The aim of this work was to find an effective protocol for in vitro propagation and to perform the in vitro polyploidization of diploid Thymus vulgaris (2n = 30) using two experimental methods based on the use of oryzalin, an antimitotic agent. The ploidy level of the obtained shoots was checked by flow cytometric analysis. The most efficient conditions for inducing polyploidy were oryzalin concentrations of 0.346 and 1.73 mg L−1 present in the medium for two weeks. The vital polyploid shoots were multiplied for further evaluation, rooting and final transfer to nonsterile glasshouse and field conditions. The chemical compositions of the essential oils (EOs)—which were obtained from dried field grown plants by steam distillation—were analyzed by gas chromatography/mass spectrometry (GC/MS). The identified substances contributed approximately 95% to the total peak area. Statistical analysis revealed that the tetraploid subclone and the diploid reference plant do not differ in total terpene content, but they do differ in the relative proportions of all the individual terpenes with the exception of α-pinene and UN5, indicating that both clones produce EOs of different quality. The obtained results showed the possibility of developing more efficient botanical insecticides based on EOs obtained from the tetraploid plants.
In 1997 and 1998 we used samples of harvested grain to verify the possibility of distinguishing 14 winter wheat genotypes and six triticale genotypes and detecting the impurity on the basis of the detection of polymorphism of prolamin kernel proteins using the methods of the PAGE ISTA. On the basis of the identity index two sister prolamin lines with different percentage of participation, which was based on the weather conditions of the year of harvest, were discovered in seven wheat genotypes (Astella, Brea, Hana, Ilona, Siria, Sofia and Šárka) and two triticale genotypes (Tornádo and KM 779). A foreign genotype was detected in the Hana and Astella varieties. The identity index of the impurity to the Astella and Hana variety (i.e. ii = 0.28 and ii = 0.20, respectively) was considerably lower. In an unknown genotype (impurity) we detected the gliadin block Gld 1B3, which is the genetic marker of rye translocation T1BL.1RS, the Sr31 gene of resistance to black rust, higher cold resistance and the marker of poor baking quality (presence of secalin genes). The results proved the potential practical application of the method of electrophoretic detection of polymorphism of prolamin proteins as markers of impurities of foreign genotypes in a seed sample.Keywords: winter wheat; winter triticale; prolamin proteins; electrophoresis; admixture In breeding work it is important to make sure to use initial seed stock that has the required properties, but also to use material of corresponding varietal (genotypic) purity. This also applies to farmers who produce and sell seeds in the following year and, last but not least, it concerns the processing industry where admixtures of an undesirable genotype have a negative effect and result in poorer quality of the end product.Both wheat (Triticum aestivum L.) and triticale (Xtriticosecale Wittmack) are self-pollinating crops with a low share of cross-pollination (4-5%). This means that most of the varieties are of the line type or they are a mixture of isogenic lines (Chloupek 2000). A number of methods can be used to detect the impurities in a sample of harvested seeds. In our case we used the polymorphism of reserve kernel proteins, i.e. the alcohol-soluble fraction -prolamins (gliadins, secalins). High polymorphism was used by many authors (Šašek and Sýkorová 1989, Metakovsky 1991, Černý et al. 1992 and is characteristic of these proteins. Compared to other variability markers they offer a number of advantages.Compared to other biochemical markers, the isoenzymes, they are not so dependent on conditions of the external environment and they are independent on the ontogenetic stage of the plant. Other markers, which are undergoing considerable development at the present time, are monitoring on the level of DNA polymorphism. However, compared to the detection of polymorphism of prolamin proteins these methods are much more expensive due to the used chemicals and technology.In terms of the above aspects, polymorphism of reserve proteins is more suitable for the detection of ...
Abstract:The polymorphism of prolamin storage proteins was studied in seed samples of 20 historical cultivars of spring barley (Hordeum vulgare L.) of Czech and Slovak origin, using polyacrylamide gel electrophoresis (PAGE). Only two samples were uniform. Most heterogeneity of prolamin patterns was observed in the oldest accessions. By means of a prolamin identity index it was possible to distinguish sister lines from admixtures within the seed samples. The obtained spectra will be used as additional descriptors for the spring barley core collection of the Collection of Genetic Resources of the Agricultural Research Institute Kroměříž, Ltd.
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