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Sonja Umićević, Marina Antić, D. Rajković, L. Sinkovič, N. Nagl, S. Goreta Ban, V. Meglič, V. Todorović et al.

Leafy kale (Brassica oleracea var. acephala L.) is one of the most important Brassicaceae species with health-promoting properties and great diversity. To investigate variability within and among accessions, 12 accessions (each accession comprised four individuals/genotypes) from the Balkan region were genotyped at 12 simple sequence repeats (SSRs) loci. The selected SSR markers originated from the genomes of B. oleracea (eight), B. rapa (two) and B. napus (two). DNA was extracted from seeds, cotyledons, shoots, and young leaves, depending on the germination energy of each seed, using magnetic extraction method. Genetic analysis included eight accessions from Serbia (24 genotypes), two accessions from Montenegro (eight genotypes), and one accession from Bosnia and Herzegovina and Croatia (with four genotypes each). Accession-specific polymorphism was found at the Ol12-FO2 and Ol10-F11a loci, where genotypes were qualitatively distinguished by horizontal agarose gel electrophoresis (binary detection). Among 12 loci, the average polymorphic information content (PIC) was 0.71, with the marker Ol13-C12 proving to be the most informative (PIC = 0.89). However, the analysis of molecular variability (AMOVA) showed 11% diversity between accessions, 19% diversity between genotypes, and 70% diversity within genotypes, explaining the molecular diversity of native Balkan leafy kale germplasm. According to the genetic structure, four genetic groups were formed with an average expected heterozygosity of 0.70 between clusters. In order to exploit the genetic diversity, it would be advisable to evaluate these accessions at phenotypic level and use their potential in breeding programmes.

S. Rašeta, M. Antic, V. Todorović

In this research the aim was to determine differences in morphological characteristics between 11 tomato accessions from the Gene Bank of the Republic of Srpska. The experiment was conducted and analysis was performed during the 2018 and 2019 seasons. A total of 16 morphological characteristics (9 quantitative and 7 qualitative) were analyzed according to International Plant Genetic Resources Institute (IPGRI) descriptors for tomato. The results showed that polymorphism (diversity) was present in all quantitative characteristics and in 6 qualitative characteristics, while only one qualitative characteristic was monomorphic (no differences between accessions). Thereby, polymorphism was present in 93.75% of morphological characteristics. Out of a total of 9 quantitative characteristics, a highly significant difference (p 0.01) was found in all characteristics except for the 1000-seed weight since this characteristic had only one value per accession measured according to the IPGRI tomato descriptors. The accessions from the Gene Bank of the Republic of Srpska have shown high diversity in all qualitative characteristics except in plant growth type, which was indeterminate in all analyzed accessions. This research provides a new insight into the research area of diversity of tomato landraces from the Republic of Srpska, which is important for further promotion and sustainable use of germplasm not only for scientific research purposes but also for national rural farmers, who are the key to preserving traditional knowledge and skills related to the cultivation and use of traditional varieties and tomato landraces.

S. Zeljković, N. Paraðiković, V. Todorović, J. D. Gidas, D. Dumanović

Mushroom production has become more popular in our environment. The most common cultivated mushroom is Agaricus bisporus. After mushrooms are harvested a large amount of used compost remains. This compost is a good material and producers used it as alternative substrate in plant production. The benefits of this compost are numerous like high content of organic matter and the rich mineral composition. The aim of this study was to determine the effectiveness of the use of spent mushroom compost (as alternative substrate) on growth and development of roots and above-ground parts of sage transplants (Salvia officinalis L.). Measurements of morphological parameters of plants (plant height, number of leaves, number of branches and plant diameter) were performed, as well as determination of fresh and dry weight of roots and above-ground parts of transplants. Application of spent mushroom substrate in the production of sage Salvia officinalis L. positively influenced growth and development as well as fresh and dry weight of roots and above-ground parts of treated plants compared to nontreated plants during transplanting growing stage. Plant height (+104%), number of leaves (+65%), number of branches (+143%), plant diameter (89%), were significantly increased by the spent mushroom compost application compared to the control - commercial substrate.

Izudin Klokić, I. Koleška, D. Hasanagić, S. Murtić, B. Bosancic, V. Todorović

ABSTRACT Plant biostimulants are commercial products based on different chemical substances used to enhance plant growth and productivity. The effects of biostimulants, Megafol and Viva, with different dominant compositions (amino acids and humic acids) on two tomato cultivars (cv. Gravitet F1 and cv. Minaret F1) exposed to conventional and low-input macronutrient nutrition were investigated. Application of both biostimulants positively affected yield at conventional nutrition. In low-input nutrition variant, biostimulant application prevented yield loss only in cv. Minaret F1. Inter-cultivar differences were noticed in fruit characteristics (total phenol content, total flavonoids content and total antioxidative capacity) where application of biostimulants leads to opposite results in the examined cultivars. We considered the possibility of biostimulant usage as an effective nutritional addition for overcoming the problem of excessive fertilisation.

