. s. l.), and carried out detailed morphological and molecular investigations for populations of Gagea spp. Nine species were identified in different altitudinal zones; one of these (Gagea almaatensis) is described as new to science. We could detect two altitudinal contact zones between closely related species: G. filiformis and G. granulosa (sect. Minimae), and G. almaatensis and G. kuraminica (sect. Gagea). Morphological and molecular investigations (ITS data and cpDNA networks) indicate ongoing hybridization of co-occurring G. filiformis into G. granulosa and putative bidirectional hybridization events between G. almaatensis and G. kuraminica.

Poor morphological and molecular differentiation in recently diversified lineages is a widespread phenomenon in plants. Phylogenetic relationships within such species complexes are often difficult to resolve because of the low variability in traditional molecular loci, as well as various other biological phenomena responsible for topological incongruence such as ILS and hybridization. In this study, we employ a Genotyping-by-sequencing (GBS) approach to disentangle evolutionary relationships within a species complex belonging to the Neotropical orchid genus Cycnoches. The complex includes seven taxa distributed in Central America and the adjacent Chocó biogeographic region, nested within a clade estimated to have first diversified in the early Quaternary. Previous phylogenies inferred from a handful of loci failed to provide support for internal relationships within the complex. Our Neighbor-net and coalescent-based analyses inferred from ca. 13,000 GBS loci obtained from 31 individuals belonging to six of the seven traditionally accepted Cycnoches species provided a robustly supported network. The resulting three main clades are corroborated by morphological traits and geographical distributions. Similarly, Maximum Likelihood (ML) inferences of concatenated GBS-loci produced results comparable with those derived from coalescence and network-based methods, albeit always with poor statistical support. The low support evident in the ML phylogeny might be attributed to the abundance of uninformative GBS loci, which can account for up to 50% of the total number of loci recovered. The phylogenomic framework provided here, as well as morphological evidence and geographical patterns, suggest that the six entities previously thought to be different species might actually represent only three distinct segregates. Our study is the first to demonstrate the utility of GBS data in phylogenomic research of a very young Neotropical plant clade (~2 Ma), and it paves the way for the study of the many other species complexes that populate the species-rich orchid family.

In this contribution new data concerning the distribution of native vascular flora in Italy are presented. It includes new records, exclusions, and confirmations to the Italian administrative regions for taxa in the generaAsplenium,Bolboschoenus,Botrychium,Chamaerops,Crocus,Galeopsis,Grafia,Helosciadium,Hieracium,Juniperus,Leucanthemum,Lolium,Medicago,Phalaris,Piptatherum,Potamogeton,Salicornia,Salvia,Seseli,Silene,Spiraea,TorilisandVicia.Rhaponticoidescalabricais proposed assynonym novumofR.centaurium. Furthermore, new combinations in the generaGalatellaandLactucaare proposed.

A new region of speciation for the genus Gagea (Liliaceae) was investigated (Bogda-Shan and Urumqi; northwestern Xinjiang, China). Two species were recorded as new for the region (G. rufidula, G. davlianidzeae); three species are described as new to science (G. angelae, G. jensii and G. huochengensis). The description of G. nigra is emendated. Sequence data (cpDNA: trnL-trnF IGS+psbA-trnH IGS, nrDNA: ITS), including representatives of all Gagea sections, were used to compare the new species with closely related taxa. A nuclear single copy gene region (pCOS At103) was analysed for representatives of the Sects. Minimae and Gagea. Network analysis of cpDNA and nDNA indicates hybridization and recent speciation in Xinjiang. ITS and pCOS At103 sequences reveal gene flow between G. davlianidzeae and G. nigra. A cpDNA haplotype network constructed from representatives of Sect. Gagea was highly informative phylogenetically. Gagea angelae and G. huochengensis, sharing gene flow, are related closely to a basal clade represented by G. ancestralis, G. xiphoidea and G. capusii, which may include the putative progenitor of all other taxa of the large Eurasian Sect. Gagea. Whereas speciation in Sect. Minimae seems to be driven mainly by hybridization, speciation in the Sect. Gagea may be influenced by both hybridization and geographical separation. We confirm the monophyly of Sects. Bulbiferae and Minimae.

The internal transcribed spacer (ITS) region (ITS1, 5.8S rDNA, ITS2) represents one of the most popular molecular markers in phylogenetics. The number of investigations revealing high degrees of intra-individual polymorphism connected with the presence of pseudogenic ITS regions is on the increase. Studies including pseudogenic ITS regions can lead to erroneous phylogenetic trees and false taxonomic conclusions. For their recognition, we focus on the 5.8S rDNA as the functional part of this region, which is also affected by degeneration processes. We outline three conserved Viridiplantae 5.8S motifs: GAATTGCAGAAwyC, TTTGAAyGCA, CGATGAAGAACGyAGC, which can be simply checked in sequence alignments. The latter 5.8S motif we also recognised in the large subunit RNA (LSU) of Escherichia coli . The utility of different methods for pseudogene detection based on easily recognisable 5.8S sequence motifs by comparison with 5.8S secondary structure reconstructions and statistical tests are discussed and illustrated with three previously published angiosperm data sets.

