Milada Čiamporová

Responses of maize (Zea mays L.) plants to copper stress—I. Growth, mineral content and ultrastructure of roots

Changes in the morphology, physiology and ultrastructure of root systems of Zea mays cv. Aris under various Cu treatments were investigated. A progressive decrease of root length and biomass with increasing Cu in nutrient solution was observed. Mineral content and distribution were markedly affected by Cu. The roots accumulated significantly higher amounts of Cu than the above ground parts. Significant reduction of root calcium and iron contents as well as extensive damage to root epidermal cells occurred at the higher Cu concentration. In the remainder of the root, the effects of Cu (80 μM) varied within the same tissues. Thus, there were cortical or stelar cells with disintegrated cytoplasm, next to cells with well preserved plasmalemma, tonoplast and cytoplasm including cell organelles. Multi-lamellar bodies inside the vacuoles indicated an increased activity in digestion of the cytoplasmic components under Cu stress. Deposits of a less dense and less compact material were found behind the plasmalemma associated with the cell wall as well as an accumulation of dense material attached to the cell walls; cells with such deposits in their walls revealed extensive cytoplasmic damage. However, root ultrastructure was affected less than the morphology and physiology. The occurrence of well preserved cells indicates that Zea mays root cells do not respond uniformly to stressful conditions and suggests the development of a resistance strategy of maize roots to Cu-toxicity.

Morphological and Structural Responses of Plant Roots to Aluminium at Organ, Tissue, and Cellular Levels

Toxic effects of aluminium are primarily root-related. This review deals with growth, morphological, and ultrastructural responses of root to aluminium, their diversity along the root axis, and in the root tissues. The cell elongation seems to be most sensitive and responsible for early inhibition of root elongation. Longer Al treatment is required to reduce cell division or to interfere with nucleic acids in the root apex. Alterations of root morphology include root thickening, disturbances of root peripheral tissues, and initiation of lateral roots closer to the root tip. Ultrastructure alterations depend strongly on position of the cells with respect to the Al source, and on their developmental stage. Cell elongation and cell ultrastructure including organisation of cytoskeleton are most sensitive within the distal part of the transition zone of the root apex. This correlates with the rate of uptake and accumulation of Al along the root apex. Recognising the diverse responses and the most sensitive sites within the root apex can help in elucidating the mechanism(s) of Al effects on plants.

The vegetation of metalliferous and non-metalliferous grasslands in two former mine regions in Central Slovakia

We investigated the composition of the vegetation in two former mining regions in Central Slovakia: Banská Štiavnica with predominant Pb-Zn contamination and Staré Hory with a very high Cu content in the soil. Old heaps rich in heavy metals are covered with specific vegetation. On the Cu-rich spoil heaps, species-poor plant communities with prevailing Agrostis stolonifera, Avenella flexuosa, Acetosella vulgaris, Arabidopsis arenosa, Silene dioica, and S. vulgaris occur. Species such as Agrostis capillaris, Acetosella vulgaris, Arabidopsis arenosa, and Thlaspi caerulescens appear frequently on Pb-Zn mine wastes. Several differences in the vegetation structure were detected between the Pb-Zn and Cu mine heaps; higher amounts of vascular plants and fewer lichen species covered the Pb-Zn mine heaps. For the Cu mine heaps, on the contrary a small number of vascular species but a high number and coverage of lichen species, especially Ceratodon purpureus and Cladonia arbuscula subsp. mitis were typical. The non-metalliferous meadows in the vicinity of the mines showed uniform structure but a higher species diversity.

