Toni Nikolić

Molecular phylogeny of the Campanula pyramidalis species complex (Campanulaceae) inferred from chloroplast and nuclear non-coding sequences and its taxonomic implications

The Campanula pyramidalis complex is a group of closely related taxa with a distribution across the Balkans, from the Gulf of Trieste in the north to the Peloponnese Peninsula in the south, with small disjunct parts of the range in the south Apennines. Although 21 taxa were described within this complex, only three, C. pyramidalis, C. versicolor, and C. secundiflora, have been generally accepted in recent synoptical taxonomic treatments. Our molecular phylogenetic analyses based on sequences of three non-coding chloroplast regions (psbA-trnH, psbZ-trnfM, trnG-trnS) as well as of nuclear ribosomal internal transcribed spacers (nrITS), lend strong support to the recognition of several lineages which only partially correspond to generally accepted taxonomic concepts. Molecular data presented in this study showed that C. pyramidalis is a polyphyletic assemblage that segregates into three distinct lineages, one of which is described here as a new species, C. austroadriatica sp. nov. The lectotype of C. pyramidalis, redefined in a strict sense, is designated. Neither C. versicolor nor C. secundiflora were found to be strictly monophyletic, but their monophyly could not be rejected. Morphological and biogeographical implications are discussed.

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.

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.

Estimating plant species occurrence in MTB/64 quadrants as a function of DEM-based variables: a case study for Medvednica Nature Park, Croatia

Abstract Croatia is among those European countries without an Atlas of Flora produced till today as a result of constant lack of greater number of active botanists and inconsistency in gathering data in the field. Recently, a standard for collection of data, based on the Central European MTB (abbreviation of German term “Mestischblatter” that stands for a sheet of topographic map) grid was proposed and tested in the field on the “Medvednica Nature Park” on Medvednica mountain near the city of Zagreb. Using the data collected in 97 MTB/64 quadrants (presence/absence of plant species), we tested the potential of estimating species occurrence at the proposed grid by models in a function of the Digital Elevation Model (DEM)-based variables, namely altitude, terrain slope, terrain aspect, and flow accumulation potential. Because of significant spatial variability of environmental factors within MTB/64 quadrants, each one was represented by descriptive statistics (median, 5-, 25-, 75- and 95-percentiles) of DEM-based variables. Thirty-seven plant species were selected arbitrarily, on the basis of their frequency in the studied area (40–60% of all quadrants). Three methods for development of predictive model were used and compared: discriminant analyses, logistic regression, and classification trees. Yielded results suggest that spatial modelling could be probably applied in flora mapping, which would optimise fieldwork. However, decreasing of mapping unit area is recommended, especially for rare species. For larger areas, inclusion of other environmental predictors (macroclimatic, lithological, landuse) in models is probably needed.

Relations between western Balkan endemic Campanula L. (Campanulaceae) lineages: Evidence from chloroplast DNA

Abstract The relationships among western Balkan and some amphi-Adriatic endemic campanulas were studied using RFLP analysis of the 1.8-kb trnT – trnF chloroplast DNA region. The results were compared to those of a comparative floral morphometry study performed earlier on a similar sample. Maximum parsimony and distance methods produced very similar results to those of the floral morphometry, both of which point to a clear separation of the taxa into three main clades. The first isophyllous, or Garganicae series, clade comprised Campanula poscharskyana, C. portenschlagiana, C. garganica, C. reatina and two C. fenestrellata-subspecies. The second “isophylloid” clade consisted of C. pyramidalis agg., C. waldsteiniana agg. and C. carpatica. The third, heterophyllous clade was very uniform, comprising undistinguishable taxa of the C. rotundifolia complex and alpine endemic C. cochleariifolia. Interestingly the Ligurian stenoendemic C. isophylla was clearly separated from the members of the ser. Garganicae, C. reatina was the most distant taxon in the Garganicae clade, the controversial species Asyneuma pichleri was nested inside the Campanula core, while Campanula carpatica was closer to the “isophylloids” than to the sect. Rapunculus Boiss. In order to resolve the taxonomic uncertainty among the closely related heterophyllous taxa more refined molecular methods, e.g. AFLP or microsatellites, should be applied.

Terrestrial Biodiversity Analyses in Dalmatia (Croatia): A Complementary Approach Using Diversity and Rarity

Here we present the methodology used for terrestrial biodiversity analysis and site selection in Phase B of the UNDP/GEF COAST project. The analysis was focused on the problem of biodiversity evaluation in four Croatian counties stretching from sea level to the highest mountain in Croatia. Data on habitats, vascular flora, and fauna (mammals, birds, reptiles, amphibians, butterflies, ground beetles, and underground invertebrates) were collected and analyzed for each of the four counties. Emphasis was given to the richness of endangered species and the rarity of endemic species. Based on the spatial analyses of habitat, fauna, and flora data, four to six areas were selected from each county and ranked according to their biodiversity importance. Overlap between areas important for richness and those important for rarity was highest for data on flora (65.5%) and lowest for data on fauna (16.7%). When different data sets were compared, the lowest overlap was between flora and fauna (17.1%) and largest between fauna and habitats (23.9%). Simultaneous overlap among all three data sets was found in just 6.5% of the overall selected areas. These results suggest that less specific data, with respect to taxa threat status, could better serve as surrogate data in estimating overall biodiversity. In summary, this analysis has demonstrated that Dalmatia is a region with a high overall biodiversity that is important in a broader European context.

