Common buckwheat (Fagopyrum esculentum Moench.) has a long history of cultivation in the large, mountainous regions of Bosnia and Herzegovina (B&H). Its commercial production is mainly based on the regionally bred variety ‘Darja’, but numerous landraces are also being grown on a smaller scale. As part of the SEEDNet (Southeast European Network on Plant Genetic Resources) project, these landraces have been collected and stored at the Gene bank of the Faculty of Agriculture and Food Sciences in Sarajevo (FAFS). To assess their utilization value, it was first important to investigate if they represent distinct landraces and to identify their genetic relationships with the most commonly grown varieties in the region (‘Darja’, ‘Goluba’ and ‘Čebelica’). Therefore, the aim of this study was to assess the genetic relationships and diversity of the common buckwheat accessions maintained at the FAFS Gene bank, as well as the value of these accessions for future breeding programs, using microsatellite markers and seventeen quantitative and fifteen qualitative morphological traits. The FCA (Factorial Correspondence Analysis) and AMOVA (Analysis of Molecular Variance) revealed that several accessions represent completely distinct landraces which clearly differentiated from the most commonly grown cultivars ‘Darja’ and ‘Goluba’. Conducted morphological analyses revealed that several of the analyzed landraces hold similar characteristics to the ones observed in ‘Darja’ and ‘Goluba’, while others possess unique traits potentially useful in breeding programs.

Traditional pear cultivars are increasingly in demand by consumers because of their excellent taste, the possibility of use in sustainable food production systems, convenience as raw materials for obtaining products of high nutritional quality, and perceived health benefits. In this study, individual sugars, organic acids, and polyphenols in the fruits of nine traditional and one commercial pear cultivar during two growing seasons were determined by HPLC. A significant influence of cultivars, growing years, and their interaction on the content of analyzed primary and secondary metabolites was determined. The commercial pear cultivar ‘Président Drouard’ and traditional cultivars ‘Dolokrahan’, ‘Budaljača’, and ‘Krakača’ had a lower content of all analyzed sugars. Overall, traditional pear cultivars had higher total polyphenols in the peel and pulp than ‘Président Drouard’, with the exception ‘Takiša’ and ‘Ahmetova’. High polyphenol content detected in ‘Budaljača’, ‘Dolokrahan’, and ‘Krakača’ shows the utilization value of traditional pear germplasm. The obtained data can serve as practical supporting data for the use of traditional pears in the neutraceutical, pharmaceutical, and food industries.

In order to fill in a decade-long information gap regarding the biological, serological and molecular data for plum tree viruses in Bosnia and Herzegovina, a three-phase study combining symptom evaluation, and serological and molecular assays with high-throughput sequencing (HTS) technology was conducted. The most frequently observed symptoms were discolorations in the form of ring patterns, bands and irregular shapes, as well as vein banding. Sharka-associated symptoms in the form of ring patterns and semicircles were prevalent. A total of 468 plum tree samples were tested by ELISA for the presence of PPV, ApMV, PDV, PNRSV, PBNSPaV, ACLSV and MLRSV. An overall infection incidence of 51.9% was detected, with PPV being the most prevalent (48.7%), followed by PDV (2.99%), PNRSV (0.21%) and mixed infections of PPV+PDV (1.71%). RT-PCR-assisted strain typing in 45 samples revealed PPV-D as the most common strain (22.22%), followed by PPV-REC (6.66%). Mixed infections of PPV-D+PPV-REC were detected (6.66%). HTS enabled the recovery of a 9743 nts long sequence of PPV-D (PPV_O7/80, MW412433), which shared the highest nucleotide and amino acid identities with isolates S13 (LC375131) from Serbia, SVN1 (LC375132) from Slovenia and N9 (LC375129) from Bulgaria. The phylogenetic analysis of the whole genome placed the isolate of the D strain in a distinctive group with the Slovenian isolate SVN1 (LC375132). In addition, the (Cter)NIb/(Nter)CP fragment of a PPV-REC isolate (MW412434) obtained in this survey formed a separate group with previously known isolates from Bosnia and Herzegovina (BOS64Pl and BOS257Pl).

