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Amaranth belongs to one of underutilized species with a potential to overcome some of the actual challenges of modern agriculture that is a part of very unstable environmental conditions. Here, two Slovak varieties of amaranth – Zobor (A. hypochondriacus × A. hybridus) and Pribina (A. cruentus) were characterized by Conserved DNA-Derived Polymorphism (CDDP), P450-based analogue (PBA) markers and Tubulin-Based Polymorphism (TBP) and by its most abundant endophytes. CDDP as well as TBP profiles showed very similar distribution of the generated fingerprints for both of the analysed varieties. PBA profiles generated the lowest value of polymorphism. A wide spectrum of amplicons was generated by all of the techniques used in the study with the length range of PCR fragments from 80 up to the 2292 bp. The variability of the endophytic composition of the Pribina variety showed the following three most abundant genera: Endobacter, Dichotomicrobium and Burkholderia. Abiotrophia, Endobacter and Limimonas were most abundant for Zobor variety.

Strawberries are popular worldwide for their nutritional benefits. Their germplasm is widely analysed by different molecular and DNA based markers. Here, two different marker techniques were used to analyze intra- and interspecies variability of 38 genotypes of two strawberry species (Fragaria x ananassa and Fragaria vesca L.). Bet v 1 based amplified polymorphism (BBAP) and conserved DNA-derived polymorphism (CDDP) utilized to obtain specific fingerprints. BBAP technique generated lower polymorphism comparing to CDDP (BBAP – 99.01%, CDDP – 99.65%), but both of the techniques separated all of the analysed strawberry genotypes. The groups of genotypes in constructed dendrograms were not clustered according the specie specificity.

Arachis hypogaea L. is an annual legume that is one of the most consumed plant species. On the other hand, it belongs to one of the most monitored clinically important allergens worldwide. The polymorphism of this species based on allergen coding genes could be useful in its characterisation, but previously, no allergen-based marker techniques have been developed for peanuts. A new type of DNA-based markers of coding regions were used to analyse the variability of 21 peanut accessions - BBAP (Bet v1 based amplicon polymorphism), PBAP (profilin based amplicon polymorphism), and VBAP (vicilin based amplicon polymorphism). All of the used technique provided polymorphic fingerprints and distinguished the analysed peanut accessions. The effectivity of these techniques corresponds to the presence of the allergen homologous sequences that are a part of the A. hypogaea genome. VBAP was the most effective in distinguishing the analysed peanut accessions when compared to the results of BBAP and PBAB. For BBAP, two of the analysed accessions provided the same fingerprinting pattern. The ability of the used markers to detect polymorphisms was comparable, with an average polymorphism information content (PIC) value of 0.47.

Apples are a popular fruit worldwide, with many health and nutritional benefits. However, this fruit is also among those that, particularly in Central and Northern Europe, are allergenic due to the Mal d 1 allergen. Mal d 1 is a homologous allergen to Bet v 1—the main pollen allergen of birch. In this study, two different approaches were used to identify the natural length polymorphism of Bet v 1 homologs in apple varieties, with the aim of characterizing their effectiveness. BBAP (Bet v 1 based amplified polymorphism) and RFLP (restriction fragments length polymorphism) profiles were characterized and compared. RFLP analysis recognizes the genetic diversity of M. domestica Mal d 1 sequences at a relatively low level. In BBAP profiles, the genetic dissimilarity was up to 50%, which appears suitable for intraspecific fingerprinting and serves as an additional method for RFLP analysis. RFLP analysis was able to distinguish some varieties that BBAP could not, such as Sonet.

DNA-based marker fingerprinting techniques are developed with the aim of their effective use in the identification or comparative studies of plant genomes. In the genus Vaccinium, DNA markers help resolve taxonomic relationships among the species included in it. In this study, the allergen coding genes and micro RNAs were used as DNA markers and applied for the characterization of the fingerprints of Vaccinium corymbosum L. and Vaccinium myrtilus L. Both techniques were found to be effective in distinguishing the analysed Vaccinium accessions and in the generating of polymorphism and in the case of Bet v 1 Based Amplicon Polymorphism and Profilin Based Amplicon Polymorphism, separated fingerprints were obtained for Vaccinium myrtilus L.

Melissa officinalis L., a well-known herb with diverse industrial and ethnopharmacological properties. Although, there has been a significant lack in the breeding attempts of this invaluable herb. This study aimed to enhance the agronomical traits of M. officinalis through in vitro polyploidization. Nodal segments were micropropagated and subjected to oryzalin treatment at concentrations of 20, 40, and 60 mM for 24 and 48 hours. Flow cytometry, chromosome counting, and stomatal characteristics were employed to confirm the ploidy level of the surviving plants. The survival rate of the treated explants decreased exponentially with increasing oryzalin concentration and duration. The highest polyploid induction rate (8%) was achieved with 40 mM oryzalin treatment for 24 hours. The induced tetraploid plants exhibited vigorous growth, characterized by longer shoots, larger leaves, and a higher leaf count. Chlorophyll content and fluorescence parameters elucidated disparities in photosynthetic performance between diploid and tetraploid genotypes. Tetraploid plants demonstrated a 75% increase in average essential oil yield, attributed to the significantly larger size of peltate trichomes. Analysis of essential oil composition in diploid and tetraploid plants indicated the presence of three major components: geranial, neral, and citronellal. While citronellal remained consistent, geranial and neral increased by 11.06% and 9.49%, respectively, in the tetraploid population. This effective methodology, utilizing oryzalin as an anti-mitotic agent for polyploid induction in M. officinalis, resulted in a polyploid genotype with superior morpho-physiological traits. The polyploid lemon balm generated through this method has the potential to meet commercial demands and contribute significantly to the improvement of lemon balm cultivation.