L. Sinkovič, B. Pipan, M. Antic, V. Todorović, S. Rašeta, V. Meglič

Grass pea belongs to the legume family and it is traditionally used as a grain for human consumption while its foliage is used for fodder. In this study morpho-agronomic evaluation and characterisation of different grass pea accessions (Lathyrus sativus L.) from Slovenia and Bosnia and Herzegovina (B&H)/Republic of Srpska was performed. A collection of 8 grass pea accessions was cultivated in Slovenia (Jablje) and Bosnia and Herzegovina/Republic of Srpska (Banja Luka) in the open field trials during the growing season of 2019. A number of quantitative and qualitative IPGRI descriptors for Lathyrus spp. concerning the vegetative growth, stem, branch, leaf, inflorescences, pods, and seeds were measured and/or visually estimated. The results of the present study will add value through the enrichment of the Lathyrus spp. collection in both countries, improving the existing data and documentation, as well as creating a certain basis for further studies of the morpho-agronomic traits of the grass pea.

Two commercial tomato cultivars were used to determine whether grafting could prevent decrease of Ca2+ concentration under salt stress conditions. The cultivars Buran F1 and Berberana F1 were grafted onto rootstock "Maxifort" and grown under three levels of the elevated soil salinity (S1 EC 3.80 dS m-1, S2 6.95 dS m-1 and S3 9.12 dS m-1). Ca2+ concentration of non-grafted plants of both examined hybrids was lower at all salinity levels in comparison to the control. In the fruits of grafted plants salt stress significantly decreased Ca2+ concentration only at the third salinity level (EC 9.12 dS m-1). The possibility of grafting tomato plants to improve influx of Ca2+ under salt stress conditions is discussed.

L. Sinkovič, B. Pipan, M. Vasić, M. Antic, V. Todorović, S. Ivanovska, C. Brezeanu, J. Šuštar-Vozlič et al.

In South-Eastern Europe, the majority of runner-bean (Phaseolus coccineus L.) production is based on local populations grown mainly in home gardens. The local runner-bean plants are well adapted to their specific growing conditions and microclimate agro-environments, and show great morpho-agronomic diversity. Here, 142 runner-bean accessions from the five South-Eastern European countries of Slovenia, Bosnia and Herzegovina, Serbia, North Macedonia and Romania were sown and cultivated in their respective countries and characterised using 28 quantitative and qualitative morpho-agronomic descriptors for Phaseolus spp. based on inflorescences, leaves, plants, pods and seeds. For each of these morpho-agronomic descriptors, the accessions can be classified into two or three specific groups. The highest correlations were observed within the fluorescence, seed and pod traits. The highest variability, at 76.39%, was between the different countries, representing different geographic origins, while the variability within the countries was 23.61%. Cluster analysis based on these collected morpho-agronomic data also classified the accessions into three groups according to genetic origins. The data obtained serve as useful genetic information for plant breeders for the breeding of new bean varieties for further studies of the morpho-agronomic traits of the runner bean.

Salinity is one of the most important ecological problems that affect irrigated agriculture in the world. Increased soil salinity inhibits plant growth through osmotic and ionic stress, but can also decrease availability of certain micronutrients. Copper is an essential metal for normal plant growth and development that participates in numerous biochemical and physiological processes and it is an essential cofactor for many metalloproteins. Copper concentration in two commercial tomato cultivars (grafted and non-grafted) was investigated under different levels of elevated soil salinity. Soil with EC 9.1 dS m-1 led to the highest copper deficiency, approximately by 37% in the non-grafted and 25% in the grafted tomato plants. The effect of a grafting technique is considered as an environmentally friendly tool for overcoming soil salinity problem.

