Protection of palak (Beta vulgaris L. var Allgreen) plants from ozone injury by ethylenediurea (EDU): Roles of biochemical and physiological variations in alleviating the adverse impacts

Ameliorative effects of ethylenediurea (N-[2-(2-oxo-1-imidazolinidyl) ethyl]-N′ phenylurea, abbreviated as EDU) against ozone stress were studied on selected growth, biochemical, physiological and yield characteristics of palak (Beta vulgaris L. var Allgreen) plants grown in field at a suburban site of Varanasi, India. Mean eight hourly ozone concentration varied from 52 to 73 ppb which was found to produce adverse impacts on plant functioning and growth characteristics. The palak plants were treated with 300 ppm EDU at 10 days after germination at 10 days interval up to the plant maturity. Lipid peroxidation in EDU treated plants declined significantly as compared to non-EDU treated ones. Significant increment in F_v/F_m ratio in EDU treated plants as compared to non-EDU treated ones was recorded. EDU treated plants showed significant increment in ascorbic acid contents and reduction in peroxidase activity as compared to non-EDU treated ones. As a result of the protection provided by EDU against ozone induced stress on biochemical and physiological characteristics of palak, the morphological parameters also responded positively. Significant increments were recorded in shoot length, number of leaves plant⁻¹, leaf area and root and shoot biomass of EDU treated plants as compared to non-EDU treated ones. Contents of Na, K, Ca, Mg and Fe were higher in EDU treated plants as compared to non-EDU treated ones. The present investigation proves the usefulness of EDU in partially ameliorating ozone injury in ambient conditions.     

Thermoluminescence as a tool for monitoring ozone-stressed plants

The effect of ozone (6 h, various concentrations from 0 to 350 ppb) on barley (Hordeum vulgare L., cv. Bomi) and tomato (Lycopersicon esculentum L., cv. Yellow Cherry) leaves was investigated in parallel by thermoluminescence (TL) and fluorescence (FL) methods. Several significant changes were found in TL glow curves measured after excitation by one single turnover flash at +2 degree C in the temperature range from 2 to 170 degree C immediately after ozone exposure. Contrary to TL, ozone induced only negligible changes in FL parameters F0, FM and Fv/FM. Measurements done 24 h after ozone exposure showed partial recovery of ozone-induced changes. The extent of recovery was not the same in different parts of TL curves. Fluorescence parameters were not significantly changed. The results demonstrate that TL parameters are more sensitive to ozone than conventially used FL parameters F0, FM and Fv/FM. Moreover, TL measurements seem to give information not only about the PSII electron transport, but also about the extent of oxidative damage and membrane lipid peroxidation. It is concluded, that TL can be a highly informative tool for monitoring the impact of ozone on plants.     

Lead tolerance and physiological adaptation mechanism in roots of accumulating and non-accumulating ecotypes of Sedum alfredii

Background, aim and scope

Lead (Pb) accumulation in soils affects plants primarily through their root systems. The aim of this study was to investigate early symptoms of the loss of membrane integrity and lipid peroxidation in root tissues and physiological adaptation mechanism to Pb in accumulating ecotypes (AE) and non-accumulating ecotypes (NAE) of Sedum alfredii under Pb stress in hydroponics.

Methods and results

Histochemical in situ analyses, fluorescence imaging, and normal physiological analysis were used in this study. Pb accumulation in roots of both AE and NAE increased linearly with increasing Pb levels (0?C200???M), and a significant difference between both ecotypes was noted. Both loss of plasma membrane integrity and lipid peroxidation in root tissues became serious with increasing Pb levels, maximum tolerable Pb level was 25 and 100???M for NAE and AE, respectively. Pb supplied at a toxic level caused a burst of reactive oxygen species (ROS) in root cells in both ecotypes. However, the root cells of AE had inherently higher activities of superoxide dismutase (SOD), guaiacol peroxidase (POD), and lipoxygenase (LOX) in control plants, and the induction response of these antioxidant enzymes occurred at lower Pb level in AE than NAE. AE plants maintained higher ascorbic acid and H₂O₂ concentrations in root cells than NAE when exposed to different Pb levels, and Pb induced more increase in dehydroascorbate (DHA), catalase (CAT), and ascorbate peroxidase (APX) in AE than NAE roots.