In order to best conserve, as well as utilize, traditional apple germplasm in Norway, an apple heritage cultivar collection was established in Ullensvang, western Norway, which aims to become the National Clonal Germplasm Repository. The establishment of the apple heritage cultivar collection was preceded by a molecular study that aimed to genotype a large number of apple accessions maintained in various ex situ sites in western and south-eastern Norway, using a rather small set of eight SSR markers. However limited, the marker set managed to identify synonyms, homonyms, and duplicates within and among the investigated collections. In this study, 171 apple accessions from the Ullensvang apple heritage cultivar collection were genotyped using a set of 20 different SSR markers. Approximately half of the accessions have been previously genotyped using eight SSR markers, enabling an assessment of whether the use of a larger marker set would yield a more accurate characterization. Based on the obtained molecular data, the apple heritage cultivar collection was determined to hold a key part of the overall genetic diversity of the Norwegian apple germplasm. Furthermore, the twelve additional SSR markers were able to differentiate several accessions groups originally thought to be synonyms, as well as to provide a more detailed insight into the genetic structure of this germplasm.

This study was conducted with the aim of developing fruit spirits by utilizing old (autochthonous) apple and pear cultivars that can be attractive to both consumers and producers. Consumers of spirits could enjoy the unique flavor, and producers could gain an opportunity for brand development. In total, eight old apple cultivars (Sarija, Žuja, Samoniklica, Prijedorska zelenika, Bobovec, Masnjača, Lijepocvjetka, and Šarenika) and three pear cultivars (Budaljača, Krakača, and Kalićanka) from Bosnia and Herzegovina were used for the spirits production and for characterizing the flavor of distillates. Golden Delicious was used as a representative of commercial apple cultivar. The aroma profile was conducted through the identification of minor volatile organic compounds (VOCs) and the sensory perception of spirits. Analysis of the VOCs was performed by gas chromatography mass spectroscopy (GC/MS) techniques after enrichment via solid-phase microextraction (SPME). Sensory evaluation was performed by 12 trained panelists. Overall, 35 minor volatile compounds were found in spirits: 13 esters, 7 alcohols, 6 acids, 5 terpenes, and 4 aldehydes. Significant differences were detected in the distribution and quantity of the VOCs, which were fruit cultivar-dependent. Spirits made from Šarenika apple cultivar showed the largest amount of all acids, especially short- and medium-chain fatty acids; however, this richness was not correlated with pleasant sensory attributes. Spirits obtained from Prijedorska zelenika and Masnjača apple cultivars had the best sensory attributes. Budeljača and Krakača pears are promising cultivars as flavoring in spirits production.

The diploid Celina/QTee® (‘Colorée de Juillet’ × ‘Williams’), one of the most promising pear cultivars developed by the Norwegian breeding program Graminor, was launched in 2010. In Norway, the flowering is medium to late, while the fruits ripen in the beginning of September. The fruits are attractive with an intense red blush (50%) on a green background. Although, ‘Celina’ is cultivated in the most climatically suitable regions for fruit cultivation, present in Norway, unfavorable environmental conditions for pear pollination can have a very negative effect on fruit set and consequent yield. The aim of this study was to determine the S-alleles of ‘Celina’, as well as its frequently used pollinizers, and, through paternity testing of ‘Celina’ seeds, give a recommendation regarding the most important pollinizers of this pear cultivar. In order to accomplish this, ‘Celina’ and its potential pollinizers were all S-genotyped. After harvest, seeds collected from ‘Celina’ fruit in 2017 and 2018 were genotyped using eleven microsatellite markers. Genomic DNA was also extracted from leaf material collected from ‘Celina’, as well as from five pear cultivars used as pollinizers in the three examined orchards, and analyzed using the same marker set. Subsequently a simple sequence repeat (SSR) database was constructed and used for gene assignment analyses with the aim of quantifying pollen donor contribution from individual pollinizers. The obtained results indicate that ‘Anna’, the only examined pollinizer that was fully cross-compatible with ‘Celina’, together with ‘Fritjof’, the genotype which had the highest flowering overlap with ‘Celina’, proved to be the most successful pollinizers across all seasons and orchards. Although both cultivars were ubiquitous in the examined orchards, either as planted trees or as branches introduced during the flowering period, they were the most abundant pollinizers in only one orchard each. It is therefore possible to conclude that pollinizer abundance has a secondary significance in pollinizer success within investigated ‘Celina’ orchards.

