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Publikacije (63)

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S. Stanivuković, Milan Žujić, Mirjana Žabić, N. Micic, B. Bosancic, G. Đurić

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.

D. Delić, B. Lolić, G. Đurić, T. Jovanović-Cvetković

In July 2015, 179 grapevine plants belonging to 16 grapevine autochthonous cultivars were assessed for sanitary status using DAS ELISA test for the presence of: Grapevine fanleaf virus (GFLV), Grapevine leafroll-associated virus 1 (GLRaV-1), Grapevine leafroll-associated virus 2 (GLRaV-2)and Grapevine leafroll-associated virus 3 (GLRaV-3). Furthermore, surveyfor the phytoplasma presence and laboratory analyses using nested-PCR/RFLP assay was conducted at the beginning of September 2015 on grapevine cultivars which were not positive in DAS ELISA test for the presence of the four viruses. Out of 179 tested plants with DAS ELISA test, 146 (81%) were positive for the presence of at least one virus. The most widespread viruses were GFLaV- 1 and GFLaV- 3 with approximately 80 % of grapevines infected. Nested–PCR/RFLP assay showed that out of 33 tested samples 2 were positive for the presence of phytoplasmas from 16SrXII group. Sanitation of infected grapevine cultivars is needed in near future.

N. Micic, G. Đurić

Molecular confirmation of the process of differentiation of meristematic tissue of plants is primarily based on histomorphological characterization of the tissues carrying these processes. For perennial plants – fruit trees, the knowledge of these processes is important to define runtime type and pomotechnical treatments but also to identify the genes responsible for determination of the apices into a generative phase of differentiation. Which apex extraction techniques will be applied depends on the structure of buds, i.e. whether an apex differentiates only into generative elements – pure flower buds, or the differentiation goes into two directions: differentiation of leaf primordium with axillar meristematic dome and differentiation of generative elements of flowers in the peak or lateral zone of an apex – mixed buds. The fruit trees of the genus Prunus have purely flower buds and for this purpose the whole apex is taken, between protection – cover leaves, on lateral positions of all collateral buds at shoot nodes. The moment of initiation of determination and the dynamics of differentiation of generative buds are different at different type of shoots on a tree. It is therefore necessary to know the shoots growth cessation time which is in correlation with apex determination. Fruit trees of the genus Prunus are ending the growth of shoots by rejection of shoot peaks, and the cast away time depends on the type, length and the position of shoots.

G. Đurić, Mirjana Žabić, Marina Rodić, S. Stanivuković, B. Bosancic, B. Pašalić

The research on 10 old and indigenous pear cultivars was conducted during 2012 and 2013 in Bosnia and Herzegovina. The following characteristics were determined: 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, vitamin C, total phenolics, total flavonoids and antioxidant activity of the fruit cell juice. On the basis of the Principal Component Analysis (PCA) of pomological fruit characteristics the studied cultivars were divided into four main groups. Based on the PCA of biochemical traits the studied cultivars can be divided into three groups. Extremely high phenolic content in the cvs Mioljnjaca, Žutica, Poljakinja, Karamut and Gradiscanka recommends them for their inclusion in a breeding programme. The cvs Mioljnjaca and Poljakinja are also characterised by large and firm fruit and since the antioxidant capacity affects the duration of fruit storage, it is expected that these traditionally grown varieties can survive longer and keep their valuable nutritional ingredients longer.

N. Micic, G. Đurić, A. Životić

The yield potential of ten plum cultivars (‘Cacak's Beauty’, ‘Cacak's Best’, ‘Cacak's Fruitful’, ‘California Blue’, ‘Elena’, ‘Hanita’, ‘Katinka’, ‘Renclode Althan’, ‘Stanley’, and ‘Top’), exhibited through the structure of buds on long bearing shoots, was analyzed under the agro-ecological growing conditions of the Banja Luka region for two fruiting seasons (2012 and 2013). Long bearing shoots with collateral buds were the most productive type of fruiting branches, which is why the structure of buds on these branches is the basis for defining specific cultural measures in order to boost yield potential. Based on the established structure of buds on the nodes along the long bearing shoots and the established yield potential, the plum cultivars evaluated were classified into 4 groups: 1) cultivars with a predisposition to have a high yield potential, without need for specific treatments in fruiting control (‘Cacak's Beauty’ and ‘Top’); 2) cultivars with a predisposition to have a high yield potential with the application of cultivar-specific cultural measures for control of fruiting (‘Cacak's Best’, ‘Elena’, and ‘Stanley’); 3) cultivars that require differentiated cultural measures in the control of yield potential for regular fruiting (‘Cacak's Fruitful’, ‘California Blue’, and ‘Katinka’) and 4) cultivars that require innovative growing systems and specific cultural treatments to control fruiting (‘Hanita’ and ‘Renclode Althan’).