The ultrastructural response of root cells to stressful conditions

Abstract Changes in the ultrastructure of root cells of Zea mays under non-lethal drought, salt, low and high temperature stresses, and of root cells of Valerianella locusta at freezing temperatures were investigated. The structural responses of cells of the same type and age, i.e. epidermis and cortex at 1.5 mm from the apex, were examined with the aim of comparing the specificity of stress effect in terms of cellular responses. Nuclei had condensed (in drought) or decondensed chromatin and highly sensitive nucleoli (in the cold, heat). ER became arranged into parallel complexes with drought and in the cold. The Golgi apparatus disintegrated only under water deficit. Mitochondrial internal structure was the most variable. However, the number of mitochondrial cristae was reduced by all types of stress indicating that reduction in energy production might contribute to the decreased growth which generally accompanies stress conditions. The reactions of ER and polyribosomes may be related to the induction of stress protein synthesis in root cells which occurs with all kinds of stresses. There is no one structural response which is unique to root cells, nor is the response of root cells uniform. The reason for such complex relationships may be that the primary effect of a stress is on cell membranes and possibly also on cell cytoskeleton or nucleus which then induced secondary or tertiary effects with similar or different structural manifestations.

Early Zn2+-induced effects on membrane potential account for primary heavy metal susceptibility in tolerant and sensitive Arabidopsis species

BACKGROUND AND AIMS Uptake of heavy metals by plant root cells depends on electro-physiological parameters of the plasma membrane. In this study, responses of the plasma membrane in root cells were analysed where early reactions to the metal ion-induced stress are localized. Three different Arabidopsis species with diverse strategies of their adaptation to heavy metals were compared: sensitive Arabidopsis thaliana and tolerant A. halleri and A. arenosa. METHODS Plants of A. thaliana Col-0 ecotype and plants of A. arenosa and A. halleri originating from natural metallicolous populations were exposed to high concentrations of Zn(2+). Plants were tested for root growth rate, cellular tolerance, plant morphology and cell death in the root apex. In addition, the membrane potential (E(M)) of mature cortical root cells and changes in the pH of the liquid culture media were measured. KEY RESULTS Primary roots of A. halleri and A. arenosa plants grew significantly better at increased Zn(2+) concentrations than A. thaliana plants. Elevated Zn(2+) concentrations in the culture medium induced rapid changes in E(M). The reaction was species-specific and concentration-dependent. Arabidopsis halleri revealed the highest insensitivity of the plasma membrane and the highest survival rate under prolonged treatment with extra-high concentrations. Plants were able to effectively adjust the pH in the control, but much less at Zn(2+)-induced lower pH. CONCLUSIONS The results indicate a similar mode of early reaction to Zn(2+), but with different extent in tolerant and sensitive species of Arabidopsis. The sensitivity of A. thaliana and a high tolerance of A. halleri and A. arenosa were demonstrated. Plasma membrane depolarization was lowest in the hyperaccumulator A. halleri and highest in A. thaliana. This indicates that rapid membrane voltage changes are an excellent tool to monitor the effects of heavy metals.

Structural aspects of bulge formation during root hair initiation

Using light and electron microscopy, the early stages of root hair initiation were investigated under control conditions and in a situation where F-actin polymerization was effectively inhibited by latrunculin B. Trichoblasts in their early stage of bulge formation possessed large vacuole traversed by cytoplasmic strands and enclosed within a narrow peripheral layer of cytoplasm. The nucleus was settled at the inner periclinal cell wall, typically opposite the site of bulge formation. Within the bulging area, dense cytoplasm and numerous ER elements, and other organelles were accumulated, together with pleiomorphic membrane-bound structures, the identity and nature of which will require further studies. These unusual structures, which were associated with the outer cell wall, contained material similar to that of the cell wall. Similar cell wall-like bodies were observed also in the cytoplasm and sometimes within vacuoles. The possible role of these novel organelles of plant cells in cell wall thinning/degradation or in the turgor pressure maintenance are discussed. Latrunculin B treatment allowed bulge formation but prevented the switch from the slow and diffuse expansion of bulge into the rapid tip-growth characteristic of the emerging root hair. Moreover, the cytoplasm of the bulging domain became extensively vacuolated and lacked abundant ER elements and other organelles including the membrane-bound structures. These results indicate important roles of F-actin in the switch from diffuse to highly polarized tip growth.

