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

ECPGR recommended simple sequence repeat loci for analyses of European plum (Prunus domestica) collections

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

Distribution and Stability of Polyphenols in Juices Made from Traditional Apple Cultivars Grown in Bosnia and Herzegovina

The present research was undertaken to investigate polyphenolic profiles of peel, pulp and juices made from two standard commercial and five traditional apple cultivars from Bosnia and Herzegovina. The main goal of the study was to monitor the distribution and changes of polyphenolic profiles through different phases of apples’ processing into cloudy and clear juices, with regard to L-ascorbic acid pretreatment. Quantitative determination of phenolic compounds was carried out by using high-performance liquid chromatography with diode-array detection. The obtained results showed that traditional cultivars, namely ‘Paradija’ and ‘Prijedorska zelenika’, displayed significantly higher content of these compounds compared to commercial ones. Flavan 3-ols and flavonol glycosides were mostly found in peels of all cultivars (21.2–44.1 and 5.40–33.3%, respectively), while phenolic acids along with flavan 3-ols were predominant in the pulp (8.20–30.8 and 5.10–13.9%, respectively). Apples’ processing into juices caused decrease (more than 90%) in the content of all polyphenols and the distribution of these compounds from fruits to final products had a negative trend, particularly evident in clear juices. The most drastic loss occurred in the flavonol glycosides and dihydrochalcones content, while chlorogenic acid displayed quite stable distribution from apples to final products due to its good solubility. Apple mash pretreatment with L-ascorbic acid had a positive impact on the preservation and retention of polyphenols.publishedVersio

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

Occurrence and Distribution Patterns of Plum Tree Viruses and Genetic Diversity of Sharka Isolates in Bosnia and Herzegovina

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).European Cooperation in Science and TechnologyPeer Reviewe

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

Genetic variability within and among generative dog rose (Rosa spp.) offsprings

Divlja ruža (Rosa canina L.) je kozmopolitska biljna vrsta koja zbog svoje prehrambene, medicinske i kozmetičke vrijednosti zaslužuje veću pažnju. Divlja ruža uzgaja se u Hrvatskoj na vrlo malim površinama, a plodovi za preradu uglavnom se uvoze. U ovom istraživanju kroz dvije vegetacije praćena su F1 potomstva (populacije) triju elitnih biljaka porijeklom iz Korčule (P1), Malešnice (P2) i Prigorja Brdovečkog (P3) porijeklom iz dvije regije (mediteranska i kontinentalna) radi procjene genetske varijabilnosti pomoloških i agronomskih svojstava. Dodatno, genetska varijabilnost unutar i između tri ispitivane populacije divlje ruže analizirana je i primjenom 6 polimorfnih SSR (Simple Sequence Repeats) markera. Najveća varijabilnost, kao i najpoželjnija agronomska svojstva ustanovljena su unutar populacije P3 dok su najniže vrijednosti istih svojstava utvrđene u populaciji P1. Genotip RC29 (P3) za većinu važnih svojstava pokazao je najviše vrijednosti. Molekularni podaci omogućili su potpuno razdvajanje populacija, ali ne i svih jedinki unutar pojedinih populacija. Selekcijom i hibridizacijom perspektivnih genotipova divljih ruža iz Hrvatske bilo bi moguće kreirati vegetativno razmnožene populacije i sorte prikladne za komercijalni uzgoj.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 Korčula, Malešnica and Prigorje Brdovečko, 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 (Simple Sequence Repeats) 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

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

Genetic Relationships and Diversity of Common Buckwheat Accessions in Bosnia and Herzegovina

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