G. Đurić, B. Lolić, M. K. Zeljković, D. Delić, M. Koprivica, M. Radulović, P. Nikolic, N. Micic et al.

Detection of viruses presence were carried out for the 225 a of pome and stone fruit trees from the collection of the Genetic Resources Institute of University of Banja Luka, located within the Botanical Garden of the University, tested by DAS-ELISA. The pome fruit trees were analyzed on presence of the following viruses: Apple Chlorotic Leaf Spot Virus (ACLSV), Apple Stem Grooving Virus (ASGV), Apple Stem Pitting Virus (ASPV) and Apple Mosaic Virus (ApMV). The stone fruits were analyzed on presence of Plum Pox Virus (PPV), Prune Dwarf Virus (PDV) and Prunus Necrotic Ring Spot Virus (PNRSV). All samples were tested serologically by DAS-ELISA. In addition to this, virus negative pear and apple accessions were tested for 'Candidatus Phytoplasma mali' and 'Candidatus Phytoplasma pyri' presence using nested-PCR/RFLP analyses.

M. Maras, B. Pipan, J. Šuštar-Vozlič, V. Todorović, G. Đurić, M. Vasić, S. Kratovalieva, A. Ibusoska et al.

In this study, genetic diversity of 119 accessions of common bean (Phaseolus vulgaris) from five former Yugoslav republics constituting the western Balkans was assessed by 13 microsatellite markers. This set of markers has proven before to efficiently distinguish between bean genotypes and assign them to either the Andean or the Mesoamerican gene pool of origin. In this study, 118 alleles were detected or 9.1 per locus on average. Four groups (i.e., Slovene, Croatian, Bosnian, and Serbian) showed similarly high levels of genetic diversity as estimated by the number of different alleles, number of effective alleles, Shannon’s information index, and expected heterozygosity. Mildly narrower genetic diversity was identified within a group of Macedonian accessions; however, this germplasm yielded the highest number of private alleles. All five germplasms share a great portion of genetic diversity as indicated by the analysis of molecular variance (AMOVA). On the basis of the scored number of migrants, we concluded that the most intensive gene flow in the region exists in Bosnia and Herzegovina. Cluster analysis based on collected molecular data classified the accessions into two large clusters that corresponded to two gene pools of origin (i.e., Andean and Mesoamerican). We found that Andean genotypes are more prevalent than Mesoamerican in all studied countries, except Macedonia, where the two gene pools are represented evenly. This could indicate that common bean was introduced into the western Balkans mainly from the Mediterranean Basin. Bayesian cluster analysis revealed that in the area studied an additional variation exists which is related to the Andean gene pool. Different scenarios of the origin of this variation are discussed in the article. Common bean (2n = 2x = 22) is the most important edible food legume for direct human consumption in Europe and in the world as it represents a valuable source of proteins, vitamins, fiber, and minerals (Broughton et al., 2003). The Andean region and Mesoamerica are distinguished as the two major centers of origin of this species, according to morphological characters (Singh et al., 1991), seed proteins (Gepts et al., 1986), isozymes (Koenig and Gepts, 1989), DNA markers (Freyre et al., 1996), and sequence data (Schmutz et al., Received for publication 10 Feb. 2015. Accepted for publication 14 Apr. 2015. This work was financially supported by FP7 Project CropSustaIn, grant agreement FP7-REGPOT-CT2012-316205, by grant No. 168/01 from the SEE-ERA.NET.PLUS FP7 Regional Programme and by grant P4-0072 from the Slovenian Research Agency. Accessions in Republic of Srpska, were collected through the National Program for Plant Genetic Resources, with a financial support by Ministry of Science and Technology of the Republic of Srpska.We are thankful toMatej Knapi c fromAgricultural Institute of Slovenia for preparing a geographic map of the western Balkans with collection sites of the studied common bean accessions. Corresponding author. E-mail: marko.maras@kis.si. 308 J. AMER. SOC. HORT. SCI. 140(4):308–316. 2015. 2014). After its domestication in the Americas, common bean promptly spread worldwide (Zeven, 1997). Introduction of this species in Europe dates to the early 16th century when Spanish and Portuguese sailors brought bean specimens to their homelands from both centers of domestication (Gepts and Bliss, 1988). During the last five centuries of cultivation, many landraces and cultivars evolved under diverse environments and farmer preferences in Europe (Zeven, 1997). Though many local cultivars were lost in the last 60 years, there are still many farmers who maintain old local landraces, which are well adapted to the pedoclimatic conditions peculiar to their limited geographical areas, and who have been exchanging their seeds with surrounding areas, mainly in local markets. The pathways of dissemination of the common bean into and across Europe were very complex, with several introductions from America, combined with direct exchanges between European and otherMediterranean countries (Papa et al., 2006). In the past two decades, phaseolin seed protein and other genetic markers have been intensively used to analyze the structure of European common bean populations and distribution of the two gene pools. A prevalence of the Andean ‘‘C’’ and ‘‘T’’ phaseolin types (76%) was first detected by Gepts and Bliss (1988), and was then confirmed by Lioi (1989) in an analysis of a large collection