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Pollen allergies are an important and relevant question in the personal life of millions of people. They are well characterized on the genomic and proteomic level of allergens per se and on medicinal level, but the very few information exists about the natural variability and background of pollen allergen transcription patterns. Here, the variability of expression of Bet v 1, Bet v 2, Cor a 1 and Cor a 2 were performed. Both of the analysed species, Betulla verrucosa as well as Corylus avellana are clinically relevant in spring pollinosis in the area of central Europe. Bet v 1 and Bet v 2 expressions were subjected to compare in pollen for a total of twelve accessions from different areas. Cor a 1 and Cor a 2 expressions were subjected to comparison in pollen for a total of thirty-one natural pollen germplasm accessions. Expression changes of Bet v 1 were changed in three different manners – in six pollen samples was the expression higher than 1.5, three pollen samples have no relevant change and in three pollen samples was the expression lower about 0.5 times. Expression changes of Bet v 2 were as follows - five of the analysed samples were expressed more than 3 times when compared to the calibrator and in all of the other samples was the expression change in the difference that was not above 0.7 times. Expression changes of Cor a 1 were very similar among themselves and only in five samples was expression relevantly higher with at least 1.1 fold change. Expression changes of Cor a 2 were very similar among themselves and only in four samples was expression relevantly higher with at least 1.4 fold change.

Arachis hypogaea, L. is an oil seed crop with a worldwide importance and the seeds are eaten at one of several stages from immature to fully ripe, raw, or cooked. The aim of the study was to analyze specific iPBS fingerprints of twenty-one accessions of peanut that were collected in the places of their natural occurrence in Bolivia and describe the existing genetic polymorphism. For genomic imprinting, three different iPBS markers were chosen - 1882, 2079, 2274 and a PCR reactions were performed. Obtained iPBS fingerprints were evaluated for the presence/absence of individual amplified loci and scored in 1/0 matrices. A Jaccard coefficient of genetic similarity was applied in UPGMA analysis for dendrogram construction. Polymorphism level was achieved in the range from 48% up to the 75% per primer. None of the iPBS markers used in the study was considered to distinguish all of the analyzed peanut accessions, but combining them in the final analysis, the level of genomic polymorphism was sufficient to clear separating of iPBS fingerprints of the collected accessions and unique iPBS loci were recorded in genomes of some of them. We found that, by selecting the appropriate iPBS markers, it is possible to characterize the peanut genome in the individual level with a specific fingerprint.

This study was designed to investigate chemical and antioxidant properties, as well as the antimicrobial and antibiofilm behaviour of Cinnamomum cassia essential oil (CCEO). MALDI-TOF MS Biotyper mass spectrometry was applied to evaluate the biofilms of Stenotrophonomonas maltophilia and Bacillus subtilis, while the antibiofilm ability of CCEO was assessed on wooden and glass surfaces. The antimicrobial activity by disc diffusion method, microdilution method, and vapour phase for two biofilm-producing bacteria and three Penicillium spp. were used. Antimicrobial and antibiofilm properties were assessed using the agar microdilution protocol. The vapour phase of Penicillium citrinum, P. crustosum, P. expansum, S. maltophilia, and B. subtilis on bread, carrot, potato, sweet potato, and apple in situ was studied. Specific molecular variations related to the biofilm formation and genetic analogies were evaluated with MSP spectra dendrograms of S. maltophilia and B. subtilis profiles were grown on different days. The results of disc diffusion and broth diffusion methods showed that CCEO was strongly effective against all tested microorganisms and the vapour phase method was effective and active against all Penicillium spp., but not strongly effective against bacteria in food preservation of food matrices.

The aim of study was to determine the antimicrobial activity of ten essential oils against ten microorganisms isolated from fish. The following essential oils were tested: Cymbopogon nardus Rendle, Citrus limon, Leptospermum petersonii Bailey, Gaultheria procumbens, Pelargonium graveolens, Citrus paradisi, Citrus aurantifolia, Litsea cubeba Pers., Citrus reticulata Blanko, Vetiveria zizanoides. The disc diffusion method and microdilution method were used for determination of antimicrobial activity. Rhodotorula spp., Aerococcus spp., Aerococcus viridans, Enterococcus faecium, Enterococcus moraviensis, Aeromonas spp., Delftia acidovorans, Pseudomonas gessardii, Yersinia spp., Yersinia ruckeri, Yersinia enterocolitica were tested in the present study. The best antimicrobial activity of Leptospermum petersonii Bailey were found against Aerococcus spp., Enterococcus faecium, E. moraviensis, Delftia acidovorans, Pseudomonas gessardii, Yersinia ruckeri and Y. enterocolitica. Litsea cubeba Pers. was effective against Delftia acidovorans, Yersinia enterocolitica, Enterococcus faecium using the disc diffusion method. The best minimal inhibition concentration (MIC 50 - 1.5 µL/mL resp. MIC 90 - 1.6 µL/mL) was found for Litsea cubeba Pers. against Aerococcus spp. and Yersinia spp. using microdilution method.

This study aimed to evaluate the microbiota, phytochemical, antioxidant profile and DNA fingerprinting of Cabernet Sauvignon grapes from Slovakia and R. North Macedonia. There were used two samples of grape berries (one sample from Slovakia and one from Macedonia). Each sample was analyzed in triplicate. The bacteria were cultivated on Plate count agar (PCA), microscopic filamentous fungi were cultivated on Malt extract agar (MEA). MALDI-TOF MS Biotyper mass spectrometry was used for the identification of microorganisms (bacteria and yeasts) and microscopic filamentous fungi with manuals. DPPH method was used to determine of antioxidant activity of grape berries. Phytochemical and antioxidant profiles were evaluated in grape berries samples. Total genomic DNA was extracted from mature grapes by GeneJET Plant Genomic DNA Purification Kit. The number of bacteria was higher in the sample of Macedonian grape (4.13 log CFU.g-1) in comparison to the grape from Slovakia as well as the number of yeasts was also higher in the Macedonian sample (2.57 log CFU.g-1). Antioxidant activity of Slovak grape berries was 0.55 mg TEAC.g-1 and of Macedonian grape, berries was 0.51 mg TEAC.g-1. Total polyphenol content was higher in grape from Slovakia (0.81 mg GAE.g-1) than in grape from Macedonia (0.77 mg GAE.g-1), while total flavonoid content was 0.57 and 0.17 mg QE.g-1 in Slovak grape and Macedonian grape, respectively. Total phenolic acid content was higher in the sample from Macedonia (0.40 mg CAE.g-1) compared to the grape from Slovakia (0.24 mg CAE.g-1). Total anthocyanin content was also higher in Macedonian grape (0.46 mg.g-1) compared to the Slovak sample (0.05 mg.g-1). The total polymorphism for all of the used primers of 87.5% was obtained for the Macedonian sample of Cabernet Sauvignon and 89.4% for the Slovak sample.

Work aimed to investigate the antimicrobial activity of medicinal plants against selected species of food industry pathogens in vitro conditions. The detection of antibacterial properties was examined by the disc diffusion method and the method of the minimum inhibitory concentrations (MIC). The cultivation of microorganisms after the 24 h was performed by disc diffusion method. Petri dishes have grown at 37 °C in which the Mueller - Hinton agar and application it to the sterile paper disc impregnated with the extract. The thickness of the resulting inhibition zone was measured with a ruler after completion of the culture. After the preparation of bacteria and extracts of certain concentrations of a subsequently added to wells microplates we use the method of the minimum inhibitory concentration (MIC) which was conducted out as the second measurement, and we took the readings absorbance spectrophotometer at 570 nm using the Glomax plate spectrophotometer. We found out, that Equisetum arvense demonstrated the largest zones of inhibition to the tested Gram-positive and Gram-negative bacteria. The greatest antimicrobial activity achieved Equisetum arvense, Urtica dioica, and Taraxacum officinale against Salmonella enterica subsp. enterica CCM 3807 and Yersinia enterocolitica CCM 5671. Equisetum arvense and Taraxacum officinale was the most effective against Escherichia coli CCM 2024 and the least effective were Tussilago farfara and Mentha piperita with using the method of minimum inhibitory concentrations.

