Sanja Kovačić

Phylogeny and Biogeography of Isophyllous Species of Campanula (Campanulaceae) in the Mediterranean Area

Abstract Sequence data from the nuclear internal transcribed spacer (ITS) were used to infer phylogenetic relationships within a morphologically, karyologically, and geographically well-defined group of species of Campanula (Campanulaceae), the Isophylla group. Although belonging to the same clade within the highly paraphyletic Campanula, the Rapunculus clade, members of the Isophylla group do not form a monophyletic group but fall into three separate clades: (i) C. elatines and C. elatinoides in the Alps; (ii) C. fragilis s.l. and C. isophylla with an amphi-Tyrrhenian distribution; and (iii) the garganica clade with an amphi-Adriatic distribution, comprised of C. fenestrellata s.l., C. garganica s.l., C. portenschlagiana, C. poscharskyana, and C. reatina. Taxa currently classified as subspecies of C. garganica (garganica, cephallenica, acarnanica) and C. fenestrellata subsp. debarensis are suggested to be best considered separate species. The molecular dating analysis, although hampered by the lack of fossil evidence, provides age estimates that are consistent with the hypothesis that the diversification within the garganica clade was contemporaneous with the climatic oscillations and corresponding sea-level changes during the late Pliocene and Pleistocene. Dispersal-vicariance analysis suggests that the garganica clade originated east of the Adriatic Sea, from where it reached the Apennine Peninsula.

Screening toxicity study in young carp (Cyprinus carpio L.) on feed amended with fumonisin B1

Fumonisin B1 (FB1) is one of several mycotoxins produced by Fusarium moniliforme, a major fungal pathogen of corn and widely spread throughout the world. FB1 produces a wide range of biological effects, some of which are specific for particular organs or species and some are common to all investigated animals. In this study we have evaluated subchronic toxicosis features in young carp (Cyprinus carpio L.) exposed to 0.5 and 5.0 mg FB1 kg−1 body weight for 42 days through nutritionally balanced diet. During the trial we observed loss of body weight in both treated groups, together with higher incidence of infective bacterial dermatological lesions erythrodermatitis cyprini (Aeromonas salmonicida subsp. nova) in the group treated with the higher FB1 dose. Several hematological parameters (erythrocyte count, platelet count) and serum chemical concentrations (creatinin, total bilirubin) and activities (aspartate aminotransferase, AST and alanine aminotransferase, ALT) were greater in the fumonisin treated groups than in the control group. Our results indicate that long-term dietary exposure to 0.5 and 5.0 mg FB1 kg−1 body weight is not lethal to young carp, but can produce adverse physiological effects. These findings also suggest that primary target organs of FB1 in the carp are kidney and liver, as it has already been observed in other animal species tested. Specifically changed red blood cell-parameters reveal that FB1 probably causes erythrocyte membrane defect or interferes with carps respiratory process.

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.

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.

Fumonisin B1 Neurotoxicity in Young Carp (Cyprinus carpio L.)

Fumonisin B1 Neurotoxicity in Young Carp (Cyprinus Carpio L.) For years scientists have suspected that the environment plays a role in neurodegenerative diseases such as Alzheimers disease, Parkinsons disease, and multiple sclerosis. Mycotoxin fumonisin B1 (FB1) is produced by several Fusarium species, mainly by Fusarium verticilioides, which is one of the most common fungi associated with corn worldwide. Fumonisins are known to cause equine leukoencephalomalacia, a disease associated with the consumption of corn-based feeds contaminated with FB1. Here we have reported chronic experimental toxicosis in one-year-old carp (Cyprinus carpio L.) receiving feed containing 100 mg kg-1 or 10 mg kg-1 of added FB1 for 42 days. We focused on fumonisin toxicity in the fish brain. After staining with hemalaun-eosin, histology of the fish brain revealed vacuolated, degenerate, or necrotic neural cells, scattered around damaged blood capillaries and in the periventricular area. These findings suggest that fumonisin, although it is a hydrophilic molecule, permeated the blood-brain barrier of young carp and had a toxic effect on neuronal cells. Neurotoksičnost fumonizina B1 u šaranske mLađi (Cyprinus carpio L.) Odavno je poznato da okoliš ima važnu ulogu u razvoju neurodegenerativnih bolesti kao što su Alzheimerova i Parkinsonova bolest te multipla skleroza. Mikotoksin fumonizin B1 (FB1) tvori nekoliko vrsta Fusariuma, najčešće F. verticillioides, koja je najučestaliji kontaminant kukuruza. Ovaj mikotoksin odgovoran je za leukoencefalomalaciju konja, mula i magaradi povezanu s konzumacijom kukuruza kontaminiranog s FB1. U ovom su radu prikazani rezultati kronične eksperimentalne toksikoze mlađi šarana (Cyprinus carpio L.) koji su u hrani primali 100 mg kg-1 i 10 mg kg-1 FB1 tijekom 42 dana. Nakon bojenja hemalaun-eozinom zabilježene su značajne histopatološke promjene na mozgu životinja uključujući vakuolizaciju, degeneraciju i nekrozu neurona, posebice u blizini oštećenih krvnih kapilara i u periventrikularnoj regiji. Ova saznanja pokazuju da FB1, kao hidrofilna molekula, prolazi kroz krvno-moždanu barijeru mladih šarana uzrokujući oštećenje neurona.