Food allergies are an increasingly common phenomenon across all age groups and can be called an epidemic of modern times. Legumes are a nutritionally attractive crop because of their high protein content and well-balanced nutritional value. However, in addition to nutritionally valuable components, they contain a relatively high amount of antinutritional factors such as glycosides, lectins, inhibitors of digestive enzymes, and antinutritional proteins, including allergens. Different genomic-based analyses of allergen-coding parts are relevant in research into legume gene resources. Here, a total of thirty different soybean varieties were analyzed for polymorphism based on the specific homologous sequences of genes for vicilin and profilin; products of both of these genes belong to allergenic molecules of this species. A total of 16 different amplicons were obtained when profilin was used as marker and 17 different amplicons were obtained when vicilin was used. Comparing both of the used techniques, vicilin provided more polymorphic profiles, but in five of analyzed varieties no amplicons were obtained. Profilin fingerprints provided a higher degree of similarity coefficients among individual varieties of the soybean. Both of used PCR-based techniques proved to be applicable for genomic-based screening of allergen homologs in the genetic resources of Glycine max L.

Background. For the Guatemalan culture, the avocado (Persea americana Mill) represents more than a crop, since it has religious, mythical, economical, and medicinal significance typical of the center of domestication of the species. Although there is speculation about the wide variability of wild avocado germplasm, there are no studies describing avocado populations. Objective. To explore wild germplasm based on agro-morphological characteristics using the standardized descriptors for avocado (Persea spp.). Methodology. The distribution of characteristics of 189 avocado trees across eight populations was evaluated using cross-tabulation and Chi-square testing. In order to properly handle qualitative and quantitative data, it was employed the factor analysis of mixed data (FAMD) and hierarchical clustering on principal components (HCPC) to analyze the relationship and variation of features within and between populations. Results. The samples showed various morphological characteristics that indicate the presence of Mexican, Guatemalan, and West Indian avocado races in Guatemala. The FAMD revealed that the fruit shape, skin color and flesh texture, and anise odor in the leaves are ideal traits for distinguishing between individual trees. At the population level, the HCPC showed considerable variance amongst the studied trees but no distinct geographic groups of the samples. Implications. Although not all wild avocados grown in Guatemala are of the highest quality, several trees of extraordinary excellence are strewn over the republic's highlands. Thus, due to the danger posed by the introduction of commercial varieties, it is advisable preserving the germplasm. Conclusion. The clustering methods revealed that the 189 avocado trees could be regrouped into three main clusters. The morphological descriptors proved to be useful for grouping trees according to known races and can therefore be used in the characterization of cultivars and wild trees whose ancestry is unknown.

Physiological and molecular marker-based changes were studied in the tissues of two-year-old Pyrus pyraster (L.) Burgsd. seedlings under salt treatment. For 60 days, 5 mL of 100 mM NaCl solution was applied to each plant per day to a cumulative volume of 300 mL in the substrate. In response to osmotic stress, the seedlings increased their water use efficiency (WUE) on day 20 of regular NaCl application and maintained a stable net photosynthetic rate (An) per unit area. Under conditions of increasing salinity, the young plants maintained a balanced water regime of the leaf tissues (Ψwl). The seedlings invested mass to their root growth (R/S), retained a substantial portion (72%) of Na+ ions in the roots, and protected their leaves against intoxication and damage. A significant decrease in the leaf gas exchange parameters (gs, E, An) was manifested on day 60 of the experiment when the cumulative NaCl intake was 300 mL per plant. The variability in the reactions of the seedlings to salinity is related to the use of open-pollinated progeny (54 genotypes) in the experiment. Lus-miR168 showed tissue- and genotype-specific genome responses to the applied stress. Polymorphic miRNA-based loci were mostly detected in the root samples on the 20th and 35th days of the experiment. The cumulative effect of the salt treatment was reflected in the predominance of polymorphic loci in the leaves. We can confirm that miRNA-based markers represent a sensitive detection tool for plant stress response on an individual level. The screening and selection of the optimal type of miRNA for this type of research is crucial. The cytochrome P450-Based Analog (PBA) techniques were unable to detect polymorphism among the control and treated seedlings, except for the primer pair CYP2BF+R, where, in the roots of the stressed plant, insertions in the amplicons were obtained. The expression ratios of cytochrome P450 in the salt-stressed plants were higher in the roots in the case of 20/100 mL and in the leaves with higher doses. The observed physiological and molecular responses to salinity reflect the potential of P. pyraster seedlings in adaptation to osmotic and ionic stress.