Recent studies reported a significant contribution of microRNAs (miRNAs) in plant response to elicitors. The aim of our study was to analyse the effect of exogenously applied plant-derived elicitor cobalt-diglycyrrhizinate on Triticum aestivum L. genome based on the functional characterization of stress associated miRNAs. Our results confirmed the impact of cobalt-diglycyrrhizinate on plant genome in a way of plant response stimulation via expression of stress-related miRNAs markers. We found that both miRNAs markers (miR168 and miR395) are involved in response to plant-based elicitor stimuli. The role of miR168 and miR395 as functional markers sensitive to applied elicitors has been confirmed. Marker miR168 is suitable for the mapping of plant genome response to native elicitor, lower concentration of an elicitor is more suitable for plant immune system stimulation and its inappropriate concentrations may be toxic to the plant. Plant genome very sensitively reacts to elicitor stimuli and the question of the appropriate concentration and the time point of elicitor application are crucial.

Ionizing radiation in environment comes from various natural and anthropogenic sources. The effect of radioactivity released after the CNPP (Chernobyl Nuclear Power Plant) on plant systems remains of great interest. Even now, more than three decades after the nuclear accident, the long-lived radionuclides represent a strong stress factor. Herein, the emphasis has been placed on analysis of genetic variability represented by activation of LTR (Long Terminal Repeat)-retrotransposons. Polymorphism in LTR-retrotransposon insertions has been investigated throughout the genome of two flax varieties, Kyivskyi and Bethune. For this purpose, two retrotransposon-based marker techniques, IRAP (Inter-Retrotransposon Amplified Polymorphism) and iPBS (inter-Primer Binding Site), have been employed. The hypothesis that chronic radioactive stress may induce mechanism of retransposition has been supported by the activation of FL9, FL11 and FL12 LTR-retrotransposons in flax seeds harvested from radioactive environment. Out of two retrotransposon-based approaches, IRAP appears to be more suitable for identification of LTR-retrotransposon polymorphism. Even though the LTR-retrotransposon polymorphism was identified, the results suggest the high level of plant adaptation in the radioactive Chernobyl area. However, it is not really surprising that plants developed an effective strategy to survive in radio-contaminated environment over the past 30 years.

The aim of this study was to identify lactic acid bacteria (LAB) in grapes, must and wines. A total amount of 90 samples including grape (n = 30), must (no = 30) and wine (no = 30) were collected from vineyards in Slovakia. LAB were used cultured on MRS agar with subsequent confirmation with MALDI-TOF mass spectrometry (Bruker Daltonics). Altogether, 904 isolates were identified. Members of the family Lactobacillaeceae were the most abundant in grape (60%), must (46%) and wine (51%). Lactobacillus, Lactococcus, Leuconostoc, Pediococcus and Weissella genera and 27 species of LAB were isolated from the examined samples. Leuconostoc mesenteroides spp. mesenteroides was the most abundant species in grape, must and wine.

Bacteria, yeasts and microscopic fungi, which are live and grow normally on the grape surfaces, are important for winemaking. The aim of this study was to investigate the microbiota of the Tokaj wines grapes of Slovak origin. The total bacterial count ranged from 2.48 log cfu/g to 4.52 log cfu/g. Malt Extract Agar (MEA) with acid base indicator bromocresol green was used for cultivation of yeast and microscopic fungi and the number of this microorganisms ranged from 1.11 log cfu/g to 4.16 log cfu/g. We found following bacterial species: Staphylococcus epidermidis, Micrococcus luteus, Paenibacillus pasadenensis and Sphingomonas sp. Identified yeasts species included Candida sorbose, Kloeckera apiculata, Metschnikowia pulcherrima, Rhodotorula glutinis, Rhodotorula mucilaginosa and Saccharomyces cerevisiae. From microscopic fungi were isolated: Alternaria sp., Arthrinium sp., Aspergillus niger, Aureobasidium pullulans, Botrytis cinerea, Cladosporium sp., Eurotium sp., Geotrichum candidum, Penicillium expansum, Penicillium sp. and Rhizopus sp. The most dominant species of Tokaj grape were Kloeckera apiculata and Metchnikowia pulcherrima.

Peach is a popular sweet fruit with a very good climate adaptation and high production. It offers many health benefits determined by its biochemical composition. However, sensitive people can be sensitised by Pru p 3 non-specific lipid transfer protein family directly or through cross reaction as a member of Bet v 1 homologues. The majority of research is focused on the protein, less data can be found in genomics or transcriptomics. This study performed RFLP analysis by chosen restricted enzymes (BfaI, MseI, NlaIII) for non-coding region of Pru p 3 (NCBI: KC311811.1) as a tool to distinguish closely related isoforms of the allergen. BfaI cut amplicon into 5 fragments corresponding to the Yulu variety in silico cleavage and polymorphism was not detected. For MseI and NlaIII polymorphisms were found in the cleavage sites, two types of restriction profiles were created for both. None of the NlaIII profiles corresponds to the restriction profile of in silico cleavage. The study confirms the varietal differences in Pru p 3 gene and supports a hypothesis that allergenicity depends on both qualitative and quantitative factors that are different and specific to each variety.

Environment around Chernobyl recoveres from nuclear accident in 1986 and provides a base for plant adaptation research. Here, the initial platform was established for molecular screening of flax genome affected by radioactive contamination. Flax (Linum usitatissimum L.) cultivated in the radio-contaminated, and remediated experimental field near Chernobyl was investigated. Specifically, two gene isoforms of FAD3 gene, FAD3A and FAD3B, were evaluated using the restriction analysis, and direct sequencing. The aim of the presented study was to identify mutations, and single nucleotide polymorphisms within the FAD3A and FAD3B genes. The investigation was performed more than 25 years after Chernobyl accident. Therefore, the effect of chronic low dose rate irradiation on flax plants was considered.

Apple allergy belongs to the most prevalent fruit allergies which in North and Central Europe is mainly attributed to cross-reaction between Bet v 1 allergen from birch pollen and Mal d 1 major apple allergen. For a long time, patients observed symptoms of unequal severity after consumption of different apple cultivars. This led scientific community to search for the basis of the cultivar-specific allergenicity. According to several studies, the amount of Mal d 1 allergen plays an important role. Currently, notable attention is mainly concentrated on genetic variability as the primary source of different allergenic potential. Mal d 1 gene family is a large family of gene isoforms and their variants differing in the primary sequence. These sequence alternations may cause changes in protein structure and potentially affect the binding capacity to IgE and thus the allergenic potential. Among many methods available to analyze genetic variability, restriction fragment length polymorphism is simple technique suitable to analyze variability of Mal d 1 allergen. This paper aims to provide a brief overview of a possible approach of interlinking genomic data (e.g. as by RFLP profiles) and clinically proven apple allergenicity.

