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The development of next-generation sequencing technology and the increasing amount of sequencing data have brought the bioinformatic tools used in genome assembly into focus. The final step of the process is genome annotation, which works on assembled genome sequences to identify the location of genome features. In the case of organelle genomes, specialized annotation tools are used to identify organelle genes and structural features. Numerous annotation tools target chloroplast sequences. Most chloroplast DNA genomes have a quadripartite structure caused by two copies of a large inverted repeat. We investigated the strategies of six annotation tools (Chloë, Chloroplot, GeSeq, ORG.Annotate, PGA, Plann) for identifying inverted repeats and analyzed their success using publicly available complete chloroplast sequences of taxa belonging to the asterid and rosid clades. The annotation tools use two different approaches to identify inverted repeats, using existing general search tools or implementing stand-alone solutions. The chloroplast sequences studied show that there are different types of imperfections in the assembled data and that each tool performs better on some sequences than the others.

With the development of next-generation sequencing technology and bioinformatics tools, the process of assembling DNA sequences has become cheaper and easier, especially in the case of much shorter organelle genomes. The number of available DNA sequences of complete chloroplast genomes in public genetic databases is constantly increasing and the data are widely used in plant phylogenetic and biotechnological research. In this work, we investigated possible inconsistencies in the stored form of publicly available chloroplast genome sequence data. The impact of these inconsistencies on the results of the phylogenetic analysis was investigated and the bioinformatic solution to identify and correct inconsistencies was implemented. The whole procedure was demonstrated using five plant families (Apiaceae, Asteraceae, Campanulaceae, Lamiaceae and Rosaceae) as examples.

Pyrethrin is a potent biopesticide, a natural mixture of six compounds (pyrethrin I and II, cinerin I and II, and jasmolin I and II), biosynthesized in plants of Dalmatian pyrethrum (Tanacetum cinerariifolium/Trevir./Sch. Bip.), a species endemic to the eastern Adriatic coast, but grown worldwide. Not many known natural insecticides encompass such superior qualities as pyrethrin—very high efficacy against a broad spectrum of pests in combination with minor adverse effects on human health and the environment. In previous decades, pyrethrin was largely replaced by its synthetic derivatives, pyrethroids. However, due to their harmful effects on various species and ecosystems, the use of pyrethrin should again take the lead. This review summarizes one century (1920–2020) of research on the properties and use of pyrethrum as a source of pyrethrins. The primary focus is on presenting its current advantages and disadvantages, toxicity on target and non-target species, biosynthesis, factors that influence the pyrethrins content in pyrethrum, comparison of different methods of their extraction and determination ; as well as its production potential and development of new products. The final goal is to present possible approaches to improve and enhance the use of this highly effective but still underused phytochemical insecticide with unique properties.

Due to the availability and affordability of large scale genotyping, identification of genomic regions affected by the natural or artificial selection in the populations of wild and domesticated plants became standard procedure in modern population genetics. Identification of these regions and their association with traits of interest helps to understand evolutionary history of the species. The structure of cultivated common bean populations strongly resembles the structure of wild populations due to occurrence of at least two independent domestication events that occurred independently in Central and South America. Long- termed spatial isolation of two gene pools preserved their differential patterns of adaptation. Basic structure in collection of Croatian common bean landraces corresponds to the general pattern: first division is between two clusters of Central and South American origin, and later than divides into two clusters identified by different phaseolin types. In this study we used the differences in allele frequencies between populations (estimated by Fst values) to identify the SNP loci under adaptive selection.

Immortelle (Helichrysum italicum /Roth./ G. Don) is a perennial plant species of the genus Helichrysum and the family Asteraceae. It is spread in the Mediterranean region inhabiting the sea coast as well as the mountains up to 2, 200 meters above sea level (Galbany-Casals et al., 2011). Immortelle is used in the cosmetic, pharmaceutical, and food industries. It is an economically important medicinal and aromatic plant in Croatia, and in recent years the demand for H. italicum essential oil has increased. This study aimed to determine the biochemical diversity of 18 immortelle populations in Croatia.

The Pannonian Plain, as the most productive region of Southeast Europe, has a long tradition of agronomic production as well as agronomic research and plant breeding. Many research institutions from the agri-food sector of this region have a significant impact on agriculture. Their well-developed and fruitful breeding programmes resulted in productive crop varieties highly adapted to the specific regional environmental conditions. Rapid climatic changes that occurred during the last decades led to even more investigations of complex interactions between plants and their environments and the creation of climate-smart and resilient crops. Plant phenotyping is an essential part of botanical, biological, agronomic, physiological, biochemical, genetic, and other omics approaches. Phenotyping tools and applied methods differ among these disciplines, but all of them are used to evaluate and measure complex traits related to growth, yield, quality, and adaptation to different environmental stresses (biotic and abiotic). During almost a century-long period of plant breeding in the Pannonian region, plant phenotyping methods have changed, from simple measurements in the field to modern plant phenotyping and high-throughput non-invasive and digital technologies. In this review, we present a short historical background and the most recent developments in the field of plant phenotyping, as well as the results accomplished so far in Croatia, Hungary, and Serbia. Current status and perspectives for further simultaneous regional development and modernization of plant phenotyping are also discussed.

Dalmatian pyrethrum (Tanacetum cinerariifolium (Trevir.) Sch. Bip) is a plant species from Asteraceae family endemic to the coastal and peri- coastal regions of Croatia, Bosnia and Herzegovina, Montenegro and Albania (Euro+Med, 2006). It grows as an herbaceous shrub with a characteristic inflorescence at the top of a stiff and erect stem consisting of white, lance-shaped, petal-like flowers at the edges and yellow disk flowers in the center (Brewer, 1968). Dalmatian pyrethrum produces pyrethrin, a mixture of six chemical compounds found primarily in the flower heads, which has insecticidal and repellent properties (Bhat, 1995). While the biochemical diversity of the species has been the subject of numerous studies focusing on both natural populations (Grdiša et al., 2013) and commercial varieties (Suraweera et al., 2020), few studies have reported on the morphological diversity of natural Dalmatian pyrethrum populations (Ban et al., 2019). The aim of this study was to morphologically characterize the natural Dalmatian pyrethrum populations based on several traits potentially related to pyrethrin production.

The development of bioinformatic solutions is guided by biological knowledge of the subject. In some cases, we use unambiguous biological models, while in others we rely on assumptions. A commonly used assumption for genomes is that related species have similar genome sequences. This is even more obvious in the case of chloroplast genomes due to their slow evolution. We investigated whether the lengths of complete chloroplast sequences are closely related to the taxonomic proximity of the species. The study was performed using all available RefSeq sequences from the asterid and rosid clades. In general, chloroplast length distributions are narrow at both the family and genus levels. In addition, clear biological explanations have already been reported for families and genera that exhibit particularly wide distributions. The main factors responsible for the length variations are parasitic life forms, IR loss, IR expansions and contractions, and polyphyly. However, the presence of outliers in the distribution at the genus level is a strong indication of possible inaccuracies in sequence assembly.