In Poland, the mountainous and hilly regions until recently were territories with unbalanced and poor diet quality, especially in early spring. Wild edible plants were or remain influential in supplementing a human diet. One of the earliest wild edible plants to appear in the growing season, characteristic of a broad swath of Europe from Ukraine to Germany, remains wild (ramsons) garlic (Allium ursinum L.). This paper assesses the impact of the vegetation site and preservation methods on the content of chlorophylls, β-carotene, vitamin C, total polyphenols, and antioxidant activity (ABTS, DPPH, and FRAP) of wild garlic leaves. In addition, the connection between its genome pattern and chemical composition was assessed. The plants were collected in three locations in the Carpathian foothills, and the reference sample was plants collected in the lowlands. The site from which the raw material was harvested has a statistically significant effect on the chemical composition of the preserved leaves of wild garlic. The profiles of the analysed components differed in the material from a different site. Wild garlic leaves from mountainous areas (400–600 m above sea level) were characterized by a higher content of the analysed ingredients than leaves obtained from lowland sites (200 m above sea level). Moreover, the material from the lowlands had a different genetic pattern from the leaves from the mountainous areas.

Citrus fruits enjoy widespread consumption globally, being among the most popular fruits. They are highly regarded for their nutritional composition, offering a range of beneficial nutrients. However, it is important to acknowledge that they can also elicit allergic reactions in sensitized individuals, which presents a contrasting aspect. The Bet v 1 cross-reacting allergen is a major birch pollen allergen, and it is the most commonly sensitizing allergen in central Europe. Bet v 1 belongs to the group of PR-10 proteins in the plant kingdom that cause various allergic reactions. The Bet v 1 allergen has a number of isoforms and homologs. These homolog genes are inherited from a common ancestor and subsequent amino acid similarity. They can cause the phenomenon of cross-reactivity in food allergies. The aim of the study was to analyze the length of polymorphism variability of the Bet v 1 homolog in orange varieties by using degenerated and nondegenerated primers. A total of eight varieties of Citrus sinensis L. Osbeck were used in the analysis. The BBAP technique (Bet v 1 based amplified polymorphism) was used to detect the length variability of fingerprints of allergen encoding genes of Bet v 1 homologs. Degenerated primer combinations and only one of the nondegenerated variants of primers provided fingerprints that were unique for every individual variety of analyzed oranges. By using other primer variants, from two up to the four varieties generated by the same BBAP profile, indicate a higher degree of Bet v 1 homolog sequential conservativity when compared to other fruit species.

En Guatemala, la producción del cultivo de papa se ve afectada por los nematodos Globodera rostochiensis y Globodera pallida. La capacidad de ambas especies para formar quistes complica su control y provoca el aumento de sus poblaciones. En Guatemala se reporta la presencia de ambas especies de nematodos por identificación morfológica, sin embargo, no se ha realizado una confirmación molecular. Este es el primer estudio para validar la presencia de ambas especies de nematodos por PCR múltiple y la determinación de la diversidad y estructura genética de las poblaciones utilizando marcadores moleculares. Se realizaron muestreos en cuatro departamentos productores de papa del país. La identificación por PCR se realizó con el cebador común ITS5 y los cebadores PITSr3 específico para G. rostochiensis y PITSp4 para G. pallida. La caracterización molecular se realizó con el marcador AFLP. Se confirmó la presencia de las dos especies de nematodos en los cuatro departamentos. Los índices de diversidad Shannon y heterocigosidad esperada revelaron mayor diversidad genética en G. rostochiensis (H = 0.311, He = 0.301) que en G. pallida (H = 0.035, He = 0.223). Los métodos NJ, DAPC y PCA exhibieron una débil estructura entre las poblaciones de ambas especies de nematodos. Los resultados sugieren un patrón de dispersión desde Quetzaltenango hacia el resto del país, atribuido a la comercialización de semilla contaminada con nematodos. Se sugiere promover programas de socialización sobre los beneficios del uso de semilla certificada, además de constantes monitoreos moleculares para un diagnóstico certero de ambas especies de nematodos.

Grape pomace is a winery industry residue of skins, seeds and stems of grapes used in agriculture for animal feed and production of food with biological activity. The aim of this study was to evaluate antimicrobial activity of Vitis vinifera L. pomace extract on growth of microscopic fungi and bacteria. The four strains of bacteria (two Gram-positive bacteria Staphylococcus aureus CCM 2461, Bacillus cereus CCM 2010; two Gram-negative bacteria Escherichia coli CCM 3988, Pseudomonas aeruginosa CCM 1959) and four yeasts strains (Candida albicans CCM 8186, Candida glabrata CCM 8270, Candida krusei CCM 8271 and Candida tropicalis CCM 8223) were studies. For detection of antimicrobial activity, the grape pomace extracts of white variety Pálava and red variety Dornfelder were used. Pálava pomace extracts were less efficient against the microorganisms tested and Dornfelder extracts were more active against Gram-positive bacteria and yeasts.

The information characteristics of AFLP and ISSR techniques were compared in the analysis of length polymorphism of gamma-treated mutant lines of Amaranthus cruentus and Amaranthus hypochondriacus x Amaranthus hybridus. Both of the techniques are widely used in the manner of universal marker system suitable for the analysis of any plant species, but differ in the time needed for the analyse and costs. Binary data matrices were used for statistical analyses. Cluster analysis was performed in UPGMA method using Euclidean distance to construct dendrograms and principal component analyses was used for generated data, too. Comparison of techniques was made on the graphs obtained from result of PCA and dendrograms that were obtained from results of cluster analysis. Matrix comparisons of Mantel test, for the correspondence of the similarity matrices was performed for the null hypothesis that there is no association between similarity matrices. In the analyzed set used in this study, ISSR primers produced 59% polymorphic bands, while AFLP primers produced 48% polymorphic bands and both of techniques were similar in their obtained information characteristics.