Vitis vinifera L. is a specie that is adapted to a very variable range of climates, from cold up to the desert one, but especially it grows in the temperate Mediterranean regions and continental areas of Europe. Grape is a widespread consumed fruit as well as processed to musts, juices or wine. The health beneficial effects of grapes and wine are very well known due to their high nutritional value and unique phytochemical composition. Despite many health protective and beneficial effects of Vitis vinifera, a part of population suffer to allergic reactions to this fruit. Allergens of wine and grapes are: endochitinases, lipid-transfer protein and thaumatin. Thaumatin is a protein having a sweet taste belonging to the PR5-like proteins. These proteins are very difersified in their functions and were described to be involved in stress responses and fruit ripening, but are expressed in healthy grape fruits in a constitutive manner and needn´t to be expressed only as a answer to the stress. Thaumatin is a minor allergen in grape, but belonging to the suspected panallergens relevant to the food cross-allergy induction, its importance is quite high. Another importance of this protein is a technological one, as reported to aggregate in wine to form a visible haze unless removed prior to bottling. In this study, expression of thaumatin-like allergen was analysed in the grapes of selected varieties. Grapes of four red varieties of Vitis vinifera, L. were obtained in the season 2017 in the Sabo winery that belongs to the Malokarpatská wine region. Fresh maturated grapes of varieties Alibernet, Cabernet Sauvignon , Frankovka modrá and Dornfelder were analysed. Expression changes of thaumatin was calculated by delta delta Ct method. Dornfelder was found as to have the lowest activity in thaumatin-like gene activity, mainly when comparing to the Cabernet Sauvignon and Frankovka modrá. Alibernet, on the other side, has the expression level of thaumatin very similar when comparing to the Cabernet Sauvignon and Frankovka modrá.

The aim of this study was clarifying the relation between genetic diversity of edible honeysuckle (Lonicera kamtschatica) and the major group of biologically active substances as total polyphenols content (TPC) including antioxidant activity (AO). Fruits of edible honeysuckle becomes more and more popular, especially in Europe. The current status of research on polyphenolic compounds in the berries of edible honeysuckle and their biological effects, including recommended utilization, are reviewed.The biological material including 14 cultivars of the edible honeysuckle ('Zoluška', 'Amfora', 'Pruhonický 44', 'Vasilijevsky', 'Moskovskaja', 'Vojtek', 'Sinoglaska', 'Altaj', 'Lipnická', 'Kamčadalka', 'Sinaja Ptica', 'Fialka', 'Modrý Triumf', and 'Leningradský velikán') originated from Czech republic (Žabcice near Brno). The content of TPC and AO were determined by location and its soil-climatic conditions and these environmental circumstances determines the RAPD profiles of analysed honeysuckle acessions, too. DPPH method was used to analyze AO and Folin-Ciocalteu method was used to determine TPC. The results of experiment showed that the highest value of AO was determined at the cultivars 'Zoluška' (81.04 mg.L-1) and the lowest was measured in 'Kamčadalka' (54.122 mg.L-1). On the contrary, the highest content of TPC was determined at the cultivar 'Kamčadalka' (51.09 mg.L-1) and the lowest value was measured at the cultivar 'Pruhonický 44' (21.65 mg.L-1). Phylogenetic trees were similar in genetic distance. The content of TPC and AO were not statistically significant in relation to cultivar. The analyzed cultivars of the edible honeysuckle were separated in 4 clusters according to used primers. In both gel images, the amplicon size ranged from 100 to 1,500 bp. We found that genetic diversity was partially related to content of total polyphenolic substances and antioxidant activity. Based on phylogenetic trees we have stated that variety 'Lipnická', 'Sinoglaska', 'Altaj', 'Leningradský velikán', 'Modrý Triumf', 'Sinaja Ptica' and 'Kamčadalka' were grouped in the similar cluster. The highest genetic distance was determined at the variety 'Vasilijevskaja' and 'Amfora'. In the same way, there were variety 'Vojtek', 'Fialka' and 'Zoluška'. © 2019 Potravinarstvo Slovak Journal of Food Sciences.

Ginkgo biloba L. is characterized by its high level of resistance to climatic conditions, diseases, and pests. In Slovakia, there is a rich collection of genetic resources of ginkgo consisting of 288 trees growing in 103 locations and providing valuable biological material for scientific research. There have been documented 45 trees of ginkgo older than 100 years (ranging from 112 to 242 years of age). Their dendrometrical parameters were recorded. For genomic imprinting, three types of microRNA-based markers were selected; highly conserved gb-miR160, moderately conserved gb-miR482 and the species-specific gb-miR75. The most efficient one can be considered the marker gb-miR482 with its genotype-unique miRNA profiles probably related to this marker functioning in the defence mechanisms of the ginkgo species. Unique miRNA loci were recorded in genomes of young ginkgo trees. We found that, by selecting the appropriate microRNA-based markers, it is possible to characterize the ginkgo genome in the context of microclimatic conditions.

Inter primer binding site (iPBS) polymorphism was investigated for common ivy (Hedera helix, L.) to obtain the knowledge on genetic diversity in this species. Actually, a very limited information exists about application of DNA markers in Hedera helix. Natural and planted European populations of ivy were analysed using an iPBS marker 5′ACCTGGCGTGCCA3′ with a total number of 238 fragments generated. Of these, 86% were polymorphic. There were determined certain attributes of this marker such as the diversity index (DI) and polymorphism information content (PIC). The value of the diversity index was 0.79 and the polymorphic information index was 0.78. The proportion of polymorphisms of the individual amplified loci ranged from 0.32% to 6.98%. Cluster analysis was performed to determine the relationships among the European ivy populations where the distribution in the dendrogram under the habitat specifity was found for the used iPBS marker. We concluded that iPBS was very efficient in analysing the genetic diversity in Hedera helix, L. and that this marker can serve as a suitable tool to find genomically specific fingerprints relevant to the factors influencing the distribution of genetic variation.

The aim of research was the identification of cow milk allergen in grapes, must, federweisser and wine by immunochemical method ELISA. Milk allergens (casein) are mostly used together with egg proteins during wine clarification. The results show that quality of calibration curves has significant importance for objective evaluation of quality detection. The degree of the variability calibration samples expressed R2 was not less than 0.9 in both calibration curves (0.9485 and 0.9659). In grape samples, concentrations of cow's milk casein below the detection limit were determined, that is recommended by the ELISA kit manufacturer. All grape samples showed casein concentration less than the value corresponding to 0 ppm standard (from 0.039 to 0.127 mg/kg). Low concentrations were recorded in three samples of must (from 0.056 to 0.077 mg/kg) as well. In case of the federweisser, the casein concentration ranged from 0.367 to 1.301 mg/kg, which is still less than 1.5 ppm standard (1.373 mg/kg). Most of the wines were found to be in the ELISA detection range. The exceptions were samples no. 3 and 9, whose absorbance was above the highest standard (45 mg/kg). These samples were then reconsidered after the first dilution, the resulting cow milk casein concentration was 67.22 mg/kg and 48.66 mg/L. Higher concentrations of this protein contained white wines (from 21.473 to 67.22 mg/L). In red wines, the milk protein concentrations ranged from 1.634 to 16.715 mg/L.

