Search

Sweet chetsnut (Castanea sativa Mill.) has been cultivated in semi-wild orchards of Sub- Mediterranean Croatia for centuries. Two such historical cultivation area are Lovran and Lovrin on the Istrian peninsula, while the third one is located on the island Cres. From these three cultivation areas, both grafted and ungrafted individual trees were sampled. Six genomic simple sequence repeat (gSSR) and nine EST-derived SSR (EST-SSR) loci were used to explore population diversity and clonality, as well as the population genetic structure. Five closely related clones have been found within the grafted group. Within the ungrafted group, many hybrids resulting from spontaneous cross-pollination between the cultivated and wild chestnut have been found. Thus, the three analyzed populations have varying levels of diversity and clonality. The analyzed semi-wild orchards defined by a diverse genetic structure represent a hotspot for further selection and could result in the creation of locally adapted, high-yielding varieties of chestnut.

We reconstructed the relationships among populations of Campanula secundiflora s.l. and closely related and geographically close populations of C. austroadriatica and C. versicolor. Based on analyses of microsatellite DNA data, the investigated populations have high overall genetic diversity and abundant allelic variation over seven investigated loci. Bayesian model-based clustering identified four clearly differentiated genetic groups of populations. The genetic differentiation was reflected by morphological differentiation, allowing us to propose a new taxonomic treatment of the constituents of C. secundiflora s.l. The populations distributed in south-western Serbia and north-eastern Montenegro were included in C. secundiflora. A new species, Campanula montenegrina sp. nov., distributed in the continental part of Montenegro and the northern part of Albania is described

Salvia brachyodon Vandas or short‐tooth sage is a clonal, narrow endemic and endangered plant species with only two known populations: on Pelješac peninsula (Croatia) and Mt. Orjen (Bosnia and Herzegovina). In order to assess the levels of its genetic diversity, 120 individuals from Pelješac peninsula population were genetically analysed using eight microsatellite markers. With intention of reaching fine‐scale intra‐population spatial genetic structure, geographic coordinates for each plant sample were recorded. Given results confirmed the highly clonal structure of this species. Spatial genetic analysis revealed large and tightly packed stands of ramets suggesting the propagation by underground stolones. For small and isolated population, such reproductive strategy is maybe an advantage in the short‐term scale, but bearing in mind that clonality can have development of inbred population with low fecundity for a consequence, it also presents a great threat. In addition, population bottleneck was also detected, confirming the high vulnerability of this population. Ecological succession and disappearance of suitable open‐type habitat are recognized as the likely causes of such dramatic reduction in population size. In contrast, large scale wild fire that has overrun this location 12 years ago is presumably population saviour. In order to protect this population, agro‐pastoral system that was once present in this location, should be encouraged and re‐ established.

Salvia officinalis L. is a perennial shrub native to eastern Adriatic coast and central and southern Appenines. Although well-known and valued for its high essential oils content, there is a significant lack of knowledge regarding its past and present native distribution. To obtain evidence of population genetic structure and phylogeography of this species, 62 natural populations sampled throughout entire distribution area were analysed using eight SSR markers. The most of the populations grouped together in accordance with their geographical position. The highest levels of genetic variability and allelic richness were observed in populations from the central and southern Adriatic. In contrast to populations from the Balkan Peninsula, Apennine populations are characterized by high differentiation levels suggesting their long- lasting genetic isolation. Based on obtained results, three glacial microrefugia were identified, two in Balkan Peninsula and one in southern Apennines. Possible dynamics and directions of post-glacial recolonization paths of S. officinalis were discussed.

Salvia officinalis L. (Dalmatian sage) is a perennial shrub native to eastern Adriatic coast and central and southern Apennines. From the Ancient times it is well-known and valued for its high essential oils content and in nowadays, its dried leaf (Salviae folium) is an authorized drug in most pharmacopoeias. To define the extent of diversity existing in 25 Dalmatian sage populations from eastern Adriatic coastal region, biochemical analysis of essential oils and DNA fingerprinting via microsatellite markers were done. Essential oils were analysed using analytical gas chromatography (GC/FID) and gas chromatothography - mass spectrometry (GC/MS) techniques to check for biochemical variability. Using multivariate analyses on the basis of eight major compounds detected, three chemotypes were distinguished: (A) cis-thujone, (B) trans-thujone and (C) camphor/β- pinene/borneol/bornyl acetate. In order to assess the amount and structure of population genetic diversity in analysed populations, eight microsatellite markers were used. A total of 186 alleles was found with a mean value of 20.13 alleles per locus. The highest values of heterozigosity and allelic richness were observed in most southern population, suggesting their high potential for possible breeding programs. In addition, levels of inbreeding and heterozygosity excess were also analysed, enabling recognition of the population characterized by unsuitable genetic structure for further selection. The implications of the given results in possible breeding programs involving S. officinalis are discussed.

Dalmatian sage (Salvia officinalis L.) and Greek sage (S. fruticosa Mill.) are two closely related species. While Dalmatian sage is naturally widespread on the Apennines and eastern Adriatic coast, Greek sage is naturally widespread in the eastern Mediterranean. The island of Vis is the only place in Croatia where S. fruticosa has been noticed. The presence of this species in Croatia, far away from its natural range, may be related to ancient Greek colonisation of the island in the fourth century BCE. From that time Greek sage probably escaped from gardens and vineyards and successfully naturalised western part of the island. Besides sharing the same habitat on the island of Vis, Dalmatian and Greek sage have the same chromosome number (2n = 14) and their flowering times overlap. The fact that the artificial hybrids between these two species are known and commercially exploited encouraged us to study spontaneous hybridization on the island of Vis. A total of 79 plant samples representing both parental species and putative hybrids were analysed using morphological and microsatellite (SSR) markers. Results of both approaches strongly supported the existence of the hybridisation events. Evolutionary, conservation and especially commercial implications of this hybridization were discussed.

