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Europski pitomi kesten plemenita je listača od koje imamo višestruku gospodarsku korist. Osim jestivih plodova i kvalitetnog drva, kestenove šume su još od davnina predstavljale važan izvor različitih sirovina. Međutim, intenzivnim korištenjem kestenovih šuma i nasada te pojavom raka kestenove kore, sredinom prošloga stoljeća došlo je do njihovog propadanja i sušenja. Iz tih je razloga u mnogim europskim zemljama pokrenut niz multidisciplinarnih projekta kojima je svrha očuvanje genskih resursa pitomog kestena i njegovog povrataka na nekadašnje značajno mjesto u ruralnim područjima i šumskim ekosustavima. U ovom radu dan je pregled najznačajnijih ­dugoročnih strategija usmjerenih na očuvanje genofonda ove plemenite listače na europskoj razini, kao i prijedlog smjernica za očuvanje genofonda ove vrste in situ i ex situ metodama na području Hrvatske., Sweet chestnut (Castanea sativa Mill.) is a noble, multi-purpose hardwood species. In addition to ­edible fruits and high-quality wood, chestnut forests have been used as a source of various resources since the antiquity. Intensive management of forests and stands, however, together with the emergence of chestnut blight, has led to their decline and dieback. As a result, in numerous European countries multidisciplinary projects were initialized, with goals of conserving the genetic resources of the ­species, as well as reestablishing its significance in rural and forest ecosystems. In this paper, an overview of the most important long-term gene conservation strategies is presented, with suggestions for ­guidelines for application of in situ and ex situ conservation methods in Croatia.

AFLP (Amplified Fragment Length Polymorphism) and SSR (Single Sequence Repeats ; microsatellites) markers are widely used in plant genetic studies and it is fair to say that the fundamental insights into plant population dynamics and structure have been gained from the analysis of either AFLP or SSR data, or both. The AFLP technique requires no prior knowledge of the genetic make-up of a species and produces a large number of polymorphic fragments that are generally scored as dominant markers. SSRs, on the other hand, are species specific, codominant, multiallelic markers. In this study, we wanted to compare some basic population genetic parameters calculated on the basis of both marker systems.

Dalmatian pyrethrum (Tanacetum cinerariifolium (Trevis.) Sch. Bip.) is an endemic species of the eastern Adriatic from the family Asteraceae. The plant produces pyrethrin, a secondary metabolite that has been shown to an effective natural insecticide with little or no adverse effects on humans. To assess genetic diversity, samples were collected from 10 natural populations of Dalmatian pyrethrum along the Adriatic coast and on the islands representing the species' range in Croatia. Genetic diversity analysis was carried out using 12 newly developed microsatellite markers. A total of 121 alleles with an average of 10.08 alleles per locus were detected in the studied populations, indicating high genetic diversity. Allelic richness (Nar) varied from 2.86 (Biokovo) to 5.02 (Zlarin). A total of 36 private alleles were observed in 10 populations with the highest number observed in Konavle (7). Evidence of bottlenecks was found only in the Biokovo population. Analysis of molecular variance showed that most of the genetic diversity can be explained by differences between individuals within populations (87.03%). Bayesian analysis of population structure revealed the existence of two gene pools, the mainland gene pool and the island gene pool. At K = 3, the Biokovo population formed a separate gene pool. Spatial analysis revealed that only a small proportion of genetic differentiation was due to isolation by distance (0.7%), while 40.6% of genetic differentiation was due to isolation by environmental distance. The results of this research will contribute to the development of future breeding programmes and hopefully commercial varieties of T. cinerariifolium to revive Dalmatian pyrethrum production in Croatia and the region. Furthermore, these results will help to improve conservation strategies for the species.

Salvia officinalis L. occupies an important place in the Albanian market for aromatic medicinal plants due to the high content and composition of its essential oil and is the most economically important species for the country. The aim of this study was to investigate the genetic diversity and structure of eight populations of Dalmatian sage from Albania, using eight microsatellite markers (SSRs) previously developed for this species. All eight- microsatellite markers yielded unique and easily detectable amplified fragments that revealed a high level of polymorphism (average of 15 alleles per locus). Overall values of expected heterozygosity (HE) ranged from 0.608 to 0.818, with a mean of 0.721. The highest gene diversity was found in northern Adriatic populations, decreasing towards the southern limit of the species range. Multilocus estimates of Wright’s inbreeding coefficient within populations (FIS) showed no significant deviation from Hardy- Weinberg equilibrium. Analysis of molecular variance showed that most of the total genetic diversity was attributable to differences between individuals within populations (83.14 %) which is to be expected due to outcrossing nature of the species. Bayesian analysis of population structure revealed the existence of two gene pools, the northern gene pool, and the southern gene pool. At K = 3, the Dibër population (the easternmost population in the sample) formed a separate gene pool. As commercial collection of Dalmatian sage in the wild has a negative impact on biodiversity, these results will form the basis for of plant genetic resource conservation and plant breeding programmes for this valuable species.

