Working activity ¹ 6. Research of functional condition of plant cells
Head of Activity: Natalija Shkute, Dr.biol., University of Daugavpils
The objectives of the activity :
- To investigate interaction of new fluorescent probes on plant cells and to analyze the possibilities of application thereof.
The list of functional responsibilities of working activity:
Tasks of the research activity:
1. To perform analysis of academic literature.
2. To investigate interaction of fluorescent probes with inner cell structures.
3. To investigate fundamental processes of plant cells (transportation of substances, cell division, differentiation, destruction etc.), using commercial and newly synthesized fluorescent probes.
4. To evaluate the possibilities of application of new fluorescent probes in quantitative cell biology researches.
At the initial stage of the activity and also in later stages, the latest academic works will be analyzed regarding corresponding researches and news since both traditional (e.g., using ELISE), and modern (using multicolor proteins) fluorescent methods of analysis in cell biology are lately undergoing rapid development. Discovery of new fluorescent probes gives a significant impulse in various directions of researches of cell biology. It is especially important to discover fluorescent probes which might be specific for pigmenting of certain parts of cells and for quantitative analysis of nucleic acids, proteins and other biologically important substances, which would not be toxic and would be usable in researches in vivo. In this sense the particular activity is very closely linked to research of the activity No. 5. Knowledge gained in this activity on binding of synthesized fluorophores with biomolecules will allow better understanding of the process of binding in cells of plants.
In the course of the work it is planned to test the types of introduction of the fluorescent probes obtained in activity No. 5 into living and fixed plant cells and protoplasts. Fluorescent and confocal methods of microscopy will be used in the research.
The possibilities of application of fluorescent probes in vivo and their impact on growth of cells and division thereof will be researched. Changes in spectral parameters of the new fluorescent probes will be compared to plant cells containing GFP.
It is planned to evaluate accrual of active oxygen in cells, depending on the stage of their development, to evaluate them with standard methods of molecular biology and, by the means of fluorescent substances obtained in activity No. 2, to determine activity of different anti-oxidant fermentative systems depending on accrual of active oxygen in the cells.
The following will also be researched:
- the programmed death of plant cells in the conditions of hyperproduction of active oxygen;
- methylation of core DNA in the conditions of hyperproduction of active oxygen;
- influence of various anti-oxidants on renewal of internal cell processes under oxidative stress.
Laboratory premises of Department of Chemistry, Institute of Ecology, G. Liberts Innovative Microscopy Centre of Daugavpils University (and also the University of Latvia for specific research) and the following academic equipment will be used in work (spectrophotometer “CECIL 1021”, spectrofluorometer “FluoroMax-3”, microscopes – confocal laser-scanning microscope Leica TSCP-5, epi-fluorescent microscope Zeiss Axioskop 40, real time PCR “AB 7300”, PCR “AB9700”, genetic analyzer “AB 310”, thermostats, refrigerators, centrifuges).
The expected outcomes of the research activity:
|
Publications in an academic magazine – 3 Reports at international conferences – 3 Participation in work seminars - 6 Promotion paper - 1 |