Staphylococcus spp. is not only a commensal bacteria but also a major human pathogen that causes a wide range of clinical infections. Recent evidence suggests that Staphylococcus has the ability to colonize the reproductive system and to affect its structure and functions. The objective of this study was to determine the chemical properties and antibacterial effects of select essential oils (EOs): Amyris balsamifera L., Boswellia carterii Birdw., Canarium luzonicum (Blume) A. Gray, Cinnamomum camphora (L.) J. Presl., Cinnamomum camphora var. linaloolifera Y. Fuita, Citrus x aurantium L., Gaultheria procumbens L., Litsea cubeba (Lour.) Pers., Melaleuca ericifolia Smith., Melaleuca leucadendra L., Pogostemon cablin (Blanco) Benth., Citrus limon (L.) Osbeck, Santalum album L., and Vetiveria zizanoides (L.) Roberty against 50 Staphylococcus spp. cultures isolated from human semen, specifically Staphylococcus aureus, S. capiti, S. epidermidis, S. haemoliticus, and S. hominis. The disc diffusion and broth microdilution methods were used to assess the antimicrobial potential and to determine the minimum inhibitory concentration (MIC) of the selected EOs. The best anti-Staphylococcus activities were found with both methods for the essential oils of C. luzonicum (Blume) A. Gray, A. balsamifera, C. camphora, and P. cabli.

The main aim of the study was to investigate the chemical composition, antioxidant, antimicrobial, and antibiofilm activity of Citrus aurantium essential oil (CAEO). The biofilm profile of Stenotrophonomonas maltophilia and Bacillus subtilis were assessed using the mass spectrometry MALDI-TOF MS Biotyper and the antibiofilm activity of Citrus aurantium (CAEO) was studied on wood and glass surfaces. A semi-quantitative composition using a modified version was applied for the CAEO characterization. The antioxidant activity of CAEO was determined using the DPPH method. The antimicrobial activity was analyzed by disc diffusion for two biofilm producing bacteria, while the vapor phase was used for three penicillia. The antibiofilm activity was observed with the agar microdilution method. The molecular differences of biofilm formation on different days were analyzed, and the genetic similarity was studied with dendrograms constructed from MSP spectra to illustrate the grouping profiles of S. maltophilia and B. subtilis. A differentiated branch was obtained for early growth variants of S. maltophilia for planktonic cells and all experimental groups. The time span can be reported for the grouping pattern of B. subtilis preferentially when comparing to the media matrix, but without clear differences among variants. Furthermore, the minimum inhibitory doses of the CAEO were investigated against microscopic fungi. The results showed that CAEO was most active against Penicillium crustosum, in the vapor phase, on bread and carrot in situ.

The aim of this study was to characterize extracts from the leaves of Ginkgo biloba L. from selected Slovakian localities in terms of the content of bioactive constituents, antioxidants and their antimicrobial properties. The results indicated that the content of antioxidants was sample-specific, and this specificity was statistically significant. Ginkgo biloba L. from the locality of Ko&scaron, ice had the best activity determined by the free radical scavenging activity (DPPH) (1.545 mg Trolox equivalent antioxidant capacity (TEAC)/g fresh matter (FM)) as well as the molybdenum-reducing antioxidant power (35.485 mg TEAC/g FM) methods. The highest content of total polyphenols (2.803 mg gallic acid equivalent (GAE)/g FM) and flavonoids (4.649 &mu, g quercetin equivalent (QE)/g FM) was also detected in this sample. All samples of G. biloba leaf extracts showed significant antimicrobial activity against one or more of the examined bacterial species, and Staphylococcus aureus subsp. aureus CCM 2461 was found to be the most susceptible (minimal inhibition concentration MIC50 and MIC90 values of 64.2 and 72.2 &micro, g/mL, respectively). Based on the results it was concluded that Ginkgo biloba L. extracts can be used as antimicrobial and antioxidant additives. Selected miRNA-based molecular markers were used to examine the environmental adaptability of Ginkgo biloba L. An almost-complete genotype clustering pattern based on locality was determined in the analysis that involved a species-specific gb-miR5261 marker. Morphologically specific exemplar, cv. Ohatsuki, was excluded.

The studies of plant bacterial endophytes, colonizing the plant tissues without any signs of diseases, are essential for understanding of ecological interactions. The aim of our study is to detect microbiological contamination and to assess the antimicrobial, antioxidant activity, total phenolic, carotenoid content, genome size, and ploidy of non-cultivated Rosa canina sampled from urban areas. Samples of Rosa canina fruits were collected in three locations in Slovakia. The highest total viable count and the Enterobacteriaceae count in fruits were 4.32 log CFU/g and 4.29 log CFU/g, respectively. Counts of the mesophilic anaerobic sporulating bacteria, Pseudomonas spp., and of the microscopic fungi and yeasts were 3.00, 2.15 log CFU/g, 3.65 log CFU/g, and 2.76 log CFU/g, respectively. Regarding the antimicrobial activity, Escherichia coli and Klebsiela oxytoca were the most sensitive species among the assayed microorganisms to the treatment with the ethanolic extracts of Rosa canina fruits. The fruits were rich in bioactive compounds, polyphenols, and carotenoids, that could be related to their antioxidant activity. Genome sizes of analyzed samples ranged from 2.3 to 2.96. DNA-based fingerprinting obtained by iPBS markers of the Rosa canina var. lapidicola Heinr. Braun., was characterized by some distinctive inserted loci. An interdisciplinary study was performed for the dog roses from different parts of Slovakia that resulted in deeper characterization of this species.

A collection of Amaranthus cruentus accessions was analysed to compare the characteristics of different dinucleotide core sequences of microsatellites repeats. ISSR analysis was performed to obtain corresponding genetic fingerprints and based on them, to construct dendrograms for every individual primer used in the study. A total of eighteen accesions were analysed. Dinucleotide sequence of (CA)6 repetition anchored by GG bases resulted in a total of 65 DNA fragments and 100-percent polymorphism. This marker has differ completely only 10 from all of the analysed genotypes. Dinucleotide sequence of (CT)8 repetition anchored by AC bases has resulted in the amplification of totally 97 fragments and 84.62 % of polymorphism. Only seven accesions were different among themselves when regarding this microsatellite core sequence. Dinucleotide sequence of (GA)6 repetition anchored by CC bases has resulted in the amplification of totally 171 DNA fragments and 100 % polymorphism. Not all the genotypes were separated based on this marker, two of them - Ames 5638 RRC 1139 and Ames 5648 RRC 1148 - possess the same profile of amplified ISSR fragments. The most distant genotype separated by all of the used primers was a landrace PI 511876 Huatle originated in Mexico.

Direct polymerase chain reaction method was use to amplify athaumatin-like sequence of Vitis vinifera L. in grapes as well as in stormy wines and musts. Thaumatin-like proteins (TLPs) of Vitis vinifera possess beside its function in abiotic and biotic stress response another one - they are able to cause protein haze in wine unless removed prior to bottling. Direct PCR is an approach where omission of DNA extraction is typical prior the amplification of the target site of plant genome. Crude extract or small pieces of plant tissues are used in the analysis directly without steps of extraction and purification of gDNA. The biological material that was used in analysis was collected during August - October 2017 in local stores and winery Sabo and comprises from cultivars Iršai, Muškát, Savignon Blanc, Svätovavrinecké, Dornfelder and Pálava. Direct PCR was performed by a cutted piece of grape tissue and a dilution buffer was use in 1:2 for stormy wine or must, respectively. Direct amplification of thaumatin-like protein sequence of Vitis vinifera was performed along with the control reactions with the primers for conserved region of plant chloroplast. Possitive amplification of thaumatin-like allergen sequence resulted in 570 bp amplicon. The most abundant amplicons were amplified in stormy wines, followed by musts and the amplicons from grapes were weaker when comparing them to others. The amplicon specificity checking of obtained PCR product of thaumatin-like allergen was performed by restriction cleavage by Psi Iand resulted in restriction amplicons of the 80 bp, 81 bp, 94 bp and 315 bp in length. Confirmation of the amplicon specificity by restriction cleavage support the potential of direct PCR to become areproducible method that will be fully applicable in routine analysis of not only plant genomes in the future, but it was demonstrated, that it works in liquids, too.&nbsp