Wine has long been under the view of researchers to test it for prospective benefits to human health. Many recent studies have claimed how moderate consumption of wine, can be beneficial for us. The aim of this study was to assess antagonistic activity and probiotic potential of the LAB strains, Lactobacillus brevis, Lactobacillus hilgardii and Lactobacillus plantarum, isolated from wine against pathogenic bacteria. The antimicrobial activity was detected with disc diffusion method against five species of Gram-negative bacteria: Escherichia coli CCM 3988, Klebsiella pneumoniae CCM 2318, Salmonella enterica subsp. enterica CCM 3807, Shigella sonei CCM 1373, Yersinia enterocolitica CCM 5671 and five Gram-positive bacteria: Bacillus thuringiensis CCM 19, Enterococcus faecalis CCM 4224, Listeria monocytogenes CCM 4699, Stapylococcus aureus subsp. aureus CCM 2461, Streptococcus pneumonia CCM 4501. These results showed antagonistic activity and probiotic potential of pathogenic bacteria isolated from wine. The cell free supernatants (CFS) from Lactobacillus brevis, Lactobacillus hilgardii and Lactobacillus plantarum inhibited the growth of pathogenic bacteria.

Stenotrophomonas maltophilia is the causative agent of nosocomial infections and is characterized by biofilm formation. Primary cell adhesion is crucial in biofilm formation and the type of contact surface has a significant effect. The aim of this work was to assess the antibiofilm activity of Melaleuca leucadendron essential oil. The changes in the biofilm profile of Stenotrophomonas maltophilia were studied using MALDI-TOF MS Biotyper on glass and wooden surfaces. The molecular differences of biofilms in different days were observed as well. The analysis of the mass spectra of S. maltophilia experimental group with Melaleuca leucadendron essential oil showed similarity of the experimental spectra and the control planktonic spectrum on the third day. The results of work proved as such MALDI-TOF MS profiling could be a useful technology for the analysis of biofilm properties.

This study was focused on monitoring of antimicrobial activity of selected medicinal plants: Artemisia vulgaris, Geum urbanum, Euphorbia peplus, Rumex hydrolapathum, Portulaca oleracea, Geranium pratense, Polygonum aviculare. The selected medicinal plants were evaluated for antimicrobial activity with disc diffusion method. Ethanolic extracts against 3 Gram-positive (G+) bacteria (Staphylococcus aureus, Enterococcus faecalis, Bacillus subtilis), 3 Gram-negative (G-) bacteria (Salmonella enterica, Pseudomonas aeruginosa, Yersinia enterocolitica) and 3 yeasts (Candida tropicalis, Candida krusei, Candida glabrata, Candida albicans) were used as model microorganisms to determine antimicrobial activity. The Portulaca oleracea extract showed the largest zone of inhibition against Staphylococcus aureus, Pseudomonas aeruginosa and Yersinia enterocolitica. Artemisia vulgaris extract formed the largest zone of inhibition against Bacillus subtilis, Enterococcus faecalis and one yeast Candida krusei. The extract from Geum urbanum and Euphorbia peplus did not create the maximum inhibition zone against any of the microorganisms that were tested.

Grapevine is a well-known and popular cultural plant that is grown on thousands of hectares worldwide. It possesses a lot of nutritional benefits but, on the other side, some of the people may suffer an allergic reaction when eating grapes or even in the case of consumption of the processed forms. Although grape allergy is not a common allergic reaction, the prevalence of this type of allergy is increasing mainly in southern European countries. One of the main proteins that has the potential to cause grapes allergy is chitinase. Here, a real-time PCR protocol was optimized to analyse the expression of chitinase in the mature grape berries. Different parameter such as analytic kit, primer annealing temperatures or cDNA dilution effects were tested. The final optimized protocol was proved for the chitinase unique isoform amplification by post melting analysis and testing of the protocol was performed for selected grape varieties.

The aim of the study was to determine antimicrobial and antioxidant activity of plant extracts against pathogenic microorganisms in vitro. Extracts of black elder (Sambucus nigra L.), stinging nettle (Urtica dioica L.), marigold (Calendula officinalis L.) and ribwort plantain (Plantago lanceolata L.) were used in experiments and tested against gram-negative bacteria: Pseudomonas aeruginosa CCM 1959, Salmonella enterica subsp. enterica CCM 3807, Yersinia enterocolitica CCM 5671, gram-positive bacteria: Enterococcus faecalis CCM 4224, Staphylococcus aureus subsp. aureus CCM 4223, Streptococcus pneumoniae CCM 4501, and against yeasts: Candida albicans CCM 8186, Candida krusei CCM 8271, Candida tropicalis CCM 8223. The evaluation of antimicrobial effects was performed with disc diffusion method and the minimum inhibitory concentration (MIC) method. Determination of antioxidant activity was performed using the spectrophotometric method. Some species of selected plants were extremely effective against individual pathogenic microorganisms. Using the disc diffusion method, the best antimicrobial activity was reported for the extract from black elder (Sambucus nigra L.) and stinging nettle (Urtica dioica L.). The best antimicrobial activity by the MIC method was found in the extract of ribwort plantain (Plantago lanceolata L.) which was effective against all gram-negative bacteria. The highest antioxidant activity was determined in the extract from black elder (Sambucus nigra L.).

