IRAP MARKERS OF PLUM TRIM RETROTRANSPOSON

The efficiency comparison of two IRAP markers of plum (Prunus domestica L.) genome was realized on nine plum genotypes. PrimerIRAP1 was derived from LTR´s (Long Terminal Repeat) 3´end of plum retrotransposon Cassandra and primer IRAP2 was designed based onLTR´s 5´end of the same retrotransposon. Orientation and length of primers were designed as the same. PCR products were amplified inoptimized conditions. By the study of electrophoreograms it have been revealed that IRAP1 primer provided 4 polymorphic levels from 10total levels and IRAP2 primer created 10 polymorphic levels from 18 total levels. In the case of IRAP2 primer there were registered uniquebands, too. Percentage content of polymorphic fragments constitutes 23% for IRAP1 and 41% for IRAP2 primer. Four plum genotypescollected from Slovak locations of their natural occurrence (Pečovská N. Ves, Torysa, Podolínec and Lipany) were evaluated based ongenetic similarity Nei, Li coefficients as the most similar by both primers

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

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 Bet v 1 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

ARPN Journal of Agricultural and Biological Science GENOME CHANGES IN MUTANT LINES OF Amaranthus AS DETECTED BY MICROSATELLITE-DIRECTED PCR

ABSTRACT Eight lines and two control genotypes of amaranth were characterized by unanchored and anchored microsatellite markers. The polymorphism of fragments length was evaluated for changes caused by γ-radiation in selected amaranth lines -Amaranthus cruenthus L. -genotype Ficha and Hybrid K-433 -the result of interspecific hybridization of species A. hypochondriacus x A. hybridus. Mutant lines were presented by the M8 generation of plants positively selected for weight of thousand seeds. The very specific changes of the mutant lines coding and noncoding regions were investigated for both the Ficha cultivar and K-433 hybrid. The obtained profiles sets confirm the active reaction of the lines to the gammaradiance treatment. The molecular differences on length polymorphism of random microsatellite markers were observed and the changes of SPAR profiles of mutant lines when compared to control genotypes were confirmed. Primer Ama-AH 5 detected interspecific and primer Ama-AH 4 revealed intra -and interspecific polymorphisms, too. Primer Ama-AH 5 distinguished only the cultivar Ficha and hybrid K-433 accessions. Primer Ama-AH 4 created more polymorphic DNA profiles and has the ability to distinguish not only Ficha cultivar and hybrid K-433, but mutant accessions among themselves

Using of AFLP to evaluate gamma-irradiated amaranth mutants

To determine which of several gamma-irradiated mutants of amaranth Ficha cultivar and K-433 hybrid are most genetically similar to their non-irradiated control genotypes, we performed amplified fragment length polymorphism (AFLP) based analysis. A total of 40 selective primer combinations were used in reported analyses. First analyses of gamma-irradiated amaranth mutant lines were done used the AFLP. In the study, primers with the differentiation ability for all analysed mutant lines are reported. The very specific changes in the mutant lines´ non-coding regions based on AFLP length polymorphism were analysed. Mutant lines of the Ficha cultivar (C15, C26, C27, C82, C236) shared a genetic dissimilarity of 0,11 and their ISSR profiles are more similar to the Ficha than those of K-433 hybrid mutant lines. The K-433 mutant lines (D54, D279, D282) shared genetic dissimilarity of 0,534 but are more distinct to their control plant as a whole, as those of the Ficha mutant lines. Different AFLP fingerprints patters of the mutant lines when compared to the Ficha cultivar and K-433 hybrid AFLP profiles may be a consequence of the complex response of the intergenic space of mutant lines to the gamma-radiance. Although a genetic polymorphism was detected within accessions, the AFLP markers successfully identified all the accessions. The AFLP results are discussed by a combination of biochemical characteristics of mutant lines and their control genotypes

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 Bet v 1 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

Detected Length Polymorphism Shows the Same Characteristics for AFLP and ISSR when Amaranth Gamma-Radiation Mutants are Evaluated

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

Endosidin 2 accelerates PIN2 endocytosis and disturbs intracellular trafficking of PIN2, PIN3, and PIN4 but not of SYT1.

We established that Endosidin2 (ES2) affected the trafficking routes of both newly synthesized and endocytic pools of PIN-FORMED2 (PIN2) in Arabidopsis root epidermal cells. PIN2 populations accumulated in separated patches, which gradually merged into large and compact ES2 aggregates (ES2As). FM4-64 endocytic tracer labeled ES2As as well. Both PIN2 pools also appeared in vacuoles. Accelerated endocytosis of PIN2, its aggregation in the cytoplasm, and redirection of PIN2 flows to vacuoles led to a substantial reduction of the abundance of this protein in the plasma membrane. Whereas PIN-FORMED3 and PIN-FORMED4 also aggregated in the cytoplasm, SYT1 was not sensitive to ES2 treatment and did not appear either in the cytoplasmic aggregates or vacuoles. Ultrastructural analysis revealed that ES2 affects the Golgi apparatus so that stacks acquired cup-shape and even circular shape surrounded by several vesicles. Abnormally shaped Golgi stacks, stack remnants, multi-lamellar structures, separated Golgi cisterna rings, tubular structures, and vesicles formed discrete clusters