The aim of this study was to isolate and identify bacteria and yeasts in different grape samples. The samples were collected in September 2017. Used 13 grape samples in this study (9 white and 4 red) were from the local Slovak winemakers. Alibernet, Irsai Oliver, Dornfelder, Blue Frankish, Feteasca regala, Green Veltliner, Pálava, Mūller Thurgau, Rhinriesling, Cabernet Savignon, Pinot Blanc, Savignon Blanc and Welschriesling. Two cultivation media were used for detection of bacteri and yeasts in grape samples. Malt extract agar base (MEA) and Tryptone Soay agar (TSA) were used for the cultivation of bacteria and yeasts. Cultivation was performed by spread plate method. Ethanol/formic acid extraction procedure was used for preparation of samples. MALDI-TOF Mass Spectrometer (Microflex LT/SH) (Bruker Daltonics, Germany) was used for identification of bacteria and yeasts. In total, 8 genera of yeasts, 8 genera of Gram-negative bacteria and 10 of Gram-positive bacteria were identified. Together 333 isolates, yeasts, Gram-negative and Gram-positive bacteria were identified.

Wheat is considered to be the world oldest and most widely grown plant. Current genetic and genomic information about wheat is extensive and contains partial knowledge of various types of markers up to the genomic sequence data. Up to date, several different types of DNA markers have been used in the mapping of the wheat genome, that were used to describe the whole genome polymorphism of various wheat varieties. In study, CDDP markers were used to analyse the length polymorphism in the genome of two drought tolerant wheat varieties and two drought susceptible wheat varieties. Polymorphism has not been obtained in relation to the type of drought resistance in the analyzed wheat varieties in the summary of the CDDP profiles. The primer combination WRKY-F and WRKY-R1 has one fragment with a constant amplicon length of 129 bp in all the analyzed varieties. For the combination of WRKY-F and WRKY-R2, the length of the amplicon shifted to 129-130 bp and to the 130-132 bp by F/R3 combination. The combinations of WRKY-F and WRKY-R2b and WRKY-F and WRKY-R3b with the exception of the Dagmar variety (WRKY-F and WRKY-2b) did not amplify amplicons of the given size. For the F/R1 primer combination, the fragments 301-304 bp, 138-140 bp and 187-192 bp were also typical. The second primer combination (WRKY-F and WRKY-R2) has generated fragments of the length of 156-158 bp and 181-186 bp in addition to the 129-130 bp fragment in all varieties. All are nonspecific. The marker system provides monomorphic profiles in all analyzed varieties and the total number of detected fragments was 81.

Apples are the most consumed fruit in the European Union with the exception of citrus fruits and apple consumption is part of a healthy and balanced diet. Although apples are recommended to consume each day, they are one of the fruits that can trigger an allergic reaction. The most important apple allergen in Central and Northern Europe is Mald 1. The aim of the study was to analyse natural restriction pattern variability of the part of Ypr 10 gene promotor in four apple varieties. All the analysed varieties resulted in different cleavage profiles for Ase I and Spe I when compared to the virtual cleavage profile of the genomic sequence stored in the public nucleotide database.

Simple Summary The aim of this study was to characterize the bacterial profile of semen collected from Holstein Friesian breeding bulls via a high-throughput sequencing approach for a 16S rRNA gene variability analysis. A total of 55 fresh semen samples of sexually mature breeding bulls were used in the study. They were gathered from Holstein Friesian breeding bulls at Slovak Biological Services in Nitra, Slovak Republic. To amplify the V4 region of the 16S rRNA bacterial gene, universal primers 515F and 806R enhanced by a 6 bp barcode identification sequence were used. The 16S rRNA high-throughput sequencing strategy was used. Two microbial clusters were identified among the analyzed samples—the first cluster was based on Actinobacteria and Firmicutes, while the second cluster contained a high prevalence of Fusobacteria. Abstract Bacterial contamination of semen is an important factor connected to the health status of bulls that may significantly affect semen quality for artificial insemination. Moreover, some important bovine diseases may be transmitted through semen. Up to now, only a very limited number of complex studies describing the semen microbiome of bulls have been published, as many bacteria are hard to cultivate using traditional techniques. The 16S rRNA high-throughput sequencing strategy allows for the reliable identification of bacterial profiles of bovine semen together with the detection of noncultivable bacterial species. Fresh samples from Holstein Friesian breeding bulls (n = 55) were examined for the natural variability in the present bacteria. Semen doses were selected randomly from Slovak Biological Services in Nitra, Slovak Republic. The most predominant phyla within the whole dataset were Firmicutes (31%), Proteobacteria (22%), Fusobacteria (18%), Actinobacteria (13%) and Bacteroidetes (12%). Samples of semen were divided into two separate clusters according to their microbiome compositions using a cording partition around a medoids analysis. Microbiomes of the first cluster (CL1) of samples (n = 20) were based on Actinobacteria (CL1 average = 25%; CL = 28%) and Firmicutes (CL1 = 38%; CL2 = 27%), while the second cluster (CL2; n = 35) contained samples characterized by a high prevalence of Fusobacteria (CL1 = 4%; CL2 = 26%). Some important indicator microbial groups were differentially distributed between the clusters.

Yacon [Smallanthus sonchifolius (Poepp. et Endl.) H. Robinson] is a plant grown worldwide originating in the Andes region. Yacon is grown for its sweet tuberous roots and leaves used for the preparation of herbal infusions. Twenty-six yacon landraces’ leaves and roots (both peeled and unpeeled) have been analysed by DART-TOF/MS. The method has been optimised and the fingerprints of the mass spectra have been statistically processed by PCA and LDA statistical analysis. The DART method has succeeded in differentiating between the yacon landraces according to their genotype and geographical origin.

Nowadays, various approaches for the precise DNA methylation analysis are known. This study applied an advanced MethyLight approach for precise detection of cytosine methylation. Chernobyl area is still contaminated by long-lived radioisotopes thus the potential risk of DNA damage by radiation remains relatively high. Plants as sessile organisms are directly affected by irradiation. For the analysis presented herein, flax seeds grown on the experimental fields near Chernobyl have been used. Cytosine methylation was quantified in the specific coding region of Fatty acid desaturase 3A gene which has an important function in fatty acid biosynthesis. Quantitative real-time MethyLight revealed fully methylated sequence in the targeted gene.

Apples are one of the oldest, most desirable and most consumed fruit in the world and in our agroclimatic conditions apples are the fruit that are also in the winter mostly consumed fresh. As apples are available all the year, they are a very important source of secondary plant metabolites. On the other hand, apples do not provide only benefits and belong to the fruit that cause an allergic reaction in a sensitized the population. Mal d 1 protein, one of the apple allergens, belongs to the PR proteins and is encoded by multiple isoforms in the genome of Malus domestica Borkh. The aim of the study was to compare the level of expression of Mal d 1.03A isoform in the pulps of apple varieties Golden and Spartan during the ripening. Expression analysis of Mal d 1.03A allergen were performed in pulps from a total of six ripening stages ranged from the early up to the physiological maturity at approximately monthly intervals. Assessment of the level of amplification of transcripts of Mal d 1.03A allergen was performed by software comparing them to a well-defined value of 1000 ng × μl-1. In the comparing analysis, the levels of amplified transcripts of Mal d 1.03A were more balanced in the first stages of ripening in the variety Spartan. A high increasing of the amount of transcripts were obtained in the first stages of ripening in the variety Golden. In Continual increasing of Mal d 1.03A transcripts was observed the middle ripening stage for both varieties. At the end of the ripening a different situation was observed in analysed apple varieties. The expression of Mal d 1.03A allergen was balanced for the variety Golden but a high increase was observed at the end of the ripening for the variety Spartan.