More than one third of European grain maize is produced in South Eastearn Europe (SEE) and utilization of historical maize material developed in SEE for its favorable alleles and diversity has long been speculated. However, molecular diversity of the SEE maize genetic material is scarce. The objectives of this study were i) to analyze diversity patterns in a large panel of densely genotyped historical accessions from SEE, ii) to compare the data with those obtained from other two European panels, and iii) to identify genomic regions that have undergone selection (selective sweeps) in response to adaptation to SEE conditions. 572 accessions of the historical inbred lines from Maize Research Institute Zemun Polje representing the SEE material were genotyped using the 600k maize genotyping Axiom array. The genotyping results were merged with two European panels DROPS and TUM. Genetic structure and diversity were analyzed using neighbor-joining cladogram, PcoA, Admixture, Structure and sNMF. To detect the selective sweep signals, Tajima’sDstatistic and RAiSD were employed. The best number of ancestral populations was K=7, whereby one of them is a subpopulation containing inbreds belong exclusively to the SEE panel. The prevalence of inbreds linked to historical US inbred lines Wf9, Oh43, Pa91 and A374 was detected in SEE. Possible soft selective sweep was detected in chromosome 2 in region harboring a gene linked to promotion of flowering FPF1. Additional scan for selective sweeps using the RAiSD methodology yielded four signals in chromosomes 5 and 6, all in gene-rich regions. Several candidates of selection were identified, influencing the plant morphology and adaptation. Our study provides the first step towards the utilization of the SEE genetic materials for use in maize breeding. Phenotypic analysis is needed for assessment of SEE accessions for favorable alleles, and identification of breeding targets.

Grass pea (Lathyrus sativus L.) is an annual legume species, phylogenetically close to pea (Pisum sativum L.), that may be infected by Fusarium oxysporum f. sp. pisi (Fop), the causal agent of fusarium wilt in peas with vast worldwide yield losses. A range of responses varying from high resistance to susceptibility to this pathogen has been reported in grass pea germplasm. Nevertheless, the genetic basis of that diversity of responses is still unknown, hampering its breeding exploitation. To identify genomic regions controlling grass pea resistance to fusarium wilt, a genome-wide association study approach was applied on a grass pea worldwide collection of accessions inoculated with Fop race 2. Disease responses were scored in this collection that was also subjected to high-throughput based single nucleotide polymorphisms (SNP) screening through genotyping-by-sequencing. A total of 5,651 high-quality SNPs were considered for association mapping analysis, performed using mixed linear models accounting for population structure. Because of the absence of a fully assembled grass pea reference genome, SNP markers’ genomic positions were retrieved from the pea's reference genome v1a. In total, 17 genomic regions were associated with three fusarium wilt response traits in grass pea, anticipating an oligogenic control. Seven of these regions were located on pea chromosomes 1, 6, and 7. The candidate genes underlying these regions were putatively involved in secondary and amino acid metabolism, RNA (regulation of transcription), transport, and development. This study revealed important fusarium wilt resistance favorable grass pea SNP alleles, allowing the development of molecular tools for precision disease resistance breeding., Financial support by Fundação para a Ciência e Tecnologia (FCT), Portugal, is acknowledged through grant PD/BD/114418/2016 (Ana Margarida Sampaio), the CEECIND/00198/2017 research contract by the stimulus of scientific employment (Carmen Santos), the research unit GREEN-IT (UID/04551/2020), and the QuaLaty project (PTDC/AGR-TEC/0992/2014). Susana de Sousa Araújo acknowledges the financial support from the NORTE 2020 throughout the I-CERES project (NORTE-01-0145-FEDER-000082), funded by the Fundo Europeu de Desenvolvimento Regional (FEDER) and project NORTE-06-3559-FSE-000103 funded by the Fundo Social Europeu (FSE). We also thank the Spanish Research Agency (AEI) project AGL2017-82907-R and PID2020-11468RB-100, the FP7 EU funding through the LEGATO project (grant agreement FP7-613551) and the project KK.01.1.1.01.0005 Biodiversity and Molecular Plant Breeding, Centre of Excellence for Biodiversity and Molecular Plant Breeding (CoE CroPBioDiv), Zagreb, Croatia.

Surina, Bostjan/0000-0002-2635-315X; Satovic, Zlatko/0000-0002-9848-1601; Liber, Zlatko/0000-0002-2370-6650; Bogdanovic, Sandro/0000-0002-1952-6059; Celep, Ferhat/0000-0003-3280-8373 WOS: 000459983900082 PubMed: 30824783 The inheritance of phenotypic, genetic and epigenetic traits in hybridization events is difficult to predict, as numerous evolutionary, ecological, and genetic factors can play a crucial role in the process of hybridization. In the middle Adriatic island of Vis, we investigated hybridization between Salvia officinalis and S. fruticosa at morphological, genetic and epigenetic levels. SSR results revealed that hybrid individuals were characterized by diploid set of chromosomes suggesting homoploid hybridization. A well-defined group that mostly comprised of F-1 generation individuals was detected. For the majority of analysed morphological characteristics, hybrids were placed in-between parental taxa, while at the same time, values of different genetic parameters were mostly higher in hybrids than in parental species. The results revealed a high contrast in the levels of phenotypic variability and epigenetic excitation between parental taxa. Environmental niche modelling confirmed that in the studied location S. officinalis experiences optimal climatological conditions, while S. fruticosa struggles with unsuitable conditions. Very low levels of gene flow between the parental species were detected. In addition, contrasting levels of epigenetic excitation in the studied groups clearly demonstrated the importance of an epigenetic response to an altered environment and confirmed the trans-generational nature of the epigenetic changes. project Biodiversity and Molecular Plant Breeding, Centre of Excellence for Biodiversity and Molecular Plant Breeding (CoE CroP-BioDiv), Zagreb, Croatia [KK.01.1.1.01.0005] This research has been funded by the project KK.01.1.1.01.0005 Biodiversity and Molecular Plant Breeding, Centre of Excellence for Biodiversity and Molecular Plant Breeding (CoE CroP-BioDiv), Zagreb, Croatia. The authors would like to thank Oleg Antonic and Martina Temunovic for helpful discussion on environmental niche modelling.

Common bean (Phaseolus vulgaris L.) exhibits a wide range of seed coat colors and this morphological trait is widely used in cultivar identification and assessment of diversity within this species. With an advancement in technology and informatics, new methods of assessing seed color are emerging in addition to traditionally used visual observation. Due to a great variety of color measuring techniques, the evaluation of the agreement between methods is needed prior to using the methods interchangeably. Seed coat color in terms of CIE L*a*b* color coordinates of 100 common bean accessions belonging to five mono- colored landraces was assessed using two methods, colorimeter and Computer vision. The percentage difference between the two methods across all samples for L* color coordinate was 5.81%, for a* color coordinate 23.32% and for b* color coordinate 44.44%. According to Bland- Altman difference plot there is a considerable lack of agreement between the two methods. However, using stepwise discriminant analysis revealed that colorimeter method correctly classified 97% of accessions into their respective landrace, while the classification success of the Computer vision was 99%.

