Phytoextraction of metals from a multiply contaminated soil by Indian mustard.

The effects of nitrilotriacetate (NTA) and citric acid applications on metal extractability from a multiply metal-contaminated soil, as well as on their uptake and accumulation by Indian mustard (Brassica juncea) were investigated. Desorption of metals from the soil increased with chelate concentration, NTA being more effective than citric acid in solubilising the metals. Plants were grown in a sandy soil collected from a contaminated field site and polluted by Cd, Cr, Cu, Pb and Zn. After 43 days of plant growth, pots were amended with NTA or citric acid at 5 mmol kg-1 soil. Control pots were not treated with any chelate. Harvest of plants was performed 1 week after chelate addition. Soil water-, NH4NO3- and DTPA-extractable Cd, Cu, Pb and Zn fractions were enhanced only in the presence of NTA. In comparison to unamended plants, Indian mustard shoot dry weights suffered significant reductions following NTA application. NTA treatment increased shoot metal concentrations by a factor of 2-3, whereas citric acid did not induce any difference compared to the control. Chromium was detected in the above-ground tissues only after NTA amendment. Due to differences in dry matter yield, a significant enhancement of metal uptake was observed in NTA-treated plants for Cu and Zn.

Nitrilotriacetate- and citric acid-assisted phytoextraction of cadmium by Indian mustard (Brassica juncea (L.) Czernj, Brassicaceae).

In a pot experiment the effects of nitrilotriacetate (NTA) and citric acid applications on Cd extractibility from soil as well as on its uptake and accumulation by Indian mustard (Brassica juncea) were investigated. Plants were grown in a sandy soil with added CdS at four levels ranging from 50 to 200 mg Cd kg(-1) soil. After 30 days of growth, pots were amended with NTA or citric acid at 10 and 20 mmol kg(-1). Control pots were not treated with chelates. Harvest of plants was performed immediately before and one week after chelate addition. Soil water-, NH(4)NO(3)- and EDTA-extractable Cd fractions increased constantly with both increasing soil metal application and chelate concentration. Shoot dry weights did not suffer significant reductions with increasing Cd addition to the soil except for both NTA treatments in which at 200 mg Cd kg(-1) a 30% decrease in dry matter was observed. Generally, following NTA and citric acid amendments, Cd concentration in shoots increased with soil Cd level. However, due to Cd toxicity, at the highest metal application rate both NTA treatments lowered Cd concentration in the above-ground parts. Compared to the control, at 10 mmol kg(-1) citric acid did not change Cd concentration in shoots, whereas NTA-treated plants showed an about 2-fold increase. The addition of chelates at 20 mmol kg(-1) further enhanced Cd concentration in shoots up to 718 and 560 microg g(-1) dry weight in the NTA and citrate treatments, respectively.

Bleaching herbicide effects on plastids of dark-grown plants: lipid composition of etioplasts in amitrole and norflurazon-treated barley leaves.

The effects of the bleaching herbicides amitrole (125 micro M) and norflurazon (100 micro M) on etioplast lipids were studied in barley plants (Hordeum vulgare L. cv. Express) grown for 7 d either at 20 degrees C or 30 degrees C in darkness. Total lipid, glycolipid and phospholipid contents of control etioplasts were increased at 30 degrees C in comparison with those at 20 degrees C. The two herbicides caused a decrease in the total lipid, glycolipid and phospholipid amounts compared to the untreated etioplasts and lowered the lipid to protein ratio. In the controls, monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG) accounted for about 66 mol% of the etioplast polar lipids, while the remainder was represented by sulphoquinovosyldiacylglycerol (SQDG) and phosphatidylglycerol (PG), in approximately equal proportions. Both amitrole and norflurazon increased MGDG at both temperatures, but decreased DGDG except with norflurazon at 30 degrees C. As a consequence, the MGDG to DGDG molar ratio was higher in the herbicide-treated etioplasts compared to the controls at both the growth temperatures. The amount of the negatively charged polar lipids SQDG and PG were decreased by treatments with amitrole at 20 degrees C and norflurazon at 30 degrees C. The two herbicides determined different responses in the fatty acid unsaturation of the individual polar lipids. Changes in the lipid composition of etioplasts and the interaction between the pigment-protein complex, protochlorophyllide-NADPH-protochlorophyllide oxidoreductase, and polar lipids are discussed.

Lipids and NADPH-dependent superoxide production in plasma membrane vesicles from roots of wheat grown under copper deficiency or excess.

The effects of in vivo copper on the lipid composition of root plasma membrane and the activities of membrane-bound enzymes, such as NADPH-dependent oxidases and lipoxygenase, were studied. Plants were grown in hydroponic culture for 11 d without Cu supply or in the presence of 50 microM Cu. Control plants were supplied with 0.3 microM Cu. Growth of roots was severely affected in the 50 microM Cu-grown plants, whereas roots grown in Cu-deficient solution did not show any difference in comparison with the control. The 50 microM Cu concentration caused an increase in the leakage of K(+) ions as well. Excess metal supply resulted in a decrease in the total lipid content of plasma membrane, a higher phospholipid amount and a reduction of steryl lipids (free sterols, steryl glycosides and acylated steryl glycosides). Cu depletion in the growth solution had only a slight effect on the plasma membrane lipid composition. In comparison with the control, only the excess of Cu caused a decrease in the lipid to protein ratio as well as a change in the phospholipid composition, with a lower phosphatidylcholine to phosphatidylethanolamine ratio. The degree of unsaturation of root plasma membranes decreased following the 0 Cu treatment and even more after the 50 microM Cu supply. Plasma membranes of wheat grown under metal deficiency and excess showed increased NADPH-dependent superoxide-producing oxidase activities, whereas membrane-bound lipoxygenase was not increased or activated due to Cu treatments. The consequences of changes in plasma membrane lipid composition and activated oxygen production as a result of Cu treatments are discussed.