The objective of this research was to induce mitotic chromosome doubling in Anemone sylvestris L. The mitosis inhibitor oryzalin was directly added to the induction medium at 1, 2, 5, 10 and 15 μM for 8, 10 or 12 weeks of cultivation. Three tetraploid plants (2n = 4x = 32), 0.8% (polyploidization efficiency), were obtained from diploid plants (2n = 2x = 16) in three treatments (1 μM for 10 weeks, 5 μM for 8 weeks and 8 μM for 10 weeks). Ploidy level was confirmed by flow cytometry. Morphological characteristics (e.g. flower diameter, total plant height, leaf area) and chlorophyll content differences between diploid and tetraploid A. sylvestris were observed together with polyphenol content and antioxidant activity. The inter primer binding sites markers were used for evaluation of polymorphism. New genotypes with different morphological and biological characteristics were obtained through somatic polyploidization. The tetraploid plants were stronger, more vigorous and had an early flowering, which is essential for its use as an ornamental plant. The iPBS analysis showed unique amplicons that can be used for the purposes of molecular identification of tetraploid plants of A. sylvestris in the future. The results demonstrate the first report of in vitro induction of tetraploids of A. sylvestris.

A plant selection system based on the phosphomannose isomerase gene (pmi) as a selectable marker is often used to avoid selection using antibiotic resistance. Nevertheless, pmi gene is endogenous in several plant species and therefore difficult to use in such cases. Here we evaluated and compared Agrobacterium-mediated transformation of Linum usitatissimum breeding line AGT-952 (without endogenous pmi gene) and Nicotiana tabacum var. WSC-38 (with endogenous pmi gene). Transformation was evaluated for vectors bearing transgenes that have the potential to be involved in improved phytoremediation of contaminated environment. Tobacco regenerants selection resulted in 6.8% transformation efficiency when using a medium supplemented with 30 g/L mannose with stepwise decrease of the sucrose concentration. Similar transformation efficiency (5.3%) was achieved in transformation of flax. Relatively low selection efficiency was achieved (12.5% and 34.8%, respectively). The final detection of efficient pmi selection was conducted using PCR and the non-endogenous genes; pmi transgene for flax and todC2 transgene for tobacco plants.

Vitis vinifera L. is adapted to a very variable range of climates but it mostly grows in the temperate regions of continental Europe. In the Slovak Republic, the Tokaj wine region is one of the territories, where wine production is concentrated. Grape is a popular fruit and when processed, it is used as musts, juices or wine. Some people may suffer from allergic reactions to grapes. Up to now, endochitinases, lipid-transfer protein, and thaumatin were identified as grape allergens. In this study, expression of chitinase and thaumatin allergen was analysed in the grapes of Tokaj region varieties – Hachat Lovelin, Tokay and Muskat Blanc. Expression changes were calculated by the delta delta Ct method. Expression differences of chitinase were found to be similar in these varieties. Thaumatin was found to be variable in its transcription.

Rapid industrial and agricultural development brought, besides the indisputable advances, also risks related to the environmental pollution. Widespread soil deterioration represents a global problem. Chernobyl area contaminated by radionuclides after nuclear accident in 1986 provides the opportunity to analyze in situ the impact of radiation on plant systems. Unlike animals and humans, plants are not able to move to another place with better living conditions. Therefore, plants are an ideal object to study adaptation to the conditions of environmental stress. Long- term exposure to the ionizing radiation causes widespread changes in plant genome and epigenome. Also, these alterations may result in changed phenotype. In particular, this review discusses the effect of ionizing radiation on genetic and epigenetic stability of plant genome., Rozvoj priemyslu a poľnohospodárstva priniesol okrem nesporných výhod aj riziká súvisiace so znečistením životného prostredia. Rádioaktívnymi prvkami kontaminovaná oblasť Černobyľu po jadrovej havárii v roku 1986 poskytuje možnosť analyzovať vplyv radiácie na rastliny v podmienkach in situ. Rastliny na rozdiel od živočíchov a človeka nie sú schopné presídliť sa na iné miesto s vyhovujúcejšími životnými podmienkami. Z tohto dôvodu sú ideálnymi objektom pre štúdium adaptácie na podmienky prostredia zmenené pôsobením stresu. Dlhodobé pôsobenie ionizujúceho žiarenia môže mať za ásledok rozsiahle zmeny v genóme a epigenóme rastlín. Tieto zmeny môžu mať za následok zmenený fenotyp. Prehľad diskutuje najmä vplyv ionizujúceho žiarenia na genetickú a epigenetickú stabilitu rastlín.

Resveratrol is commonly found in food and drinks, including red wine and grapes. Grape extracts have a potent antimicrobial activity in vitro. The antimicrobial activity of plant extracts is the base of their potential application in food preservation agents, pharmaceuticals, cosmetics, alternative drugs and natural therapies. The aim of our study was to evaluate the antimicrobial activity of resveratrol and Blue Frankish pomace extract against Grampositive and Gramnegative bacteria as well as yeasts from the genus Candida. Six bacterial strains (three Grampositive bacteria Staphylococcus aureus CCM 2461, Enterococcus faecalis CCM 4224 and Listeria monocytogenes CCM 4699; three Gramnegative bacteria Escherichia coli CCM 3988, Pseudomonas aeruginosa CCM 1959 and Salmonella enteritidis subsp. enteritidis CCM 4420) and three yeast strains (Candida albicans CCM 8186, Candida krusei CCM 8271 and Candida tropicalis CCM 8223) were evaluated using the antimicrobial assay. Pure resveratrol and grape pomace extracts of red variety Blue Frankish were used. Our results show that resveratrol and red grape pomace extract have a very good antimicrobial activity against Grampositive bacteria when compared with Gramnegative bacteria and yeasts.

Effective and reliable methods of identification of Pseudomonas species are important for the characterization of microorganisms. Freshwater ecosystems are an important source of Pseudomonas species, including those pathogenic to fish and humans. The aim of the present study was to compare the identification conducted with MALDI-TOF MS Biotyper and 16S rDNA sequencing of fish-borne Pseudomonas spp. Altogether, 13 different Pseudomonas spp. were isolated from freshwater fish. Phylogenetic analysis showed a clear taxonomic placement only for 13 out of 15 Pseudomonas isolates. Accordance of identification method was found only in 6 out of 15 isolates. The human pathogenic Pseudomonas spp. were not found in our study, indicating that the fish could be considered as safe for consumption. The present study revealed a high discriminatory power of the mass spectra investigation and 16S rDNA gene sequencing technology for the identification of Pseudomonas spp. associated with freshwater fish.