Landraces represent valuable genetic resources for breeding programmes to produce high-yielding varieties adapted to stressful environmental conditions. Although the common bean (Phaseolus vulgaris L.) is an economically important food legume for direct human consumption worldwide, common bean production in Croatia is based almost exclusively on landraces and there is no common bean breeding program. Information on phaseolin type and results of population structure and genetic diversity obtained by analysis of SSR and SNP markers, in combination with the morphological characterization of 174 accessions of 10 common bean landraces (morphotypes), enabled thorough classification of accessions. The accessions were classified into phaseolin type H1 (“S”) of Mesoamerican origin and phaseolin types H2 (“H” or “C”) and H3 (“T”) of Andean origin. By applying distance- and model-based clustering methods to SSR markers, the accessions were classified into two clusters at K = 2 separating the accessions according to the centres of origin, while at K = 3, the accessions of Andean origin were further classified into two clusters of accessions that differed in phaseolin type (H2 and H3). Using SNP markers, model-based analysis of population structure was performed, the results of which were consistent with those of SSR markers. In addition, 122 accessions were assigned to 14 newly formed true-type morphogenetic groups derived from three different domestication events: (1) Mesoamerican (H1A) (“Biser”, “Kukuruzar”, “Tetovac”, “Trešnjevac”), (2) Andean—indeterminate type (H2B1) (“Dan noć”, “Sivi”, “Puter”, ”Sivi prošarani”, “Trešnjevac”) and (3) Andean—determinate type (H3B2) (“Bijeli”, “Dan noć”, “Puter”, “Trešnjevac”, “Zelenčec”). The rest of the accessions could represent putative hybrids between morphogenetic groups. The differences between the true-type groups of accessions were further analysed based on nine quantitative traits, and the subsets of traits that best distinguish among centres of origin (A: Mesoamerican, B: Andean) and genetic groups (H1A, H2B1, H3B2) were proposed.

This study examines genetic diversity and structure of a Croatian garlic germplasm collection using 13 simple sequence repeat (SSR) markers. A total of 71 alleles were observed across 64 accessions representing 3 Croatian regions (Istria, Dalmatia and continental Croatia) and 16 foreign landraces, with an average of 5.46 alleles per locus. Among the 80 accessions analysed, 61 distinct multilocus genotypes (MLG) were identified, of which 51 represented unique genotypes and the remaining accessions were divided into 10 MLG groups, comprising potential duplicates or redundant genotypes. Model-based Bayesian and hierarchical UPGMA clustering approaches revealed five major groups within the collection which partially correlated with geographical origin. The analysis of molecular variance (AMOVA) showed that the majority (87.71%) of the total molecular diversity is within the Croatian groups of accessions, even though a significant share (12.29%) of diversity derived from genetic diversity among groups. These results support regional structuring, as well as the existence of significant diversity within local populations. This study is the first comprehensive report on an extensive evaluation of genetic resources of garlic maintained by Croatia with the aim of setting the course for future preservation strategies with particular emphasis on the value of diversity in the context of climate change both on macro and micro levels.

Sweet pepper (Capsicum annuum L.) is one of the most important vegetable crops in the world because of the nutritional value of its fruits and its economic importance. Calcium (Ca) improves the quality of sweet pepper fruits, and the application of calcite nanoparticles in agricultural practice has a positive effect on the morphological, physiological, and physicochemical properties of the whole plant. The objectives of this study were to investigate the effect of commercial calcite nanoparticles on yield, chemical, physical, morphological, and multispectral properties of sweet pepper fruits using a combination of conventional and novel image-based nondestructive methods of fruit quality analysis. In the field trial, two sweet pepper cultivars, i.e., Šorokšari and Kurtovska kapija, were treated with commercial calcite nanoparticles (at a concentration of 3% and 5%, calcite-based foliar fertilizer (positive control), and water (negative control) three times during vegetation). Sweet pepper fruits were harvested at the time of technological and physiological maturity. Significant differences were observed between pepper cultivars as well as between harvests times. In general, application of calcite nanoparticles reduced yield and increased fruit firmness. However, different effects of calcite nanoparticles were observed on almost all properties depending on the cultivar. In Šorokšari, calcite nanoparticles and calcite-based foliar fertilizers significantly increased N, P, K, Mg, Fe, Zn, Mn, and Cu at technological maturity, as well as P, Ca, Mg, Fe, Zn, Mn, Cu, and N at physiological maturity. However, in Kurtovska kapija, the treatments increased only Ca at technological maturity and only P at physiological maturity. The effect of treatments on fruit morphological properties was observed only at the second harvest. In Šorokšari, calcite nanoparticles (3% and 5%) increased the fruit length, minimal circle area, and minimal circle radius, and it decreased the fruit width and convex hull compared to the positive and negative controls, respectively. In Kurtovska kapija, calcite nanoparticles increased the fruit width and convex hull compared to the controls. At physiological maturity, lower anthocyanin and chlorophyll indices were found in Kurtovska kapija in both treatments with calcite nanoparticles, while in Šorokšari, the opposite effects were observed.

Neven (Calendula officinalis L.) je poznat po svojim ljekovitim svojstvima kao i primjeni u medicinskoj, kemijskoj, kozmetičkoj i prehrambenoj industriji. Prilagođen je umjerenoj klimi te mu je optimalna temperatura klijanja 20 °C. Klijanje sjemena i rani rast sadnica su najosjetljiviji stadiji na abiotske stresove, kao što su nepovoljna temperatura i suša, a predsjetveni tretmani sjemena pozitivno utječu na svojstva klijavosti ili kvalitetu sjemena i sadnica u stresnim uvjetima. S obzirom da se sjetva nevena obavlja čim se uspostave minimalni uvjeti, a tada temperature i količine oborina mogu biti znatno niže od optimalnih, cilj ovog istraživanja bio je utvrditi utjecaj predtretmana na svojstva klijavosti sjemena nevena u uvjetima niskih temperatura i suše (-0,8 MPa PEG 6000). Laboratorijskim pokusom istražen je utjecaj temperature (14 °C, 18 °C i 22 °C) na klijavost sjemena nevena uz predtretmane dH2O i GA3 (250 ppm) u trajanju od 24 h. Netretirano sjeme je predstavljalo kontrolu. Utvrđeno je da sjeme nevena ne klije u uvjetima suše bez obzira na predsjetveni tretman i na temperaturu pri kojoj je klijalo. Ni temperatura niti predtretman sjemena ne utječu značajno na broj proklijalih sjemenki nevena, ali utječu na brzinu klijanja., Marigold (Calendula officinalis L.) is known for its medicinal properties and is used in the medical, chemical, cosmetic, and food industries. It is adapted to a temperate climate and its optimum germination temperature is 20 °C. Seed germination and early seed ling growth are the most sensitive to abiotic stresses such as unfavourable temperatures and drought, while seed priming has a positive effect on germination properties of seed and seed ling quality under stressful conditions. Since sowing of marigolds is done as soon as the minimum conditions are established, when temperature sand precipitation canbes ignificantly lower than optimal, the aim of this study was to determine the impact of seed priming on the germination properties of marigold seeds under low temperature and drought (-0.8 MPa PEG 6000). The effect of temperature (14 °C, 18 °C, and 22 °C) on germination of marigold seeds with treatments containing dH2O and GA3 (250 ppm) for 24 h was studied under laboratory conditions. Untreated seed represented control. Marigold seeds did not germinate under drought conditions, regard less of seed priming treatments and temperatures. It was also found that neither temperature nor seed priming significantly affected the number of germinated marigold seeds, but did affect the germination rate.

