Katarína Ražná

Changes in expression of BetV1 allergen of silver birch pollen in urbanized area of Ukraine.

The aim of the study was to determinate the level of expression of silver birch allergen Betv1 in pollen samples from different Ukraine areas by RT-qPCR SYBR Green assay. Protocol for quantifying the expression of Betv1 allergen was developed when testing of three housekeeping genes—cyclophylin, alpha-tubulin and transcription factor CBF1. Samples from urbanized area was analysed by real-time PCR when a sample from forest growth conditions was used as a calibrator. Real-time PCR based quantifying of Betv1 provides a useful method for rapid and sensitive analyses of this silver birch allergen. Our results show higher expression levels in samples from central parts of urbanized area as housing estates when compared to the samples from borders of the urbanized area.

Variability of Corylus avellana, L. CorA and profilin pollen allergens expression.

Corylus avellana is the source of inhalant allergies induced by hazel pollen as well as food allergies induced after ingestion of hazelnuts. In this study, real-time PCR approach was used to analyse expression of hazel pollen allergens on the molecular level. Relative quantity of hazelnut allergens Corylus avellana, L. CorA and Corylus avellana, L. pollen profiling in samples from different Ukraine areas were determining and comparing. Differences among the levels of both analysed allergen transcripts were found for hazel CorA and profillin. In both cases, the expression within the urbanized growth conditions was higher when compared to the sample from village area. The average expression for CorA was 0.84 times higher than for profilin and the results are very variable depending on the place of growth. Expression levels here were within the range of 2.957 up to the 52.936. Profilin expression was the highest in the sample from the polluted place of growth—cement plant area with the value of 52 times higher when compared to the sample from the village area. In this study, comparison of expression levels of hazel CorA and profiling pollen allergens was performed for the first time. Real-time PCR assay developed in this study proved the sensitivity for detection of the changes of the hazel pollen allergens expression levels and could benefit labs by fast and reproducible detection method of these allergens.

SODIUM AZIDE INDUCED MORPHOLOGICAL AND MOLECULAR CHANGES IN PERSIMMON (DIOSPYROS LOTUS L.)

Sodium Azide Induced Morphological and Molecular Changes in Persimmon (Diospyros Lotus L.) Persimmon Diospyros lotus L. is an underutilized species, used as rootstock for Japanese persimmon (Diospyros kaki Thunb.) in many European countries. It has a potential for ornamental uses due to its diploid genome that suggests an easy mutation induction. This study was aimed at unravelling the effects of mutagenic sodium azide (in concentrations of 0.1% and 0.5%) on morphological characters (plant height, maximal leaf blade length, maximal leaf blade width and leaf petiole length) and molecular relations of treated accessions, based on AFLP (amplified fragment length polymorphism). In the M1 generation, the analysed characters of two-year old plants differed among the treatments. Sodium azide influenced persimmon stem and leaves in the opposite way, with increase up to 31.57% in plant height and decrease up to 30.33% in leaf petiole length. Changes in morphological characters were significant, that implies the importance of azide influence on the persimmon. Most genotypes treated with 0.1% azide were without significant differences among them and clustered together. The results suggest that the higher azide concentration of 0.5% is more suitable for persimmon seeds azide treatment to form genotypes, significantly different from the control, at the molecular level.

ISSR markers as a tool to distinguish Idt and SSS populations of Zea mays L

Maintaining genetic diversity for crop improvement and improving the quality of genetic resource management is both an inevitable part of nowadays maize breeding. ISSR markers brings the potential of finding a marker system to discriminate Iowa Stiff Stalk Synthetic and Iodent Reid populations of Zea mays, L. and on the base on coefficients of genetic distance and UPGMA grouping appropriate maize genotypes for the best potential for hybridization can be selected. The work presents the potential of ISSR markers in analysis of length polymorphism when a specific ISSR pattern can be used for fast screening of differences among maize genotypes and based of these differences lines can be used in breeding programms in a very specific mode.

Multiple haploids, triploids, and tetraploids found in modern-day “living fossil” Ginkgo biloba

Ginkgo biloba, the last extant representative of a lineage of Mesozoic gymnosperms, is one of the few seed plants with an exceptionally long (~300 Myr) evolutionary history free of genome-wide duplications (polyploidy). Despite this genome conservatism, we have recently found a viable spontaneous tetraploid Ginkgo sapling during routine screening of several plants, demonstrating that natural polyploidy is possible in Ginkgo. Here we provide a much wider flow cytometry survey of ploidy in some European Ginkgo collections, and own seedlings (>2200 individuals and ~200 cultivars). We found a surprisingly high level of ploidy variation in modern-day Ginkgo and documented altogether 13 haploid, 3 triploid, and 10 tetraploid Ginkgo plants or cultivars, most of them being morphologically distinct from common diploids. Haploids frequently produced polyploid (dihaploid) buds or branches. Tetraploids showed some genome size variation. The surveyed plants provide a unique resource for future Ginkgo research and breeding, and they might be used to accelerate the modern diversification of this nearly extinct plant lineage.Genetics: Mutant Ginkgos occur with surprising frequencyThe genetic abnormality, polyploidy, which underlies some unusual varieties of Ginkgo biloba, is more common than previously thought. Genome-wide duplications called polyploidies occur repeatedly in the evolution seed plants. The ancient Ginkgo tree, widely cultivated for its esthetic and medicinal properties, is one of the few seed plant species with a very long polyploidy-free evolutionary history. Yet, when Petr Smarda, of Masaryk University, in Brno, Czech Republic, and colleagues screened more than 2200 Ginkgo individuals and around 200 cultivars, they found 13 haploids, 3 triploids, and 10 tetraploids. These had unusual characteristics such as being shorter than normal and having especially small or large leaves. Improved understanding of the genetic mutations underlying these varieties could facilitate efforts to preserve them and help accelerate diversification of this ancient and nearly extinct plant lineage.