Essential oils are applied for many purposes in pharmaceutical, cosmetic, agriculture, and food industries for their beneficial properties. Due to their bioactive compounds essential oils have an anti-inflammatory, analgetic, cardiovascular or anticancer potential. They can be used as an antioxidant and inhibitor of pathogenic microorganisms in order to improve food quality and extend the shelf life of the food products. The aim of the study was to characterise antioxidant and antimicrobial activity of Citrus aurantium var. bergamia essential oil from Slovak company. Antioxidant capacity of Citrus aurantium var. bergamia essential oil was determined by DPPH radical scavenging method at 54.11%. Antimicrobial activity of Bergamot essential oil was the most effective against yeasts C. krusei and C. tropicalis with inhibition zones 12.67 and 9.33 mm, respectively. The bacteria P. aeruginosa and B. subtilis were also vulnerable where inhibition zones ranged 8.67 and 8.33 mm, respectively.

Cryopreservation is a technique, commonly used for a long-term storage of male gametes as an effective way to preserve their longevity and vitality. During cryopreservation and following the thawing process, sperm cells are exposed to thermal stress, which may lead into disruptions of their morphological structures and a subsequent functional dysfunction. The objective of our study was to monitor HSP90 (heat shock protein 90) gene expression in cryopreserved bovine spermatozoa. HSP90 is one of the main heat-related cellular proteins, which belongs into chaperone protein family. It is involved in the formation of protein complexes, which stabilize proteins against heat stress and aid in the protein degradation. For the experimental purposes were used semen samples from 12 sexually mature Holstein-Friesian bulls. The samples were divided into two identical fractions, one fraction was cryopreserved and kept in liquid nitrogen at -196°C, while the other fraction was processed as non-cryopreserved fresh semen. The sperm motility rate was evaluated using CASA (Computer Assisted Semen Analysis). Quantification of isolated RNA was performed by real-time PCR and the expression of the HSP90 gene was assessed with the Western blot technique. For a proper analysis, we compared cryopreserved sperm samples with fresh ones. The data from our study indicate a reduction in the sperm motility (P

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.

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.

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.

Cadmium (Cd) is a heavy metal that can cause damage to living organisms at different levels. Even at low concentrations, Cd can be toxic to plants, causing harm at multiple levels. As they are unable to move away from areas contaminated by Cd, plants have developed various defence mechanisms to protect themselves. Hyperaccumulators, which can accumulate and detoxify heavy metals more efficiently, are highly valued by scientists studying plant accumulation and detoxification mechanisms, as they provide a promising source of genes for developing plants suitable for phytoremediation techniques. So far, several genes have been identified as being upregulated when plants are exposed to Cd. These genes include genes encoding transcription factors such as iron-regulated transporter-like protein (ZIP), natural resistance associated macrophage protein (NRAMP) gene family, genes encoding phytochelatin synthases (PCs), superoxide dismutase (SOD) genes, heavy metal ATPase (HMA), cation diffusion facilitator gene family (CDF), Cd resistance gene family (PCR), ATP-binding cassette transporter gene family (ABC), the precursor 1-aminocyclopropane-1-carboxylic acid synthase (ACS) and precursor 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) multigene family are also influenced. Thanks to advances in omics sciences and transcriptome analysis, we are gaining more insights into the genes involved in Cd stress response. Recent studies have also shown that Cd can affect the expression of genes related to antioxidant enzymes, hormonal pathways, and energy metabolism.

This study aimed to characterize the bacterial profiles and their association with selected semen quality traits among two chicken breeds. Thirty Lohmann Brown and thirty ROSS 308 roosters were selected for semen quality estimation, including sperm motility, membrane and acrosome integrity, mitochondrial activity, and DNA fragmentation. The oxidative profile of the semen, including the production of reactive oxygen species (ROS), antioxidant capacity, protein, and lipid oxidation, were assessed as well. Moreover, the levels of pro-inflammatory cytokines, including tumor necrosis factor alpha (TNF-α), interleukins 1 and 6 (IL-1, IL-6) and C-reactive protein, as well as the concentrations of selected antibacterial proteins (cathelicidin, β-defensin and lysozyme) in the seminal plasma were evaluated with the enzyme-linked immunosorbent assay. The prevailing bacterial genera identified by the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry were Citrobacter spp., Enterococcus spp., Escherichia spp. and Staphylococcus spp. While the bacterial load was significantly higher in the ROSS 308 line (p < 0.05), a higher number of potentially uropathogenic bacteria was found in the Lohmann Brown roosters. Antimicrobial susceptibility tests revealed a substantial resistance of randomly selected bacterial strains, particularly to ampicillin, tetracycline, chloramphenicol, and tobramycin. Furthermore, Lohmann Brown ejaculates containing an increased proportion of Escherichia coli presented with significantly (p < 0.05) elevated levels of TNF-α and IL-6, as well as ROS overproduction and lipid peroxidation. Inversely, significantly (p < 0.05) higher levels of β-defensin and lysozyme were found in the semen collected from the ROSS 308 roosters, which was characterized by a higher quality in comparison to the Lohmann Brown roosters. In conclusion, we emphasize the criticality of bacteriospermia in the poultry industry and highlight the need to include a more complex microbiological screening of semen samples designated for artificial insemination.