Basil is one of the most widespread aromatic and medicinal plants, which is often grown in drought- and salinity-prone regions. Often co-occurrence of drought and salinity stresses in agroecosystems and similarities of symptoms which they cause on plants complicates the differentiation among them. Development of automated phenotyping techniques with integrative and simultaneous quantification of multiple morphological and physiological traits enables early detection and quantification of different stresses on a whole plant basis. In this study, we have used different phenotyping techniques including chlorophyll fluorescence imaging, multispectral imaging, and 3D multispectral scanning, aiming to quantify changes in basil phenotypic traits under early and prolonged drought and salinity stress and to determine traits which could differentiate among drought and salinity stressed basil plants. Ocimum basilicum “Genovese” was grown in a growth chamber under well-watered control [45–50% volumetric water content (VWC)], moderate salinity stress (100 mM NaCl), severe salinity stress (200 mM NaCl), moderate drought stress (25–30% VWC), and severe drought stress (15–20% VWC). Phenotypic traits were measured for 3 weeks in 7-day intervals. Automated phenotyping techniques were able to detect basil responses to early and prolonged salinity and drought stress. In addition, several phenotypic traits were able to differentiate among salinity and drought. At early stages, low anthocyanin index (ARI), chlorophyll index (CHI), and hue (HUE2D), and higher reflectance in red (RRed), reflectance in green (RGreen), and leaf inclination (LINC) indicated drought stress. At later stress stages, maximum fluorescence (Fm), HUE2D, normalized difference vegetation index (NDVI), and LINC contribute the most to the differentiation among drought and non-stressed as well as among drought and salinity stressed plants. ARI and electron transport rate (ETR) were best for differentiation of salinity stressed plants from non-stressed plants both at early and prolonged stress.

The high-biodiversity phenomenon of the Balkan Peninsula is a result of the past and present topographic, climatic and geological conditions, as well as human impact. The Dinaric Alps, as a part of the Balkan Peninsula, harbour a high number of endemic species, some of them comprising a small number of populations, which are often endangered. We investigated spatial distribution, community composition, site ecology, genetic diversity and conservation of Degenia velebitica, a stenoendemic species of the north-western Dinaric Alps. Our results showed that D. velebitica is nowadays restricted to three localities with the area of occupancy of 48, 560 m2, harbouring approximately 37, 000 individuals. Degenia velebitica stands are differentiated into three well- characterized, floristically homogenous syntaxa, very distinct from the surrounding vegetation, suggesting their azonal occurrence and restriction to spatially highly fragmented microsites exposed to stormy winds. Spatial distances of populations, differences in D. velebitica community composition and site ecology are mirrored in genetic variation patterns of the populations, such as high-frequency down- weighted marker values in the north-western populations and high gene diversity in the south-eastern group of populations. The fact that we could not find a single D. velebitica individual at the locus classicus indicates the existence of the ever-growing fragmentation. The analyses of genetic structure using AFLP data recognized two main genetic groups of populations as evolutionary significant units that should be considered when planning protection measures. According to our IUCN Red List reassessment, D. velebitica should be treated as a critically endangered species that requires immediate conservation actions.

Campanula fenestrellata subsp. fenestrellata and C. fenestrellata subsp. istriaca belong to the morphologically and phylogenetically well established amphi-Adriatic group Campanula ser. Garganicae. The two morphologically differentiated endemic taxa are distributed on the north-eastern Adriatic islands and coast. In order to study the relationships among C. fenestrellata s.l. populations and to gain insight into genetic basis of taxonomic separation established on morphology, we examined amplified fragment length polymorphisms (AFLPs) as well as nuclear and plastid DNA sequences (ITS and trnL–trnF). With the sequence data, no structure and no distinction between the two subspecies were observed. The AFLPs separated populations geographically and inferred high number of most likely clusters. The northern and the southernmost populations were assigned to uniform separate clusters indicating the longer isolation of respective populations, while the admixed populations were evident in the central area where the two taxa geographically overlap, indicating the presence of gene flow. Distinct fragmentation patterns and the existence of several temporally continuous microrefugia during the Quaternary climatic oscillations likely promoted strong genetic differentiation within the species range, while secondary postglacial contacts are reflected in observed admixture. Although the two taxa exhibited differences in ecological preferences, niche conservatism was detected. Overall, phylogeographic reconstruction of C. fenestrellata s.l. populations highlights the importance of microrefugia in shaping the intraspecific genetic variation as well as of melting-pot areas in rearranging such variation as a result of a more recent admixture.

Salvia officinalis is a perennial species, native and endemic to the Western Balkans and the Apennine Peninsula. Due to its medicinal and aromatic properties, it is used in pharmaceutical, cosmetic, and food industries. The main objectives of the study were to infer the genetic structure of S. officinalis populations in the northern and central parts of the eastern Adriatic coast, to detect the phylogeographical barriers among the putative microrefugia and to assess the genetic diversity among the resulting ancestral clusters. Twenty-five populations were assessed using amplified fragment length polymorphism markers. High polymorphism and high diversity within populations were typical for this outcrossing long-lived species. The Fitch– Margoliash tree based on Nei’s genetic distance matrix showed that most of the populations tended to group in accordance with the geographical position of their collecting sites. Spatial analysis of the genetic diversity revealed a typical pattern of isolation by distance. Very low overall among-population differentiation and detection of only three private alleles indicate that there has been high gene flow among populations. By using Bayesian Analysis of Population Structure on population level, two distinct ancestral clusters were obtained. It is likely that these two ancestral clusters were separated for a longer period by Pleistocene glaciation, although the subsequent fast recolonization resulted in diminished genetic differences. High rarity of northern and southern populations of the investigated area indicates that S. officinalis presumably survived in both northern and southern microrefugia and expanded from there resulting in secondary contact zones, characterized by lower rarity and equal genetic diversity.

Arbuscular mycorrhizas (AM) can improve phosphorus (P) nutrition and could serve as an environmentally friendly approach for sustainable crop production under P-limiting conditions. The objectives of this study were to assess the effect of AM on different physiological traits and to quantify the responsiveness of different basil (Ocimum basilicum L.) cultivars to AM under low P availability. The basil cultivars &lsquo, Genovese&rsquo, &lsquo, Sweet Basil&rsquo, Dark Opal&rsquo, and &lsquo, Erevanskii&rsquo, were inoculated (AMI) using Rhizophagus irregularis. Photochemical efficiency and gas exchange were measured on AMI and non-inoculated (AMC) plants and, at harvest, the shoot biomass, shoot P concentration, root morphological traits, frequency of mycorrhizas in the roots (F%), and extent of root colonization (M%) were determined. Significant differences in F% and M% were found among the examined cultivars, with the highest found in &lsquo, Dark Opal&lsquo, and the lowest in &lsquo, Erevanskii&lsquo, AMI reduced the shoot biomass and increased the shoot P concentration as well as other examined root traits in &lsquo, whereas it did not affect those traits in &lsquo, indicating differences in responsiveness to AM. AMI positively affected the gas-exchange parameters in all examined cultivars, probably due to the increased sink capacity of a bigger root system and/or AM structures within the roots.