Discussion and conclusion

Results indicate that histochemical in situ analyses of root cell death and lipid peroxidation under Pb short-term stress was sensitive, reliable, and fast. Higher tolerance in roots of accumulating ecotype under Pb stress did depend on effective free oxygen scavenging by making complex function of both constitutively higher activities and sensitive induction of key antioxidant enzymes in root cells of S. alfredii.     

Uptake and translocation of metals in Spinacia oleracea L. grown on tannery sludge-amended and contaminated soils: effect on lipid peroxidation, morpho-anatomical changes and antioxidants

The plants of Spinacia oleracea L. grown on contaminated soil (CS) and different amendments of tannery sludge (TS) have shown high accumulation of metals in its edible part. The accumulation of toxic metal (Cr) in the leaves of the plants grown on CS was recorded as 40.67 microgg(-1)dw. However, the leaves of the plants grown on 100% TS have accumulated about two times (70.80 microgg(-1)dw) higher Cr than the 10% TS (31.21 microgg(-1)dw). Among growth parameters, the root length was more affected at 90 d than the shoot length, number of leaves and leaf area. The study of scanning electron micrographs showed 29.31% increase in stomatal length in the leaves of the plants grown on CS as compared to garden soil (GS), which served as control, however it decreased in the plants grown on higher amendments of TS. The decrease in MDA content at initial period of exposure and lower amendment was recorded in the leaves, whereas, significant increase (>10% TS onward) was observed with increase in tannery sludge ratio at 90 d as compared to GS. A coordinated increase in all the studied antioxidants (cysteine, non-protein thiol, ascorbic acid, carotenoid contents) was found up to 75 d of growth. At 90 d, most of the antioxidant decreased as compared to 75 d causing oxidative stress as evidenced by increased level of lipid peroxidation and decreased chlorophyll and protein contents. Maximum increase of 181.43% in MDA content and maximum decrease of 53.69% in total chlorophyll content was recorded in the leaves of the plants grown on 100% TS after 90 d of growth. The plants grown on CS have shown an increase in shoot length, number of leaves, leaf area, photosynthetic pigments and protein contents and in all the studied antioxidants. Thus, these plants are able to combat stress involving defense mechanism, resulting in healthy growth of the plants. The results are well coordinated as there is no change in the MDA content as compared to the plants grown on GS. In view of high Cr accumulation in edible part of S. oleracea grown on CS after irrigation with tap water, it is not advisable to use these plants for edible purposes. Summing up, it is recommended that the level of metals in the edible part should be checked instead of healthy growth as deciding parameter for consumption. It is demonstrated through this study that metal enriched plants have detoxification mechanism and grow well on organic matter enriched contaminated soil.     

Chlorophyll Content of Soybean Foliage in Relation to Seed Yield and Ambient Ozone Pollution

A field test was conducted to determine if ozone pollution adversely affected the chlorophyll content and seed yield of soybean. Eight soybean cultivars were grown to maturity in test plots in central New Jersey; one-half of the plots was treated with an antioxidant (ethylenediurea) to protect the plants from the effects of ambient ozone and one-half was left untreated. Periodic chlorophyll measurements revealed no significant difference between EDU-treated and untreated plots during the major part of plant growth. The absence of a yield effect predicated on the normal chlorophyll content was corroborated by actual total seed measurement. Our results did not support predictive models that forecast a significant yield reduction from a 7-h seasonal mean of 0.058 ppm 0₃, but agreed with results obtained previously in Maryland and Georgia.     