Cultural Extension Of Taxodium In The Territory Of Slovakia And Its Genetic-Genomical Notes

CULTURAL EXTENSION OF TAXODIUM IN THE TERRITORY OF SLOVAKIA AND ITS GENETIC-GENOMICAL NOTES Mňahončáková Erika*, Ražná Katarína, Hrubík Pavel Botanic garden, Slovak University of Agriculture in Nitra, Tr. Andreja Hlinku 2, 949 76 Nitra, Slovakia Department of Genetics and Plant Breeding, Faculty of Agrobiology and Food Resources, Slovak University of Agriculture in Nitra, Slovakia Emeritus professor, Dunajská 16, 949 11 Nitra, Slovakia

The Impact of Multifunctional Crop Rotation on the Yield of Milk Thistle Fruits in the Years 2012 – 2015

Milk thistle can be an annual, rarely a biennial medicinal plant. Polyfactorial field experiment was established during the vegetation period of the years 2012–2015. Three evaluated factors were as follows: crop residues – intercrop – fertilization. Milk thistle was integrated to four crop rotation design with following order of crops: 1. common pea 2. winter wheat 3. milk thistle 4. maize. Milk thistle was subjected to the experimental treatments as follows: 1. K – straw of forecrop removed from the field, R – straw incorporated into soil, 2. M – white mustard as a freezing-out intercrop, B – no intercrop, 3. O – no fertilization, F – with fertilization, 4. experimental year (2012, 2013, 2014, 2015). The recalculated yield of Silybi mariani fructus ranged in the amount from 297 kg*ha-1 (RMO treatment – with crop residues, with intercrop and without the use of fertilizers, year 2015) to 745 kg*ha-1 (KMF treatment – without crop residues, with intercrop and the use of fertilizers, year 2013). Statistical testing of individual factors found highly significant effect of year and highly significant effect of fertilization, and statistically inconclusive effect of sowing intercrop and ploughing crop residues. In the experiment was found a statistically significant difference between yield on the treatments without fertilization and the use of mineral fertilizers.

Analysis of miRNA polymorphism during the selected developmental processes of flax

MicroRNAs represent small non-coding RNAs that play important role in regulating gene expression under various biotic and abiotic stresses as well as in different developmental stages of plants. They are involved in wide variety of biological and metabolic pathways of plants. The research was focused on the potential of selected miRNA-based molecular markers (miR156 and miR168) to map the polymorphism level of flax genome in the selected developmental stages (flower bud - flowering – boll development) as a reflection of the activity of specific miRNAs in these flax organs and tissues. The miRNA polymorphism was evaluated on 8 flax genotypes by miRNA-based molecular marker assays and data were supported by the morphology measurements on buds, flowers, petals and developing bolls. Extent of PCR amplification of miRNA fragments ranged from 40 bp to 200 bp (miR168a-F/miR-R primer pair) or 35 bp to 120 bp (miR156b-F/miR-R primer pair), respectively. MiRNAbased primers amplified in total 196, respectively 158 miRNA loci. Based on the results can be concluded that the representation of miR168 loci have increased according to developmental stages ascending (stage of bud, flower and boll). In overall miRNA156b loci profile in all three analyzed developmental stages is possible to observe the increase of miRNA loci amplification almost in all genotypes. Taking into account the genetic background of genotypes, based on the peak analysis profile of amplified miRNA loci, implemented by GeneTools software, was possible to identified unique loci in linseed and intermediate genotypes in individual developmental stages. Have been found out that only the genotype Bolley Golden differ from other genotypes in both analysis (molecular and morphological) if the variability of miR168 loci number in the flower bud growing stage was compared to the variability in trait petals colour in bud.

Genotyping of Flax Genetic Resources by Mirna-Based Molecular Markers and Morphology

Abstract MicroRNAs (miRNAs) are a class of non-coding RNAs about 20-24 nucleotides long. They play an important role in the gene regulation at the post-transcriptional level. They affect the plant genome response to environmental stress. The miRNA-based molecular markers is type of functional markers reported in very few plants. However, the information connected to the evaluation of genotypes by this type of markers within a single species is missing. Considering the stability, polymorphism, functionality and transferability potential of miRNA-based markers, the research was conducted to apply selected types of them (miR156b, miR408a and the combined type of miR156b/miR408a) for the genotyping analysis of eight flax genotypes of different origin together with the morphology analyses. A total of 145 miRNA loci were identified, of which 19 were unique. The highest numbers of miRNA loci (57) and unique fragments (9) as well as the highest percentage of polymorphism and the extent of polymerase chain reaction (PCR) amplification of miRNA fragments have been observed with the combination of miR156b-F and miR408-F markers. By means of the miRNA markers has been recorded the unique profile of the miRNA loci for individual accessions. The morphology study has shown that the genotypes are the same in the expression of selected morphological traits despite the different use and different places of origin. However, we have identified an interface between some of morphological traits and miRNA-based markers for genotyping the genetic resources of flax. By mutually linking these two types of markers, we were able to determine unique genotypes of flax.