Arbuscular mycorrhizas (AM) can improve phosphorus (P) nutrition and could serve as an environmentally friendly approach for sustainable crop production under P-limiting conditions. The objectives of this study were to assess the effect of AM on different physiological traits and to quantify the responsiveness of different basil (Ocimum basilicum L.) cultivars to AM under low P availability. The basil cultivars ‘Genovese’, ‘Sweet Basil’, ‘Dark Opal’, and ‘Erevanskii’ were inoculated (AMI) using Rhizophagus irregularis. Photochemical efficiency and gas exchange were measured on AMI and non-inoculated (AMC) plants and, at harvest, the shoot biomass, shoot P concentration, root morphological traits, frequency of mycorrhizas in the roots (F%), and extent of root colonization (M%) were determined. Significant differences in F% and M% were found among the examined cultivars, with the highest found in ‘Dark Opal‘ and the lowest in ‘Erevanskii‘. AMI reduced the shoot biomass and increased the shoot P concentration as well as other examined root traits in ‘Genovese’ and ‘Erevanskii’, whereas it did not affect those traits in ‘Dark Opal’ and ‘Sweet Basil’, indicating differences in responsiveness to AM. AMI positively affected the gas-exchange parameters in all examined cultivars, probably due to the increased sink capacity of a bigger root system and/or AM structures within the roots.

The intensive exchange and diffusion of plant material over the whole Mediterranean Basin including the East Adriatic region, has contributed to the confusion in denomination of olive varieties, and formation of a large number of synonyms and homonyms. In the present study, we report on clonal diversity and genetic relationships among a total of 190 olive trees sampled along the East Adriatic coast in Slovenia (SVN), Croatia (HRV) and Montenegro (MNE). Samples belonged to the six East Adriatic olive varieties with homonymic denomination: ‘Istrska belica’ (SVN), ‘Bjelica’ (HRV), ‘Žutica’ (MNE), ‘Crnica’ (SVN), ‘Crnica’ (HRV), and ‘Crnica’ (MNE). Results showed polymorphism in all twelve microsatellite markers (DCA3, DCA9, DCA11, DCA14, DCA16, DCA18, EMO03, EMO90, GAPU101, GAPU103A, GAPU71B and UDO99-019), revealing a total of 83 alleles. A total of 36 different genotypes in the panel of 190 samples were found. Genetic profiling of six homonymic East Adriatic olive varieties revealed four different varieties: 1) ‘Istrska belica’ (SVN), 2) ‘Bjelica’ (HRV)/‘Žutica’ (MNE), 3) ‘Crnica’ (HRV)/‘Crnica’ (MNE) and 4) ‘Crnica’ (SVN). Detailed clonal and genetic relationship analyses demonstrated genetic uniformity of ‘Istrska belica’ (SVN), negligible low diversity of ‘Crnica’ (SVN), and moderate diversity of ‘Crnica’ (HRV)/‘Crnica’ (MNE). Polyclonal property was assigned to only ‘Bjelica’ (HRV)/‘Žutica’ (MNE) variety. High genetic similarity between ‘Bjelica’ (HRV)/‘Žutica’ (MNE), and ‘Crnica’ (HRV)/‘Crnica’ (MNE), with different distribution of clones within them, reflects the past selection and propagation processes in Croatia and Montenegro. For the first time, international synonymy of ‘Žutica’ (MNE) and ‘Bjelica’ (HRV), and homonymy of ‘Istrska belica’ (SVN) and ‘Bjelica’ (HRV)/‘Žutica’ (MNE), as well as ‘Crnica’ (SVN) and ‘Crnica’ (MNE, HRV) were reliably confirmed with DNA markers.

Almond-leaved pear (Pyrus spinosa Forssk., Rosaceae) is a scientifically poorly researched and often overlooked Mediterranean species. It is an insect-pollinated and animal-dispersed spiny, deciduous shrub or a small tree, with high-quality wood and edible fruits. The aim of the study was to assess the phenotypic diversity of almond-leaved pear in the eastern Adriatic region. The examination of phenotypic diversity was based on a morphometric analysis of 17 populations using ten phenotypic traits of leaves. Varieties of multivariate statistical analyses were conducted to evaluate the within- and among-population diversity. In addition, the Mantel tests were used to test the correlations between geographic, environmental, and phenotypic differences among populations. High phenotypic variability was determined both among and within the studied populations. Leaf-size-related traits proved to be the most variable ones, in contrast to more uniform leaf shape traits. Furthermore, three groups of populations were detected using multivariate statistical analyses. The first group included trees from northern- and southernmost populations characterized by high annual precipitation. However, the trees from the second and third group were highly overlapped without a clear geographical pattern. In addition, we revealed that both environmental and geographical interactions proved to be responsible for the patterns of phenotypic variation between almond-leaved pear populations, indicating significant isolation by environment (IBE) and isolation by distance (IBD) patterns. Overall, our results provide useful information about phenotypic diversity of almond-leaved pear populations for further conservation, breeding, and afforestation programs.

Ocimum basilicum L. exhibits a great variety of cultivars grown for various purposes. The aim of this research was to analyse the chemical composition of 85 O. basilicum accessions cultivated throughout the world using gas chromatography (GC/FID and GC/MS). All accessions were previously categorized into six morphotypes based on 24 morphological traits: True basil, Small-leaf, Lettuce-leaf, Purple basil A, Purple basil B and Purple basil C morphotype. A total of 77 volatiles were observed, with seven of them being in concentrations higher than 10% in at least one accession (1,8-cineole, linalool, linalool acetate, methyl chavicol, eugenol, trans-methyl cinnamate, trans-α-bergamotene). Based on the essential oil composition of 85 O. basilicum accessions, we propose the intraspecific characterization of O. basilicum into five chemotypes: (A) High-linalool, (B) Linalool/trans-α-bergamotene, (C) Linalool/methyl chavicol, (D) Linalool/trans-methyl cinnamate and (E) High-methyl chavicol chemotype. Groups based on morphological and biochemical traits do not necessarily coincide with one another. Due to great morphological and biochemical diversity, both morphological and chemical characterizations are crucial for the effective management of germplasm collections to facilitate the use of the accessions in future breeding programs.

Modern maize breeding programs gave rise to genetically uniform varieties that can affect maize's capacity to cope with increasing climate unpredictability. Maize populations, genetically more heterogeneous, can evolve and better adapt to a broader range of edaphic–climatic conditions. These populations usually suffer from low yields; it is therefore desirable to improve their agronomic performance while maintaining their valuable diversity levels. With this objective, a long-term participatory breeding/on-farm conservation program was established in Portugal. In this program, maize populations were subject to stratified mass selection. This work aimed to estimate the effect of on-farm stratified mass selection on the agronomic performance, quality, and molecular diversity of two historical maize populations. Multilocation field trials, comparing the initial populations with the derived selection cycles, showed that this selection methodology led to agronomic improvement for one of the populations. The molecular diversity analysis, using microsatellites, revealed that overall genetic diversity in both populations was maintained throughout selection. The comparison of quality parameters between the initial populations and the derived selection cycles was made using kernel from a common-garden experiment. This analysis showed that the majority of the quality traits evaluated progressed erratically over time. In conclusion, this breeding approach, through simple and low-cost methodologies, proved to be an alternative strategy for genetic resources’ on-farm conservation. published

Sideritis scardica Giseb. is a subalpine/alpine plant species endemic to the central part of the Balkan Peninsula. In this study, we combined Amplified Fragment Length Polymorphism (AFLP) and environmental data to examine the adaptive genetic variations in S. scardica natural populations sampled in contrasting environments. A total of 226 AFLP loci were genotyped in 166 individuals from nine populations. The results demonstrated low gene diversity, ranging from 0.095 to 0.133 and significant genetic differentiation ranging from 0.115 to 0.408. Seven genetic clusters were revealed by Bayesian clustering methods as well as by Discriminant Analysis of Principal Components and each population formed its respective cluster. The exception were populations P02 Mt. Shara and P07 Mt. Vermio, that were admixed between two clusters. Both landscape genetic methods Mcheza and BayeScan identified a total of seven (3.10%) markers exhibiting higher levels of genetic differentiation among populations. The spatial analysis method Samβada detected 50 individual markers (22.12%) associated with bioclimatic variables, among them seven were identified by both Mcheza and BayeScan as being under directional selection. Four bioclimatic variables associated with five out of seven outliers were related to precipitation, suggesting that this variable is the key factor affecting the adaptive variation of S. scardica.

