The Choice of Suitable Conditions for Wheat Genetic Transformation

Wheat is one of three most important cereals worldwide. Its production rises every year. There is a possibility to improve quantitative and qualitative parameters by biolistic method of transformation. The process of introduction of desired gene into the wheat genome and plant regeneration is affected by many factors. To identify the suitable conditions, selection system, the influence of donor, plant environment and the regeneration capacity of wheat genotypes were tested. The scutella of immature embryos served as the initial explants. Plant regeneration was achieved by 6 out of 11 genotypes tested. The highest values were reached by the cultivar Ilias. The effect of donor, plant environment was evaluated by two genotypes. Plants from growth chamber appeared to be better source of explants compared to plants grown in the natural conditions. The selection system was optimized as follows: regeneration medium in the dark and subsequently in photoperiod conditions (DR) with 5 mg/l of phosphinotricin (PPT), shoot induction medium (DS) with 7 mg/l of PPT

Arbuscular Mycorrhizal Fungi – Their Life and Function in Ecosystem

Arbuscular mycorrhizal fungi living in the soil closely collaborate with plants in their root zone and play very important role in their evolution. Their symbiosis stimulates plant growth and resistance to different environmental stresses. Plant root system, extended by mycelium of arbuscular mycorrhizal fungi, has better capability to reach the water and dissolved nutrients from a much larger volume of soil. This could solve the problem of imminent depletion of phosphate stock, affect plant fertilisation, and contribute to sustainable production of foods, feeds, biofuel, and raw materials. Expanded plant root systems reduce erosion of soil, improve soil quality, and extend the diversity of soil microflora. On the other hand, symbiosis with plants affects species diversity of arbuscular mycorrhizal fungi and increased plant diversity supports diversity of fungi. This review summarizes the importance of arbuscular mycorrhizal fungi in relation to beneficial potential of their symbiosis with plants, and their function in the ecosystem

Elicitation of Hyoscyamine Production in <i>Datura stramonium</i> L. Plants Using Tobamoviruses

Datura stramonium L. produces tropane alkaloids, and the hyoscyamine is dominant among them. Hyoscyamine is produced by hairy root cultures in vitro derived from native plants or plants with the genetically modified biosynthetic pathway for hyoscyamine. A common procedure is extraction from cultivated plants. Elicitors for increased production can be used in both cases. Live viruses are not well known for use as elicitors, therefore, D. stramonium plants grown in soil were artificially infected with the tobamoviruses Pepper mild mottle virus (PMMoV), Tomato mosaic virus (ToMV), and Tobacco mosaic virus (TMV). Differences in the content of hyoscyamine were between capsules and roots of infected and non-infected plants. Elicitation increased content of hyoscyamine in capsules 1.23–2.34 times, compared to the control. The most effective viruses were PMMoV and ToMV (isolate PV143), which increased content to above 19 mg/g of fresh weight of a capsule. The effect of each virus elicitor was expressed also in hyoscyamine content in roots. Elicited plants contained 5.41–16.54 times more hyoscyamine in roots compared to non-elicited plants. The most effective elicitor was ToMV SL-1, which raised production above 20 mg/g fresh weight of roots. It has been shown that tobamoviruses can be used as biotic elicitors

Formation of Potential Heterotic Groups of Oat Using Variation at Microsatellite Loci

An evaluation of polymorphism at the microsatellite loci was applied in distinguishing 85 oat (Avena sativa L.) genotypes selected from the collection of genetic resources. The set of genotypes included oats with white, yellow, and brown seeds as well as a subgroup of naked oat (Avena sativa var. nuda Koern). Variation at these loci was used to form potential heterotic groups potentially used in the oat breeding program. Seven from 20 analyzed microsatellite loci revealed polymorphism. Altogether, 35 microsatellite alleles were detected (2–10 per locus). Polymorphic patterns completely differentiated all genotypes within the subgroups of white, brown, and naked oats, respectively. Only within the greatest subgroup of yellow genotypes, four pairs of genotypes remained unseparated. Genetic differentiation between the oat subgroups allowed the formation of seven potential heterotic groups using the STRUCTURE analysis. The overall value of the fixation index (Fst) suggested a high genetic differentiation between the subgroups and validated a heterotic grouping. This approach can be implemented as a simple predictor of heterosis in parental crosses prior to extensive field testing or development and implementation of more accurate genomic selection

Introduction of a synthetic Thermococcus-derived α-amlyase gene into barley genome for increased enzyme thermostability in grains

Background: The enzymes utilized in the process of beer production are generally sensitive to higher temperatures. About 60% of them are deactivated in drying the malt that limits the utilization of starting material in the fermentation process. Gene transfer from thermophilic bacteria is a promising tool for producing barley grains harboring thermotolerant enzymes. Results: Gene for α-amylase from hydrothermal Thermococcus, optimally active at 75–85°C and pH between 5.0 and 5.5, was adapted in silico to barley codon usage. The corresponding sequence was put under control of the endosperm-specific promoter 1Dx5 and after synthesis and cloning transferred into barley by biolistics. In addition to model cultivar Golden Promise we transformed three Slovak barley cultivars Pribina, Levan and Nitran, and transgenic plants were obtained. Expression of the ~50 kDa active recombinant enzyme in grains of cvs. Pribina and Nitran resulted in retaining up to 9.39% of enzyme activity upon heating to 75°C, which is more than 4 times higher compared to non-transgenic controls. In the model cv. Golden Promise the grain α-amylase activity upon heating was above 9% either, however, the effects of the introduced enzyme were less pronounced (only 1.22 fold difference compared with non-transgenic barley). Conclusions: Expression of the synthetic gene in barley enhanced the residual α-amylase activity in grains at high temperatures. Keywords: Amylopectin, Amylose, Beer, Fermentation, Gene transfer, Glutenin, Hordeum, Promoter, Seed-directed expression, Synthetic gene, Thermotolerant bacteri

Molecular and Biological Characterisation of Turnip mosaic virus Isolates Infecting Poppy (Papaver somniferum and P. rhoeas) in Slovakia

In recent years, the accumulated molecular data of Turnip mosaic virus (TuMV) isolates from various hosts originating from different parts of the world considerably helped to understand the genetic complexity and evolutionary history of the virus. In this work, four complete TuMV genomes (HC9, PK1, MS04, MS15) were characterised from naturally infected cultivated and wild-growing Papaver spp., hosts from which only very scarce data were available previously. Phylogenetic analyses showed the affiliation of Slovak Papaver isolates to the world-B and basal-B groups. The PK1 isolate showed a novel intra-lineage recombination pattern, further confirming the important role of recombination in the shaping of TuMV genetic diversity. Biological assays indicated that the intensity of symptoms in experimentally inoculated oilseed poppy are correlated to TuMV accumulation level in leaves. This is the first report of TuMV in poppy plants in Slovakia