Morphological evaluation of common bean diversity in Bosnia and Herzegovina using the discriminant analysis of principal components (DAPC) multivariate method

In order to analyze morphological characteristics of locally cultivated common bean landraces from Bosnia and Herzegovina (B&H), thirteen quantitative and qualitative traits of 40 P. vulgaris accessions, collected from four geographical regions (Northwest B&H, Northeast B&H, Central B&H and Sarajevo) and maintained at the Gene bank of the Faculty of Agriculture and Food Sciences in Sarajevo, were examined. Principal component analysis (PCA) showed that the proportion of variance retained in the first two principal components was 54.35%. The first principal component had high contributing factor loadings from seed width, seed height and seed weight, whilst the second principal component had high contributing factor loadings from the analyzed traits seed per pod and pod length. PCA plot, based on the first two principal components, displayed a high level of variability among the analyzed material. The discriminant analysis of principal components (DAPC) created 3 discriminant functions (DF), whereby the first two discriminant functions accounted for 90.4% of the variance retained. Based on the retained DFs, DAPC provided group membership probabilities which showed that 70% of the accessions examined were correctly classified between the geographically defined groups. Based on the taxonomic distance, 40 common bean accessions analyzed in this study formed two major clusters, whereas two accessions Acc304 and Acc307 didn’t group in any of those. Acc360 and Acc362, as well as Acc324 and Acc371 displayed a high level of similarity and are probably the same landrace. The present diversity of Bosnia and Herzegovina’s common been landraces could be useful in future breeding programs

GENETIC PURITY ASSESSMENT OF COMMON BUCKWHEAT VARIETY ‘DARJA’ WITH THE USE OF SSR MOLECULAR MARKERS

In order to assess the genetic purity of common buckwheat variety ‘Darja’ which is the most commonly produced variety of this crop in Bosnia and Herzegovina, 10 SSR markers have been used. Five samples have been collected from different production regions in B&H (Breza, Nisici Plateau, Ustikolina, Bihac and Bosanska Krupa) and compared to the reference ‘Darja’ sample obtained from an ex situ seed collection from Slovenia. Seven out of ten primer pairs used managed to amplify SSR alleles. Analyses of molecular variance (AMOVA) showed a significant differentiation between the reference and all analyzed ‘Darja’ samples. Furthermore, the factorial correspondence analysis revealed a clear differentiation between the reference and ‘Darja’ samples from the most known production regions of common buckwheat in B&H clustering four out of five analyzed samples very close together. The most divergent one among the analyzed samples was the one from Ustikolina. Genetic purity of varieties of all of cross pollinated species produced in Bosnia and Herzegovina is questionable due to the general use of farm-saved seeds

Genetic structure of apple accessions maintained ex situ in Bosnia and Herzegovina examined by microsatellite markers

In order to identify possible mislabeling of the apple accession maintained ex situ in Srebrenik and to gain insight into the genetic structure of the conserved germplasm, 14 accessions from the collection were genotyped using 10 SSR (Simple Sequence Repeats) markers. Obtained SSR profiles were then added to an existing database constructed for previously characterized 24 traditional and 13 international, reference apple cultivars maintained at the same collection. Bayesian analysis implemented in the STRUCTURE program grouped 42 out of 51 analyzed apple accessions (38 traditional and 13 international) into three RPPs (reconstructed panmictic populations) with probability of membership qI higher than 75%. Almost all international, reference cultivars grouped in RPP3, whereas traditional B&H cultivars from the Srebrenik collection grouped in all three RPPs. Large and significant differentiations between all three individual RPPs were detected through the analyses of molecular variance and confirmed with FCA (factorial correspondence analyses). NJ cluster analysis, based on the Bruvo genetic distance, revealed that out of 38 traditional B&H apple cultivars, analyzed in the study, ‘Ljepocvjetka’, ‘Bobovec’ and ‘Bobovec J’ grouped closest to the international reference cultivars. Available date indicates that unlike a large number of B&H apple cultivars which were introduced during the reign of the Ottoman Empire, ‘Ljepocvjetka’ and ‘Bobovec’ were probably introduced at a later date. Cluster analyses also enabled the detection of one synonym and three homonyms within the collection. In four cases, previously conducted identification based on phenotypic analyses was confirmed by genetic analyses. Results of the structure analyses indicate a heterogeneous genetic structure of the analyzed accessions. This characteristic of the B&H apple germplasm could be useful for future breeding programs

Effects of Fertilization on Vegetative Growth, Yield and Fruit Quality of Black Chokeberry (Aronia melanocarpa Elliot)