Lentil is an important cool season food legume. Vascular wilt incited by Fusarium oxysporum f.sp. lentis (FOL) is the most important disease of lentil worldwide. Knowledge of pathogen genetic structure is crucial to develop effective control strategies. In this study, the genetic structure of a collection of FOL isolates from Iran, Syria and Algeria was analysed using SSR markers. Eight markers were developed in this study and constitute a valuable resource for future molecular studies in FOL. Our results showed that there is a high molecular variation within regions, with isolates from North West Iran showing the highest gene diversity. Variation among regions also exists, with Iranian populations differing significantly from non-Iranian ones, having some private alleles. The clustering of isolates was well in agreement in both distance-based and model-based approaches suggesting the presence of seven ancestral FOL lineages, being three present exclusively in Iran while the others were distributed across all the regions. These results suggest that Iran could be a regional center of origin for FOL., This research was carried out based on PhD educational mission No. 3972/200-28/1/1389 of first author from Agricultural Research, Education and Extension Organization (AREEO) of Iran and financially supported by Project AGL2014-52871-R co-financed by FEDER.

The red raspberry (Rubus idaeus L.) is one of the most important fruit species in Serbia; there are currently approximately 11000 ha under cultivation. Wild red raspberry populations can be an important source of genes for breeding new raspberry varieties. This study was carried out to determine the genetic variability of wild populations, as well as their relationship with the most common cultivated cultivars using amplified fragment length polymorphism (AFLP) markers. In this study were included 128 individuals from seven wild populations and seven cultivar specimens from a nursery ('Mekeer', 'Willamette', 'Polka', 'Polana', 'Loganberry', 'Tayberry', 'Black Jewel'). Four AFLP combinations yielded a total of 247 polymorphic bands in 135 R. idaeus specimens. Analysis of AFLP markers showed that the cultivars are clearly distinct from wild populations. Population P4 (Mt. Ozren) had the highest gene diversity, the highest number of private markers and the highest rarity index. Analysis of molecular variance among and within wild populations showed that the most of the genetic diversity was attributable to differences between individuals within populations. A Mantel test showed that differences in altitude play the crucial role in structuring of the genetic diversity of red raspberry populations. Altitude contributed more to the population differentiation than spatial distance. Populations of wild raspberry from the territory of Serbia are characterized by high diversity, and should be protected and used as a resource in selection and breeding processes for the production of new varieties, with improved properties in terms of better yield and stronger resistance to external factors.

Campanula versicolor is a constituent of the Campanula pyramidalis complex, distributed in the central and southern Balkan Peninsula with a small disjunct range in SE Italy. The taxonomy of the broadly defined C. versicolor was controversial in the past due to high morphological variability of the populations across the range. We used microsatellite DNA data along with morphometric analyses on an extensive population sampling covering the entire range to reconstruct the relationships among populations of the intricate C. versicolor s.l. Based on the results three distinct entities can be distinguished, circumscribed here on the subspecies level, with newly established combinations: C. versicolor subsp. versicolor, C. versicolor subsp. korabensis and C. versicolor subsp. tenorei. We propose a new taxonomic treatment of C. versicolor s.l. with description, distribution and habitat data for each of the subspecies and a key for their identification.

Ascochyta blight is one of the most damaging pea diseases. Resistance to this disease in pea is quantitative, being governed by several genes with minor effect. Knowledge of the genes controlling resistance would allow their pyramiding and tracking in breeding programs. In previous studies, a number of QTLs associated with resistance to this disease have been identified. Complementarily, genes differentially expressed in resistant reactions have been identified. However, the actual genes controlling resistance, underlying these QTLs are unknown. Previously, genes with a putative involvement in defense and located into QTLs associated with resistance to P. pinodes, have been postulated as candidate genes. This study wanted to go a step forward, being the first report of candidate genes involved in defense that, besides being located in a genomic region controlling resistance, are also differentially expressed in resistant reactions. With this aim, in this study ten genes previously shown to be induced after infection in the resistant accession P665 were selected and mapped in the RIL population P665 × Messire, previously used to identify QTLs for resistance to this disease. In addition, another gene, that according to other pea maps, could be located into a QTL associated with resistance in this RIL population, was also mapped. Single-marker analysis revealed that five candidate genes showed a significant correlation with resistance traits, being also located in a genomic region showing an increased LOD for the corresponding trait. Furthermore, two of them were in the 2-LOD interval of QTLs associated with resistance traits.

Selection for wheat (Triticum aestivum L.) grain quality is often costly and time-consuming since it requires extensive phenotyping in the last phases of development of new lines and cultivars. The development of high-throughput genotyping in the last decade enabled reliable and rapid predictions of breeding values based only on marker information. Genomic selection (GS) is a method that enables the prediction of breeding values of individuals by simultaneously incorporating all available marker information into a model. The success of GS depends on the obtained prediction accuracy, which is influenced by various molecular, genetic, and phenotypic factors, as well as the factors of the selected statistical model. The objectives of this article are to review research on GS for wheat quality done so far and to highlight the key factors affecting prediction accuracy, in order to suggest the most applicable approach in GS for wheat quality traits.

The main objective of this research was to assess the genetic diversity of 5 natural field elm populations in Croatia. 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 the sampled populations is currently negligible. However, one population displayed a significant excess of heterozygosity, implying a genetic bottleneck. The existence of a very clear genetic differentiation between the continental and the Mediterranean populations of Ulmus minor in Croatia was noticed.

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. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2016, 180, 64–88.

By performing a high-resolution spatial-genetic analysis of a partially clonal Salvia brachyodon population, we elucidated its clonal architecture and seedling recruitment strategy. The sampling of the entire population was based on a 1 &times, 1 m grid and each sampled individual was genotyped. Population-genetic statistics were combined with geospatial analyses. On the population level, the presence of both sexual and clonal reproduction and repeated seedling recruitment as the prevailing strategy of new genets establishment were confirmed. On the patch level, a phalanx clonal architecture was detected. A significant negative correlation between patches&rsquo, sizes and genotypic richness was observed as young plants were not identified within existing patches of large genets but almost exclusively in surrounding areas. The erosion of the genetic variability of older patches is likely caused by the inter-genet competition and resulting selection or by a random die-off of individual genets accompanied by the absence of new seedlings establishment. This study contributes to our understanding of how clonal architecture and seedling recruitment strategies can shape the spatial-genetic structure of a partially clonal population and lays the foundation for the future research of the influence of the population&rsquo, s clonal organization on its sexual reproduction.

Essential oils from seven populations of Dalmatian sage from the North of Albania were extracted and analyzed according European Pharmacopeia 7.0. The yield differed from 1.95 to 2.75% v/w. The GC-FID-MS analysis of the sage oil revealed 42 constituents. Camphor (20.50-29.74%), followed by α-thujone (19.87-24.29%), 1,8-cineole (6.88-12.64%) and β-thujone (4.00-9.14%) were predominant constituents in all analyzed populations, representing 55.70-68.00% of the total components. All of the analyzed sage populations from northern Albania comply with ISO 6571/2008 standard and European Pharmacopeia 7.0. for Dalmatian sage essential oil content, while only four populations met the requirements of the ISO 9909/1997 standard for Dalmatian sage essential oil’s composition.

