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The phenomenon of male sterility has often been observed in investigations on the role of histone H1 in regulation of morphogenetic and cytological processes in transgenic tobacco plants. These changes were accumulated by disturbances in flower development, consisting in lengthening of the pistil style in relation to stamen heads. This prevented pollination and production of seeds. As similar abnormalities occurred also in the present investigations (depending on combination, the sterility% was 84.4 to 19.9, at only 8.1 in the control), the main problem of our investigations was an attempt to explain their reasons. It is commonly known that one of the conditions for formation of fertile pollen is the properly functioning tapetum. Here, we carried out observations of ultrastructure of anther tapetum control cells in respect of abnormalities which occurred during microsporogenesis of transgenic plants with inactivated expression of two major (A, B) and two minor (C, D) histone H1 variants. The investigations were carried out on the following groups of plants: (1) control group with a full set of histone variants (K), (2) with inactivated A and B variants (-AB); (3) with inactivated A, B, C and D variants (-ABCD), (4) with inactivated C and D variants (-CD). It was found that tapetal development was normal in all the investigated groups of plants, and the sequence of changes was similar as in the control. However, certain ultrastructural differences appeared when tapetum functioned as secretory tissue, and in the degeneration phase. In tapetal cell cytoplasm, with participation of rER, lipid bodies were formed, which, having penetrated to the cell surface and to locules, took part in formation of pollen grain sporoderm. Both in the control and in the remaining combination, excluding -ABCD, these bodies looked similar: they were grey, homogenous and surrounded by black jagged deposits. In -ABCD plants, these bodies were more translucent, slightly rarefied, and not surrounded by the deposits. Moreover, in -CD plants, large lipid deposits were frequently observed between remainders of degraded tapetal cells. They did not occur in the control and the remaining combinations.
The effects of stress conditions in a polluted environment on the reproductive processes of plants were studied for two successive years. Vicia cracca is a component of the spontaneous green belt at the base of a copper processing post-flotation waste reservoir (Żelazny Most, Legnica-Głogów Copper Basin, Silesia, Poland). Plants from the site were compared with plants derived from seeds harvested from the contaminated site which grew in non-polluted soil on an experimental plot near Cracow. Almost all of the studied flowers showed degenerative tapetal processes in ~50% of the anthers, starting simultaneously in various stages of meiotic divisions in the pollen mother cells. At the tetrad stage nearly 50% of the anthers contained wholly degenerated tetrads. Disturbances in meiosis increased the amount of degenerated pollen grains. The proportion of potentially functional pollen grains was 47% in 1997 and 56% in 1998; in the control material the corresponding proportions were 87% and 84% (1998 and 1999). Various kinds of developmental disturbances and degenerative processes eliminated some of the ovules from the seed production. The percentages of ovules forming seeds was 56% and 59% in successive years (85% and 87% in the control). Most of the disturbances and degeneration can be attributed to the combined negative impacts of specific environmental factors.
The cytological differentiation of the anthers’ tapetum in the tetraploid cytotype of Ranunculus fluitans Lam. (2n = 32) in relation to that in diploid cytotype of this species (Turała-Szybowska 1984) was studied. In the tetraploid plants the first acytokinetic mitosis is obligatory and leads to the formation of binucleate tapetal cells. Two further mitotic divisions do not comprise all cells. Contrary to the diploid cytotype, in the tetraploid these two mitoses most frequently, are not inhibited at prophase but may proceed further. The mitotic disturbances in the tapetal cells, like the bridges between two chromosome groups and the fusion of metaphase or anaphase plates result in the formation of a number of polyploid nuclei of irregular shapes. In the final stage, in different binucleate tapetal cells the nuclei evaluated as diploid, tetraploid or octoploid occur.
We examined the ultrastructure of plastids during the development and degradation (programmed cell death) of the secretory tapetum in Ornithogalum virens, a species with a pollen coat. Plastids present in meristematic tapetum cells differentiate into amyloplasts and function as amyloplasts during the period of tapetal activity. They undergo two cycles of starch synthesis and hydrolysis before entering the degradation phase in which their structure disintegrates and plastoglobules form, preceding pollen coat formation. We showed that during these processes the plastids are nonrandomly distributed in tapetal cells and that the spatial relation between the plastids, cell nuclei and ER depends on the stage of tapetum and pollen development. We also showed that polar distribution of the plastids and polar localization of starch hydrolysis activity characterized the phase of intensive tapetal secretion.
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