I. Koleška, D. Hasanagić, V. Todorović, S. Murtić, Ivana Maksimović

1 University of Banja Luka, Faculty of Agriculture, Banja Luka, Republic of Srpska, Bosnia and Herzegovina 2 University of Banja Luka, Faculty of Natural Sciences and Mathematics, Banja Luka, Republic of Srpska, Bosnia and Herzegovina 3 University of Sarajevo, Faculty of Agricultural and Food Sciences, Sarajevo, Bosnia and Herzegovina 4 University of Novi Sad, Faculty of Agriculture, Novi Sad, Serbia

Murtic S., Oljaca R., Koleska I., Karic L., Todorovic V. (2018): Response of cherry tomato seedlings to liquid fertiliser application under water stress. Hort. Sci. (Prague), 45: 22–28. The aim of this study was to examine the impact of different liquid fertilisers on selected physiological parameters in order to evaluate the drought tolerance of cherry tomato seedlings. The following physiological parameters were investigated: total phenolic and flavonoid content, total antioxidant capacity and proline content of leaf extracts. Total phenolic and flavonoid content were determined using the Folin-Ciocalteu and aluminium chloride colorimetric methods, respectively. The ferric-reducing/antioxidant power (FRAP assay) was used to measure the total antioxidant capacity, while proline content was evaluated according to the method of Bates. The contents of proline, total phenolics and flavonoids were significantly higher in the leaves of cherry tomato seedlings exposed to water stress, which suggests that the higher synthesis of these substances by plants represents an important defence mechanism of drought tolerance. The results also indicate that the application of all the used fertilisers in accordance with the manufacturer’s instructions can significantly increase the content of phenol compounds and total antioxidant capacity of plants under normal growth conditions, thus improving survival under subsequent stress.

Djordje Moravčević, J. G. Varga, N. Pavlović, V. Todorović, M. Ugrinović

A great number of spring garlic populations are present in Serbia. It is a very heterogeneous biological material. To date, it has been little studied. Therefore, the aim was to study its production and chemical characteristics. Eighteen populations of spring garlic were examined. Most of the production of this garlic in Serbia is based on them. They were examined in Zminjak, which is located in the central Macva (Serbia). The method of field experiments was used. The research lasted for three years. The results comprise two groups of parameters. The first group contains the results which show the most important productive characteristics (weight of bulb, number of cloves per bulb, bulb yield). The second group consists of the parameters for chemical characteristics of the investigated populations (dry matter, etheric oil, and allin). In this case, the populations demonstrated a significant influence, and their stability was also very conspicuous. Most of the investigated populations of spring garlic can serve as a good resultant material for obtaining new cultivars. The populations can be classified as follows: SG-18, SG-25, SG-28, SG-16, SG-29 (yield), SG-21, SG-26 (technological quality) and SG-29, SG-18, SG-16 (foodstuff and pharmaceutical quality). They can remain as populations, but it would be significantly better to create new cultivars by using them.

B. Pašalić, V. Todorović, I. Koleška, B. Bosancic, Nikolina Đekić

Tomato ( Lycopersicon esculentum , Mill) is vegetable which is relatively resistant to salt concentration in growing substrate. However, research has shown that salinity stress causes changes in the quality of the fruit, which indirectly affects the yield. So the aim of this research was to determine the change in color of ripening fruits, the amount of sugar and acid in the fruit as indicators of the quality of the fruit. Tests were conducted in a greenhouse on tomato cv. Buran F1, where were two varieties of seedlings, grafted and non grafted, grown in the control substrate (EC = 1.7 dS/m) and substrate with EC = 6.8 dS/m. The quality of fruit is observed by a difference in the maturing of the first fruit of the first and second branches of tomatoes, and through the concentration of sugar and acid in the fruit, depending on the position of the plants. The obtained results show that salinity induces higher sugar content and total acidity in grafted and non-grafted tomato fruits. Also salinity leads to faster ripening of grafted and non-grafted tomato.

I. Koleška, D. Hasanagić, V. Todorović, S. Murtić, Izudin Klokić, N. Parađiković, B. Kukavica

ABSTRACT Plant biostimulants are substances which have the capacity to modify physiological processes in plants in a way that provides potential benefits to growth, development or stress response. Effects of biostimulant application on two tomato hybrids (Ombeline F1 and Bostina F1) submitted to reduced nitrogen, phosphorus and potassium (NPK) nutrition aiming at prevention of oxidative stress generation as well as yield and fruit quality loss were investigated in this study. According to obtained results, foliar applied Viva® biostimulant decreased superoxide dismutase (SOD, EC 1.15.1.1) and peroxidase (POD, EC 1.11.1.7) activity in tomato leaves even when recommended NPK nutrition was reduced at 40%. Fruit quality parameters (total soluble solids, total acidity, ascorbic acid and lycopene content) and yield were also maintained in reduced macronutrient fertilization when biostimulant was added. Combination of biostimulant with reduced NPK fertilizer enabled stability of cell homeostasis in tomato plants and their better adaptation to stress conditions. The possibility of biostimulant being used as environmental friendly tool in the reduction of mineral fertilizers without negative consequences regarding yield and fruit quality was discussed.

S. Zeljković, U. Šušak, N. Parađiković, J. D. Gidas, Monika Tkalec, V. Todorović

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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