Plants have developed a range of strategies for resisting environmental stresses. One of the most common is the synthesis and deposition of callose, which functions as a barrier against stress factor penetration. The aim of our study was to examine whether callose forms an efficient barrier against Pb penetration in the roots of Lemna minor L. exposed to this metal. The obtained results showed that Pb induced callose synthesis in L. minor roots, but it was not deposited regularly in all tissues and cells. Callose occurred mainly in the protoderm and in the centre of the root tip (procambial central cylinder). Moreover, continuous callose bands, which could form an efficient barrier for Pb penetration, were formed only in the newly formed and anticlinal cell walls (CWs); while in other CWs, callose formed only small clusters or incomplete bands. Such an arrangement of callose within root CWs inefficiently protected the protoplast from Pb penetration. As a result, Pb was commonly present inside the root cells. In the light of the results, the barrier role of callose against metal ion penetration appears to be less obvious than previously believed. It was indicated that induction of callose synthesis is not enough for a successful blockade of the stress factor penetration. Furthermore, it would appear that the pattern of callose distribution has an important role in this defence strategy.
The content of lead in greening etiolated barley leaves remained the same, regardless the time of incubation of excised leaves in the presence of lead ions (8 -24 h). The lead deposits have not been detected within mesophyll cells, but were found in intercellular spaces of mesophyll, in guard cells and in cuticle coveting stomata. This suggests that lead may be transported in the leaves via transpiration stream. Lead reduced the content of chlorophyll, especially chlorophyll b content and the average number of grana, whereas in the presence of kinetin the content of chlorophyll increased. In the combined treatment (lead + kinetin) kinetin diminished the inhibitory effect of lead on the chlorophyll content. The number of chloroplasts in mesophyll cells remained unchanged after lead treatment, whereas kinetin alone or applied together with. lead increased the average chloroplasts number. The thylakoids system in chloroplasts of kinetin and kinetin + lead treated plants was similar to that observed in control, although the grana number was smaller. Both lead and kinetin increased the content of condensed chromatin in nuclei.
The main sites of lead entering the protonemata were spores with broken cell walls and apical cells of protonemata. The lead ions could enter the symplast probably by endocytosis but we can not exclude diffusion as the way of entering. Lead deposits inside a protoplast were observed after 2 h of lead application. After 12 h, Pb deposits were commonly present in all cell structures surrounded by a membrane. Some disturbances in ultrastructure of organelles were found after 8 h. The most frequent were: change of endoplasmatic reticulum configuration from linear to concentric one in many cases surrounding other organelles (e.g. mitochondria) and large and numerous plastoglobuli in chloroplasts. Longer treatment of protonemata with lead (24 h or 48 h) caused degeneration of organeUes and even death of the cell.
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