Effect of iron on lipid peroxidation, and enzymatic and non-enzymatic antioxidants and bacoside-A content in medicinal plant Bacopa monnieri L

The effect of Fe was investigated in medicinally important plant, Bacopa monnieri L. and the response on malondialdehyde (MDA) content, superoxide dismutase (SOD), peroxidase (POD) and ascorbate peroxidase (APX) was found different in roots and leaves of the metal treated plants. Iron induced stress was observed as indicated by high level of lipid peroxidation, being more steep increase in leaves than roots. In roots, SOD activity was found to increase in metal treated plants except 80 and 160 microM at 72 h, whereas, it decreased in leaves except 10 and 40 microM after 48 h as compared to their respective controls. Among H2O2 eliminating enzymes, POD activity increased in roots, however, it decreased in leaves except at 10 and 40 microM Fe after 48 h as compared to control. At 24 and 48 h, APX activity and ascorbic acid content followed the similar trend and were found to increase in both parts of the metal treated plants as compared to their respective controls. The level of cysteine content in the roots increased at initial period of exposure; however, no marked change in its content was noticed in leaves. In both roots and leaves, non-protein thiol content was found to increase except at higher metal concentrations at 72 h. The data of proline content have shown significant (p<0.01) increase at 40 microM onwards in both part of the plants after 48 and 72 h. Correlation coefficient was evaluated between metal accumulations with various parameters and also between different antioxidant parameters with MDA. Since the level of bacoside-A (active constituent) content in metal treated plants increases, therefore, it is advisable to assess the biological activity of the plants before using for medicinal purposes, particularly in developing countries.     

Effect of Nitrogen Nutrition on the Response of Tobacco to Ozone in the Atmosphere

Experiments were conducted to determine the effect of nitrogen nutrition on the response of tobacco to ozone in the atmosphere. With Nicotiana rustica L., a noncommercial tobacco, plants receiving an “optimum” nitrogen supply were found to be more susceptible to ozone injury than those receiving deficient or luxury supplies. Furthermore, it was found on analyses of the third oldest tobacco leaf that, while these “optimum” plants showed little or no increase in respiration as a result of ozone in concentrations too low to produce visible injury, plants grown with deficient or excess nitrogen showed a significant increase in respiration under these same conditions. The increase in respiration was accompanied by a comparable and equally significant decrease in carbohydrate con-lent in the latter plants which was not apparent in plants at the “optimum” nitrogen supply, as well as an increase in total nitrogen, protein-nitrogen, and amino-acid nitrogen. In conclusion, it is suggested that the carbohydrate reserve built up in plants grown with deficient nitrogen nutrition serves as protection against the ozidative potential of ozone. Likewise, a decrease in carbohydrate content below that of “optimum” plants provides protection in plants grown with an overabundance of nitrogen fertilization, though possibly by a more complex mechanism.     

Effects of ethylenediurea and ozone on the antioxidative systems in beans (Phaseolus vulgaris L.)

To study the biochemical mechanism of EDU protection against ozone injury, peroxidase, ascorbate-dependent peroxidase, and catalase activities, and the contents of ascorbic acid, dehydroascorbic acid, malondialdehyde and soluble protein were measured in Phaseolus vulgaris L. cv. Lit exposed to ozone and ethylenediurea (EDU) in open-top chambers. Plants not treated with EDU showed foliar bronzing due to ozone, while EDU-treated plants were not affected. EDU application modified the reaction of biochemical parameters to ozone. Soluble protein content was elevated by EDU. Peroxidase activity increased with ozone exposure in untreated plants only, while ascorbate-dependent peroxidase activity was lower in EDU treated plants. Catalase activity decreased in EDU-untreated plants. The ratio of ascorbic acid to dehydroascorbic acid was significantly increased in EDU treated plants. These results suggest that EDU might induce ascorbic acid synthesis and therefore provide the plant with a very potent antioxidant. Or the content of hydrogen peroxide was reduced due to other unknown processes and caused a delay in foliar senescence, regardless of whether these processes were ozone-induced or due to natural aging processes.     

Variation in ozone sensitivity among clones of Betula pendula and Betula pubescens

Forty clones of Betula pendula and 6 clones of Betula pubescens, originating from southern and central Finland, were ranked in order of ozone sensitivity according to visible injuries, growth and leaf senescense under low ozone exposure. The plants were fumigated in natural climatic conditions using an open-air exposure system during two growing seasons. Control plants were grown under ambient air, and the elevated-ozone exposures were 1.6x the ambient in 1994 and 1.7x the ambient in 1995. The differences in ozone sensitivity among clones were large. Ozone tolerance was related to thicker leaves and higher stomatal density as compared to sensitive clones. Ultrastructural ozone-induced symptoms were found in chloroplasts of sensitive clones. Increased number of visibly injured leaves on fumigated plants was correlated with reduced leaf formation, foliage area, shoot dry wt and number of stomata, and increased yellowing of leaves. The results suggest that a considerable proportion of birch trees, showing high sensitivity to ozone, are at risk if ambient ozone exposures increase.     