Black chokeberry (Aronia melanocarpa Elliot) is a deciduous fruit shrub from Rosaceae family that does not have a long cultivating tradition in Bosnia and Herzegovina. Therefore, the aim of this research was to determine the effect of variety, quantity and combination of fertilizers on characteristics of vegetative growth, yield and fruit quality of black chokeberry of Nero cultivar, in agro - ecological conditions of the Una – Sana Canton (Bosnia and Herzegovina). The experimental site fertilized with the combination of mineral and organic nitrate fertilizers gave the best results in morphometric characteristics of fruit, yield and vegetative characteristics of growth. Fruits from the control site had the highest values when it comes to the most of the following characteristics: dry matter content, ash, total acidity, vitamin C and total phenolic content. Statistical analyses showed that factors of quantity and the manner of fertilization influence significantly most of examined parameters in this research (P ≤ 0.05)

GENETIC IDENTITY OF RASPBERRY ‘POLANA’ PLANTLINGS EXAMINED USING MICROSATELLITE MARKERS

Raspberry cultivars are clonally propagated and therefore all plants belonging to a single cultivar represent the same genotype. Cultivar integrity of raspberry plantlings placed on the market in Bosnia and Herzegovina (B&H) is based on examining of morphological traits, which is not a reliable tool for genetic identification. In this study plantlings declared as cultivar ‘Polana’ were genotyped using seven microsatellites, in order to gain preliminary insight into the genetic integrity of raspberry plantlings marketed in B&H. Plant tissue (leaves) from 10 raspberry plants were randomly sampled from a batch of plantlings sold by major fruit nursery in Bosnia and Herzegovina. Along with these samples, four reference cultivars with confirmed identity (‘Polka’, ‘Autumn Bliss’, ‘Heritage’ and ‘Polana’) were also included in the study. Seven primer pairs amplified 31 alleles, or on average 4.4 alleles per locus. UPGMA cluster analysis, based on the Jaccard similarity coefficient, revealed that among the ten samples declared as ‘Polana’ plantlings only five were genetically identical to any of the other samples. The cluster analyses also exposed that none of the ten samples declared as ‘Polana’ seedlings were in fact identical or even closely related to the ‘Polana’ reference cultivar or any of the other reference cultivars. These findings clearly show that the genetic identity of primocane raspberry plantlings , currently sold in Bosnia and Herzegovina, needs to be tested using objective and reliable methods rather than simple morphologic observation

Genetic structure and diversity of European chestnut (Castanea sativa Mill.) populations in western Balkans: On a crossroad between east and west

European chestnut (Castanea sativa Mill.) is highly valued in the western Balkans as a source of timber and fruit, but also as an important source of nectar and pollen for the production of honey. In this study, four chestnut populations, covering a major portion of the western Balkans, and a reference population from the northern Italy were examined using 21 microsatellite markers. The highest genetic diversity was detected within the populations geographically closest to the Italian Peninsula, which also displayed the highest level of admixture with the samples from Italy. The strongest genetic differentiation was noted among the southern and eastern chestnut populations from Bosnia and Herzegovina (B&H) (Gst=12.05%). This pronounced differentiation is probably caused by the genetic adaptations to notably different climatic conditions present in the south (Mediterranean climate) and east (Continental climate) of B&H. The clear genetic differentiation of the southern and eastern B&H chestnut populations from the Italian population, determined by pairwise Gst, FCA and Bayesian Structure analyses, indicates that these populations most likely originated from independent shelter zones (refugia), after the last glaciation period. Based on these results we propose a presence of an introgression zone in the northwestern Balkans, established through gene flow from the Italian and the Balkan Peninsula. The obtained insights into the structure of all analyzed populations will significantly contribute towards establishing a regional conservation and utilization strategy for European chestnut in western Balkans

Assessment of genetic relationships among common buckwheat (Fagopyrum esculentum Moench) varieties from Western Balkans using morphological and SSR molecular markers

In order to assess the genetic relationships and diversity among common buckwheat from Western Balkans, three varieties produced under the names ‘Čebelica’, ‘Darja’ and ‘Goluba’ were evaluated with a set of 10 SSR and 32 morphological markers. Eight of ten primer pairs used managed to amplify SSR alleles, in average 7 alleles per locus. Analysis of molecular variance (AMOVA) revealed that only 2.6% of the total diversity was attributed to the differences among ‘Darja’ and ‘Goluba’. The largest percentage of variance between varieties was detected among ‘Goluba’ and ‘Čebelica’ (fCT = 0.136; p < 0.001). Factorial correspondence analysis also revealed a clear differentiation between these two varieties. Results of hierarchical clustering based on morphological data of the three analyzed common buckwheat varieties were not in a complete accordance with the results obtained through genetic analyses, as it displayed a much larger difference between ‘Darja’ and ‘Čebelica’, than between ‘Goluba’ and ‘Ĉebelica’. However, the differentiation among the analyzed varieties based on SSRs and Euclidean distances, calculated using morphological data, was the same. Namely, both approaches identified ‘Čebelica’ as the most divergent material among the analyzed varieties. The results of the genetic characterization indicate that the purity of varieties of cross pollinated species produced in Western Balkans is questionable due to use of seed material of unverified origin, as well as the general use of farm saved seeds