Antić, M., Zeljković, M. K. & Đurić, G. (2020). Diversity assessment of wild cherry germplasm by using RAPD markers. Bulg. J. Agric. Sci., 26 (2), 404–408 The value of wild relatives of cultivated plants for food safety is widely recognized, but there is still a lack of knowledge about the diversity that exists and how this diversity can be used to improve cultivated plants. The genetic integrity of the wild fruit species is threatened by habitat loss due to increased fragmentation and degradation of the forest and the hybridization with the cultivated varieties. Prunus avium L. have been listed on a EUFORGEN priority lists for development of conservation strategies. Investigations of the genetic diversity and structure of local populations are required to determine the most suitable conservation policies for this species at different scales. In this study the RAPD markers were used in order to evaluate genetic similarity between 31 wild cherry trees from 13different test polygons. The test polygons are placed in Forest park Starčevica close to Banja Luka, Bosnia and Herzegovina. Primers OPA-04, OPA-07, OPA-14, OPA-15, OPA-17, and OPG-10 are used to analyse wild cherry trees. High degree of polymorphism is determined between analysed wild cherry trees which imply that in Forest Park Starčevica are present different genotypes of wild cherry.

According to the oral tradition, four to five degrees of kinship back, at the locality of the village of Batočići, the hamlet of Bare, the municipality of Čajnice – Republic of Srpska in BiH, there was an appearance of a tree with atypical sympodial graft in the spiral or alternating position of annual growth with a different tendency of negative heliotropism. This tree reached a height of up to 20 m and dried at the age of about 170 years. According to the oral tradition, after a longer period, between two and three degrees of kinship backward, the appearance of new single beech trees with the same characteristics in the formation of habit was observed. These new trees appeared successively in a relatively narrow space around the first tree, which for this reason was called the "Mother of Fairy Beech". Morphological characteristics of tree habit in this beech population conform with the variety of "tortuous beech" Fagus sylvatica var. – tortuosa, as well partly conforming to the variety of "weeping beech" Fagus sylvatica var. pendula. In this research, the question of inheritance of irregular sympodial branching with a tendency of negative heliotropism in the population of "Fairy Beech from Čajniče" has been raised, regardless of the characterization of the varieties. Thus, in the population of Fairy Beech from Čajniče, which makes about 40 trees in relatively close surroundings, four trees aged over 30 years have been identified in order to determine whether their tree form is reproduced by vegetative cloning and generatively, by seed. The research was conducted in the period from 2014 to 2016 at the Institute for Genetic Resources of the University of Banja Luka. The results show that the form of habit of all four parent trees are conveyed by branching as well as by seed, which proves that there was a mutation of the first tree, i.e. the mother of Fairy Beech from Čajniče, which is transmitted to the offspring.

Fruit germplasm plays an important role in the glob a agrobiodiversity and is a source for both a direct use of fruit genetic resou rces as well for fruit breeding programme. The ex-situ field collection is still the main way to successf ully conserve fruit germplasm. Thirty pear accessions fr om Bosnia and Herzegovina were characterized during three years in the ex situ collection maintained by the Institute of Genetic Resources of the University of B anja Luka. The following characteristics were determined: flowering time, harv est maturity time and global tree architecture. The obtained results showed that 53.3% of pear accessions flowered during all three years, 40% of them during two years and 6.7% of them during only one year. The most present tree archite tur form was upright. According to the harvest maturity time, extremely ea rly ccessions were the most represented, than early, medium and very early pear accessions. Conserved pear germplasm in this ex situ collection represents a valuable material for dire ct use and future breeding programmes.