Stem injection of Populus nigra with EDU to study ozone effects under field conditions

EDU or ethylenediurea (N-[2-(2-oxo-1-imidazolidinyl)ethyl]-N'-phenylurea) has been used in experiments to assess ozone effects on vegetation under field conditions because it provides protection against oxidative damage. Tests have mainly been conducted on crop plants, but for woody species only few reports have provided evidence that it can be used in long-term experiments. In this study we tested the technique of stem injection of EDU to study the effects of ozone exposure on Populus nigra cv. Wolterson over one growing season. Cuttings of Populus nigra were grown in pots in the field and between mid-July and early September plants were repeatedly injected with EDU solution (5 mg/plant) or with water at 14-day intervals. Significant differences were found between EDU- and water-injected plants: water-treated plants had more foliar injury, more chlorotic leaves, and shedding of leaves started earlier, suggesting EDU was effective in preventing visible ozone injury and acceleration of senescence. Photosynthetic rates, measured for one leaf age, showed no differences but were mostly higher for the EDU-treated plants. At the end of the growing season diameter increment was 16% higher and there was a non-significant trend for above-ground biomass to be increased by 9% for the EDU-treated plants. This experiment has provided evidence that for this clone serious ozone damage occurs at relatively low concentrations and that EDU can provide protection against visible injury, as well as against longer term growth reductions.     

Copper and zinc induction of lipid peroxidation and effects on antioxidant enzyme activities in the microalga Pavlova viridis (Prymnesiophyceae)

The metal-induced lipid peroxidation and response of antioxidative enzymes have been investigated in the marine microalga Pavlova viridis to understand the mechanisms of metal resistance in algal cells. We have analyzed superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) activities and glutathione (GSH) contents in microalgal cells grown at different concentrations of copper and zinc. In response to each metal, lipid peroxidation was enhanced with the increase of concentrations, as an indication of the oxidative damage caused by metal concentration assayed in the microalgae cells. Exposure of P. viridis to the two metals caused changes in enzyme activities in a different manner, depending on the metal assayed: after copper treatments, total SOD activity was enhanced, while it was reduced after zinc exposure. Copper and zinc stimulated the activities of CAT and GSH whereas GPX showed a remarkable increase in activity in response to copper treatments and decrease after zinc treatments. These results suggest that an activation of some antioxidant enzymes was enhanced to counteract the oxidative stress induced by the two metals.     

Translocation of metals and its effects in the tomato plants grown on various amendments of tannery waste: evidence for involvement of antioxidants

A field experiment was conducted to study the effect of different amendments of tannery sludge on physiological and biochemical parameters of tomato plant (Lycopersicon esculentum L. Mill). The accumulation of metals (Cr, Fe) in different parts of tomato plants grown on tannery sludge amended soil increased in a concentration and duration-dependent manner. The accumulation of both the metals was found lowest in the fruits of the plant. The statistical analysis of the results showed an increase in chlorophyll and protein contents in lower sludge amendment ratio at all exposures followed by a decrease at highest (100%) sludge amendment ratio. Lipid peroxidation enhanced in both root and leaves of sludge grown plants of tomato at all the sludge amendments and exposure periods, which is evidenced by increased malondialdehyde content, however the maximum increase was found in the roots (43.63%) and leaves (56.66%) of the plant grown on 100% tannery sludge at 60 d, over respective controls. The level of antioxidants, cysteine, non-protein thiol and ascorbic acid increased in the sludge grown plants of tomato to cope up with stress induced by the excess amount of the heavy metals present in the tannery sludge. The maximum increase was found in cysteine content (75.53% in the leaves), non-protein thiol content (92.68% in the roots) and ascorbic acid content (29.66% in the roots) of the plant at 75% tannery sludge after 30 d. The tomato plants were found well adopted for minimizing damage induced by reactive oxygen species, when grown on tannery sludge amendments in the present study.     

