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Differences in susceptibility to infestation by spruce spider mite of 3 investigated spruce species (Picea glauca ‘Conica’, P. pungens and P. omorika) may be due to features of anatomical and morphological structure of needles. In P. omorika, showing some resistance to the spruce spider mite, we noted lamellar structure of epidermal cell walls and extensive supporting tissues (hypodermis and sclerenchyma fibres) at the whole circumference of the needle.
Imbibed immature zygotic embryos of Gentiana punctata L. were cultured on MS (Murashige and Skoog) medium consisting of 4.5 µM dicamba, 0.54 µM NAA (naphtaleneacetic acid), 8.88 µM BAP (6-benzylaminopurine) and 0.43 mM AS (adenine hemisulfate). The primary response of expiants consisted in thickening of the subcotyledon and hypocotyl root (HR) zone. Cotyledons and the seminal root did not show any response. Ultrastructural analysis of the initial stages of callus formation revealed numerous changes in cells of expiants. Dedifferentiation of the explant tissues was associated with separation of cells resulting from thickening and folding of walls, destruction of plasmodesmata, and enlargement of intercellular spaces. At the same time, the number of lipid bodies decreased and starch appeared. Indicative of changes in 3-day cultures, the first cell divisions were observed to occur in the HR zone, including cells of the primary cortex, endodermis and pericycle. The dividing cells contained small vacuoles, large, centrally located, layered nuclei with vacuolated nucleoli, amyloplasts with starch, lipid bodies, numerous active Golgi structures, mitochondria and rough endoplasmic reticulum. Actively dividing cells formed callus tissue in which three zones of cells could be distinguished after 14 days of culture: (I) outer (starch) layer, (II) middle layer with actively at dividing small cells, and (III) inner layer containing large vacuolated cells. As the result of cell divisions, at about the fourth week of culture the starch zone formed meristematically active centers of small cells, with dense cytoplasm and large amounts of starch. Among them were small cellular complexes consisting of three cells, with the cell wall structure typical for pre-embryos. By the fifth week of culture, numerous globular and early heart-shaped somatic embryos which formed cotyledons were observed, and further mature somatic embryos showing conversion ability.
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.
Badania wykazały zmiany zarówno jakościowe, jak i ilościowe w zawartości związków fenolowych w liścieniach i osi zarodka dojrzewających nasion rzepaku.
The influence of an Uncaria tomentosa extract on the development of Capsicum plants grown in green-house conditions was examined. The effect of the treatment was investigated with microscopic techniques (light and electron microscope) in leaves from three levels of control plants and plants after treatment with the extract added to the soil in doses of 0.4 and 16 mg/ml (200 ml per pot/plant). In control leaves, changes typical of the subsequent phases of normal development were observed: nuclear chromatin became slightly condensed, plastoglobuli of chloroplasts increased in number and size, intragranal thylakoids were somewhat dilatated. In addition to such commonly occurring changes, some symptoms typical of pepper were observed in the ontogenesis of the examined plant: an increased number of spherical electron-dense deposits in vacuoles, an increased number of peroxisomes, the occurrence of numerous paracrystalline structures in chloroplasts of mature leaves, and, starting in mature leaves, expulsion of plastoglobuli from chloroplasts. After the treatment, most of those changes, leading to ageing, occurred much earlier and were more distinct. Chloroplasts, already in the youngest examined leaves, showed dilatation of intergranal thylakoids, which intensified with aging of the leaves and degradation of grana in the oldest leaves. Starch grains decreased in size and number and plastoglobuli became large. Vesiculation of ground cytoplasm in all leaves was stronger than in the control. No paracrystalline structures in chloroplasts or expulsion of plastoglobuli were observed. Another unusual phenomenon was the disappearance of spherical electron-dense deposits in the central vacuoles of cells. Those observations suggested that U. tomentosa extract enhanced the natural ontogenesis of Capsicum annuum leaves, by accelerating and enhancing the typical characteristics of ageing, and, additionally, it changed the structure and physiology of cells.
As continuation of investigations in to the mechanism of the role of the H1 histone, which is a crucial protein component chromosomes of all eukaryotes, transgenic tobacco plants with different levels of the H1 histone variants were examined. Tobacco has six sequential variants of the H1 histone: two major ones (H1A and H1B), constituting ca. 90% of all H1, and four minor ones (H1C, H1D, H1E and H1F), occurring in very small quantities. The following groups of plants were examined: K - control group with a full set of histone variants; -AB -with the A and B variants removed; -ABCD - with the A, B, C and D variants removed; and -CD - with the C and D variants removed. The analysis of microsporogenesis in those plants, based on preparations squeezed in acetoorcein, revealed the asynchronous course of meiosis in -AB and -ABCD plants, occurrence of chromosomal aberration, and, consequently, the formation of sterile pollen grains (accordingly: 84,4% and 81,4%). In -CD plants, the percentage of aberration and sterile pollen grains was similar to the control material. Electron microscope observations of microsporogenesis showed ultrastructural changes. In -AB and -ABCD plants, a major portion of the pollen grains were degraded. The smallest number of degraded pollen grains, in comparison with the control, was found in the -CD group.
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