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.

The Chrysopogono grylli-Nerietum oleandri association in Croatia as compared with other Rubo ulmifolii-Nerion oleandri communities (Tamaricetalia, Nerio-Tamaricetea) in the Mediterranean

Abstract A new association, Chrysopogono grylli-Nerietum oleandri, situated in the south of Croatia, is proposed. According to its floristic structure, physiognomy and ecology, it belongs to the Rubo ulmifolii-Nerion oleandri alliance (Tamaricetalia, Nerio-Tamaricetea). It is the only association with Nerium oleander in Croatia. This community has developed along a temporary watercourse as a vegetation exclave (extrazonal stand) within the climazonal evergreen forest vegetation of the Oleo-Ceratonion alliance. The association has a thermophilous character and a large number of companions from the Quercetalia ilicis and Scorzonero-Chrysopogonetalia communities. This report compares the floristic composition of this association with others of the Rubo ulmifolii-Nerion oleandri alliance from other Mediterranean areas. The Croatian association is most similar to the Spartio-Nerietum oleandri association from the Ionian side of Calabria (south Italy). The syntaxonomic status of the Montenegrin association Andropogono distachyi-Nerietum oleandri was determined, and the status of the stands with Nerium oleander is discussed.

Vegetation of Croatia: Phytosociological classification of the high-rank syntaxa

Abstract Croatia is among the most ecologically diverse and floristically rich countries in Europe, with a great variety of communities. The vegetation elaboration according to the standard central European method was initiated in Croatia at the beginning of the 20th century. In previous overviews of Croatian vegetation, the number of classes and alliances was underrepresented in relation to the country’s floristic richness. Furthermore, the level of knowledge and the amount of available data varied greatly among the various types of vegetation. The aims of this paper are mainly to compile a stabile syntaxonomic list of classes, orders and alliances dominated by vascular plants in Croatia and to adjust Croatian vegetation to the new European syntaxonomic system (EuroVegChecklist). It introduces a consistent description of high-rank syntaxa in Croatian. In conclusion, the vegetation of Croatia comprises 66 classes, 121 orders and 201 alliances. The number of syntaxa shows vegetation diversity that is rather high compared to most other European countries; this is related to the high floristic richness and endemism. The list points out the obvious problems and gaps in our knowledge of vegetation in Croatia and can serve as a baseline for the future vegetation studies.

Composition and ecology of the Quercus coccifera L. communities along the eastern Adriatic coast (NE Mediterranean)

The aim of this study is to classify and describe all plant communities with Quercus coccifera covering the entire eastern Adriatic coast and islands from north Croatia to south Albania, and to relate their species composition, chorotypes and life forms to environmental factors using Pignatti ecological indicator values. From total 70 phytosociological relevés, we identified and described four floristically and ecologically distinctive vegetation communities (two new proposed subassociations, one association and stand each) using TWINSPAN and the Braun-Blanquet classification scheme. In Croatia and Montenegro, Q. coccifera is forming macchia within the Fraxino orni–Quercetum cocciferae pistacietosum lentisci. Quercus coccifera occurs only sparsely in south Croatia as a shrubland within Fraxino orni–Quercetum cocciferae nerietosum oleandri subassociation or macchia within the Erico arboreae–Arbutetum unedonis association. Despite the difference in biogeographic position and bioclimates, low shrubby Albanian Q. coccifera stands are more closely related to the Q. coccifera communities from the western Mediterranean. Eastern Adriatic communities appear exclusively within the Querceta ilicis vegetation zone and spread within the meso-Mediterranean belt. They nevertheless are an important part of the regions natural heritage and management plans must ensure that all forms of land are used in a sustainable way.

Syntaxonomy of Arundo stands along the eastern Adriatic coast

Based on the diagnostic morphological characters of the Arundo taxa, and after both field and herbarium observation, three species (A. micrantha, A. plinii and A. donax) were found along the eastern Adriatic coast. Altogether, 50 phytocoenological relevés from Arundo stands were collected in Croatia, Montenegro and Albania between 2010 and 2013. Three associations with a predominance of Arundo species were determined: Arundinetum plinianae, Arundinetum micranthae and Arundini donacis–Convolvuletum sepium. Among them, Arundinetum micranthae is described and proposed as a new association, belonging to the Artemisietea vulgaris class. It occurs frequently along the coastal belt between the cities of Split and Dubrovnik in South Croatia. The syntaxonomy of the associations is discussed and comparison with similar communities from Italy is made. The vegetation and floristic structure of the associations is discussed as a potential basis for biodiversity conservation programmes and for evaluating the state of these ecosystems in the future.