Elevated atmospheric ozone increases concentration of insecticidal Bacillus thuringiensis (Bt) Cry1Ac protein in Bt Brassica napus and reduces feeding of a Bt target herbivore on the non-transgenic parent

Sustained cultivation of Bacillus thuringiensis (Bt) transgenic crops requires stable transgene expression under variable abiotic conditions. We studied the interactions of Bt toxin production and chronic ozone exposure in Bt cry1Ac-transgenic oilseed rape and found that the insect resistance trait is robust under ozone elevations. Bt Cry1Ac concentrations were higher in the leaves of Bt oilseed rape grown under elevated ozone compared to control treatment, measured either per leaf fresh weight or per total soluble protein of leaves. The mean relative growth rate of a Bt target herbivore, Plutella xylostella L. larvae was negative on Bt plants in all ozone treatments. On the non-transgenic plants, larval feeding damage was reduced under elevated ozone. Our results indicate the need for monitoring fluctuations in Bt toxin concentrations to reveal the potential of ozone exposure for altering dosing of Bt proteins to target and non-target herbivores in field environments experiencing increasing ozone pollution.     

Lead induced changes in antioxidant metabolism of horsegram (Macrotyloma uniflorum (Lam.) Verdc.) and bengalgram (Cicer arietinum L.)

One-month old horsegram (Macrotyloma uniflorum (Lam.) Verdc. cv VZM1) and bengalgram (Cicer arietinum L. cv Annogiri) were exposed to different regimes of lead stress as Pb(NO3)2 at 0, 200, 500 and 800 ppm concentrations. The extent of oxidative damage as the rate of lipid peroxidation, antioxidative response and the accumulation of lead in roots and shoots of both plants were evaluated after 12 days of lead stress. Lead (Pb) treated plants showed increased levels of lipid peroxidation as evidenced from the increased malondialdehyde content coupled with the increase in the activities of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), glutathione reductase (GR), glutathione S-transferase (GST) compared to control (untreated) plants. Lead stress caused significant changes in the activity of antioxidative enzymes. The effect of lead was found to be concentration dependent. Higher concentration of lead (800 ppm) resulted 2- to 3-fold increase in SOD, catalase and peroxidase activities, 3- to 5-fold increase in GR activity and 3- to 4-fold increase in GST activity in roots and leaves of both horsegram and bengalgram plants. Lead stress caused a significant increase in the rate of peroxidation as showed in the levels of malondialdehyde content in roots and leaves of both plant species. Horsegram registered lower Pb accumulation than bengalgram, however localization of Pb was greater in roots than leaves in both plants. In general, lipid peroxide levels and antioxidative enzyme activities were higher in horsegram than bengalgram and also more in roots than leaves which best concordance with the lead contents of both the plants and organs. These results suggest that Pb toxicity causes oxidative stress in plants and the antioxidative enzymes SOD, CAT, POD, GR, GST could play a pivotal role against oxidative injury.     

Nickel accumulation and its effect on growth,physiological and biochemical parameters in millets and oats

With the boom in industrialization, there is an increase in the level of heavy metals in the soil which drastically affect the growth and development of plants. Nickel is an essential micronutrient for plant growth and development, but elevated level of Ni causes stunted growth, chlorosis, nutrient imbalance, and alterations in the defense mechanism of plants in terms of accumulation of osmolytes or change in enzyme activities like guiacol peroxidase (POD), catalase (CAT), and superoxide dismutase (SOD). Ni-induced toxic response was studied in seedlings of finger millet, pearl millet, and oats in terms of seedling growth, lipid peroxidation, total chlorophyll, proline content, and enzymatic activities. On the basis of germination and growth parameters of the seedling, finger millet was found to be the most tolerant. Nickel accumulation was markedly lower in the shoots as compared to the roots, which was the highest in finger millet and the lowest in shoots of oats. Plants treated with a high concentration of Ni showed significant reduction in chlorophyll and increase in proline content. Considerable difference in level of malondialdehyde (MDA) content and activity of antioxidative enzymes indicates generation of redox imbalance in plants due to Ni-induced stress. Elevated activities of POD and SOD were observed with high concentrations of Ni while CAT activity was found to be reduced. It was observed that finger millet has higher capability to maintain homeostasis by keeping the balance between accumulation and ROS scavenging system than pearl millet and oats. The data provide insight into the physiological and biochemical changes in plants adapted to survive in Ni-rich environment. This study will help in selecting the more suitable crop species to be grown on Ni-rich soils.