Studying the hazel flowering was carried out in agro-ecological conditions of the Banja Luka in the period from December to March during 2010/2011 and 2011/2012. The study was conducted on 13 hazel cultivars in two orchards. In the first orchard, the following four cultivars were observed: ‘Tankoljuskasti’, ‘Multiflora’, ‘Merveille de Bollwiller’and ‘Tonda Gentile Romana’, whilst the second orchard included the following: ‘Hall’s Giant’, ‘Gustav’s Zellernuss’, ‘Istrian Round’, ‘Avellana’, ‘Romai’, ‘Apolda’, ‘Ludolph’s Zellernuss’, ‘Imperiale de Trebizonde’and ‘Jean’s’. Phenological observations of the development of flowering phenophase with the aforementioned cultivars were monitored every three days in both years. In 2010/2011 the blooming of male and female inflorescence of the observed hazel cultivars lasted from late December to mid-March in total. The presence and pollen germination was proven to be a limiting factor for a successful pollination and fruit set in 2011. Namely, the pollen of cultivars: ‘Hall’s Giant’, ‘Istrian Round’, ‘Ludolph’s Zellernuss’, ‘Avellana’, ‘Imperiale de Trebizonde’ and ‘Romai’ did not germinate, and with ‘Gustav’s Zellernuss’, pollen germination was very low (7,22%). In ‘Apolda’, ‘Jean’s’, ‘Merveille de Bollwiller’, ‘Tonda Gentile Romana’, ‘Multiflora’ and ‘Tankoljuskasti’ pollen germination ranged between 28,57-81,69%. In 2011/2012 the flowering of male and female inflorescence of the observed hazel cultivars started 30 to 45 days later than the previous year i.e., female inflorescence of all the observed cultivars bloomed in early and mid-March, except in ‘Tankoljuskasti’, in which female inflorescence bloomed in late January and early February. Male inflorescence of ‘Tankoljuskasti’, ‘Hall’s Giant’ and ‘Gustav’s Zellernuss’ bloomed in the second half of January and early February, and in all other cultivars in mid-March. Pollen germination in 2012 proved to be a limiting factor for ‘Apolda’ and ‘Romai’, as their pollen did not germinate, as well as for ‘Hall’s Giant’ and ‘Jean’s’ where pollen germination was below 5%. For other cultivars, pollen germination was very good and ranged from 37,24 (‘Merveille de Bollwiller’) to 73,97% (‘Multiflora’). Based on the analysis of dynamics of blooming and pollen germination of 13 hazel cultivars in two years, the monitoring of pollen germination and strategy of artificial pollination must be adopted as a basic approach to control hazel fertility in the conditions of the Banja Luka region.

A large number of grapevine cultivars have functionally female flowers. The causes of male sterility in these cultivars in the spheres of microsporogenesis and pollen morphology are only partially tackled. This study examines microsporogenesis and pollen functional ability in two most economically important indigenous grapevine cultivars in Bosnia and Herzegovina by the evaluation of permanent histological sections and the use of a scanning electron microscope. When observed at the cytogenetic and embryological level, microsporogenesis in cvs. Blatina with sterile pollen and Žilavka with fertile pollen shows regularity, they coincide and there are no differences in the sequence of differentiation events, except in their timing and duration. In ‘Žilavka’, the development of pollen in the anther locule at all sequential events, including pollen release from the tetrad, formation of the structural elements of the exine, physiological and morphological preparations for pollen to be dispersed and its release from the anthers, progresses normally with typical – vital cytogenetic characteristics maintained. In ‘Blatina’, during pollen development in the anther locule upon release from the tetrads and during the formation of the structural elements of the exine, uncontrolled coverage of pollen by the exine occurs, ultimately resulting in the formation of inaperturate pollen – morphologically sterile pollen grains.

The aim of this research is to examine the morphological and pomological characteristics of the wild pear population in the northwestern part of Bosnia and Herzegovina with the purpose of allocating genotypes for breeding programs of varieties and rootstocks. During 2012 and 2013, the study of the wild pear characteristics was carried out on the localities of: Banja Luka, Manjaca, Potkozarje and Kozara. This research included morphological characterization of vegetative organs and pomological characterization of the fruits of all the selected genotypes. The shape of leaves in all the examined genotypes was elongated. The study of wild pear populations in the Banja Luka region has shown that wild pears are characterized by mostly smaller fruits, rounded form, while a few genotypes had fruits of elongated and flattened shape. Fruit weight varied from 7.44 g to 21.47 g and 73% of genotypes had fruit weight of over 10 g.