Rosa canina, L. is widely used for medicinal purposes as well as in food industry where it is a valuable source, bioactive compounds and natural colorants. Actually, no specific method is recommended as suitable one for DNA extraction from rose hips. The aim of the study was to compare three commercial and three non-commercial methods to extract total genomic DNA from rose hips hyphanthium. Four methods are based on the precipitation in principle and two methods are based on resin-binding. Extracted DNA was proved for the effectivity in following PCR. In total, six different DNA isolations was performed for differently heat processes rose hips - fresh hyphanthium, 2-weeks frozen hyphanthium, dried hyphanthium (50 °C) and boiled hyphanthium (100 °C). The amplification parameters of 500 bp chloroplast gene amplicon were evaluated. Obtained amounts of extracted DNA was very variable not only for every individual method used but for individual treatment of samples, too. In general, non-commercial method provided higher amount of extracted DNA, but the A260/280 ratio was lower. When regarding the processing treatment of the samples, high differences were found among the samples untreated by heat and those that were dried or boiled for three of the used extraction methods. All the samples were positive for amplification, but different amounts of amplified product were obtained. The comparison of data for concentrations of extracted DNA and concentrations of amplified product showed large differences when regarding the achieved purity of DNA in extraction.

Received: 2016-06-25 | Accepted: 2016-07-29 | Available online: 2016-12-22 http://dx.doi.org/10.15414/afz.2016.19.04.144-149 The most suitable method of total DNA extraction still remains the crucial step for many plant species, although there are many different protocols and commercial kits for DNA isolation. In this study, five different extraction protocols were analysed to find out the most appropriate method for DNA extraction from Hedera helix L. This species has numerous medical and pharmaceutical uses and is also characterized by antioxidant effects on human body. In spite of its wide medical utilization, it belongs to those plant species, where the genomic information is very limited. Comparing of different protocols resulted in the yield of extracted DNA that has ranged from 6.3 to 487 ng μl-1. The purity of extracted DNA has ranged from 1.4 up to 2.0 A260/A280. All the extraction methods used in this study were evaluated not only in term of quantity and purity of DNA but also its functionality in the restriction endonuclease digestion and polymerase chain reaction based downstream analysis was performed. Keywords: DNA extraction, CTAB, SDS,commercial kit, Hedera helix L. References ABU-ROMMAN, S. (2011). Comparison of methods for isolating high quality DNA from sage (Salvia officinalis). Journal of Medicinal Plants Research, vol 5, no. 6, pp. 938-941 BALANSARD, G. et al. (1980). Douvicidal and Antifungal Activities of α-Hederin Extracted from Hedera helix leaves. Planta Medica, vol. 39, p. 234. BARRA, M. et al. (2012). Simple and robust DNA extraction mehod for the large-scale analysis of genotypes containing high polyphenolic content, such as landraces of Solanum tuberosum and Zea mays. Plant breeding, genetic and genetic resources, vol. 39, no. 3, pp. 593-601. BEDIR, E. et al. (2000). Triterpene Saponins from the Fruits of Hedera helix. Phytochemistry, vol. 53, pp. 905-909. CLARKE, M.M. et al., (2006). Prevalence of different horticultural taxa of ivy (Hedera spp., Araliaceae) in invading populations. Biological Invasions, vol. 8, pp. 149–157. COTA-SANCHEZ, H. J. et al. (2006). Ready-to-Use DNA Extracted with a CTAB Method Adapted for Herbarium Specimens and Mucilaginous Plant Tissue. Plant Molecular Biology Reporter, vol. 24, no. 2, pp.161-167. CRESPIN, F. et al. (1995). Identification of 3O-beta-D-Glucopyranosyl Hederagenin from the Leaves of Hedera helix. Fitoterapia, vol. 66, pp. 47. DABO, S. M. et al. (1993). A method for the isolation of nuclear DNA from cotton (Gossypium) leaves. Analytical Biochemistry, vol. 210, pp. 34-38. DELLAPORTA, S. L. et al. (1983). A Plant DNA Minipreparation: Version II. Plant Mol. Biol. Rep., vol. 1, pp. 19-21. DEMEKE, T. and JENKINS, R. (2010). Influence of DNA extraction methods, PCR inhibitors and quantification methods on Real time PCR assay of biotechnology-derived traits. Analytical and Bioanalytical Chemistry, vol. 396, pp. 1977-1990. DOOSTY, B. et al. (2012). Comparative Analysis and Optimization of Different DNA Extraction Protocols in Satureja khuzistanica. International Journal of Biology, vol. 4, no. 4,pp. 112-116. ELIAS, R. et al. (1991). Triterpenoid saponins from the leaves of Hedera helix. Journal of Natural Products, vol. 54, pp. 98-103. GREEN, A. F. et al. (2011). Phylogeny and Biogeography of Ivies ( Hedera spp., Araliaceae), a Polyploid Complex of Woody Vines. Systematic Botany, vol 36, no. 4, pp. 1114-1127. HUAQIANG, T. et al. (2013). Comparative Analysis of Six DNA Extraction Methods in Cowpea (Vigna unguiculata L.Walp). Journal of Agricultural Science, vol 5, pp. 82-90. IVERSEN, J. (1944). Viscum, Hedera and Ilex as climatic indicators. Geologiska Foreningers Stockholm Forhandlinger, vol. 66, pp. 463–483. JOBES, D. V. et al. (1995) Plant DNA isolation: A Method to Efficiently Remove Polyphenolics, Polysaccharides, and RNA. Taxon, vol. 44, pp. 379-386. KALENDAR, R. et al. 2010. iPBS: a universal method for DNA Fingerprinting and retrotransposon isolation. Theoretical and Applied Genetics, vol. 121, pp. 1419–1430. KRAJEWSKA, A. (1981). Phytochemical Studies of Hedera helix Herbage, Flowers and Fruits. Herba Polonica, vol. 27, pp. 303-309. LADE, B.D. et al., (2014). Efficient genomic DNA extraction protocol from medicinal rich Passiflora foetida containing high level f polysacharide and polyphenol. SpingerPlus, vol. 3, p. 457. LUTSENKO, Y. et al. (2010). Hedera helix as a medicinal plant. Herba Polonica, vol. 56, pp. 83- 96. MAJESTERSAVORNIN, B. et al. (1991). Saponins of the ivy plant. Planta Medica, vol. 57, p. 260- 262.

Hedera helix, L. is a specie that is used for its ornamental and medicinal properties widely. In spite of its very good biochemical characterization, the knowledge about the DNA variability is very limited and no DNA markers were used to analyse the genomic variability of the ivy populations, up to date. Here, a direct PCR approach was used to evaluate a cytochrome P450 sequences based polymorphism by PBA technique. A set of 4 samples from Slovakia, 2 samples from Czech Republic, 2 samples originated in Poland, 2 samples from Croatia, 2 samples from Scotland and samples from Germany, Austria and Spain were used in analysis. Using a set of three PBA primers and their combinations, the number of amplified fragment levels obtained by individual primer combinations was for both of them 11 and the obtained polymorphism was 91% or 100% respectivelly. The PCoA analysis was performed and the genotypes belonging to a different geographic localities were grouped together.

MicroRNAs (miRNAs) are a class of 19 - 24 nucleotide long non-coding RNAs derived from hairpin precursors, regulating various biological, metabolic and developmental processes at the post-transcriptional level. Many of the known miRNAs are evolutionary conserved across diverse plant species and function in the regulatory control of fundamentally important biological processes. It is known that exogenous plant miRNAs specifically target approximately 30% of protein-coding genes in mammals. The research was focused to analyze the occurrence of selected families of miRNAs (miR156, miR168 and miR171) in less used species but nutritionally important plant food resources (flax and medlar) and medicinal plant (milk thistle). The analyses were done by two individual approaches, by (a) miRNA-based molecular markers - as a novel type of functional markers and (b) qualitative Real-Time PCR. The expression pattern of selected miRNAs was analyzed depending on various plant tissues and developmental stages. Results have confirmed the significance and reliability of novel type of markers based on miRNA molecules as well as the species-specific and tissues-specific expression patterns of plants miRNAs. Significant polymorphism profile of miR156b was detected in various flax tissues of genotypes varying in the content of alpha-linolenic acid. Conclusions indicate that the variable behavior of the miRNA molecules, depending on various factors, may reflect the variability of the gene expression regulation of the human genome. The exploitation of the background of miRNA functioning within different species and plant tissues will help us to understand the molecular machinery as well as the regulatory mechanisms involved in the expression of miRNAs in plants and consequently in human genome.