     

Response to ambient ozone of two white clover (Trifolium repens L.cv. "Regal") clones, one resistant and one sensitive, grown in a Mediterranean environment

Two clones of white clover (Trifolium repens L.) differing in ozone tolerance were grown in southern Italy during 1997 and 1998 to study the effects of ambient ozone exposure on yield, leaf morphology and water use. Ambient ozone levels were high in both years with values exceeding the threshold for leaf injury reported in the literature. In both years ozone injury was observed on the sensitive clone (NC-S) but not on the resistant one (NC-R), and leaf and stolon dry matter production was significantly lower in NC-S than in NC-R. However, it cannot be excluded that other factors, such as high temperature, interacted with the effect of ozone on biomass production. The clones differed in morphological characteristics. Lower total leaf area in NC-S plants was due to a smaller number of leaves per plant, but the average area per leaf was higher in NC-S. Specific leaf weight and net assimilation rate were higher in the more productive clone (NC-R). Cumulative plant water use was higher in NC-R in each growth period because of the larger leaf area; by contrast, water use per unit leaf area was higher in NC-S, indicating higher leaf conductance to water vapour. The results suggest that ozone significantly reduces the yield of sensitive white clover plants under well-watered conditions, and that the difference in ozone tolerance between clover clones is related to differences in leaf morphology and water use.     

Tropospheric ozone decreases biomass production in radish plants (Raphanus sativus) grown in rural south-west Sweden

Potted plants of radish (Raphanus sativus L., cv. Cherry Belle) were grown in the ambient air for 5 weeks, with or without the application of a soil drench of the anti-ozonant ethylenediurea (EDU). The 24-h mean ozone concentration during the experimental period was 31 nl l(-1). Towards the end of the experiment two ozone episodes, with maximum concentrations around 70 and 115 nl l(-1), occurred. No visible injury that could be attributed to ozone was observed on any of the plants. Shoot and hypocotyl biomass were significantly lower in the non-EDU-treated plants than in the EDU-treated plants. The non-EDU-treated plants had a 32% lower hypocotyl biomass and a 22% lower shoot biomass. The shoot:hypocotyl ratio of the non-EDU-treated plants was higher than that of the EDU-treated plants, although the difference was not statistically significant. EDU treatment increased the leaf area and decreased the chlorophyll content of the leaves. These differences were, however, not statistically significant. It is suggested that the ambient rural ozone climate in southern Sweden has the potential to decrease biomass production in Cherry Belle radishes in the absence of visible injury.     

The relationship between EDU pre-treatment and C2H4 evolution in ozonated pea plants

A soil drench of [Formula: see text] (EDU) (150 ppm) applied to 'Progress No. 9' pea plants 24 h before an acute ozone exposure (0.25 ppm, 4 h) completely protected the foliage from visible symptoms normally induced by the pollutant. In the absence of ozone, EDU-treated plants were found to emit the same amount of C(2)H(4) as plants not treated with EDU. Based on this evidence, EDU-induced tolerance to ozone could not have been attributed to the prevention of an interaction between ethylene and ozone (sensu Mehlhorn and Wellburn). In the presence of ozone, EDU-treated plants did not emit the burst of C(2)H(4) that normally occurs (sensu Craker), extending the observation that EDU-treated plants do not exhibit the adverse physiological responses normally caused by ozone. The classic C(2)H(4) biosynthesis inhibitor aminoethoxyvinylglycine (AVG) did not prevent ozone phytotoxicity, although it significantly reduced ethylene emission from the ozonated tissue.     