A set of nine Simple Sequence Repeat (SSR) loci, approved by the ECPGR Prunus working group, are proposed as a standard set for genotyping European plum accessions. These loci show sufficient reliability in spite of problems caused by hexaploidy. Polymorphism in the loci is high and enables differentiation between unique plum accessions as well as analyses of genetic grouping and overall genetic structure. A set of seven reference accessions are described. A compiled dataset with allelic information for 165 accessions is presented. Genetic structure reveals three different K-values (2, 4 and 9) demonstrating a major dichotomy between Prunus insititia-related accessions and cultivars belonging to Prunus domestica sensu stricto, as well as differentiation among minor subgroups defined by pomological traits and geographical origin.

The genotyping of European fruit tree collections has helped to identify synonyms, determine parentage, reveal key specimens in the collections and provide information on the development of modern cultivars from one or several progenitors. However, studies on European plum Prunus domestica L. accessions have been lagging behind, mainly because of the hexaploid chromosome number. In this co-operative study, 104 accessions conserved by 14 partners across Europe were phenotyped for 20 descriptors, and genotyped for 8 SSR loci together with 8 reference cultivars. Based on the descriptors and additional information supplied by the partners, as well as the scientific and horticultural literature, each accession was assigned to one of six pomological groups; (1) egg plums sensu lato (E), (2) prunes of the French d’Agen type (P/A), (3) prunes of the Central-Southeast European Zwetschen type (P/Z), (4) greengages (G), (5) mirabelles (M) and (6) bullaces, damsons and var. pomariorum (D/B). A MANOVA conducted on descriptor data revealed significant differentiation among the pomological groups as well as a geographic impact on the differentiation of local plum accessions in Europe. SSR data showed that two trios and seven pairs of genotypes had very similar allele profiles and possibly are genetically identical in spite of different accession names. An AMOVA indicated sparse genetic differentiation when accessions were grouped according to geographic origin whereas significant differences were obtained among pomological groups. A Bayesian analysis of genetic structure, as well as a discriminant analysis of principal components (DAPC), further revealed levels of similarity among and within the different pomological groups, suggesting that egg plums sensu lato (E) and greengages (G) can be referred to subsp. domestica while damsons and bullaces (D/B) but also Central-Southeast European prunes (P/Z) show more affinity to subsp. insititia. The small and possibly heterogeneous groups with mirabelles (M) and prunes of the d’Agen type (P/A) take an intermediate position suggesting a hybridogenic origin.

European plum cultivars (Prunus domestica L.) are hexaploid and partially self-fertile or self-sterile requiring compatible pollinizers with overlapping bloom times. Therefore, inter-planting of different pollinizer cultivars is recommended. In order to identify successful pollinizers of the plum cultivars ‘Edda’, ‘Opal’ (self-fertile), ‘Jubileum’, ‘Reeves’, ‘Mallard’, ‘Avalon’, ‘Cacanska Lepotica’ (self-fertile), and ‘Valor’, 60 fruits per cultivar were collected from nine orchards in 2017 and 2018, all of which were located in Ullensvang, western Norway. DNA extraction was subsequently conducted from the obtained embryos, followed by genetic characterization using seven microsatellite markers. Tissue samples from all possible pollinizers were collected during the summer of 2017 and the same DNA approach was conducted. Results showed that ‘Opal’ was the most successful pollinizer among the investigated plum cultivars. The main exception was ‘Cacanska Lepotica’, which consistently displayed very high level of self-pollination. The most successful foreign pollinizer of ‘Opal’ was ‘Mallard’. However, in more than two thirds of embryos extracted from ‘Opal’ fruits self-fertilization was determined. ‘Reeves’ was identified as the most successful pollinizer among embryos collected from ‘Valor’. Among the five cultivars (‘Edda’, ‘Jubileum’, ‘Reeves’, ‘Mallard’, and ‘Avalon’) that did not display self-pollination, the pollinizer success rate of ‘Opal’, ranged from 36.5% (‘Mallard’) to 93.5% (‘Edda’) in 2017, while in 2018 this rate ranged from 43.5% (‘Jubileum’ and ‘Reeves’) up to 96.5% (‘Edda’). Overall, genotyping embryos using SSRs (simple sequence repeats) proved an effective method in determining the success rate of individual pollinizers among European plum cultivars.

