Ivana Rešetnik

Molecular phylogeny and systematics of the Lilium carniolicum group (Liliaceae) based on nuclear ITS sequences

The Lilium carniolicum group consists of several taxonomically dubious taxa endemic to the European flora. Internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA (nrDNA) were used to clarify both the delineation of, and relationships among, taxa in the group as well as to provide insight on the phylogenetic position of the group within the genus. Maximum parsimony, maximum likelihood and Bayesian analyses were in general agreement, with all taxa in the group being very closely related, and the entire group being monophyletic. L. pyrenaicum and L. pomponium are placed at the basal position in the group, while L. chalcedonicum is shown to be more closely related to L. carniolicum than previously thought. Our analyses suggested that L. albanicum and L. jankae are distinct from L. carniolicum, while no evidence was found to support the same separation for L. bosniacum.

Phylogenetic relationships in Brassicaceae tribe Alysseae inferred from nuclear ribosomal and chloroplast DNA sequence data.

Numerous molecular systematic studies within Brassicaceae have resulted in a strongly improved classification of the family, as morphologically defined units at and above the generic level were often found to poorly reflect phylogenetic relationships. Here, we focus on tribe Alysseae, which despite its size (accounting for about 7% of all species) has only received limited coverage in previous phylogenetic studies. Specifically, we want to test phylogenetic hypotheses implied by current tribal and generic circumscriptions and to put diversification within tribe Alysseae into a temporal context. To this end, sequence data from the nrDNA ITS and two plastid regions (ndhF gene, trnL-F intergenic spacer) were obtained for 176 accessions, representing 16 out of 17 currently recognized genera of the tribe, and were phylogenetically analysed, among others, using a relaxed molecular clock. Due to large discrepancies with respect to published ages of Brassicaceae, age estimates concerning Alysseae are, however, burdened with considerable uncertainty. The tribe is monophyletic and contains four strongly supported major clades and Alyssum homalocarpum, whose relationships among each other remain uncertain due to incongruences between nuclear and plastid DNA markers. The largest genus of the tribe, Alyssum, is not monophyletic and contains, apart from A. homalocarpum, two distinct lineages, corresponding to sections Alyssum, Psilonema, Gamosepalum and to sections Odontarrhena and Meniocus, respectively. Clypeola, whose monophyly is supported only by the plastid data, is very closely related to and possibly nested within the second Alyssum lineage. Species of the genus Fibigia intermingle with those of Alyssoides, Clastopus, Degenia, and Physoptychis, rendering Fibigia polyphyletic. The monotypic genera Leptoplax and Physocardamum are embedded in Bornmuellera.

Plant species diversity of Adriatic islands: An introductory survey

Abstract The European Mediterranean region is one of the worlds major centres of biodiversity. Research on plant species diversity in this region has neglected the area along the eastern part of the Adriatic basin in comparison to the other Mediterranean areas. The main aim of this study was to focus on this neglected area, by supplying data which is at present lacking in order to discuss the species-area relationship (SAR), floristic richness and endemism of the Adriatic islands and coast. Floristic data for 106 Croatian islands collected by different authors were integrated, systematised and presented in a form usable by other researchers worldwide. The Power (Arrhenius) function was used for modelling (by non-linear regression) the SAR. Residuals around the regression curve (as indicators of floristic diversity without the influence of island area) were calculated for each island. The proportion of endemics in the total island flora varies from 0% up to 28.6%, and 17.5% for narrow endemics. Floristic richness of the broader region was estimated (on the basis of SAR extrapolation) on 1807 species for all Croatian islands, and on 2797 species for the entire Mediterranean area in Croatia.

Disentangling relationships among the diploid members of the intricate genus Knautia (Caprifoliaceae, Dipsacoideae)