Chromium induced lipid peroxidation in the plants of Pistia stratiotes L.: role of antioxidants and antioxidant enzymes

In the plant, Pistia stratiotes L., the effect of different concentrations of chromium (0, 10, 40, 80 and 160 microM) applied for 48, 96 and 144 h was assessed by measuring changes in the chlorophyll, protein, malondialdehyde (MDA), cysteine, non-protein thiol, ascorbic acid contents and superoxide dismutase (SOD), ascorbate peroxidase (APX) and guiacol peroxidase (GPX) activity of the plants. Both in roots and leaves, an increase in MDA content was observed with increase in metal concentration and exposure periods. In roots, the activity of antioxidant enzymes viz. SOD and APX increased at all the concentrations of Cr at 144 h than their controls. The GPX activity of the treated roots increased with increase in Cr concentration at 48 and 96 h of exposures, however, at 144 h its activity was found declined beyond 10 microM Cr. The level of antioxidants in the roots of the treated plant viz. cysteine and ascorbic acid was also found increased at all the concentrations of Cr at 144 h than their respective controls, however, an increase in the non-protein thiol content was recorded up to 40 microM Cr followed by decrease. The chlorophyll content decreased with increase in Cr concentrations and exposure periods. However, the protein content of both roots and leaves were found decreased with increase in Cr concentrations at all the exposure periods except an increase was recorded at 10 microM Cr at 48 h. In Cr treated plants, the no observed effect concentration (NOEC) and lowest observed effect concentration (LOEC) for leaves chlorophyll and protein contents were 40 and 80 microM Cr, respectively after 48 h exposure while NOEC and LOEC for root protein content were 10 and 40 microM, respectively after 48 h. The analysis of correlation coefficient data revealed that the metal accumulation in the roots of the plant was found positively correlated with antioxidant parameters except SOD after 48 h of exposure, however, negatively correlated with most of all the parameters studied at 144 h in both part of the plant. It may be suggested from the present study that toxic concentrations of Cr cause oxidative damage as evidenced by increased lipid peroxidation and decreased chlorophyll and protein contents. However, the higher levels of enzymatic and non-enzymatic antioxidants suggest the reason for tolerating higher levels of metals.     

Brassinosteroid alleviates phenanthrene and pyrene phytotoxicity by increasing detoxification activity and photosynthesis in tomato

The present study was carried out to investigate the effects of exogenously applied 24-epibrassinolide (BR) on growth, gas exchange, chlorophyll fluorescence characteristics, lipid peroxidation and antioxidant systems of tomato seedlings grown under different levels (0, 10, 30, 100 and 300 μM) of phenanthrene (PHE) and pyrene (PYR) in hydroponics. A concentration-dependent decrease in growth, photosynthetic pigment contents, net photosynthetic rate (Pn), stomatal conductance (Gs), maximal quantum yield of PSII (Fv/Fm), effective quantum yield of PSII (Φ_PSII), photochemical quenching coefficient (qP) has been observed following PHE and PYR exposure. By contrast, non-photochemical quenching coefficient (NPQ) was increased. PHE was found to induce higher stress than PYR. However, foliar or root application of BR (50 nM and 5 nM, respectively) alleviated all those depressions with a sharp improvement in the activity of photosynthetic machinery. The activities of guaicol peroxidase (GPOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR) as well as content of malondialdehyde (MDA) were increased in a dose-dependent manner under PHE or PYR treatments. Compared with control the highest increments of GPOD, CAT, APX, GR and MDA by PHE/PYR alone treatments were observed following 300 μM concentration, which were 67%, 87%, 53%, 95% and 74% by PHE and 42%, 53%, 30%, 86% and 62% by PYR, respectively. In addition, both reduced glutathione (GSH) and oxidized glutathione (GSSG) were induced by PHE or PYR. Interestingly, BR application in either form further increased enzymatic and non enzymatic antioxidants in tomato roots treated with PHE or PYR. Our results suggest that BR has an anti-stress effect on tomato seedlings contaminated with PHE or PYR and this effect is mainly attributed by increased detoxification activity.