The increasing number of pseudogenic internal transcribed spacer (ITS) regions (ITS1, 5.8S rDNA, ITS2) reported for plant taxa raises questions—for example, concerning the proportion of functional copies. Our application of quantitative PCR (qPCR) with genomic DNA under stringent conditions revealed that

Crocus heilbronniorum is described as a new species from Fethiye, Mugla province (SW Turkey). Diagnostic morphological characters, a full description and detailed illustrations are provided based on the type specimen and wild specimens. Morphologically, C. heilbronniorum is close to Crocus atrospermus , but the newly described species differs by its indistinct corm tunic neck, the presence of teeth on rings, basal leaf number and narrower leaves, star-like flowers, longer filaments, almost half length shorter anthers, and style always longer than stamens. Furthermore, Crocus heilbronniorum possesses a different chromosome number (2 n = 2 x = 12) with regard to C. atrospermus (2n = 10). In order to clarify the phylogenetic position of this species within Series Lyciotauri , data on ITS region sequences are also reported.

Most flowers display distinct colour patterns comprising two different areas. The peripheral large-area component of floral colour patterns attracts flower visitors from some distance and the central small-area component guides flower visitors towards landing sites. Whereas the peripheral colour is largely variable among species, the central colour, produced mostly by anthers and pollen or pollen mimicking floral guides, is predominantly yellow and UV-absorbing. This holds also for yellow flowers that regularly display a UV bull's eye pattern. Here we show that yellow-flowering Crocus species are a noticeable exception, since yellow-flowering Crocus species-being entirely UV-absorbing-exhibit low colour contrast between yellow reproductive organs and yellow tepals. The elongated yellow or orange-yellow style of Crocus flowers is a stamen-mimicking structure promoting cross-pollination by facilitating flower visitors' contact with the apical stigma before the flower visitors are touching the anthers. Since Crocus species possess either yellow, violet or white tepals, the colour contrast between the stamen-mimicking style and the tepals varies among species. In this study comprising 106 Crocus species, it was tested whether the style length of Crocus flowers is dependent on the corolla colour. The results show that members of the genus Crocus with yellow tepals have evolved independently up to twelve times in the genus Crocus and that yellow-flowering Crocus species possess shorter styles as compared to violet- and white-flowering ones. The manipulation of flower visitors by anther-mimicking elongated styles in Crocus flowers is discussed.

Crocus musagecitii is described as a new species. Diagnostic morphological characters, a full description and detailed illustrations are provided on the basis of the type specimen and wild specimens. Morphologically, C. musagecitii is close to Crocus biflorus subsp. pseudonubigena . Crocus musagecitii differs from C. biflorus subsp. pseudonubigena by the lack of stripes or narrow purplish tongue on outside of outer tepals, wider tepals, and homogenously yellow anthers. In order to clarify the phylogenetic position of this species within the Crocus adamii species complex, we sequenced the internal transcribed spacer region (ITS: ITS1 + 5.8SrDNA + ITS2) and 5’ external transcribed spacer (ETS) of the nuclear ribosomal DNA (rDNA). A phylogenetic tree obtained by Bayesian phylogenetic inference is given. Phylogenetic analyses revealed that the new taxon is close to C. munzurensis. Crocus musagecitii differs from its phylogenetically closest relative C. munzurensis by the corm tunics ( C. musagecitii : coriaceus; C. munzurensis : membranous), the number of leaves ( C. musagecitii : up to 8; C. munzurensis : up to 4) and non-hairy leaf margins.

Hybridization seems to play an important role in speciation of Gagea Salisb., a genus which is characterised by polyploid taxa lines and in which diploids (2n = 24) appear only to be common in basal sections. Hybrid detection was applied utilising direct and cloning nrDNA ITS data (ITS1, 5.8S rDNA, ITS2) combined with neighbour and ribotype networks and discussed in connection with previously published cpDNA, morphological and karyological data of the authors. We have evidence of the hybrid origin of taxa within the section Gagea (G. pomeranica, G.megapolitana) and the monophyletic clade of sections Didymobulbos and Fistulosae (G. microfistulosa, G. polidorii, G. cf. bohemica). Morphologically and karyologically differentiated Gageamegapolitana and G. pomeranica, adapted to synanthropic habitats, represent both hybrids of G. pratensis × G. lutea. Gagea microfistulosa represents a hybrid of G. villosa × G. fragifera; Gagea polidorii could represent the reverse hybrid. G. glacialis is also closely related to the latter complex.

The internal transcribed spacer (ITS) region (ITS1, 5.8S rDNA, ITS2) represents the most widely applied nuclear marker in eukaryotic phylogenetics. Although this region has been assumed to evolve in concert, the number of investigations revealing high degrees of intra-individual polymorphism connected with the presence of pseudogenes has risen. The 5.8S rDNA is the most important diagnostic marker for functionality of the ITS region. In Mammillaria, intra-individual 5.8S rDNA polymorphisms of up to 36% and up to nine different types have been found. Twenty-eight of 30 cloned genomic Mammillaria sequences were identified as putative pseudogenes. For the identification of pseudogenic ITS regions, in addition to formal tests based on substitution rates, we attempted to focus on functional features of the 5.8S rDNA (5.8S motif, secondary structure). The importance of functional data for the identification of pseudogenes is outlined and discussed. The identification of pseudogenes is essential, because they may cause erroneous phylogenies and taxonomic problems.