Fatma Jemal

Interactions of zinc and cadmium toxicity in their effects on growth and in antioxidative systems in tomato plants (Solarium lycopersicum)

The interaction between zinc and cadmium was investigated in tomato plants (Solanum lycopersicum). Ten-day-old seedlings were treated with 10 micromol/L CdCl2 associated to different concentrations of ZnCl2 (10, 50, 100, and 150 micromol/L). Zn supply clearly reduced Cd accumulation in leaves and simultaneously increased Zn concentration. Cd induced oxidative stress in leaves as indicated by an increase in thiobarbituric acid-reactive substances (TBARS) level and chlorophyll breakdown. Furthermore, compared with control, Cd-treated plants had significantly higher activities of superoxide dismutase (SOD, EC 1.15.1.1), whereas, catalase (CAT, EC 1.111.1.6), ascorbate peroxidase (APX, EC 1.11.1.11), and glutathione reductase (GR, EC 1.6.4.2) activities were significantly suppressed by Cd addition. Zn supplementation, at low level, restored and enhanced the functional activity of these enzymes (SOD, CAT, APX and GR) as compared to Cd-alone-treated plants. The beneficial effect of adequate Zn level on Cd toxicity was confirmed by a significant decrease in TBARS level and restoration of chlorophyll content. However, when Zn was added at high level in combination with Cd there was an accumulation of oxidative stress, which was higher than that for Cd or excess Zn alone treatments. These results suggested that higher Zn concentrations and Cd are synergistic in their effect on plant growth parameters and oxidative stress.

Analysis of in vivo chlorophyll fluorescence spectra to monitor physiological state of tomato plants growing under zinc stress

The effects of zinc (Zn) on plant chlorophyll fluorescence were investigated in 10-day-old tomato (Solanum lycopersicum) seedlings subjected for 7 days to a series of zinc (10, 50, 100 and 150μM) applied via the nutrient solution. The chlorophyll fluorescence spectra of leaves were recorded in the spectral region 650-800nm using the spectroscopic technique of ultraviolet light emitting diode induced fluorescence spectroscopy (UV-LED IFS). These spectra have been used to analyze the effect of several doses of zinc on the photosynthetic activities of tomato plants. The fluorescence intensity ratios (FIR) at the two maxima (F(690)/F(735)) of control as well as treated tomato plants were calculated by evaluating curve-fitted parameters using a Gaussian spectral function. The variable chlorophyll fluorescence decrease ratio (R(Fd)) values were determined from the fluorescence induction kinetics curves recorded at 690nm and 735nm. In addition, Zn accumulation in plants, plant growth, photosynthetic pigments content and malondialdehyde level (MDA, an index of lipid peroxidation) were also estimated. The results indicated that the plants treated with 10μM of zinc exhibited better growth, however, higher concentrations of zinc were harmful for plants. Excess Zn induced a decrease in the R(Fd) values, which was associated with a strong decline of the total chlorophylls content and an increase of MDA level. The total chlorophylls content decline could also be followed via an increase of the chlorophyll fluorescence ratio F(690)/F(735).

Glutathione and phytochelatin contents in tomato plants exposed to cadmium

The effect of cadmium on growth and contents of glutathione (GSH) and phytochelatins (PCs) were investigated in roots and leaves of tomato plants (Lycopersicon esculentum Mill. cv. 63/5 F1). The accumulation of Cd increased with external Cd concentrations and was considerably higher in roots than in leaves. Dry mass production decreased under Cd treatment especially in leaves. In both roots and leaves, exposure to Cd caused an appreciable decline in GSH contents and increase in PCs synthesis proportional to Cd concentrations in the growth medium. At the same Cd concentration, PCs production was higher in roots than in leaves. The implication of glutathione in PC synthesis was strongly suggested by the use of buthionine sulfoximine (BSO). The major fraction of Cd accumulated by tomato roots was in the form of a Cd-PCs complex.

Spectroscopic studies of photosynthetic responses of tomato plants to the interaction of zinc and cadmium toxicity.