Th e paper presents the results of the analysis of pomological-technological characteristics of 13 hazel cultivars present in production plantations in the area of Bosnia and Herzegovina (BIH). Th rough previous analysis of pomological characteristics of fruit in several hazel plantations it was noted that there was a possibility of erroneous cultivar denomination. Th ese fruits are sold at the domestic market as hazels for consumption, as well as for confectionary and cosmetic industry. Due to the increasing interest of producers in BIH for hazel cultivation and the uncontrolled spread of seedlings by producers through the off shoots from the existing production plantations, a possibility has been created to establish a plantation with erroneous assortment and thus an inadequate cultivar composition, particularly in terms of pollination and fertilization. Th erefore, a comparison was carried out of pomological characteristics of fruit and the content of oil in the kernel of 13 hazel cultivars from older plantations in BIH with cultivars from the Collection of Nuts in Maribor owned by the Biotechnical Faculty of Ljubljana. Based on the analysis conducted, a deviation of fruit characteristics of some cultivars was determined, compared to the standard features of cultivars by which names they were labelled. Cultivars labelled by the producer during the collection of samples as: ‘Istrian Round’, ‘Tonda di Giff oni 2’, ‘Hall ś Giant’ and ‘N.N.1’ were identifi ed and they fully corresponded with their characteristics to the following cultivars: ‘Istrian Long’, ‘Mortarela’, ‘Fertile de Coutard’ and ‘Hall ś Giant’. Th e research also showed that the fruits of the examined cultivars in BIH according to their pomological properties are as good as the fruits of cultivars grown in other areas.

During spring 2016, a survey was carried out in Bosnian-Herzegovinian (BiH) and Montenegrin (MNE) fig orchards, germplasm collection plots and outdoor gardens, to investigate the presence of unreported fig viruses possibly present in both countries, i.e. Fig leaf mottle-associated virus 2 (FLMaV-2), Fig latent virus 1 (FLV-1), Fig cryptic virus 1 (FCV-1), Fig fleck-associated virus (FFkaV) and Fig badnavirus 1 (FBV-1); as well as those previously reported, i.e. Fig leaf mottle-associated virus 1 (FLMaV-1), Fig mild mottle-associated virus (FMMaV) and Fig mosaic emaravirus (FMV). A total of 84 fig samples (49 from BIH and 35 from MNE) were collected and tested by PCR/RT-PCR using sets of virus-specific primers. Results showed that FBV-1 was the prevailing virus with all samples (100%) infected, followed by FLMaV-1 (54% ) , FMV (35%), FMMaV (7%), FFkaV (6%) and FLMaV-2 (1%); whereas FLV-1 and FCV-1 were not detected. Excluding the FBV-1 detection, 35% of tested trees were infected with at least one other virus. Sequence analyses of PCR/RT-PCR fragments obtained from different viruses showed that FBV-1 was the least variable (0.9% of nucleotides divergent) compared with FLMaV-1 (15.7% sequence variation), FLMaV-2 (17.4%), FMMaV (14.9%), FMV (16.9%) and FFkaV (14.3%). Phylogenetic trees constructed with obtained sequences, together with their homologues retrieved from the Genbank database, showed distinct separation of the BiH and MNE isolates from those of different origins, in particular for FFkaV and FMV; whereas for closteroviruses (FLMaV-1, FLMaV-2 and FMMaV), there was no distinction between the isolates. This is the first report on sequence analyses of fig viruses in this geographical region, and of the presence of FBV-1 in BiH and MNE, and of FLMaV-2 and FFkaV.

The assessment of quality of old apple varieties was made in order to preserve and use these germplasm in the area of Bosnia and Herzegovina. The characterization was done for ten old varieties of apples selected on the basis of good characteristics of sensory analysis. The following parameters were considered: fruit weight, fruit length and width, stalk length and width, fruit flesh firmness; soluble solids content and total dry matter content of the fruit juice; pH, titratable acidity (TA), vitamin C, total phenolics, total flavonoids and antioxidant potential evaluated by DPPH radical scavenging assay. Statistical analysis was performed separately for sensory, pomological and biochemical characterization (in two consecutive years) by analysis of variance. Only traits such as weight, width and height of the fruit were the biggest in ‘Vinjugara’ which had a low content of titratable acidity whereby it could be singled out as an acceptable variety for consumers. ‘Đedovaca’ had the highest content of phenols (1711.8 mg GAE/100 g of fruit weight), which is in complete relationship with good free radical scavenging activity (11.1 mg of fresh fruit/ml). ‘Kolacara’ and ‘Gospoinjaca’ were distinguished as particularly interesting for good nutritional characteristics, primarily the content of phenols, flavonoids and free radical scavenging ability, but also for suitable storage characteristics, °Brix content and firmness. Due to favourable pomological and biochemical characteristics these varieties were recommended to be included in the breeding program that will result in the enrichment of germplasm of the apples from these areas.