Ultrastructure of meristematic epidermal cells of maize root under water deficit conditions

SummaryStructural components of meristematicZea mays primary root epidermal cells were observed after osmotic stress of the nutrient medium (12.5 atm.) and after rehydration. After non-lethal osmotic stress (24 hours), aggregation of nuclear chromatin, parallel ER arrangement, and reduction of mitochondrial cristae were found. Increased number of Golgi cisternae indicated vesicle production inhibition, and microtubules were absent in treated cells, although plastid structure remained unchanged. After lethal osmotic stress (48 hours), fragmentation of cytoplasmic membranes and a more severe structure damage of all cellular components occurred. Structure of the nucleus, mitochondria, Golgi apparatus and microtubules reappeared in the cells after rehydration. The only new feature of these cells was occurrence of smooth ER, which may indicate that the ER system has acquired a different function in regenerated cells.

The effect of polyethylene glycol-induced water stress on the maize root apex

Following a 24-h exposure to a solution of polyethylene glycol 4 000 of a —12.66 bar osmotic potential the roots of maize ceased growing. The inhibition of growth was conditioned by the inhibition of cell elongation and division. The elongation of cells was substituted by their radial enlargement which took place both in the peripheral and central root parts. The cells either did not divide at all, or sporadic mitoses still occurred in the roots. The meristematic cells treated were highly vacuolized, chromatin condensation being observed in their nuclei. In contrast to growth processes, differentiation was stimulated: the formation of the secondary wall in protoxylem elements occurred at a shorter distance,i.e. 1 500–2 400 µm from the apex, in comparison with 4000–5 000 µm in the control, this evidently being caused not only by the inhibition of growth, but also by the capacity of cells to differentiate more rapidly. The changes induced by a 24-h exposure to water stress were of a reversible nature; however, a 48-h treatment brought about irreversible changes.

Accumulation of pathogenesis-related proteins in barley leaf intercellular spaces during leaf senescence

Accumulation of the pathogenesis-related (PR) proteins localised in intercellular spaces of barley primary leaves, chlorophyll content, structure of chloroplasts, and photosynthesis were examined during natural and in vitro induced leaf senescence (cultivation of whole plants in the dark or detached leaves under nutrient deficiency). Some of PR proteins accumulated during natural senescence, but their accumulation pattern was different from those of pathogen-induced as well as during in vitro-induced senescence, which indicate different molecular bases of these processes. Photosynthetic rate and chlorophyll content indicate that natural senescence of barley primary leaves began from 15th day after sowing. In 35-d-old first leaves, the chloroplasts showed typical characteristics of senescence as significant decrease of size, greater grana, and prominent plastoglobuli. The chloroplasts contained more grana under in vitro induced senescence and they had reduced length in the dark. Correspondingly, accumulation of PR proteins was detectable on about the 15th day but the content of some PR proteins increased in later stages of senescence.

Metal uptake, antioxidant status and membrane potential in maize roots exposed to cadmium and nickel

Root growth of the seedlings of maize cultivars Premia and Blitz exposed to 2 μM cadmium (Cd), nickel (Ni) or both metals acting simultaneously (Cd + Ni) for 72 h was significantly reduced but not ceased. The effect was more pronounced in the seedlings of the cv. Blitz. The heavy metals (HMs) contents increased significantly in the roots. Simultaneous application of metals had an antagonistic effect on either Cd or Ni uptake in Premia but not in Blitz. In control roots the contents of ascorbic acid (AsA) and dehydroascorbic acid (DHA) were lower and gluthatione (GSH) content was higher in Premia than in Blitz. A decrease of AsA content was induced by all metal treatments in Premia but only by Cd + Ni in Blitz while an increase was induced by single metals in this cultivar. All metal treatments increased DHA contents in both cultivars. GSH content decreased significantly in Premia treated with Cd or Cd + Ni, and in Blitz treated with Ni. Unlike the contents of AsA, DHA and GSH, the increased metal concentrations in root cells did not affect the membrane potential (EM). The changes in antioxidant contents depended on both, maize genotypes and HMs treatments. Nevertheless, the results indicated a role of antioxidative system in minimizing the effects of oxidative stress and protecting cell membranes in both maize cultivars.