The genus Knautia (Caprifoliaceae, Dipsacoideae) encompasses 40-60 species mainly distributed in western Eurasia, with highest species diversity in the Alps and the Balkan Peninsula. It is traditionally regarded as one of the taxonomically most challenging European genera due to the widespread occurrence of polyploidy, the high incidence of hybridisation and the maintenance of morphologically intermediate forms. A prerequisite for assessing the complex spatiotemporal diversification of a polyploid group is a comprehensive hypothesis of the phylogenetic relationships among its diploid members. To this end, DNA sequence data (nrDNA ITS and plastid petN(ycf6)-psbM) combined with AFLP fingerprinting were performed on 148 diploid populations belonging to 35 taxa. Phylogenies obtained by maximum parsimony and Bayesian analyses were used to test the monophyly of the genus and its three sections Trichera, Tricheroides and Knautia, to provide insights into its evolutionary history and to test previous hypotheses of inter- and intrasectional classification. Both nuclear and chloroplast datasets support the monophyly of Knautia and its three sections, with ambiguous placement of K. cf. degenii. The majority of species belong to the nearly exclusively perennial section Trichera (x=10). Within section Trichera all markers revealed largely unresolved phylogenetic relationships suggesting rapid radiation and recent range expansion. In addition, extensive sharing of plastid haplotypes across taxa and wide geographic ranges of plastid haplotypes and ribotype groups were observed. The molecular data are partly at odds with the traditional informal grouping of taxa within section Trichera. Whereas the traditional groups of K. dinarica, K. drymeia and K. montana can be maintained, the new, smaller and well supported Midzorensis and Pancicii Groups as well as the SW European Group are separated from the heterogeneous traditional K. longifolia group. The former groups of K. arvensis, K. dalmatica, K. fleischmannii and K. velutina are clearly polyphyletic. Their diploid members have to be rearranged into the Xerophytic Group, the Carinthiaca Group, and the Northern and Southern Arvensis Groups. The annual sections Tricheroides (x=10) and Knautia (x=8) with only a few taxa are resolved in the ITS and plastid trees on long branches as early diverging lineages within the genus.

Heteroploid Knautia drymeia includes K. gussonei and cannot be separated into diagnosable subspecies

PREMISE OF THE STUDY Knautia drymeia is a morphologically variable, diploid and tetraploid temperate forest understory species distributed in southeastern Europe and adjacent areas. The species is an excellent system to explore the influence of polypoidy on taxonomic delineations, the role of hybridization among genetically distant populations in polyploid evolution, and the impact of glacial refugia on the evolution of polyploids. METHODS Amplified fragment length polymorphism fingerprinting and multivariate analyses of morphological characters were performed on 57 populations spanning the distribution area of K. drymeia. K-means clustering, comparison of in-silico tetraploids and observed tetraploids, and a phylogeographic analysis using relaxed random walks were used to explore the genetic structure within the diploids, to infer the origin of the tetraploids and to reconstruct range expansion through time. Further, we contrasted the morphology and genetic groups with current taxonomy and evaluated the status of the tetraploid Apennine endemic K. gussonei and the intraspecific taxa of K. drymeia. KEY RESULTS The genetic structure was strongly geographically correlated and yielded four genetic groups; K. gussonei was inseparable from K. drymeia. Distributions of diploid lineages are suggestive of glacial refugia in the northwesternmost and southeastern Balkan Peninsula. Polyploids originated at least two times, as autopolyploids and probably additionally also as allopolyploids. Morphological divergence corresponded with neither genetic groups nor current taxonomy. CONCLUSIONS Genetic and morphometric data confirmed neither divergence of K. gussonei nor recognition of subspecies within K. drymeia. We therefore propose treating K. drymeia as a morphologically and genetically variable species without infraspecific taxa.

Campanula Skanderbegii: Molecular and Morphological Evidence of a New Campanula Species (Campanulaceae) Endemic to Albania

Abstract A new species of Campanula from Central Albania, named C. skanderbegii, belonging to isophyllous Campanula ser. Garganicae is illustrated and described. Morphologically and phylogenetically it is well differentiated from other species of this series. It is morphologically similar to C. portenschlagiana from Central Dalmatia (Croatia) based on corolla shape. Phylogenetic analyses, maximum parsimony, maximum likelihood, and Bayesian inference, based on nuclear ITS and chloroplast trnL—trnF data support C. skanderbegii as a clearly distinct taxon within the isophyllous species of Campanula. A distribution map for C. skanderbegii and a key to all species of Campanula ser. Garganicae are provided. Because of its rarity and highly restricted distribution, C. skanderbegii should be included in the IUCN red list as a critically endangered species.