Protein dynamics in thylakoids of the desiccation-tolerant plant Boea hygroscopica during dehydration and rehydration.

Plants of Boea hygroscopica F. Muell were dehydrated to 9% relative water content (RWC) by withholding water for 26 d, and afterward the plants were rehydrated. Leaves were taken from control plants after 7, 12, and 26 d from the beginning of dehydration, and after 6 and 48 h from rehydration. The RWC decreased by 80% during dehydration, but the leaves regained RWC with rehydration. Dehydrated plants showed lesser amounts of proteins, lipids, and chlorophyll, all of which increased following rewatering. The lipid-to-protein ratio, which decreased during dehydration, returned to control level after 48 h of rehydration. Thylakoid lipids were more unsaturated when RWC reached the value of 9%. EPR measurements of spin-labeled proteins showed the presence of three different groups of proteins with different mobility in thylakoid membranes. The rotational correlation time of groups 1 and 2 increased with dehydration and decreased upon rehydration, whereas group 3 showed little changes. Desiccation did not cause thylakoid swelling or breakage, but the membrane system assemblage showed changes in thylakoid stacking. After 48 h of rehydration the membrane system recovered completely the organization of the fully hydrated state, showing several well-defined and regularly distributed grana.

Changes in plasma-membrane lipid composition: a strategy for acclimation to copper stress.

Wheat seedlings were grown hydroponically in the presence of 50 microM Cu2+. The copper stress resulted in plasma-membrane (PM) changes of the root cells as altered lipid composition, a decreased phosphatidylcholine (PC)/phosphatidylethanolamine (PE) ratio from 0.7 to 0.3, a decreased fatty acyl unsaturation and a decrease in the lipid/protein ratio. Membrane vesicles made from total lipid extracts of isolated PMs of wheat grown under copper excess showed a remarkably low permeability to polar molecules like glucose, as compared with the control, and no difference in proton permeability. Permeability studies of vesicles of plasma-membrane lipids, which were selectively modified by addition of specific lipids such as PC and PE, were also performed. The results are discussed with emphasis on the role of the increased PE proportion.

Superoxide and hydroxyl radical generation, and superoxide dismutase in PSII membrane fragments from wheat.

Illumination of photosystem II (PSII) membrane fragments of wheat (Triticum durum Desf. cv. Adamello) gave rise to both O2*- and *OH radicals adducts of the novel spin trap 5-(diethoxyphosphoryl)-5-methyl-1-pyrroline-N-oxide (DEPMPO). With time, *OH became predominant displaying the conversion of O2*- into *OH. An intrinsic activity of superoxide dismutase (SOD) was found in PSII. Photoreduction of nitroblue tetrazolium (NBT) by PSII membrane fragments was induced by the addition of sodium azide and hydrogen peroxide. Western blotting of PSII proteins showed that a 29 kDa protein was recognised by an antibody against chloroplastic Fe-SOD from water lily. An increased formation rate of O2*- was observed in damaged PSII where the SOD activity decreased following a treatment with a free radical-generating system. Damage in PSII consisted also in a decrease in chlorophyll and in carotenoids as well as in a change in the lipid : chlorophyll : protein ratio. Upon treatment a decrease in the unsaturation of PSII membrane fragments was also monitored together with a degradation towards more saturated molecular species of monogalactosyldiacylglycerol.

Lipid Composition and Protein Dynamics in Thylakoids of Two Wheat Cultivars Differently Sensitive to Drought.

Two wheat (Triticum durum Desf.) cultivars with different sensitivities to drought were either grown under regular irrigation or subjected to water deficit by withholding water for 14 d. Water-stressed plants of both cultivars underwent similar decreases in leaf water potential, but the drought-tolerant cultivar showed higher relative water content and turgor. Neither osmotic nor elastic adjustment mechanisms appeared to be active under the conditions described here. Thylakoids isolated from the stressed, drought-tolerant wheat showed an increase in lipid-to-protein ratio, in comparison with the control, whereas this ratio remained unchanged in the sensitive wheat. In both cultivars, water deficit determined different rearrangements in the composition of the thylakoid individual polar lipids, but their unsaturation level remained unaffected with the exception of monogalactosyldiacylglycerol. In the drought-sensitive cultivar, an accumulation of free fatty acids together with a reduction in polar lipid amount was observed. Electron paramagnetic resonance measurements of spin-labeled proteins of stressed plants from the sensitive cv Adamello showed a higher spin label rotational correlation time together with lower sulphydryl group and mobile proteic portion levels, in comparison with the control. In the tolerant cv Ofanto, the first two parameters changed to a lesser extent following water depletion, and the mobile proteic portion was not altered.