No abstract

We developed an alternative method of staining cell nuclei and chloroplast nucleoids of algal cells using SYBR Green I (the fluorescent dye used commonly for detecting dsDNA in agarose and polyacrylamide gels as an alternative to highly mutagenic ethidium bromide and for DNA staining of viruses and bacteria followed by flow cytometry, digital image analysis or real-time PCR), which enabled routine staining in vivo. Cells do not need to be fixed or treated chemically or physically before staining, thus the shape, size and position of DNA-containing structures are not affected. The fluorescence signal is sharp and reproducible. Examples of application of the method are shown in color microphotographs for representatives of eukaryotic algae from the taxa Chlorophyta, Rhodophyta and the prokaryotic Cyanophyta. The method is also useful for studying progress of the cell cycle in algal cells dividing by multiple fission, as shown by observation of changes in nuclear number during the cell cycle of the green alga Chlamydomonas reinhardtii and Scenedesmus quadricauda. Staining with SYBR Green I can be recommended as a fast, safe and efficient method for the detection of DNA-containing structures in vivo.

No abstract

The effect of cadmium on growth processes (accumulation of RNA, proteins and cell volume), cell cycle reproductive events (DNA replication, mitosis, protoplast fission and daughter-cell formation) and the regulatory activity of histone H1 kinases were monitored in synchronized cultures of the chlorococcal alga Scenedesmus quadricauda. Distinct dosage-dependent inhibitory effects of cadmium ions were found in individual growth and reproductive processes. At concentration of about 60 mumol/L CdCl2, the growth processes were slowed down after about half of the cell cycle but the cells grew to the same or larger size than did untreated cells. At higher concentration, the growth became progressively inhibited, being completely blocked above 240 mumol/L. Total RNA accumulation was the most sensitive growth process. Each of the reproductive events was a target for cadmium ions with increasing sensitivity in the following order: DNA replication, mitosis, protoplast fission and daughter cell formation. Throughout the entire experiment, the activity of "mitosis-specific" histone H1 kinases was negligible in the cadmium (60 mumol/L CdCl2) treated cultures, whilst that of the control culture varied, peaking just prior to nuclear divisions. The activity of "growth-associated" histone H1 kinases was not affected by cadmium ions. No effect was found if cadmium was present during the precommitment period. The longer the period in the presence of cadmium, the stronger inhibition of reproductive events.