Genetic Diversity and Demographic History of Wild and Cultivated/Naturalised Plant Populations: Evidence from Dalmatian Sage (Salvia officinalis L., Lamiaceae)

Dalmatian sage (Salvia officinalis L., Lamiaceae) is a well-known aromatic and medicinal Mediterranean plant that is native in coastal regions of the western Balkan and southern Apennine Peninsulas and is commonly cultivated worldwide. It is widely used in the food, pharmaceutical and cosmetic industries. Knowledge of its genetic diversity and spatiotemporal patterns is important for plant breeding programmes and conservation. We used eight microsatellite markers to investigate evolutionary history of indigenous populations as well as genetic diversity and structure within and among indigenous and cultivated/naturalised populations distributed across the Balkan Peninsula. The results showed a clear separation between the indigenous and cultivated/naturalised groups, with the cultivated material originating from one restricted geographical area. Most of the genetic diversity in both groups was attributable to differences among individuals within populations, although spatial genetic analysis of indigenous populations indicated the existence of isolation by distance. Geographical structuring of indigenous populations was found using clustering analysis, with three sub-clusters of indigenous populations. The highest level of gene diversity and the greatest number of private alleles were found in the central part of the eastern Adriatic coast, while decreases in gene diversity and number of private alleles were evident towards the northwestern Adriatic coast and southern and eastern regions of the Balkan Peninsula. The results of Ecological Niche Modelling during Last Glacial Maximum and Approximate Bayesian Computation suggested two plausible evolutionary trajectories: 1) the species survived in the glacial refugium in southern Adriatic coastal region with subsequent colonization events towards northern, eastern and southern Balkan Peninsula; 2) species survived in several refugia exhibiting concurrent divergence into three genetic groups. The insight into genetic diversity and structure also provide the baseline data for conservation of S. officinalis genetic resources valuable for future breeding programmes.

Campanula aureliana (Campanulaceae), a new species from Albania

Campanula is a species-rich genus with high variability of the morphological traits, controversial taxonomic treatments within the Mediterranean basin as a species diversity center. One of the monophyletic groups in the genus is the Campanula series Garganicae Trinajstić distributed in the amphi-Adriatic and Ionian region. The group as currently delimited encompasses 11 taxa, mostly with highly restricted distributional ranges. In the present study plants previously named as Campanula garganica Ten. var. albanica Markgr. are described and illustrated as an isophyllous species from central Albania, for which name Campanula aureliana Bogdanović, Rešetnik, Brullo & Shuka is proposed. The results of phylogenetic analyses based on nuclear ITS and chloroplast trnL–trnF data support C. aureliana as a clearly distinct taxon within the Campanula ser. Garganicae. Its description is given, and diagnostic morphological and SEM seed micromorphological characters of closely related species are compared and discussed. Morphologically C. aureliana shows close affinity with C. fenestrellata Feer, mainly for the habit, shape and size of the corolla, but differs in a lot of significant features. Information on the ecology and conservation status of the newly described species is presented.

Taxonomic revision of the genus Fumana (Cistaceae) in Croatia

Abstract Taxonomic and herbarium revision of the genus Fumana (Dunal) Spach (Cistaceae) coupled with field surveys in Croatia yielded several interesting new data. Two overlooked and poorly known species, Fumana scoparia Pomel and Fumana laevis (Cav.) Pau, are confirmed for Croatian flora. The existence of Fumana ericoides (Cav.) Gand. has not been confirmed and all actual Croatian populations are ascribable to morphologically closely related Fumana ericifolia Wallr. Particular attention is paid to the confusion between the two taxa in the Mediterranean flora, and morphological features for their distinction are specified. Consequently, seven taxa are currently regarded as being present in Croatia: Fumana arabica (L.) Spach, F. ericifolia Wallr., Fumana laevipes (L.) Spach, F. laevis (Cav.) Pau, Fumana procumbens (Dunal) Gren. et Godr., F. scoparia Pomel and Fumana thymifolia (L.) Spach ex Webb. Each taxon is described and diagnostic characters, such as the indumentum type and seed morphology are given. An identification key, original line drawing for F. scoparia and distribution maps for all species of the genus Fumana occurring in Croatia are provided.

Taxonomic notes on Puccinellia teyberi (Poaceae), a critical species of Croatian flora

Puccinellia teyberi Hayek (Poaceae) is a critical species of the Croatian flora, described for the first time as Atropis rupestris Teyber from two small islets of central Adriatic Sea. Nomenclature, lectotypification, morphology, karyology, leaf anatomy, palynology, ecology and conservation status of this taxon are examined. According to these data, it must be treated as a distinct species, taxonomically related to P. convoluta and P. festuciformis. Besides, it represents a neoendemic apomictic species, confined to rocky costal places and having a punctiform distribution.