Milan Chňapek

Molecular analysis of buckwheat using gene specific markers

Buckwheat ( Fagopyrium esculentum ) is a pseudo-cereal which has spread troughout the world and nowadays it represents cultural, economic and nutritionally important pseudocereal . It´s enviromentally friendly, characterized by high fiber, routine, protein and B vitamins, and is general-purpose. The goal of the present study was to analyze 17 genotypes of buckwheat by using 7 SCoT markers. In total, 52 fragments were detected, of which 38 were polymorphic. The average number of polymorphic fragments was 5.43. The most polymorphic fragments were detected in SCoT 26 and SCoT 29 markers, and the average percentage of polymorphism was 73.36 %. SCoT 29 reached the highest percentage of polymorphism (87.5 %) and SCoT 36 was lowest (60 %). The DI values ​​ranged from 0.625 (SCoT 36) to 0.887 (SCoT 26) and the average DI value was 0.749. The average PIC value was 0.729 with PIC values ranging from 0.386 (SCoT 36) to 0.831 (SCoT 26). To determine the genetic diversity of 17 genotypes of the buckwheat, a dendrogram was created using the hierarchical cluster analysis. The genotypes were divided into two major clusters (I and II). Cluster I was divided into three other subgroups. Sixteen genotypes were included in cluster I and the genotype of Madawaska (USA) was genetically the farthest in cluster II. Genetically the closest were the varieties of Ballada (Russia) and Bamby (Austria). Used SCoT markers were sufficiently polymorphic, were able identify and differentiate chosen set of buckwheat genotypes.

Comparison of nutritional and technological quality of winter wheat (Triticum aestivum L.) and hybrid wheat (Triticum aestivum L. x Triticum spelta L.)

Wheat is one of the most important food crop in the world. Gluten proteins represent major protein fraction of the grain and are responsible for unique properties of the dough. The aim of this study was to analyze one genotype of the winter wheat (Triticum aestivum L.) and one genotype of hybrid wheat (Triticum aestivum L. x Triticum spelta L.) based on protein polymorphism and to compare their nutritional and technological quality. According to Kjeldahl and Golenkov, content of total nitrogen and protein fraction composition of analyzed samples were determined. Higher content of albumins and globulins (26.39%) was detected in hybrid wheat genotype PS Lubica with total nitrogen 2.02% and crude protein content 11.49%. Winter wheat genotype Elinor showed higher content of prolamins and glutelins (70.68%) and higher coefficient of nutritional quality (91.79%) than analyzed hybrid wheat. Higher total content of amino acids was determined in hybrid wheat genotype PS Lubica (122.02 mg*g-1 dry weight). For the separation of glutenin protein subunits, SDS-PAGE method by ISTA was used. On the basis of molecular weight, glutenins were separated in SDS-PAGE into HMW-GS (Elinor 10.7%; PS Lubica 3.78%) and LMW-GS (Elinor 58.4%; PS Lubica 67.28%). Electrophoretic profile of genotype Elinor was 0, 7+9, 5+10 with Glu Score 7 and composition of HMW-GS in genotype PS Lubica was 0, 20, 2+12 with Glu Score 4. According to the results, better nutritional and technological quality was proved in winter wheat genotype Elinor. According to identical results in 5 biological replicates, hybrid wheat genotype PS Lubica was not recommended for bread-making.

Lunasin detection in coloured wheat genotype

Lunasin is a biologically active protein, composed of 43 amino acid residues. There has been proven many health-promoting effects of lunasin peptide. The most important health benefits include: anti-hypertension, antioxidant activity, cancer prevention or therapy. It was also demonstrated anti-inflammation, hypocholesterolemic activity, anti-obesity and immunomodulation. The focus of our research is to summarize the discovery, characterization and biological activities of lunasin, which will provide a reference for the future development and utilization of lunasin, and a basis for exploring the underlying mechanisms of these health-beneficial functions. Lunasin was first isolated in 1987 at Niigata University School of Medicine in Japan, during the screening of protease inhibitors from soybean seeds. It was subsequently found in other beans, grains and herbal plants, including wheat, barley, rye, triticale. Concentration of lunasin is ranging from 0.013 to 70.5 mg protein lunasin/g of protein. Big step forward in the understanding of the lunasine operating mechanism in the fight against cancer has arisen after study on cloning of the soybean lunasin gene and subsequent transfection into mammalian cells which led to the discovery that the lunasin gene can disrupt mitosis and induce chromosome breakage, ultimately leading to cell apoptosis. The main goal of our work was to evaluate collection of wheat with unusual grain colour for presence of lunasin gene. DNA was extracted by commercial kit and lunasin gene was detected by PCR reaction. Our results showed presence of lunasin gene detected by 3 combinations of 2 sets of primer pair and indicated lunasin peptide presence in cereal grains. These findings are necessary to confirmed by proteome analysis. Normal 0 21 false false false CS JA X-NONE

Polymorphism of proteins in selected slovak winter wheat genotypes using SDS-PAGE

Winter wheat is especially used for bread-making. The specific composition of the grain storage proteins and the representation of individual subunits determines the baking quality of wheat. The aim of this study was to analyze 15 slovak varieties of the winter wheat (Triticum aestivum L.) based on protein polymorphism and to predict their technological quality. SDS-PAGE method by ISTA was used to separate glutenin protein subunits. Glutenins were separated into HMW-GS (15.13%) and LMW-GS (65.89%) on the basis of molecular weight in SDS-PAGE. At the locus Glu- A1 was found allele Null (53% of genotypes) and allele 1 (47% of genotypes). The locus Glu-B1 was represented by the HMW-GS subunits 6+8 (33% of genotypes), 7+8 (27% of genotypes), 7+9 (40% of genotypes). At the locus Glu-D1 were detected two subunits, 2+12 (33% of genotypes) and 5+10 (67% of genotypes) which is correlated with good bread-making properties. The Glu – score was ranged from 4 (genotype Viglanka) to 10 (genotypes Viola, Vladarka). According to the representation of individual glutenin subunits in samples, the dendrogram of genetic similarity was constructed. By the prediction of quality the results showed that the best technological quality was significant in the varieties Viola and Vladarka which are suitable for use in food processing.