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1

An improved method for the cell cycle synchronization of Vicia faba root meristem cells

100%
Polit J. T.
Acta Physiologiae Plantarum
|
2008
|
tom 30
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nr 3
Plant root meristem cells divide asynchronously which makes biochemical analysis of cell cycle regulation particularly difficult. In the present article a high level of cell cycle synchronization in Vicia faba root meristems was obtained by using a rich medium (HNS), special culture conditions and a double-block method with replication inhibitor—hydroxyurea (HU). Two HU concentrations were tested and different periods of the first and the second synchronization, and of cycle recommencement between the first and the second blockage. The level of synchronization was estimated on the basis of 3-thymidine labeling indices, mitotic, and phase indices and indices determining the percentage of G1 and G2 cells, which were identified by cytophotometric measurements of DNA content in individual nuclei. The highest level of cell cycle synchronization was obtained after double treatment of meristems with 1.25 mM HU (18 and 12 h) separated by 6-h incubation in HNS without HU. During the second postincubation in HNS in subsequent hours: 4, 7, 10, 11, over 90% of cells in the S phase, nearly 70% in G2 phase, 86% in mitosis, and nearly 70% in G1 phase were received, respectively. The use of 2.5 mM HU in a similar experimental procedure caused disturbed divisions.
2
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Sucrose uptake and transport are inhibited by okadaic acid during regeneration of the sugar-starved Vicia faba root meristem cells

63%
Polit J. T., Zabka A.
BioTechnologia. Journal of Biotechnology Computational Biology and Bionanotechnology
|
2013
|
tom 94
|
nr 3
3
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DNA replication stress-induced biphasic nuclear structures in Allium cepa root meristem cells

51%
Zabka, Polit J. T., Maszewski J.
BioTechnologia. Journal of Biotechnology Computational Biology and Bionanotechnology
|
2013
|
tom 94
|
nr 3
4
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IAA and BAP affect protein phosphorylation-dependent processes during sucrose-mediated G1 to S and G2 to M transitions in root meristem cells of Vicia faba

51%
Polit J. T., Maszewski J., Rosiak M.
Acta Societatis Botanicorum Poloniae
|
2004
|
tom 73
|
nr 1
In carbohydrate-starved root meristems of Vicia faba subsp. minor, the expression of two Principal Control Points located at the final stages of the G1 (PCP1) and G2 (PCP2) phases has been found to be correlated with a marked decrease of protein phosphorylation within cell nuclei, nucleoli and cytoplasm. Adopting the same experimental model in our present studies, monoclonal FITC conjugated antibodies that recognize phosphorylated form of threonine (αTPab-FITC) were used to obtain an insight about how the indole-3-acetic acid (IAA), benzyl-6-aminopurine (BAP), and the mixture of both phytohormones influence the time-course changes in an overall protein phosphorylation during sucrose-mediated PCP1→S and PCP2→M transitions. Unsuspectedly, neither IAA, BAP, nor the mixture of both phytohormones supplied in combination with sucrose did up-regulate protein phosphorylation. However using the block-and-release method, it was shown that root meristems of Vicia provided with sucrose alone indicated higher levels of αTPab-FITC. Contrarily, phytohormones supplied in combination with sucrose induced apparent decline in phosphorylation of cell proteins, which - when compared with the influence of sucrose alone - became increasingly evident in time. Thus, it seems probable, that a general decline in the amount of αTPab-FITC labeled epitopes may overlay specific phosphorylations and dephosphorylations governed by the main cell cycle kinases and phosphatases.
5

DGAT2 revealed by the immunogold technique in Arabidopsis thaliana lipid bodies associated with microtubules

45%
Kwiatkowska M., Stepinski D., Poplonska K., Wojtczak A., Polit J. T.
Folia Histochemica et Cytobiologica
|
2012
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tom 50
|
nr 3
6

Immunogold method evidences that kinesin and myosin bind to and couple microtubules and actin filaments in lipotubuloids of Ornithogalum umbellatum ovary epidermis