El fósforo (P) es un elemento esencial en la producción agrícola, pero debido a su compleja dinámica en el suelo, solo una pequeña cantidad es aprovechable para las plantas, ya que la mayoría del P se encuentra en formas insolubles, especialmente, en suelos Andisoles de origen volcánico. Los microorganismos con capacidad solubilizadora de fósforo (MSF) son una alternativa para transformar el P a formas solubles y aprovechables por las plantas; además de brindar múltiples beneficios ambientales. Este trabajo identificó y evaluó in vitro, aislados nativos de Pseudomonas fluorescens Mingula, obtenidos de regiones guatemaltecas con suelos Andisoles que limitan la producción agrícola por la alta fijación de P. Se realizaron cultivos in vitro de la bacteria en medio National Botanical Research Institute's phosphate growth (NBRIP), con fosfato tricálcico Ca3(PO4)2 como fuente de P insoluble y se midió el índice de solubilización de fósforo (ISF). Un total de 35 aislados de P. fluorescens fueron identificados y confirmados por PCR específico. El análisis de relaciones genéticas con el marcador AFLP, mostró dos grupos: el grupo A incluyó a los aislados con ISF mayores a 1.75, mientras el grupo B incluyó a aquellos con ISF menor a 1.75. La comparación de ISF entre los aislados y departamentos, demostró diferencia estadísticamente significativa (p < .001), con el aislado Pf_33 como más eficiente. Debido al potencial de solubilización de los aislados nativos del grupo genético A (ISF > 1.75), estos se recomiendan para futuras investigaciones que determinen su respuesta a condiciones de campo y estrategias para el desarrollo de biofertilizantes.

Melissa officinalis L. is well known for its lemon-scented aroma and various pharmacological properties. Despite these valuable properties, the genes involved in the biosynthetic pathways in M. officinalis are not yet well-explored when compared to other members of the mint family. For that, gene expression studies using quantitative real-time PCR (qRT-PCR) are an excellent tool. Although qRT-PCR can provide accurate results, its accuracy is highly reliant on the expression and stability of the reference gene used for normalization. Hence, selecting a suitable experiment-specific reference gene is very crucial to obtain accurate results. However, to date, there are no reports for experiment-specific reference genes in M. officinalis. Therefore, in the current study, ten commonly used reference genes were assessed for their suitability as optimal reference genes in M. officinalis under various abiotic stress conditions and different plant organs. The candidate genes were ranked based on BestKeeper, comparative ΔCt, geNorm, NormFinder, and RefFinder. Based on the results, we recommend the combination of EF-1α and GAPDH as the best reference genes to normalize gene expression studies in M. officinalis. On the contrary, HLH71 was identified as the least-performing gene. Thereafter, the reliability of the optimal gene combination was assessed by evaluating the relative gene expression of the phenylalanine ammonia lyase (PAL) gene under two elicitor treatments (gibberellic acid and jasmonic acid). PAL is a crucial gene involved directly or indirectly in the production of various economically important secondary metabolites in plants. Suitable reference genes for each experimental condition are also discussed. The findings of the current study form a basis for current and future gene expression studies in M. officinalis and other related species.

The aim of the study was to analyse selected aspects of the natural variability of selected varieties of Vitis vinifera. Grapevine is one of the most popular and desirable crops in the world due to the great tradition of wine production, but grape extracts also have a wide range of pharmaceutical effects on the human body. It is important to identify different varieties for the conservation of genetic resources, but also for commercial and cultivation purposes. The variability of conserved DNA-derived polymorphism profiles, as well as microbial characteristics, were analysed in this study. Six different varieties of Vitis vinifera L. were used in the study: Cabernet Savignon, Chardonney, Welschriesling, Weisser Riesling, Gewurztramines and Gruner Veltliner. Genetic polymorphism was analysed by CDDP markers for WRKY genes. Polymorphic amplicon profiles were generated by all primer combinations used in the study. Gruner Veltliner and Welschriesling were the most similar, with a similarity value at 0.778. Microbiological quality of grape and antimicrobial activity against Gram-positive and Gram-negative bacteria and yeasts were analysed further. The plate diluting method for microbial quality and the disc diffusion method for antimicrobial activity were evaluated. The number of total count of bacteria ranged between 3.12 in Cabernet Sauvignon to 3.62 log cfu/g in Grűner Veltliner. The best antimicrobial activity showed Gewurztramines against Salmonella enterica, Yersinia enterocolitica, Pseudomonas aeroginosa, Staphylococcus aureus, Listeria monocytogenes, Candida albicans, Candida krusei, and Candida tropicalis. The best antimicrobial activity was found against Enterococcus faecalis in variety Weisser Riesling.

A plant microbiome is an important factor in plant growth, stress resistance, health status, and consumer quality and safety. The rhizosphere microbiome evolves in a negotiation between microbial communities that inhabit soil and plant root tissue. In this study, the rhizosphere and root internal tissue microbiome of six varieties of lettuce were analyzed in normal conditions and under salinity stress. The metabarcoding analysis used 16S rRNA gene and ITS2 region sequencing. The microbiomes of root samples were significantly less diverse with different members of the community compared to those of the rhizosphere. A significant effect of lettuce variety was found on the diversity index for bacteria and fungi. Varieties formed very different communities of bacteria in roots. Pseudomonas, Herbaspirillum, Mycobacterium, potentially pathogenic Enterobacter, and other genera were more prevalent in certain varieties. Salinity stress had a significant negative impact on bacterial diversity and community composition, whereas the diversity of fungi has not changed significantly, and the fungal community has changed less than the bacterial one. Changes were more evident in varieties that were more resistant to salinity stress than in sensitive varieties.

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´.