Most of the common bean (Phaseolus vulagris L.) production in Croatia is based on local landraces, grown by small-scale farmers in low input production systems. The landraces are commonly known by their traditional names based primarily on seed coat pattern and colour. In order to assess the genetic diversity of Croatian common bean landraces, 174 accessions were analysed using a DNA marker for phaseolin type, microsatellite markers and a DArTseq method of SNP genotyping. The information on phaseolin type and the results of the distance- and model-based analysis of population structure coupled with the morphological characterization of the accessions enabled the thorough classification of the accessions, that was, for the most part, congruent. Out of 174 accessions, 135 were assigned to 14 newly formed morpho-genetic groups originating from three distinct domestication events: (1) Mesoamerican, (2) Andean - indeterminate type, and (3) Andean - determinate type. The rest of accessions could represent putative hybrids among morpho-genetic groups including the hybrids among the gene pools. This study would be a first step into a genome-wide association mapping using a panel of Croatian common bean landraces. In conjunction with a detailed data on bioactive nutrient content, a whole genome association mapping analysis will be carried out to identify quantitative trait loci (QTLs) related to bioactive nutrient content.

To understand the role of epigenetic processes in plant adaptation, existing epigenetic variation of natural plant populations has been assessed and compared to genetic variation. Twenty-five Dalmatian sage (Salvia officinalis L.) populations, each consisting of 20 to 25 plants, were analysed using epigenetic Methylation-Sensitive Amplification Polymorphism markers (MSAP) and genetic AFLP markers. Epigenetic and genetic diversity was compared by analysing within-population diversities and pairwise genetic differentiation between populations. The importance of epigenetic processes in divergence and adaptation of Dalmatian sage was analysed by comparing the number of outlier epiloci/loci under divergent selection and by the assessment of the association between epiloci/loci and bioclimatic variables. The identification of FST outlier epiloci/loci was carried out using a Bayesian method while the spatial analysis was used to test the probability of presence of an epiallelic/allelic variant given the environmental conditions of the sampling locations.

In plants, epigenetic variations based on DNA methylation are often transmitted across generations. Epigenetic processes may cause significant heritable variation in phenotypic traits without changing the sequence and thus increase considerably the evolutionary potential of plants in response to abiotic and biotic stress. Thus, heritable epigenetic variation could influence the course of evolution in plants, as it can affect the processes of adaptation and divergence through selection of stable epigenetic variants without involvement of genetic variation. In order to understand the true importance of epigenetic processes, existing epigenetic variation of natural plant populations have been assessed and compared to genetic variation. Three Croatian endemic Salvia species including (1) Salvia officinalis L., (2) Salvia brachyodon Vandas, and (3) Salvia x auriculata Mill. have been used as a study system. The species were chosen for having contrasting range sizes and genetic diversity heavily influenced by different phenomena: clonality (S. brachyodon) and interspecies hybridization (S. x auriculata). S. officinalis is widely distributed along the eastern Adriatic coast while S. brachyodon is a stenoendemic plant of a very narrow range size. S. x auriculata is a natural hybrid between S. officinalis and S. fruticosa. Preliminary results based on the analysis of 25 S. officinalis populations showed that the proportion of epigenetic variance attributable to differences among populations was similar to the proportion of the among-population component of genetic variance. Systematic patterns of epigenetic variation were observed in relation to a number of bioclimatic variables related to amount of precipitation at sampling sites. The investigation of clonal S. brachyodon revealed the existence of substantial epigenetic variation among genetically identical plants (clones). Finally, hybrid S. x auriculata exhibited higher epigenetic diversity in comparison to parental species.