The in vivo chlorophyll (Chl) fluorescence spectra of Solanum lycopersicum leaves were recorded in the spectral region 650-800nm using a spectroscopic method based on ultraviolet light emitting diode induced fluorescence spectroscopy (UV-LED IFS). These spectra have been used to analyze the interactive functions of cadmium (Cd(2+)) and zinc (Zn(2+)) on photosynthetic activities of S. lycopersicum plants. The fluorescence intensity ratios (F(690)/F(735)) of the chlorophyll bands at 685 and 730nm were calculated by evaluating curve fitted parameters using a Gaussian spectral function, for control as well as treated plants. The fluorescence induction kinetics (Kautsky effect) was also measured on dark adapted intact plant leaves at the chlorophyll bands for determining the variable chlorophyll fluorescence decrease ratio (R(Fd) values) and the stress adaptation index (Ap). In addition, metal accumulation in plants, plant growth and photosynthetic pigments content were estimated. It was found that the R(Fd)(690), R(Fd)(730) and Ap values decreased whereas the F(690)/F(735) ratio increased in the case of 10μM Cd(2+) treated plants, indicating an impairment of the photosynthetic efficiency. Zn(2+) supplementation, at low concentration (10 and 50μM), in combination with Cd(2+) protect the photochemical functions. However, the high Zn(2+) concentration exacerbated the negative effects of Cd(2+) and showed a severe decrease of R(Fd)(690), R(Fd)(730) and Ap values compared to Cd(2+) alone. It is seen that F(690)/F(735) ratios are strongly correlated with chlorophyll contents. The results demonstrate the usefulness of F(690)/F(735), Ap and R(Fd) values in determining the potential photosynthetic activity of an intact attached leaf in a non-destructive way.

Cadmium and copper genotoxicity in plants

Heavy metal contamination in soils is easily transmitted to human through plants via the food chain. A major concern is to understand the plant response to heavy metal soil contamination to develop phytoremediation. Two plant models have been investigated in our study, the tomato, which is of agronomical importance, Arabidopsis, which is used as a model for molecular genetics. Heavy metal toxicity is described to induce oxidative stress linked to oxidation of proteins and membrane lipids but also to alterations of DNA damage response. We have investigated the metabolic response of cadmium and copper in parallel in both plant models and analyzed the transcriptional response of Arabidopsis RNR genes coding for isoforms of ribonucleotide reductase, an essential enzyme involved in DNA synthesis. Both Cd and Cu had a dose-dependent effect on plant growth. We also observed a rapid increase of catalase activity upon Cd or Cu treatments in tomato and Arabidopsis. At the transcriptional level, treatment with Cd resulted in a biphasic induction of two RNR genes in Arabidopsis; the first induction peak could be paralleled to the increase of the catalase activity.

The effect of cadmium on lipid and fatty acid biosynthesis in tomato leaves

This research aims to examine the effect of cadmium uptake on lipid composition and fatty acid biosynthesis, in young leaves of tomato treated seedlings (Lycopersicon esculentum cv. Ibiza F1). Results in membrane lipids investigations revealed that high cadmium concentrations affect the main lipid classes, leading to strong changes in their composition and fatty acid content. Thus, the exposure of tomato plants to cadmium caused a concentration-related decrease in the unsaturated fatty acid content, resulting in a lower degree of fatty acid unsaturation. The level of lipid peroxides was significantly enhanced at high Cd concentrations. Studies of the lipid metabolism using radioactive labelling with [1-14C]acetate as a major precursor of lipid biosynthesis, showed that levels of radioactivity incorporation in total lipids as well as in all lipid classes were lowered by Cd doses. In total lipid fatty acids, [1-14C]acetate incorporation was reduced in tri-unsaturated fatty acids (C16:3 and C18:3); While it was enhanced in the palmitic (C16:0), palmitoleic (C16:1), stearic (C18:0) and linoleic (C18:2) acids. [1-14C]acetate incorporation into C16:3 and C18:3 of galactolipids [monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG)] and some phospholipids [phosphatidylcholine (PC) and phosphatidylglycerol (PG)] was inhibited by Cd stress. Our results showed that in tomato plants, cadmium stress provoked an inhibition of polar lipid biosynthesis and reduced fatty acid desaturation process.

Detection of photosynthetic activity under cadmium stress by measurement of the red and far-red chlorophyll fluorescence

In this work it is shown that, with a new spectroscopic method based on ultraviolet light emitting diode-induced fluorescence spectroscopy (UV-LED IFS), the photosynthetic activity of an intact attached leaf can be measured in a non-destructive way within a few seconds. The effects of cadmium (Cd) toxicity on the photosynthetic activity of tomato plants (Solanum lycopersicum) were studied using various CdCl2 (cadmium chloride) concentrations. The Cd treatment decreases the photosynthetic activity, measured via a decline of the variable chlorophyll (chl) fluorescence decrease ratio (R Fd) values, calculated at 690 and 730 nm. The chl fluorescence ratios of intensity (FIR) and band area were also calculated. It was found that the R Fd(690) and R Fd(730) values decreased whereas the FIR increased from 1 μM Cd treatment and this behaviour is enhanced with the increasing of Cd concentrations. At 10 and 25 μM Cd, the decrease in R Fd values was associated with a strong decline in the chlorophyll-a + chlorophyll-b (chl-a + b). The chl-a + b decline could also be followed via an increase of the chl fluorescence ratio F 690/F 730. Our study demonstrates that the UV-LED IFS is a promising technique for early detection of Cd stress (from 1 μM) in tomato plants, before visual symptoms appear.