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There are many unsolved taxonomic problems at the intraspecific level in the genus Thalictrum (Ranunculaceae), which may be clarified using molecular systematics methods. No molecular systematics methods have been applied yet to the genus Thalictrum; this study analyzes different leaf tissue preservation and DNA isolation techniques, and the applicability of RAPD. A modified DNA isolation procedure using solution of laundry detergent as a detergent buffer system was the most suitable, especially for dealing with large samples. Since the use of differently preserved leaf tissues simultaneously with fresh leaf tissue may improve sampling in taxonomic research, and because the RAPD technique is sensitive to different factors, the possible drawbacks of using such tissues in RAPD analyses were checked. Of the four preserved leaf tissues, only DNA from silica gel-preserved leaf tissue gave suitably reliable RAPD results to be used with fresh leaf tissue in more extensive taxonomic research. Differently preserved leaf tissues are very problematic starting materials for simultaneous use with fresh leaf tissue in the same RAPD analysis. If differently preserved leaf tissues are to be used and reliable results are to be obtained, research techniques similar to those used in this paper should be applied.
Although intraspecies researches within the black pine (Pinus nigra Arnold) have a long tradition, the intraspecies taxonomy, classification and chorology are still unclear. Among the numerous reasons that have caused this situation the most important are: the absence of a study that would completely cover the whole range of this species, the impossibility of connection of results of the existing detailed studies of certain areas, and the high variability of traits which have been used so far. Since the characteristics of the molecular systematic techniques could make possible the research free of the mentioned shortages, the intention of this study was to determine the relationships among nine populations of black pine using the random amplified polymorphic DNA (RAPD). The obtained results were compared to the recent results of the morphological and anatomical analysis of the leaves of the same populations. The RAPD results clearly divided the Croatian populations from populations of Austria (subsp. nigra) and Turkey (subsp. pallasiana), while among Croatian populations, as in previous study, the existence of several groups (subsp. illyrica, subsp. dalmatica and transitional population between them) was noticed. It is assumed that the optimisations conducted in this study will finally make possible estimating the relationships on the level of the whole range of the black pine and the classification based on molecular traits that are probably less dependent on environmental influences than it has been the case with the characteristics mostly used so far.
Employing the volumetric method by use of a Hirst sampler, a total of 71,286 pollen grains, as many as 94.20% of them allergenic, were recorded in the air samples from the city of Zagreb during the 2002 pollen season. Among identified pollen of 35 plant species/genera/families, 23 were allergenic: Taxus/Juniperus, Alnus sp., Fraxinus sp., Betula sp., Corylus sp., Poaceae, Urticaceae, Artemisia sp., Ambrosia sp., Carpinus sp., Castanea sp., Chenopodiaceae, Salix sp., Populus sp., Ulmus sp., Juglans sp., Quercus sp., Platanus sp., Fagus sp., Plantago sp., Pinus sp., Picea sp. and Abies sp. The pollen of these plants also cause the majority of pollinosis in Europe. Study results and the pollen calendar designed for the 2002 pollen season for the City of Zagreb provide useful data for allergologists to reach an accurate diagnosis. The calendar also provides timely information on airborne pollen types and air concentrations for individuals with pollen hypersensitivity, thus allowing them to adjust their daily activities so as to minimize their contact with allergens and improve their quality of life both at home and at work.
Plant regeneration via somatic embryogenesis and organogenesis was achieved in leaf base and ovary culture of the Croatian endemic Iris adriatica Trinajstić ex Mitić. Callus induction from leaf base explants occurred in the dark on three media with MS mineral solution containing 4.52 μM dichlorophenoxyacetic acid (2,4-D), 4.83 μM naphthaleneacetic acid (NAA), 0.46 μM kinetin (Kin), 5% sucrose and 200 mg L-1 casein hydrolysate. The media differed only in vitamin and/or proline content. Calli from ovary culture were achieved on MS medium containing 45.25 μM 2,4-D. The mean percentage of callus induction from leaf base explants was 18.9%, with no significant differences between media, and 27.3% from ovary sections. All embryogenic calli were formed on MS media containing 0.45 μM 2,4-D, 4.44 μM benzyladenine (BA) and 0.49 μM indole-3-butyric acid (IBA) under low light intensity (25 μE m-2s-1). Transfer of embryogenic calli to hormone-free medium enabled the development of mature somatic embryos on the surface of 6.0% of induced calli produced from leaf base explants and 4.0% of those from ovary sections. Genotype had the main effect on plant regeneration efficiency in Iris adriatica
The aim of the study was to determine the onset, duration and termination of the ragweed pollen season; intradiurnal, daily and monthly pollen variation, and possible differences in the occurrence and concentration of ragweed pollen according to sampling sites between 2002 and 2003. The study was conducted at three sampling sites in central Croatia over two pollen seasons (2002 and 2003) using the volumetric method of sampling (Hirst type sampler). In 2003, the ragweed pollen season was by 43% longer and the percentage proportion of ragweed pollen by 3.4% greater in comparison with 2002. The total ragweed pollen count and number of days with ragweed pollen concentration greater than 30 pollen grains per m3 air showed a declining tendency from East to West in both seasons. The intradiurnal peak concentration occurred between 10.00-14.00. The air concentration of ragweed pollen decreased with temperature decline and precipitation. Results of the study provided useful information to individuals allergic to ragweed pollen thereby allowing them to adjust their outdoor activities to avoid contact with the allergen.
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