Centaurea ragusina L. (Asteraceae) is a Dalmatian narrow endemic species that grows as typical chasmophyte mostly on inaccessible vertical sea cliffs. It’s represented by two subspecies: typical C. ragusina subsp. ragusina with pinnatifid basal leaves which is distributed from north to south Dalmatia (Dugi Island to Konavle), while C. ragusina subsp. lungensis has mainly entire basal leaves, present only on the islands of Dugi and Kornati archipelago. We used amplified fragment length polymorphisms (AFLPs) to access the genetic diversity, differentiation and structure of populations across its entire distribution range (on 210 individuals from 22 populations). The aims were to test taxonomic validity of existence of two subspecies, to investigate genetic structure, to search for variations in the cpDNA sequences using trnL-trnF and rpL32-trnL markers and to quantify the genetic diversity within and among populations of C. ragusina. The results obtained from analyses of molecular variance (AMOVA) showed that 36.09% of genetic variation can be attributed to differences among populations. Mantel test revealed positive correlation between genetic and geographical distance between pairs of populations. Our study implies that most previous taxonomic infra- specific concepts of C. ragusina were artificial and according to results obtained by BAPS and STRUCTURE there is no clear genetic evidence that support existence of subsp. lungensis as a separate taxon from typical subsp. ragusina. On total of 60 samples, 16 different chloroplast haplotypes were detected. Haplotypes network indicate very fragmented distribution of C. ragusina that can be consequence of evolutionary history and colonization of vertical rocky cliffs during the Pleistocene sea level changes. Recommendations for conservation purposes of C. ragusina based on the patterns of genetic diversity were proposed.

One of the main challenges in molecular ecology and population genetics is the detection of genes, or genomic regions, that have been targeted by natural selection. The aim of this study was to quantify the relative contribution of natural selection in shaping the genetic variation observed among Dalmatian sage (Salvia officinalis L.) populations. Genetic diversity of 25 Dalmatian sage populations from Croatia and Bosnia and Herzegovina, each consisting of 20 to 25 plants, was assessed using AFLP markers. Two alternative methods for the identification of FST outlier loci have been used: the frequentist methods implemented in Mcheza and the Bayesian method implemented in BayeScan. Moreover, the spatial analysis method as implemented in Samada was used to compute multiple univariate logistic regressions to test the probability of presence of an allelic variant for a polymorphic marker given the environmental conditions of the sampling locations. The climate data for the sampling locations were obtained from the WorldClim database. The ecological characteristics were described using 19 bioclimatic variables representing the annual trends, seasonal variations and extremes in temperature and precipitation. The comparison of the outlier marker loci detected by the three approaches could help to identify the factors that are responsible for the observed spatial structuring of genetic diversity in Dalmatian sage populations.

The aim of the research was to assess genetic diversity among basil (Ocimum basilicum L.) accessions held at the Collection of Medicinal and Aromatic Plants (University of Zagreb, Faculty of Agriculture, Department of Seed Science and Technology) as a part of the National Bank of Plant Genes. Seventy-eight basil accessions belonging to five basil morphotypes (True basil, Small-leaf, Lettuce-leaf, Purple basil A and Purple basil B) were analysed using AFLP markers. Sixteen AFLP primer combinations yielded a total of 2, 449 polymorphic markers. Analysis of molecular variance (AMOVA) revealed that most of the genetic diversity was attributable to differences among accessions within morphotypes, while the remaining variability was almost equally distributed among morphotypes within groups of morphotypes classified according to anthocyanin coloration on stems (‘green’ vs. ‘purple’ morphotypes) and among the groups. Bayesian analysis of genetic structure resulted in assignment of the investigated accessions to two clusters. Most of the accessions belonging to ‘green’ morphotypes were assigned to Cluster A, while the Cluster B included most of the accessions belonging to ‘purple’ morphotypes. The separation of ‘green’ vs. ‘purple’ morphotypes was confirmed further by principal co-ordinate analysis (PCoA) and the neighbour-joining cluster analysis, both based on pairwise Dice’s distance matrix between accessions. The assessment of the genetic diversity of the basil accessions is the first step for the future association mapping studies aimed at identifying markers associated with main essential oil compounds.