45%
Kwiatkowska M., Polit J. T., Poplonska K., Stepinski D., Wojtczak A.
Acta Physiologiae Plantarum
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2013
|
tom 35
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nr 06
Lipotubuloids in ovary epidermis of Ornithogalum umbellatum which are a domain of cytoplasm containing a lot of lipid bodies, microtubules and actin filaments, ribosomes, endoplasmic reticulum as well as scarce mitochondria, microbodies, dictyosomes, autolytic vacuoles, exhibit progressive-rotary motion. The immunogold method demonstrated that microtubules and actin filaments of lipotubuloids might be connected with one another by myosin and kinesin. It was supposed that collaboration of motor proteins with actin filaments and microtubules makes autonomic high peripheral speed rotary motion of lipotubuloids in epidermis cells possible. Moreover, myosin was also detected in Golgi bodies in lipotubuloid. In lipotubuloids, the immunogold method demonstrated immunosignals after the use of an antibody to dynein light chains but spectroscopy mass analysis showed that in O. umbellatum epidermis lacked dynein heavy chains.
7

Microtubule heterogeneity of Ornithogalum umbellatum ovary epidermal cells: non-stable cortical microtubules and stable lipotubuloid microtubules

45%
Kwiatkowska M., Stepinski D., Polit J. T., Poplonska K., Wojtczak A.
Folia Histochemica et Cytobiologica
|
2011
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tom 49
|
nr 2
8

Localization sites of nuclear envelope SUN2-like proteins in root meristem cells of Allium cepa under hydroxyurea-induced DNA replication stress

45%
Zabka A., Winnicki K., Polit J. T., Bernasinska J., Maszewski J.
Acta Physiologiae Plantarum
|
2015
|
tom 37
|
nr 08
Regardless of the DNA replication stress induced by low concentration of hydroxyurea (HU), root apical meristem cells of Allium cepa keep growing, and some of them override the DNA damage checkpoint mechanisms initiating either premature or an abnormal mitotic chromosome condensation. Prolonged incubation of onion seedlings with HU results in an increased level of immunodetectable proteins sharing epitopes with SUN2, one of the highly conserved elements linking nuclear envelope (NE) to the cytoand nucleoskeletal structures. In addition toNE, phragmoplast and cell plate, our observations extend an array of subcellular compartments at which SUN2-like proteins (SUN2-LPs) are localized. These include cortical preprophase band of microtubules, centromeric regions of ana- and telophase chromosomes, and nuclear bodies (SUN2-NBs) polarly localized in interphase nuclei according to Rabl’s configuration. SUN2- NBs (distinct from fibrillarin-rich Cajal bodies) colocalize with late-replicating areas of heterochromatin and are thought to represent clustered centromeres. Three-dimensional spatial analysis of SUN2-NBs suggests their connections with NE. An enhanced expression and additional localization sites of SUN2-LPs may be correlated with a considerable reprogramming of cellular functions triggered in response to prolonged HU treatment.
9
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Cooperation of motor proteins with microtubules in generations of lipotubuloid's motion in Ornithogalum umbellatum ovary epidermis

45%
Kwiatkowska M., Polit J. T., Poplonska K., Stepinski D., Wojtczak A.
BioTechnologia. Journal of Biotechnology Computational Biology and Bionanotechnology
|
2013
|
tom 94
|
nr 3
10

Inhibition of germination and early growth of rape seed (Brassica napus L.) by MCPA in anionic and ester form

39%
Polit J. T., Praczyk T., Pernak J., Sobiech L., Jakubiak E., Skrzypczak G.
Acta Physiologiae Plantarum
|
2014
|
tom 36
|
nr 03
MCPA (4-chloro-2-methylphenoxy) acetic acid is a common synthetic auxin used as a herbicide. The purpose of this study was to determine the effects of four new forms of MCPA being the herbicidal ionic liquids (HILs) with MCPA as an anion and two previously known formulations (potassium–sodium salt and 2-ethylhexyl ester) on seed germination and seedling development of winter oilseed rape (Brassica napus). Rape plants are susceptible to MCPA and volunteers can be a big problem in crop rotation. Seedling fresh weight and root length were quantified, mitotic activity, as well as lipid, starch, hydrogen peroxide and polyphenol contents were assessed by light and fluorescence microscopy and the computer-aided cytophotometer. In primary roots mitotic activity was almost completely inhibited under the influence of herbicides, cell elongation zones and root hair zones were significantly reduced, and a characteristic bolded root segment formed just above a meristem. In contrast to the traditional salt formulation the new HILs were weak inducers of hydrogen peroxide synthesis, but were potent stimulators of the synthesis of phenolic compounds and storage as well as emergency substances such as lipids and starch. All tested forms of MCPA caused strong phytotoxic effect on winter rape seedlings, but the tested HILs were more effective.
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