The objective of the current study was to assess the efficiency of oryzalin in inducing polyploids in Callisia fragrans (Lindl.) Woodson by in vitro polyploidization. Shoot tips were subjected to Murashige and Skoog (MS) medium containing oryzalin at concentrations 1, 5, and 10 μM for 4 and 8 weeks. Further, the ploidy levels of the plants were confirmed using flow cytometry and chromosome counting. Among all treatments, six tetraploid plants (2n = 4x = 24) were obtained after 8 weeks in MS medium containing 5 μM oryzalin. Upon ex vitro transfer, tetraploid plants were morphologically distinct compared to diploid plants. The size of the leaf and flower increased significantly and nearly doubled when compared to the mother diploid plant. Further, inductively coupled plasma–optical emission spectrometry showed that tetraploid plants exhibited significantly higher sodium, iron, and calcium content, and the potassium content was increased by 100%. Molecular analysis utilizing iPBS and CDDP markers was tested for the first time in C. fragrans to assess the variation between tetraploid and diploid genotypes. Both the markers generated three major clusters, indicating a clear distinction between diploid, tetraploid, and the mixoploid genotypes. In conclusion, in vitro polyploidization using oryzalin could effectively induce polyploids in this and related species. Additionally, the results obtained in this study will provide a basis for future breeding opportunities in this species.

The nuclear reactor accident in Chernobyl, Ukraine, resulted in effects both locally and farther away. Most of the contaminated areas were the agricultural fields and forests. Experimental fields were established near Chernobyl—radioactively contaminated fields localized 5 km from Chernobyl Nuclear Power Plant as well as the remediated soil that is localized directly in the Chernobyl town. Two flax varieties growing under chronic exposition to ionizing radiation were used for this study—the local Ukrainian variety Kyivskyi and a commercial variety Bethune. The screening of the length polymorphism generated by transposable elements insertions were performed. All known types of common flax transposon, retrotransposons and iPBS approach were used. In the iPBS multiplex analyze, for the Kyivskyi variety, a unique addition was found in the seeds from the radioactive contaminated field and for the Bethune variety, a total of five amplicon additions were obtained and one deletion. For the TRIM Cassandra fingerprints, two amplicon additions were generated in the seeds from radioactive contaminated fields for the Bethune variety. In summary, the obtained data represent the genetic diversity between control and irradiated subgroups of flax seeds from Chernobyl area and the presence of activated transposable elements due to the irradiation stress.

Oyster mushroom (Pleurotus ostreatus) is still one of the most cultivated edible and medicinal mushrooms. Despite its frequent cultivation around the world, there is currently just a little information available on the variability of strains in terms of the content of β-glucans in them. This work presents an extensive study of 60 strains in terms of the content of α-glucans and β-glucans in their caps and stipes. The authenticity of the production strains based on an analysis of the variability of their genome by CDDP (Conserved DNA-derived polymorphism) markers was confirmed, whereas identical CDDP profiles were identified between samples 45, 89, 95, and 96. Genetic variability of the analyzed production strains showed a high polymorphism and effective discriminative power of the used marking technique. Medium positive correlations were found among the CDDP profiles and β-glucan content in the group of strains that generated the same CDDP profiles, and low negative correlation was found among these profiles in the group of low β-glucan content strains. For the determination of glucans content, Mushroom and Yeast analytical enzymatic kit (Megazyme, Bray, Co. Wicklow, Ireland) were used. The results clearly showed that the stipe contains on average 33% more β-glucans than the cap. The minimum detected β-glucan content in the stipe was in strain no. 72, specifically 22%, and the maximum in strain no. 43, specifically 56%, which after the conversion represents a difference of 155%. From the point of view of β-glucan content, the stated strain no. 43 appears to be very suitable for the commercial production of β-glucans under certain conditions.

El aguacate es un cultivo de gran consumo a nivel mundial, localizándose su lugar de origen en la región de la Sierra Nevada en California hace ocho a cinco millones de años. En Guatemala se ha reportado una alta diversidad genética mediante caracterizaciones morfológicas, pero debido al comportamiento de polinización cruzada e hibridaciones interraciales, estas caracterizaciones presentan complicaciones y no es posible detallar la riqueza genética total. Los marcadores moleculares son una herramienta útil para este tipo de estudios al estar exentos de efectos ambientales y enfocarse en las diferencias a nivel del ADN. Este estudio analizó la diversidad genética del aguacate nativo guatemalteco colectado en siete poblaciones ubicadas en ocho departamentos del país, empleando el marcador molecular AFLP. Los datos fueron analizados mediante los índices de estructura poblacional que permitieron detallar el grado de diversidad genética de la especie, la cual presenta un alto grado de diferenciación a nivel de individuos (Ht= 0.1933, Hw = 0.1872). Este resultado sugiereuna alta taza de migración, reflejado a través de la similitud genética entre las poblaciones (Hb = 0.0061)y resultando en un alto grado de mezcla de los materiales presentes en el país. La escasa estructura poblacional es atribuida al proceso de selección, al sistema de polinización cruzada, el intercambio de semillas y al proceso de domesticación al cual aún esta sometido el aguacate. Finalmente, se sugiere el resguardado del recurso fitogénetico, así como una caracterización agromorfológica complementaria para establecer una adecuada estrategia de aprovechamiento en programas de mejoramiento genético.

Direct polymerase chain reaction method was use to amplify a thaumatin-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 I and 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 a reproducible 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.