The settlement of Posidonia oceanica (L.) Delile along the eastern coast of the Adriatic is fairly recent taking into account that during the peak of the last glaciation, 18.000 years before present, the sea level was approximately 120 m lower than today. In the Adriatic Sea P. oceanica inhabits the narrow zone in the coastal sea from the surface to approx. 40 m of depth. In order to assess the levels of genetic diversity and to investigate inter- and intra-population genetic structure of P. oceanica meadows along the eastern Adriatic coast, a total of 268 samples were collected from 15 localities. After DNA extraction, 9 nuclear polymorphic microsatellites loci (Po15, Po4-3, Po5, Po5-39, Po5-49, Poc5, Poc26, Poc42, Poc45) were analysed. The results supported the expectation of high clonality and low genetic diversity levels of this species. This could be explained by predominant clonal reproduction and vegetative dispersal in contrast to sexual reproduction and seed dispersal of this species in the Adriatic. The results are discussed in respect to eastern Adriatic geography, consequent climatic and ecological differences and the dominant sea current flow.

Sideritis scardica Griseb. is an endangered plant species, endemic to the Balkan peninsula. Aerial parts of Sideritis scardica Griseb. are used in traditional medicine as a herbal tea for treatment of lung and gastrointestinal disorders, antimicrobial infections and rheumatic disorders. The plants are extensively gathered from natural resources and due to often overexploitation many wild populations are seriously threatened. Therefore, the design of conservation strategies is crucial, thus requiring detailed analysis of the level and distribution of genetic diversity. For this purpose 12 natural S. scardica populations from Greece and FYR of Macedonia were analyzed using Amplified fragment length polymorphism (AFLP) markers. Analysis of molecular variance (AMOVA) indicated that the most genetic variation was found within populations (65.02%). Gene diversity ranged from 0.088 (Mt. Kozuf ; MKD) to 0.123 (Mt. Vermio ; GR). Populations tended to group together according to their geographical origin as revealed by distance and model based methods. Isolation by distance analysis indicated that 52.2% of population differentiation could be explained with isolation by distance. The obtained results provide basic information for the efficient conservation strategy development for S. scardica and introduction into agricultural cultivation.

This study, conducted in Croatia, encompassed for the first time ever populations of the field elm that are solely natural. We studied the genetic diversity and assessed recent bottleneck events and the population structure of 5 natural field elm populations from continental and Mediterranean Croatia, using 5 microsatellite markers. The study results suggest that the observed populations are characterized by a satisfactory amount of heterozygosity, and that the impact of the Dutch elm disease on the amount of genetic diversity in sampled populations is currently negligible. As genetic diversity in forest tree species is slowly diminishing, in the long term there is a danger of the depletion of genetic resources. The danger of inbreeding within populations is low, and the majority of the populations are in Hardy-Weinberg equilibrium. The appearance of bottleneck, initiated by anthropogenic habitat destruction, has been established in only one population. The values of Nei’s standard genetic distance, as well as the values of the genetic differentiation index between pairs of observed populations are indicative and suggest, together with the performed factor correspondence analysis and unrooted neighbor-joining tree, the existence of a very clear genetic differentiation between the continental and the Mediterranean populations of the field elm in Croatia.

Dalmatian pyrethrum (Tanacetum cinerariifolium /Trevir./ Sch. Bip.) is a perennial plant species endemic to the east coast of the Adriatic Sea. Secondary metabolite of Dalmatian pyrethrum is a natural insecticide pyrethrin. Pyrethrin is effective on a broad range of insect pest and low in toxicity to humans and worm blooded animals. Amplified fragment length polymorphism (AFLP) markers were used to investigate genetic diversity within and among 25 wild populations of Dalmatian pyrethrum. Six primer combinations produced 985 bands of which 935 (94.9%) were polymorphic. Analysis of the molecular variance (AMOVA) revealed that the most of the AFLP diversity was distributed among individuals within populations (85.89%). Bayesian analysis of population structure enabled identification of two dominant genetic units. Spatial analysis of genetic diversity indicated that 3.72% of genetic differentiation resulted from isolation by distance, while 12.55% of genetic differentiation among populations followed the pattern of isolation-by-adaptation. The results have shown that wild Dalmatian pyrethrum populations are valuable source of genetic diversity that could be exploited in future plant breeding programs. Moreover, the results will have great contribution in the development of adequate conservation strategies.

