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

The Influence of Pesticides and Biopesticides on Morphological Changes in Raspberry (Rubus ideaus L. „POLKA“) Infected with the Bacterium Agrobaterium tumefaciens

Proizvodnja maline (Rubus idaeus L.), kao jedna od najperspektivnijih grana voćarstva u Bosni i Hercegovini, mogla bi biti trajno ugrožena zbog sve češćega pojavljivanja bolesti bakterioznoga tumora korijena, čiji je uzročnik Agrobacterium tumefaciens. Cilj ovoga rada bio je ispitati morfološke promjene na sadnicama mali ne izazvane bakterijom A. tumefaciens, kao i veličinu i brojnost tumora u zavisnosti od primijenjenoga biološkog ili kemijskog tretmana. U radu je korišten referentni soj Agrobacterium tumefaciens MTCC 431 (ATCC 15955). Morfometrijski parametri na sadnicama maline izmjereni su uz pomoć programa ImageJ 1.48v. Za mjerenje dijametra guka korišteno je binokularno povećalo pri uvećanju od 20x. Statistička analiza rezultata podrazumijevala je analizu glavnih komponenata na osnovi korela cijske matrice u računalnome programu R v. 3.4.0. Sadnice maline koje su inokuli rane bakterijom A. tumefaciens imale su niži potencijal rasta u odnosu na kontrolnu skupinu. Soj K1026 Rhizobium rhizogenes kao aktivna tvar biopesticida pozitivno je utjecao na rast zaraženih biljaka maline suzbijajući infekciju. Primijenjeno kemijsko sredstvo na bazi bakrova (I) oksida pokazalo je nešto manji učinak na intenzitet pro mjena uzrokovanih A. tumefaciens u odnosu na biološko sredstvo. U ovome je radu zabilježen i niži intenzitet promjena kod istodobne primjene biopesticida i fungicida u odnosu na pojedinačne tretmane.The production of raspberry (Rubus idaeus L.) as one of the most promising branches of fruit growing in Bosnia and Herzegovina could be permanently endangered due to the increasing incidence of crown gall disease caused by Agrobacterium tumefaciens. The aim of this paper was to examine the morphological changes on raspberry seedlings caused by the bacterium A. tumefaciens, as well as the size and number of tumors depending on the applied biological or chemical treatment. The strain Agrobacterium tumefaciens MTCC 431 (ATCC 15955) was used. Morphometric parameters on raspberry seedlings were measured using the ImageJ 1.48v program. A binocular magnifier at 20x magnification was used to measure the the gook diameter. The computer program R c. 3.4.0 was used for a statistical analysis of results and an analysis of main components. It was observed that the raspberries inoculated with A. tumefaciens had a lower growth potential when compared to the control line. Rhizobium rhizogenes strain K1026, as an active substance of biopesticide, had a positive effect on the growth of infected raspberry plants, reducing bacterial infection. The applied chemical agent based on copper (I) oxide demonstrated lesser impact on the intensity of A. tumefaciens infection when compared to the biological agent. In this paper, a lower intensity of changes in raspberry plants was observed when the biopesticides and fungicides were applied simultaneously, in comparison with the individual treatments

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

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

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 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

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

Water shortage and optimal pattern of field cropping cultivation: Addressing economic and environmental concerns in Qazvin Plain, Iran

peer reviewedIran is currently suffering from a severe water shortage. Frequent droughts, along with over-abstraction of surface and groundwater, have pushed the country's water situation to crisis levels. To deal with this issue, changing the cultivation pattern has been suggested. The planners stated that other environmental objectives would be achieved by implementing changes to the cultivation pattern along with reducing water consumption. Concerning this issue, the present study aimed to identify the optimal water-saving cultivation pattern in the field cropping's subsector in Qazvin province. This province is particularly important in the production of field cropping, which has caused the groundwater level to drop. Therefore, 246 farmers were selected using a simple two-stage random sampling method, and the information was collected in 2019. According to the experts' ideas, a 15% increase in gross profit, a 10% decrease in agricultural water consumption, job stability, and a 15% decrease in chemical fertilizers and pesticides were considered the optimal goals. The findings showed that through the optimal allocation of the inputs to Low Water-Demand (LWD) and High Water-Demand (HWD) crops, the farmers could not only save water and different types of fertilizers and pesticides but also achieve their objectives in terms of their desired profit. Therefore, designing a system of allocation and distribution, establishing a standardization system, and monitoring and controlling farmers' behavior concerning the use of inputs can achieve the goal concerning farmers' income and goals for protecting water resources and the environment

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&amp;H). Its commercial production is mainly based on the regionally bred variety &lsquo;Darja&rsquo;, 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 (&lsquo;Darja&rsquo;, &lsquo;Goluba&rsquo; and &lsquo;&#268;ebelica&rsquo;). 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 &lsquo;Darja&rsquo; and &lsquo;Goluba&rsquo;. Conducted morphological analyses revealed that several of the analyzed landraces hold similar characteristics to the ones observed in &lsquo;Darja&rsquo; and &lsquo;Goluba&rsquo;, while others possess unique traits potentially useful in breeding programs