Relations of the western Balkan endemic Campanula L. (Campanulaceae) lineages based on comparative floral morphometry

Abstract In the north-eastern quadrant of the Mediterranean Region many endemic Campanula (Campanulaceae) taxa and lineages arose, of which the Isophylla group and the Rotundifolia complex (sect. Heterophylla), essentially separated on the basis of their leaf shapes, and some of the endemic western Balkan, Dinaric lineages (i.e. ‘isophylloid’ aggregates Waldsteiniana and Pyramidalis) are members. The aim of this study is to revise the mutual morphological floral relations of these endemic lineages, on the basis of 14 taxa, 33 populations and 409 flower samples, and measure simple flower traits and their ratios. For data interpretation, a phenetic approach using multivariate analyses was applied, as a first attempt in understanding biometrical floral relations between the similarly distributed Campanula lineages, and to provide a fundamental background for the cladistic and molecular analyses that will follow. Two ‘natural’Campanula groups, isophyllous/isophylloid and heterophyllous, are identified according to their floral properties. The floral pattern of Campanula waldsteiniana and C. tommasiniana are clearly separated, by the corolla shapes, into isophyllous and heterophyllous, respectively, while C. pyramidalis follows the isophyllous pattern, but is plainly recognizable.

Simultaneous RP‐HPLC‐DAD Separation, and Determination of Flavonoids and Phenolic Acids in Plantago L. Species

A rapid reversed‐phase (RP) high‐performance liquid chromatography method was developed and applied for simultaneous separation, and determination of flavonoids and phenolic acids in eight Plantago L. taxa (P. altissima L., P. argentea Chaix, P. coronopus L., P. holosteum Scop. ssp. depauperata Pilger, P. holosteum ssp. holosteum, P. holosteum ssp. scopulorum (Degen) Horvatić, P. lagopus L., and P. maritima L.) growing in Croatia. Chromatographic separation was carried out on Zorbax Eclipse XDB‐C18 using gradient elution with a H2O (pH 2.5, adjusted with CF3COOH) and MeCN mixture at 30°. The contents of analyzed phenolic compounds (% of the dry weight of the leaves, dw) varied among examined species: rutin (max. 0.024%, P. argentea), hyperoside (max. 0.020%, P. lagopus), quercitrin (max. 0.013%, P. holosteum ssp. holosteum), quercetin (max. 0.028%, P. holosteum ssp. scopulorum), chlorogenic acid (max. 0.115%, P. lagopus), and caffeic acid (max. 0.046%, P. coronopus). Isoquercitrin was detected only in P. argentea (0.020%), while isochlorogenic acid content was below limit of quantification in all investigated species. Multivariate analyses (UPGMA and PCA) showed significant differences in contents of investigated polyphenolic compounds between different Plantago taxa. Accordingly, investigated substances might be employed as chemotaxonomic markers in the study of the complex genus Plantago.

An evaluation of floristic diversity in Medvednica Nature Park (northwestern Croatia)

Abstract Mount Medvednica (northwestern Croatia), mostly consisting of slates and marls, rises to the north, above the Croatian capital of Zagreb, from 300 to 1,035 m a.s.l. A floristic research was carried out in the area of Medvednica Nature Park, which consists mainly of the western part of the mountain. On the whole, 1,352 taxa of vascular plants were recorded from all sources (field observations, literature and herbaria). Out of these, 48 taxa are strictly protected, 228 are protected, 67 taxa are considered threatened according to the IUCN categories, and 9 taxa are endemic for the Croatian flora. The life-form analysis of the investigated flora shows the predominance of hemicryptophytes, followed by therophytes and geophytes, while the chorological analysis has evidenced the prevalence of Euro-Asiatic, South European and cosmopolite geographical groups. The biological/chorological cross-spectrum was laid out by considering the percent of every life-form for each geographical element. In order to evaluate the floristic richness of the studied area, the α-index was calculated and then compared with the α-indices of similar regions in Croatia and several European countries. A floristic diversity map of Medvednica Nature Park is presented here for the first time.

Invasive alien plants in Croatia as a threat to biodiversity of South-Eastern Europe: Distributional patterns and range size

During the analysis of alien and invasive flora of Europe, as a threat to biodiversity, data for Croatia were missing. The aim of our research was to analyse distributional patterns and range size of all invasive alien plants (64) for the state area (57,000 km(2)). They were detected on 49% of the state territory, averaging five taxa per 35 km(2). The greatest number of invasive plants (>30 per grid cell) was recorded in the major urban centres, increasing in the south-east direction and reflecting positive correlation with temperature and negative with altitude. The most endangered areas are in the Mediterranean region, especially on islands. The number of invasive plants increased with habitat diversity and almost 75% of all sites with invasive plants are located within a few habitats with direct anthropogenic influence. The results should provide a reliable regional and global basis for strategic planning regarding invasive alien plants management.