Since its development in the mid-1990s the amplified fragment length polymorphism (AFLP) markers have been widely used in genetic studies because of the rapidity and ease with which a high number of polymorphic markers can be generated. Genetic variation is generally represented by the presence or absence of amplified DNA fragments, whose signals behave as dominant markers like in case of a number of other marker systems such as random amplified polymorphic DNA (RAPD), inter simple sequence repeat (ISSR) or directed amplification of minisatellite DNA polymerase chain reaction (DAMD-PCR). Each locus is thus less informative than a typical multi-allelic microsatellite (single sequence repeat ; SSR) locus, although the large number of AFLP markers available across the genome and their largely random distribution balance this drawback. Moreover, the use of dominant markers in population genetic studies is hampered by the lack of complete genotypic information. Recent advances in statistical methods make analyses of population diversity and structure based on dominant marker data possible although some problems of bias cannot be completely eliminated. Two basic approaches include the 'band-based' methods (direct study of AFLP band presences or absences on individual level) and 'frequency-based' methods (estimation of allelic frequencies from dominant biallelic data on population level). Genetic diversity within populations can be estimated using the Shannon's information index (Shannon, 1948 ; Lewontin, 1972) based on frequencies of band presence/absence at each marker within the population. Shannon’s information index can be also used to measure the total diversity (HT) as well as the average intrapopulation (HP) diversity and the proportions of diversity within (HP/HT) and among populations [(HT–HP)/HT] can be obtained. Pairwise genetic distances among individuals can be calculated from binary (presence/absence) data using Dice's coefficient (Dice, 1945) which takes into account only positive matches between two individuals and is therefore unaffected by potentially homoplasic absent bands. Subsequently, Dice's distance matrix can be used as an input for cluster analysis (using UPGMA or neighbour-joining method), principal co-ordinate analysis (PCoA) and the analysis of molecular variance (AMOVA). The analysis of molecular variance (AMOVA) can be used to partition the total variance into within and among population variance component. The variance components can be tested statistically by non-parametric randomisation tests. The simplest method for the estimation of allele frequencies from dominant data is the square-root procedure which uses the inbreeding coefficient (known a priori) and the square root of the frequency of null homozygotes (i.e. of band absences) to calculate the frequency of the null allele. However, a Bayesian method with non-uniform prior distribution of allele frequencies (Zhivotovsky, 1999) gives better estimates of marker allele frequencies. Genetic diversity and population genetic structure can be computed following the treatment of Lynch and Milligan (1994). Expected heterozygosity or Nei's gene diversity (Nei, 1973) can be calculated for each population and the total gene diversity (HT), the average gene diversity within populations (HW), the average gene diversity among populations in excess of that observed within populations (HB), and finally Wright's FST can be obtained. The marker allele frequencies can be used for as an input for cluster analysis on population level. Yet another Bayesian method developed by Holsinger et al. (2002) can be used in order to estimate FIS (inbreeding coefficient) from dominant marker data. The estimates of FIS often seem to be unreliable and have to be regarded with great caution. Finally, the model-based clustering program STRUCTURE (Pritchard et al., 2000) can be used to estimate the underlying population structure using individual multilocus genotypes. This Bayesian method enables identification of clusters of genetically similar individuals without prior knowledge of their population affiliation. In this approach, it is assumed that there are K populations contributing to the gene pool of the sampled populations. Spatial genetic structure can be analysed by a number of methods including spatial pattern analysis of genetic diversity (Hardy, 2003 ; based on kinship coefficient for dominant markers among individuals), isolation by distance analysis (Rousset, 1997 ; based on FST matrix among populations) and Bayesian spatial modelling of genetic population structure (Corander et al., 2008).

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.

AFLP markers were used for genetic diversity analysis of 40 basil accessions representing eight species (O. americanum L., O. basilicum L., O. champechianum Mill., O. x citriodorum Vis., O. gratissimum L., O. kilimandscharicum Baker ex Gürke O. minimum L., O. tenuiflorum L.). Genetic relationships were determined by Neighbor Joining cluster analysis based on Dice's distance matrix between accessions. High bootstrap support values for the branches separating species were observed as well as grouping of morphologically similar accessions within the O. basilicum cluster. The position of O. minimum accession within the O. basilicum cluster indicate that at genomic level O. minimum is closely related to other taxa of O. basilicum. O. minimum should be considered as a variety of O. basilicum, although it possesses a limited number of dwarfing genes giving it clearly distinguishable morphological features. The accessions of O. americanum and O. x citriodorum grouped together with two accessions of O. basilicum var. purpurascens. The explanation could be the hybrid origin of O. x citriodorum (= O. basilicum x O. americanum) as well as of these two accessions of var. purpurascens (= O. basilicum var. purpurascens s. stricto x O. x citriodorum). AFLP markers can contribute in resolving existing problems concerning basil taxonomy at both inter- and intra-specific levels.