Hedera helix is a plant that is valued for its medicinal properties and on the other hand criticized for its invading potential. Beside its medicinal use, a wide utilization as decorative plant is typical for this specie. Up-to date only a few molecular data are available for Hedera helix genome, thus the aim of the study was to evaluate the utilization of different DNA markers for the its further population studies purposes. In total, eight different DNA marker techniques were used for the PCR analysis – iPBS, BARE1 IRAP, Cassandra IRAP, Tst1 IRAP, HACRE 1 IRAP, RAPD, ISSR and miRNA. In total, 38 different iPBS primers were tested with the best results for primers 2152, 2270, 2079, 1899 and 2033. In the case of RAPD primers, all of them worked in the PCR excellent. For ISSR, only a 42 % of the used primers provided results that vere evaluated as very good or excellent ISSR profile. The most successful results were obtained by microsatelite repetitions (GA)8T, (CTG)3GC and (TG)10. A novel type of DNA markers – miRNA provided results that are promising for Hedera helix, L. population studies for their further utilization.

Plants are considered one of the toughest organisms in the world because of their ability to survive under the harsh conditions. Chernobyl offers a unique environment to study plant adaptation. Two experimental fields were established in the area near Chernobyl to closely study the effect of radioactive contamination on flax. Radioactivity severely impacts plant systems. However, in terms of DNA methylation, chronic radiation stress may manifest differently than acute irradiation. The methylation pattern of the fatty acid desaturase 3A gene was investigated using a combination of two techniques, bisulfite sequencing and methylation-sensitive high resolution melting. Using bisulfite sequencing, the gene-specific DNA methylation patterns between flax seeds cultivated in a remediated and radioactively contaminated environment were identified. Subsequent quantification of DNA methylation revealed partial demethylation in the coding sequence of the target gene. Flax seeds harvested from the Chernobyl area after 6 subsequent years of cultivation showed clear signs of adaptation to the environment.

The aim of study was to isolate and identify the gut bacteria of Apis mellifera and to evaluate antagonistic effect of the bacteriota against Paenibacillus larvae, which causes American foulbrood (AFB) in honeybees. The dilution plating method was used for the quantification of selected microbial groups from digestive tract of bees, with an emphasis on the bacteriota of the bees&rsquo, intestines. Bacteria were identified using mass spectrometry (MALDI-TOF-MS Biotyper). Overall, five classes, 27 genera and 66 species of bacteria were identified. Genera Lactobacillus (10 species) and Bacillus (8 species) were the most abundant. Gram-negative bacteria were represented with 16 genera, whereas Gram-positive with 10 genera. Delftia acidovorans and Escherichia coli were the most abundant in the digestive tract of honey bee. Resistance to a selection of antimicrobials was assessed for the bacterial isolates from bee gut and confirmed against all antimicrobials included in the study, with the exception of cefepime. Lactobacillus spp., especially L. kunkeei, L. crispatus and L. acidophilus. showed the strongest antimicrobial activity against P. larvae, the causal pathogen of AFB. Antimicrobial activity of essential oils against isolated bacteria and two isolates of P. larvae were assessed. Application of a broad selection of plant essential oils indicated that Thymus vulgaris had the highest antimicrobial activity against P. larvae.

The aim of this study was to compare the natural variability of the expression of class I chitinase gene in grapes of Vitis vinifera L. varieties as well as investigate their antimicrobial activity. Further, the effect of antimicrobial activity in chitinase expression was analyzed. Matured berries of thirteen grape varieties were used in the study&mdash, Alibernet, Dornfelder, Cabernet Sauvignon, Blaufr&auml, nkisch, Sauvignon Blanc, Welschriesling, Weisser Riesling, Irsai Oliver, Pinot Blanc, P&aacute, lava, M&uuml, ller-Thurgau, Grűner Veltliner, and Feteasca Regala, grown in the Small Carpathians wine region of Slovakia, Vrbov&eacute, Chitinase I gene expression was found to be similar in most of the analyzed varieties with the except of two varieties, where the expression of chitinase I was significantly higher&mdash, Pinot Blanc and Savignon Blanc. By comparing the varieties between the lowest and highest expression fold change, they were statistically different in the level of its chitinase I gene expression. Grape pomace extracts of analyzed Vitis vinifera L. were effective against Gram-positive, Gram-negative bacteria and yeasts, but a weak correlation tendency was found between chitinase I gene expression and antimicrobial activity, only against Staphylococcus aureus subsp. aureus, Candida albicans, and Candida glabrata.

This study aimed to analyze the genetic diversity of selected cultivars and clones of Lonicera sp. (edible honeysuckle) from two countries with different climatic and soil conditions. Accordingly, the determination of total polyphenols content (TPC) and total antioxidant activity (TAA) in fruits of Lonicera sp. in 2018&ndash, 2019 was performed. Applied statistical methods evaluating TPC and TAA were: correlation and regression, ANOVA, and agglomerative hierarchical clustering. The neighbor-joining method and gel images analysis (PyElph software, Pavel and Vasile, Bucharest, Romania) were used for genetic analysis. The average TPC of all cultivars and clones was 5.32 mg L&minus, 1 in the year 2018 and 3.78 mg L&minus, 1 in 2019. The average TAA values of all samples in 2018 were 8.24 mg L&minus, 1, and 9.27 mg L&minus, 1 in 2019. Statistically significant correlations between years and cultivars and clones were found. Based on the lengths and number of fragments by random amplified polymorphic DNA (RAPD)-PCR&rsquo, s, it was found that &lsquo, clone 5&rsquo, and &lsquo, clone 7&rsquo, were genetically close to the grown cultivars from the Czech Republic. Therefore, edible honeysuckle fruits are a valuable source of important nutrients for the human body and its health.

Phytoremediation efficiency depends on the ability of plants to accumulate, translocate and resist high levels of metals without symptoms of toxicity. This study was conducted to evaluate the potential of grain amaranth for remediation of soils contaminated with Cd. Three grain amaranth varieties, &ldquo, Pribina&rdquo, (A. cruentus), &ldquo, Zobor&rdquo, (A. hypochondriacus x A. hybridus) and Plainsman (A. hypochondriacus x A. hybridus) were tested under different level of Cd (0, 5, 10 and 15 mg/L) in a hydroponic experimental treatment. All could be classified as Cd excluders or Cd-hypertolerant varieties able to grow and accumulate significant amounts of Cd from the hydroponic solution, preferentially in the roots. Under the highest level of Cd exposure, qRT-PCR expression analysis of five stress-related genes was examined in above- and below-ground biomass. The results show that the Cd concentration significantly increased the mRNA level of chitinase 5 (Chit 5) in amaranth roots as the primary site of metal stress. The involvement of phytochelatin synthase (PCS1) in Cd detoxification is suggested. Based on our findings, we can conclude that variety &ldquo, is the most Cd-tolerant among three tested and can be expected to be used in the phytomanagement of Cd loaded soils as an effective phytostabiliser.