Reverse transcriptase transcribes the cDNA based on its previous extraction and standardization. Reverse transcription step is considered to be critical in the workflow of quantification of transcribed genes. The aim of the study was to extract total RNA by different methods and to analyse the results of the subsequent reverse transcription reaction when different commercial RT kits were used to process RNA extracted from pulp of matured peach fruit. Mature peach pulp was used in the study. The fruit of variety Vistarich was collected in summer 2017 in the orchard of Dvory nad Žitavou. Two RNA extraction methods, TRIzol® Reagent and GeneJET Plant RNA Purification Kit, were tested in to determine the suitable method for peach fruit RNA extraction. Three different cDNA reagent sets were used to transcribe 115 ng/500 ng total RNA or 11 ng/115 ng, respectively. Both variants of the primers, random hexamers as well as oligo (dT) 18, were used to anneal the target mRNA of Pru p 3 allergen following the manufacturer instructions. No specific effect was obtained in the case of peach fruit when using ethanol-extracted tissue treatment and the effect of the used extraction method was more significant. The A260/230 ratios were similar for three from four tested methods. In the case of these three methods, the A260/A230 ratios for all the extracted samples were higher than 1.9 which indicates high purity without contamination by polyphenols and polysaccharides. The specificity of obtained amplicons was proved by restriction cleavage using Tse I restriction endonuclease. This provided the cleavage of the 179 bp long product in all amplicons. Working with mature fruit meet a specific situation in the field of RNA extraction and subsequently all the downstream applications. That is, why choosing the most fitting methods and kits is a crucial step. Here, the method for the semi-quantitative analysis of the Pru p 3 allergen expressions was set up in the way that will be directly applicable for Pru p 3 expression analyses.

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.

Bacterial contamination of semen is an often overlooked, yet important, factor contributing to decreased sperm vitality. Understanding the impact of bacterial presence on sperm structural integrity and functional activity may assist the development of effective strategies to prevent, or manage, bacteriospermia in the breeding practice. The aim of this study was to describe the bacterial profiles of ram semen (n = 35), and we also focused on the associations between bacteriospermia, sperm structure, and function, as well as oxidative and inflammatory characteristics of semen. For a better insight, the samples were divided into three groups, according to the breeds used in the study: native Wallachian (NW), improved Wallachian (IW), and Slovak dairy (SD) breeds. The results showed a significantly lower motility and membrane integrity in the NW group in comparison to the IW and SD groups, which was accompanied by a significantly higher concentration of leukocytes, increased reactive oxygen species (ROS) generation, and subsequent oxidative insults to the sperm lipids and proteins. Accordingly, the NW group presented with the highest bacterial load, in which Staphylococcus and Escherichia were the predominant representatives. The Pearson correlation analysis uncovered positive relationships amongst the bacterial load and leukocytospermia (r = 0.613), the extent of lipid peroxidation (r = 0.598), protein oxidation (r = 0.514), and DNA fragmentation (r = 0.638). Furthermore, positive correlations were found between the bacterial load and pro-inflammatory molecules, such as the C-reactive protein (r = 0.592), interleukin 1 (r = 0.709), and interleukin 6 (r = 0.474), indicating a possible involvement of the immune response in the process of bacteriospermia. Overall, our data indicate that ram semen quality may be equally affected by the bacterial load and diversity. Furthermore, we can assume that the presence of bacteria in ejaculates triggers inflammatory processes, causes ROS overproduction, and, thereby, contributes to alterations in the sperm structure, while at the same time compromising the fertilization ability of male gametes.

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.

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.

The purpose of this study was to investigate changes in TPCs, acid value and peroxide value as well as fatty acids composition in edible oils during french fries production. Lower TPCs content was found in rapeseed oil (3.3%) and the threshold (24%) was achieved on the fourth day. The total time for the deterioration of deep-frying rapeseed oil was 23½ hours. On the contrary, in fresh sunflower oil at the first day was TPCs content 5.5% and the limit of 24% was reached on the third day. The total time for the deterioration of deep-frying sunflower oil was 17½ hours. The results indicated significant differences (

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.

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.

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.

The identification of yacon, a medicinal plant, in tea mixtures by rapid Polymerase Chain Reaction (PCR) and the Direct Analysis in Real Time coupled with Time-of-Flight Mass Spectrometry (DART/TOF-MS) method were evaluated. Three tea products and a pure yacon tea were analysed using the molecular method PCR, concretely the intraspecific variation of the internal transcribed spacer (ITS) regions of rDNA and the DART method coupled with TOF-MS. The results show the reliability of PCR and restriction cleavage of the ITS as a combined approach to confirm the presence of yacon in herbal tea mixtures. Three fragments of approximately 700, 408, and 235 bp in length are present when yacon is a part of the herbal tea mixture. The Principal Component Analysis (PCA) based on the fingerprints of the complete Total Ion Current (TIC) mass spectra shows sufficient separation of herbal teas with and without yacon leaves. The reported methods are technically rapid and can be used as an effective tool for the purposes of yacon identification or authentication.

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.

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 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—Alibernet, Dornfelder, Cabernet Sauvignon, Blaufränkisch, Sauvignon Blanc, Welschriesling, Weisser Riesling, Irsai Oliver, Pinot Blanc, Pálava, Müller-Thurgau, Grűner Veltliner, and Feteasca Regala, grown in the Small Carpathians wine region of Slovakia, Vrbové. 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—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.