U Podravini se jasno razlikuju sastojine bijele i crne johe, no ponegdje se mogu naći obje vrste u istim sastojinama. Cilj diplomskog rada je bio potvrditi ili osporiti postojanje hibridnih jedinki bijele i crne johe u Podravini, morfometrijskim i molekularnim pristupom. Morfometrijski pristup se temeljio na mjerenju i usporedbi 10 parametara lista, a molekularni na provedbi analize 12 mikrosatelitnih lokusa. U istraživanju su analizirane 102 jedinke iz pet populacija. U crne johe je zapažena veća genetička, a u bijele johe nešto veća morfološka raznolikost. Hibridno podrijetlo je dokazano za samo tri jedinke iz populacije bijele johe. Upotrebom Bayesovske analize populacijske strukture na molekularnim podacima bilo je moguće procijeniti razinu introgresije genoma crne johe u hibridnim jedinkama. Hibridne jedinke se razlikuju od roditeljskih vrsta po većoj površini plojke, dužini plojke mjerenoj od osnove do mjesta najveće širine te po dužini peteljke. Različitim antropogenim zahvatima na području nizinskih šumskih ekosustava u Podravini nepovratno su promijenjene stanišne prilike pa osobine hibrida bijele i crne johe kao što su veća otpornost na sušu i na bolesti korijena, manji zahtjevi za kvalitetom tla te dobre fizičko-tehničke osobine drva, čine ovu svojtu prihvatljivim izborom za obnavljanje šuma u Podravini. In the Podravina region, white and black alder stands are clearly distinguished, although sporadically both species can be found in the same stand. Aim of this graduation thesis was to confirm or deny the existence of black and white alder hybrid individuals in the Podravina region by morphometric and molecular approaches. The morphometric approach was based on the measurements and comparison of 10 leaf parameters, while the molecular one was based on the analysis of 12 microsatellite loci. The study analyzed 102 individuals from the five populations. In the black alder was observed a higher genetic while in the grey alder slightly higher morphological diversity. Hybrid origin has been proven only for three individuals in a grey alder population. Bayesian analysis of population structure on molecular data allowed the assessement of the level of introgression of the genome of black alder in hybrid individuals. Hybrid individuals differ from the parental species by the larger surface of the leaf blade, the length of the blade from the base to the point of the maximum leaf width and by the length of the petiole. Various anthropogenic interventions in the area of lowland forest ecosystems in Podravina region irreversibly changed the habitat conditions and because of that the characteristics of the hybrid of the grey and black alder, such as higher resistance to the drought and the root diseases, lower requirements for soil quality and good physical and technical characteristics of the wood, make this taxon an acceptable choice for the regeneration of the forests in the Podravina region.

Genetička raznolikost jedan je od najvažnijih elemenata prilagodbe svih živih bića na neprestane promjene u okolišu. Intenzivno iskorištavanje kestenovih šuma i bolest poznata kao rak kestenove kore ostavili su trajne negativne posljedice na populacije europskog pitomog kestena. Svaka spoznaja o genetičkoj varijabilnosti europskog pitomog kestena izravno doprinosi razvoju učinkovitijih planova zaštite i gospodarenja ovom vrstom. Cilj ovog diplomskog rada je bio usporediti genetičku raznolikost između i unutar tri kontinentalne (Bilogora, Medvednica, Moslavačka gora) i tri primorske populacije (Poreč, Buje, Cres) pitomog kestena u Hrvatskoj analizom devet SSR i devet EST-SSR mikrosatelitnih biljega. Dobiveni rezultati nisu potvrdili jasnu genetičku diferencijaciju kontinentalnih i primorskih populacija pitomog kestena u Hrvatskoj. Svojom genetičkom raznolikošću populacije Medvednica, Bilogora, Moslavačka gora i Buje ukazivale su na prirodno podrijetlo, dok su populacije Poreč i Cres ukazivale na jako veliki antropogeni utjecaj u njihovom formiranju. Protok gena iz nasada lovranskih maruna u prirodnu populaciju Cres moguće je objašnjenje specifične genetičke raznolikosti ove populacije, dok bi populacija Poreč mogla biti kombinacija gena talijanskih prirodnih populacija i/ili kultivara, istarskih prirodnih populacija i lovranskog maruna. U odvojenim SSR i EST-SSR analizama svi dobiveni populacijsko-genetički parametri bili su gotovo isti pa se u budućim istraživanjima preporučuje analiza ujedinjenih podataka. Genetic diversity is one of the most important elements in adapting of all living beings to the continual changes in the environment. The intensive exploitation of chestnut forests and the disease known as chestnut blight have left negative effects on the populations of European chestnut. Any knowledge of the genetic variability of the chestnut directly contributes to the development of more effective conservation and management plans for this species. The aim of this thesis was to compare genetic diversity between and within three continental (Bilogora, Medvednica, Moslavačka gora) and three coastal populations (Poreč, Buje, Cres) of European chestnut in Croatia using nine SSR and nine EST-SSR microsatellite markers. The obtained result did not confirm the clear genetic differentiation between continental and coastal populations of European chestnut in Croatia. Due to their genetic diversity, the populations of Medvednica, Bilogora, Moslavačka Gora and Buje indicated their natural origin, while the populations of Poreč and Cres indicated a very large anthropogenic influence during their formation. Gene flow from Lovran Marron plantations to the natural population of Cres is a possible explanation for the specific genetic diversity of this population, while the population Poreč could be a combination of the genes of Italian natural populations and / or cultivars, Istrian natural populations and Lovran Marron. In the separate SSR and EST-SSR analyzes, all obtained population-genetic parameters were almost the same, so the recommendation is that in the future data analysis should be applied on the combined data set.