A DNA marker-based study in European plum was performed within the project “Identification of a representative set of Prunus domestica accessions of European origin, well documented and characterized, to be included into the AEGIS system (PRUNDOC)”. A total of 46 local plum accessions from 10 European countries (Belgium, France, Germany, Greece, Italy, Latvia, Norway, Serbia, Slovakia and Sweden) were analysed using SSR loci. In addition, seven reference cultivars (Bistrica, Hanita, Mirabelle de Nancy, Reine Claude Violette, Stanley, Valor and Victoria) were analysed for standardization of allele sizes. The following nine primers were used; PacA33 is an EST-SSR developed in apricot, BPPCT039, BPPCT007, BPPCT014, BPPCT034, BPPCT040, UDP96 and UDP98 were developed from genomic peach DNA while CPSCT026 was developed from genomic DNA of Japanese plum Prunus salicina. None of the 46 investigated local plum accessions were identical, nor were any of them identical to any of the 7 reference cultivars. Genetic similarity among accessions was examined using Jaccard's similarity coefficients. The obtained dendrogram showed that the plum accessions did not group in a pattern corresponding to their country of origin.

Three commercial apple cultivars ('Jonagold', 'Granny Smith' and 'Idared') and the local apple cultivar ('Prijedorska Zelenika') from Bosnia and Herzegovina were analysed by HPLC-MS for the content of phenolic compounds in peel and pulp as well content of individual sugars and organic acids. Catechin, (-)-epicatechin, chlorogenic acid, caffeic acid, quercetin 3-O-xyloside, quercetin 3-O-arabinoside, quercetin 3-O-rhamnoside, quercetin 3-O-rutinoside, quercetin 3-O-galactoside and quercetin 3-O-glucoside were identified in apple peel and (-)-epicatechin, chlorogenic acid and caffeic acid in apple pulp at all observed cultivars. The total sugars content of analysed apple cultivars ranged between 91.61 and 105.45 g kg-1 FM, while the total organic acids content was from 5.70 to 15.05 g kg-1 FM. The levels of total organic acids and sugars, glucoce/fructose ratio and sugar/acid ratio were affected by cultivars. The mean content of total phenolic compounds was between 427.92 and 1457.95 mg kg-1 FM in peel and from 113.58 to 439.83 mg kg-1 FM in pulp and depending upon the cultivars. The presented data clearly demonstrated that traditional apple cultivar ('Prijedorska Zelenika') had significantly higher individual phenolic compounds in pulp in comparison to the commercial cultivars, i.e., 'Idared', 'Jonagold' and 'Granny Smith' and with respect of that 'Prijedorska Zelenika' is recommended as raw material for cloudy juice processing.

Dog rose (Rosa canina L.) is a cosmopolitan plant species that due to its nutritional, medical and cosmetic values deserves more attention. Dog roses are cultivated in Croatia on very small scale and most of the rose hip used in the domestic processing industry is imported. In this study, three dog rose F1 progeny populations developed from selected plants collected in Korcula, Malesnica and Prigorje Brdovecko, originating from two climatic regions (P1 - Mediterranean and P2 and P3 - continental) have been studied as a part of a field experiment for two growing seasons, in order to assess the level of genetic variability for pomological and agronomic traits. In addition, genetic variation within and among examined three dog rose populations has been assessed using 6 polymorphic SSR markers. The highest variability, as well as the most desirable agronomic traits were found within the P3 population, while the lowest ones were present in the P1 population. Most desirable properties and highest values for commercially important traits were detected in the genotype RC29 (P3). Molecular data was sufficient to separate three studied populations but not all individuals within the populations. Through selection and hybridization of perspective genotypes of dog roses from Croatia, it would be possible to create clonally propagated varieties suitable for commercial cultivation.