Short-toothed sage (Salvia brachyodon Vandas) is Illyrian-Adriatic endemic species. It is reliably known to grow only in two localities: at the summit Sv. Ilija on Pelješac peninsula, Croatia and near the village Vrbanj on Mt. Orjen at the border between Bosnia and Herzegovina and Montenegro. In order to study population diversity of short-toothed sage, the samples from both localities have been analysed using 16 morphological traits and eight microsatellite primers previously developed for Dalmatian sage (Salvia officinalis L.) and successfully cross- amplified in short-toothed sage. The morphological analysis revealed that two populations could be discriminated using four morphological traits: inflorescence length, glandular sessile trichome on a flower peduncle, calyx length and calyx appendices length. The short-toothed sage tends to form lax cushions that can cover up to few dozen square meters, thus suggesting clonal reproduction via stolones. The molecular analysis confirmed that the clonal propagation is frequent in short- toothed sage given that out of 180 individual plant samples, only 98 were of unique multilocus genotype.

Microsatellite markers and comparative morphometry were used to distinguish the subtaxa within the Anthylis vulneraria species complex in Croatia and neighbouring countries. Analyses were carried out on 445 samples from 58 different populations. Neighbour-Joining method, based on Nei's distance, as well as the Neighbor-Net diagram, indicates that corolla colour is important distinguishing character. Existence of two dominant genetic units was found. Data showed that microsatellite markers do not discriminate the subtaxa within the A. vulneraria species complex. According to the morphometrical analysis, the most efficient characters to distinguish subtaxa of A. vulneraria were corolla colour, calyx colour and hairiness, flower length, calyx width, bract length, terminal leaflet length of rosette leaf, stem hairiness and uppermost internode length and stem height ratio. The subspecies alpestris was well distinguished from others by its smoke-grey calyx, while the subspecies carpatica, polyphylla and polyphylla 2 were not separated among themselves. In the area of investigation, the subspecies pulchella most probably forms intermediates to ssp. alpestris. The subspecies maura is well distinguished from other red flowered taxa by its longer corolla and the evenly distributed leaves along stem. In Croatia the subspecies praepropera forms three different geographical groups: (i) Istrian and North and Central Dalmatian group, (ii) South Dalmatian group, (iii) remote Adriatic islands group.

Eleven polymorphic microsatellite markers were developed from common sage (Salvia officinalis L). The variability of specific microsatellite regions was assessed in wild population from the eastern Adriadic coast. All eleven primer pairs showed single-locus amplification with a total of 142 alleles and observed heterozygosity ranged from 0.300 to 0.833.

Dalmatian sage is a perennial shrub native of the northern coastal region of the Mediterranean. In Croatia, Dalmatian sage is often gathered from the wild and commercialized as medicinal and aromatic plant. The assessment of genetic diversity is a starting point for its introduction into plant breeding programmes and agricultural production since commercial gathering could have a negative impact on biodiversity conservation. The aim of this study was to assess the amount and structure of population genetic diversity, the geographic differentiation, the occurrence of demographic bottlenecks and the ecotypic divergence. For this purpose five microsatellite markers were developed from enriched genomic DNA libraries and successfully applied for the assessment of genetic diversity in ten Dalmatian sage populations collected in Croatia.

Ljekovita kadulja (Salvia officinalis L.) je višegodišnji polugrm kamenjarskih staništa i jedna od najznačajnijih autohtonih vrsta u flori istočne jadranske obale i otoka. Uz pomoć osam mikrosatelitnih biljega analizirana je genetička raznolikost i populacijska struktura 23 populacije iz Hrvatske te dvije populacije iz Bosne i Hercegovine. Utvrđeno je da je većina populacija u Hardy-Weinbergovoj ravnoteži, dok je kod šest populacija utvrđen signifikantan manjak heterozigota. Južne populacije pokazuju najveću gensku raznolikost kao i alelno bogatstvo, koje se prema sjeveru postupno smanjuje. Vrijednosti genetičke diferencijacije su niske i ukazuju na visoku razinu protoka gena. Analizom molekularne varijance utvrđeno je da je većina ukupne genetičke raznolikosti uzrokovana unutarpopulacijskom raznolikošću. Bayesovske analize populacijske strukture ukazuju na postojanje riječnih barijera u smjeru sjeveroistok-jugozapad (Zrmanja, Cetina) i planinskih masiva u smjeru sjeverozapad-jugoistok (Velebit, Biokovo). Utvrđeno je postojanje izolacije uslijed udaljenosti. Ne postoje jasni znakovi da su populacije prošle kroz nedavno genetičko usko grlo. Dalmatian sage (Salvia officinalis L.) is a perennial subshrub growing in rocky habitats and represents one of the most significant autochthonous species in flora of eastern Adriatic coast and islands. Genetic diversity and population structure of 23 populations from Croatia and two from Bosnia and Herzegovina were analyzed using eight microsatellite markers. The majority of populations were in Hardy-Weinberg equilibrium while significant deficit of heterozygotes was detected in six populations. Southern populations showed the highest values of gene diversity and allelic richness which gradually decreased towards north. Values of genetic differentiation were low indicating high levels of gene flow. Analysis of molecular variance showed that most of total genetic diversity was attributable to within population variation. Bayesian analyses of population structure revealed the existence of riverine barriers in the northeast-southwest direction (Zrmanja, Cetina), as well as mountain ranges (Velebit, Biokovo) in the direction of northwest-southeast. A significant pattern of isolation by distance was observed. There was no clear indication of recent genetic bottlenecks.