Kada istražujemo genetičku raznolikost masline moramo u vidu imati postojanje tri tipa maslina: divlje masline (oleaster), sjemenjaci (olivaster) i kultivirane masline. U ovom istraživanju prikupljeni su uzorci tri populacije divljih maslina s otoka Paga i Lastova, poluotoka Pelješca i 15 najčešćih hrvatskih kultivara. Provedena je analiza 12 mikrosatelitnih lokusa. Izračunati su različiti genetički parametri i upotrebljene različite statističke metode kao što su: informacijski sadržaj polimorfizma, prosječan broj alela po populaciji, alelno bogatstvo, broj privatnih alela, primijećena i očekivana heterozigotnost, koeficijent samooplodnje, genetska diferenciranost, izračun matrice genetičke udaljenosti, izrada NJ stabla, Bayesovska analiza skupina na temelju modela te faktorska analiza korespondencije. Jedinke u populaciji Pelješac određene su kao olivasteri, a ne kao prave divlje masline (oleasteri). Pravim divljim populacijama su se pokazale samo populacije Pag i Lastovo. Kultivari Piculja i Mrčakinja nisu kultivari, nego prave divlje masline koje zbog obilne i redovite cvatnje služe u maslinicima kao obilan izvor peluda tijekom sezone oprašivanja, signifikantan koeficijent samooplodnje (FIS) u populaciji Pag ukazuje da je ova divlja populacija, vjerojatno zbog smanjenja veličine, prošla kroz genetičko usko grlo (eng. bottleneck). Dobiveni rezultati naglašavaju potrebu očuvanja prirodnih populacija divljih maslina kao rezervata rijetkih i vrijednih alela u oplemenjivačkim programima. When we research the genetic diversity of olives, we have to keep in mind the existence of three types of olives: wild olives, feral forms and cultivated olives. In this study, the samples of three wild olive populations from the islands of Pag and Lastovo, the peninsula Pelješac and 15 most common Croatian cultivars were collected. An analysis of 12 microsatellite loci was performed. Various genetic parameters were calculated and different statistical methods were used such as: polymorphism information content, average number of alleles per population, allelic richness, number of private alleles, observed and expected heterozygosity, inbreeding coefficient, genetic differentiation, calculation of genetic distances, creation of NJ tree, Bayesian model-based cluster analysis and factorial correspondence analysis. The individuals of the population Pelješac were characterized as feral forms not as true wild olives (oleaster). Wild origin has only been confirmed to the populations Pag and Lastovo. The cultivars Piculja and Mrčakinja were not cultivars but true wild olives, which, due to their abundant and regular flowering, serve as an abundant source of pollen during pollination season in the olive groves. The significant FIS coefficient in the Pag population indicates that this wild population, probably due to a decrease in size, has undergone a genetic bottleneck. The obtained results underlined the need to conserve natural populations of wild olives as reserves of rare and valuable alleles in breeding programs.

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.