The mixture of six components (pyrethrin I and II, cinerin I and II, and jasmolin I and II) constituting natural insecticide pyrethrin is responsible for the insecticidal activity of Dalmatian pyrethrum (Tanacetum cinerariifolium /Trevir./ Sch. Bip.). In order to obtain deeper knowledge of its insecticidal potential, the extraction of mentioned components from plant samples was carried out for the first time, by matrix solid-phase dispersion. The influence of different factors such as the type of sorbent, sorbent-sample ratio, the elution solvent and its volume, addition of co-sorbent and drying agent, affecting extraction yield was evaluated. The pyrethrin components were extracted under optimized conditions: florisil (0.5 g) as the sorbent, mixed with 0.25 g of pyrethrum flower heads, 5 mL of acetone-ethyl acetate (1:1, v/v) as the elution solvent and 0.4 g of Na2SO4 as the drying agent. All six components in the extracts were qualitatively and quantitatively determined by HPLC-DAD. Analytical method provides high degree of linearity with r2≻0.9974, except for pyrethrin II (0.9961). Also, the method showed good precision (intra- and inter-day) with a relative standard deviation below 14 %, and recoveries mainly above 80 %. In order to demonstrate the applicability of the developed method, three Dalmatian pyrethrum natural populations from Croatia were analyzed.

Three genes for resistance to Erysiphe pisi, named er1, er2 and Er3 have been described in pea so far. er1 gene is located in pea linkage group VI, while er2 gene has been mapped in LGIII. SCAR and RAPD markers tightly linked to Er3 gene have been identified, but the position of these markers in the pea genetic map was unknown. The objective of this study was to localize Er3 gene in the pea genetic map. Towards this aim, the susceptible pea cv. Messire (er3er3) and a resistant near isogenic line of Messire (cv. Eritreo, Er3Er3) were surveyed with SSRs with known position in the pea map. Three SSRs were polymorphic between ‘‘Messire’’ and ‘‘Eritreo’’ and further surveyed in two contrasting bulks formed by homozygous Er3Er3/er3er3 individuals obtained from a F2 population derived from the cross C2 (Er3Er3) 9 Messire (er3er3). A single marker, AA349, was polymorphic between the bulks. Subsequently, other ten markers located in the surrounding of AA349 were selected and analysed in Er3Er3 and er3er3 plants. As a results, another SSR, AD61, was found to be polymorphic between Er3Er3 and er3er3 plants. Further linkage analysis confirmed that SSRs AA349 and AD61 were linked to Er3 and to the RAPD and SCAR markers previously reported to be linked to this gene. Er3 gene was located in pea LGIV at 0.39 cM downstream of marker AD61. The location of Er3 gene in the pea map is a first step toward the identification of this gene.

The long tradition of common bean (Phaseolus vulgaris L.) cultivation in Croatia has resulted in the development of many landraces, which create a valuable basis for screening different physiological traits. Studying gas exchange parameters could provide valuable information that could be useful in creating new crop germplasm with enhanced agronomic attributes. Variability of gas exchange parameters, such as the net photosynthetic rate (A), transpiration rate (E), stomatal conductance (gs), intercellular CO2 concentration (Ci), light use efficiency (LUE), and water use efficiency (WUE) were studied on the collection of 226 common bean accessions and the relationship of the gas exchange parameters with the origin and morphological traits was tested. The highest variability among all landraces was found for the WUE and gs, and lowest for the A and LUE traits, indicating high potential of these traits to be used for selection and breeding programs. Accessions of the Mesoamerican origin had higher A and LUE and lower Ci values compared to accessions of the Andean origin. Accessions with the determinate growth had higher gs and Ci values and lower A, LUE and WUE values compared to accessions with the indeterminate growth. The variance components analysis showed that the growth type has a more pronounced effect on gs, Ci and WUE, whereas the origin has a stronger impact on A and LUE. The principal component analysis (PCA) showed that A separates accessions according to their origin, whereas Ci separates accessions according to their growth type. Accessions with higher Ci had lower A, indicating possibly higher rates of carboxylation per active site on the enzyme and/or higher regeneration capacity of Ribulose-1, 5-bisphosphate (RuPB) in the Calvin cycle in accession with high A.

Pre-breeding and elite breeding are two steps in creating high yielding sunflower hybrids that differ in well established procedures and selection methods. However, a methodology that bridge efficient use of introgression lines as product of pre-breeding procedures and their crossing to elite inbreed lines, is not yet very well established. Therefore, the development of cost- and time-efficient methods for the determination of best parent heterosis and the use of best inbred lines in crosses with introgression lines for obtaining high- yielding and stable hybrids is highly desirable. In this regard, sixteen Cytoplasmic Male Sterile (CMS) inbred lines (A) derived from four heterogeneous interspecific lines originating from three annual: H. debilis silvestris (DEB-SIL), H. praecox runyoni (PRA- RUN), H. deserticola (DES) and one perennial H. resinosus (RES) wild species were evaluated. Seven agronomic traits were measured over a period of 2 years and 38 DNA loci were analysed, in order to compare four different methods for the estimation of best parent heterosis (BPH). New inbred lines were characterized by Principal Component Analysis (PCA) of morphological traits and Principal Coordinate Analysis (PCoA) of molecular marker data. Line × tester mating design was used to evaluate General Combining Ability (GCA), while Genetic Distance (GC) estimated by markers was evaluated as a predictor of BPH by Locally Weighted Sequential Smoothing (LOESS). Analysis of combining ability is one of the most important tools breeders use to identify superior inbred lines on the basis of their performance in hybrid combinations. Results obtained in this research show that PCA of morphological and PCoA of molecular marker data on parental lines are generally in agreement with GCA effects for examined traits. GD versus BPH relationships indicate that intermediate to high GD between parental lines was optimal for best heterotic effects of most traits. In this study, we show that the combination of the PCA of morphological data, PCoA of molecular marker data and GD between parental lines is fast and affordable, giving the most important information for parental choice of introgression and elite lines in sunflower breeding programs.

Sea fennel (Crithmum maritimum L.) is a perennial plant tolerant to the soil salinity and could be used as an alternative culture on salty soils. For successful sea fennel cultivation, it is necessary to test germinability of the seed. In this study, germination tests have been carried out on a population of sea fennel from Lopar, island of Rab, Croatia. Th e aim was to examine the eff ect of seed priming on seed germination, radicle length, radicle surface area, average diameter of radicle and radicle volume of sea fennel under diff erent salt concentrations. Seeds were primed with sodium chloride (NaCl) (50 mM) or distilled water (dH2O) during seven days aft er which its germinability and early seedling growth was tested on the germination paper treated with diff erent concentrations of NaCl (50, 100 and 150 mM) and dH2O as control. Seeds that were primed with dH2O and 50 mM NaCl solution showed the better performance than nonprimed seeds ; priming alleviated negative eff ects of low NaCl concentration.