from Italy, Greece, and Cyprus (66% in total), by Logozzo et al. (2007) for a broad European collection (76%), and by others for Portuguese and Spanish genotypes (Rodino et al., 2001, 2003). Similar distribution of Andean and Mesoamerican genotypes has also been observed in phaseolin and molecular marker analyses at a regional scale (Angioi et al., 2009; Limongelli et al., 1996; Piergiovanni et al., 2000; Sicard et al., 2005; Su star-Vozli c et al., 2006). Moreover, several studies have focused on hybridization between the Andean and Mesoamerican gene pools in Europe. This phenomenon was first evidenced in the Iberian Peninsula by analyzing phaseolins, allozymes, and morphological characters (Rodino et al., 2006; Santalla et al., 2002), and later by inter-simple sequence repeat and simple sequence repeat (SSR) markers from both the chloroplast and nuclear genomes of European genotypes (Angioi et al., 2009, 2010; Sicard et al., 2005). Information on genetic diversity of common bean in the western Balkans that encompasses former Yugoslav republics (i.e., Slovenia, Croatia, Bosnia and Herzegovina, Macedonia, and Serbia) is scarce. In this region, common bean represented a very important food in the human diet for centuries. Until World War II, this crop was grown on large areas (>1 million ha) in the field often together with maize (Zea mays). In the second half of the last century new cultivars of both maize and common bean were introduced into crop production, and the old cropping system was abandoned, which subsequently, lead to a great reduction of the areas covered by beans ( 120,000 ha). Different approaches for assessing diversity at the molecular level are presently available. Microsatellites have been considered as the reference markers for cultivar fingerprinting in common bean because they are codominant, widely distributed in the genome, highly polymorphic, and highly repeatable (Powell et al., 1996; Yu et al., 1999). In this study, the genetic diversity of common bean from the western Balkans was assessed by SSR markers. A total of 13 markers that proved in previous studies (Maras et al., 2006, 2013) to be highly polymorphic and as efficient as amplified fragment length polymorphism markers in distinguishing common bean genotypes according to their gene pool of origin (Maras et al., 2008)were employed. The collectedmolecular data allowed us to: 1) examine the relationships among the accessions and the organization of common bean genetic variation in the western Balkans, 2) identify the original gene pool (Andean or Mesoamerican) of the studied plant material, and 3) clarify the bean dissemination process in the western Balkans. Materials and Methods PLANT MATERIAL. A total of 119 common bean landraces from national gene banks of five former Yugoslav republics were used in this study (Table 1; Fig. 1). These included 25 accessions from Bosnia and Herzegovina, 18 from Croatia, 28 from Macedonia [former Yugoslav Republic of Macedonia (FYROM)], 30 from Serbia, and 18 from Slovenia (passport data of the accessions are available upon request). Out of 18 Slovene accessions included, 14 of them have already been assessed for genetic diversity and phaseolin type in our previous studies (Maras et al., 2013; Su star-Vozli c et al., 2006) and were used here as a reference material for the determination of gene pool of origin of the other 105 accessions. DNA EXTRACTION. Total DNA was extracted from bulked leaf material of 10 plants of each accession using BioSprint15 DNA Plant Kit (Qiagen, Germantown, MD) and MagMax Express Magnetic Particle Processor (Life Technologies, Grand Island, NY) following manufacturer’s instructions. Integrity and quality of DNA were evaluated by electrophoresis on 1.0% agarose gels. Concentrations of DNA samples were determined with a fluorometer (DyNA Quant 200; Hoefer, Holliston, MA). MOLECULAR ANALYSES. Thirteen SSR loci developed by Metais et al. (2002) andGaitan-Solis et al. (2002)were employed (Table 2). Amplification reactions were performed with a Veriti Thermal Cycler (Life Technologies) in 10-mL reaction mixtures. Each reaction contained 1 · polymerase chain reaction (PCR) buffer, 2 mM MgCl2, 200 mM nucleoside triphosphates, 0.25 mM unlabeled right primer, 0.25 mM labeled left primer, 0.5 U of Taq DNA Polymerase (Biotools, Madrid, Spain), and 20 ng of genomic DNA. Loci were amplified using a profile of initial denaturation at 95 C for 3 min, followed by 30 cycles of strand denaturation at 94 C for 30 s, primer annealing at 47 to 62 C for 30 s, DNA extension at 72 C for 30 s, and final extension at 72 C for 4min. Fluorescently labeled PCR products were mixed with formamide and internal size standard GeneScan350 ROX (Life Technologies) and genotyped on the 3130xl Genetic Analyzer (Life Technologies). DATA ANALYSES. For each SSR marker, alleles of different sizes were scored. Basic statistics, including observed number of alleles, expected heterozygosity, polymorphic information content (PIC), and probability of identity (PI) were calculated in Identity 1.0 (Wagner and Sefc, 1999) and MicrosatelliteToolkit (Park, 2001). The number of total, effective, and private alleles and alleles with frequency over 5% were calculated for each of the five groups of accessions using GenAlEx 6.1 (Peakall and Smouse, 2006). The same software was used for the estimation of Shannon’s information index and expected heterozygosity of overall loci in single groups of accessions. A

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