The essential oils of 25 populations of Dalmatian sage (Salvia officinalis L.) from nine Balkan countries, including 17 indigenous populations (representing almost the entire native distribution area) and eight non-indigenous (cultivated or naturalized) populations were analyzed. Their essential-oil yield ranged from 0.25 to 3.48%. Within the total of 80 detected compounds, ten (β-pinene, 1, 8-cineole, cis-thujone, trans-thujone, camphor, borneol, trans-caryophyllene, α-humulene, viridiflorol, and manool) represented 42.60 to 85.70% of the components in the analyzed essential oils. Strong positive correlations were observed between the contents of trans-caryophyllene and α-humulene, α-humulene and viridiflorol, and viridiflorol and manool. Principal component analysis (PCA) on the basis of the contents of the ten main compounds showed that four principal components had an eigenvalue greater than 1 and explained 79.87% of the total variation. Performing cluster analysis (CA), the sage populations could be grouped into four distinct chemotypes (A–D). The essential oils of 14 out of the 25 populations of Dalmatian sage belonged to Chemotype A and were rich in cis-thujone and camphor, with low contents of trans-thujone. The correlation between the essential-oil composition and geographic variables of the indigenous populations was not significant ; hence, the similarities in the essential-oil profile among populations could not be explained by the physical proximity of the populations. Additionally, the southeastern populations tended to have higher EO yields than the northwestern ones

oBackground and Aims Wild olive (Olea europaea subsp. europaea var. sylvestris) is important from an eco- nomic and ecological point of view. The effects of anthropogenic activities may lead to the genetic erosion of its genetic patrimony, which has high value for breeding programmes. In particular, the consequences of the introgres- sion from cultivated stands are strongly dependent on the extent of gene flow and therefore this work aims at quanti- tatively describing contemporary gene flow patterns in wild olive natural populations. oMethods The studied wild population is located in an undisturbed forest, in southern Spain, considered one of the few extant hotspots of true oleaster diversity. A total of 225 potential father trees and seeds issued from five mother trees were genotyped by eight microsatellite markers. Levels of contemporary pollen flow, in terms of both pollen immigration rates and within-population dynamics, were measured through paternity analyses. Moreover, the extent of fine-scale spatial genetic structure (SGS) was studied to assess the relative importance of seed and pollen disper- sal in shaping the spatial distribution of genetic variation. oKey Results The results showed that the population under study is characterized by a high genetic diversity, a relatively high pollen immigration rate (0o57), an average within-population pollen dispersal of about 107 m and weak but significant SGS up to 40 m. The population is a mosaic of several intermingled genetic clusters that is likely to be generated by spatially restricted seed dispersal. Moreover, wild oleasters were found to be self- incompatible and preferential mating between some genotypes was revealed. oConclusions Knowledge of the within-population genetic structure and gene flow dynamics will lead to identify- ing possible strategies aimed at limiting the effect of anthropogenic activities and improving breeding programmes for the conservation of olive tree forest genetic resources.

“VASO” is a Portuguese participatory maize breeding project (1984), where several maize landraces such as “Pigarro” have been selected both by a farmer’s (phenotypic recurrent selection) and a breeder’s approach (S2 lines recurrent selection). The objectives of this study were to determine the phenotypic and genotypic responses to participatory selection using these two different approaches, to clarify to which extent both selection methods preserve genetic diversity, and conclude what is the preferred method to apply in sustainable farming systems. The results, obtained via ANOVA, regression nalyses and molecular markers, indicate that for both selection methods, genetic diversity was not significantly reduced, even with the most intensive breeder’s selection. Although there were some common outputs, such as the determinated versus indeterminated ears, cob and ear weight ratio per ear and rachis 2, specific phenotypic traits evolved in opposite directions between the two selection approaches. Yield increase was only detected during farmer selection, indicating its interest on PPB. Candidate genes were identified for a few of the traits under selection as potential functional markers in participatory plant breeding.

The sweet chestnut (Castanea sativa Mill.) is a widely spread and important multipurpose tree species in the Mediterranean area, which has played an important role in human history. Natural events, such as glaciations, and human influence played significant roles in the distribution and genetic makeup of the sweet chestnut. In order to better understand how natural and human-mediated past events affected the current genetic diversity and structure of the sweet chestnut, we analysed populations from Central Europe and the western part of the Balkan Peninsula, utilizing ten polymorphic nuclear microsatellite markers. The study revealed the existence of three genetically and, to a large extent, geographically distinct and well-defined groups of sweet chestnut populations. Two not entirely separated groups of populations were detected in the northern part of the studied area and one in the southern. Our results indicate that the genetic structure of sweet chestnut populations in Central Europe and the western part of the Balkan Peninsula is the result of both natural colonization events and significant and lengthy human impact. Furthermore, it has been proven that the gene flow between cultivated/grafted trees’ and wild chestnut stands can influence their genetic structure. However, our results reveal that cultivated-to-wild introgression in the sweet chestnut is dependent on the close proximity of chestnut orchards and naturally occurring populations.

Common bean is one of the most cultivated and consumed grain legumes worldwide, showing a high level of genetic diversity. Here is presented a detailed review of development and mapping of simple sequence repeats (SSRs, microsatellite markers) in the common bean. In the last 25 years, common bean has been the subject of numerous genetic studies, in which the identification and use of SSRs were conducted, and lead to the development of genetic maps. First genetic maps of common bean have been developed in the 1990s and were based on different molecular markers, and included domestication genes and important agronomic traits. Later, SSRs allowed the genetic mapping of more narrow crosses that are often of interest in plant breeding. Most genetic maps have been correlated with the core map established in the recombinant inbred population BAT93 x Jalo EEP558, and includes different markers, RFLP (restriction fragment length polymorphism), RAPD (random amplified polymorphic DNA), AFLP (amplified fragment length polymorphism), and SSRs in particular. More than 2,000 SSR markers are available for the common bean and they are an important tool to evaluate the genetic diversity of common bean landraces. SSRs are also useful to evaluate intra-specific diversity within the genus Phaseolus., Grah je jedna od najviše uzgajanih i najviše korištenih zrnatih mahunarki u svijetu koja ima visoku razinu genetske raznolikosti. U ovom radu je prikazan detaljan pregled razvoja i kartiranja mikrosatelitnih biljega (SSR, ponavljajuće jednostavne sekvence) kod graha. U posljednjih 25 godina, grah je bio predmetom brojnih genetskih istraživanja u kojima je provedena identifikacija i upotreba SSR-ova što je dovelo do razvoja genetskih karata. Prve genetske karte graha su razvijene u 1990-ima i bile su bazirane na različitim molekularnim biljezima, a uključivale su gene koji su povezani uz udomaćenje i važna agronomska svojstva. Kasnije su SSR-ovi omogućili genetsko kartiranje križanaca koji su često od interesa u oplemenjivanju bilja. Većina genetskih karata povezana je s osnovnom genetskom kartom uspostavljenom na rekombinantnoj inbred liniji BAT93 x Jalo EEP558, koja uključuje različite biljege kao što su RFLP (polimorfizam dužine restrikcijskih ulomaka), RAPD (nasumično umnožena polimorfna DNA), AFLP (polimorfizam dužine umnoženih ulomaka) te SSR-ove. Više od 2 000 SSR biljega je dostupno za grah i oni su važan alat za procjenu genetske raznolikosti tradicijskih kultivara graha. SSR-ovi su također korisni za procjenu unutarvrsne raznolikosti unutar roda Phaseolus.