Effects of exogenous salicylic acid on manganese toxicity, element contents and antioxidative system in cucumber

The effects of salicylic acid (SA) on manganese (Mn) toxicity in cucumber plants (Cucumis sativus L.) were studied by investigating the symptoms, plant growth, lipid peroxidation, antioxidative enzymes and antioxidants. Excess Mn caused serious chlorosis and inhibited the growth of cucumber plants, and dramatically increased accumulation of Mn in both shoots and roots, furthermore, inhibited the absorption of Ca, Mg and Zn. Addition of SA decreased the transport of Mn from roots to shoots, alleviated the inhibition of Ca, Mg and Zn absorption induced by excess Mn, reduced the toxicity symptoms and promoted the plant growth. The accumulation of reactive oxygen species (ROS) significantly increased in cucumber leaves exposed to excess Mn, and resulted in the lipid peroxidation, which was indicated by accumulated concentration of thiobarbituric acid-reactive substances (TBARS). Addition of SA significantly decreased the level of ROS and lipid peroxidation. Activities of antioxidant enzymes showed different changes, addition of SA inhibited catalase (CAT) and ascorbate peroxidase (APX) activities, while increased activities of superoxide dismutase (SOD), peroxidase (POD), dehydroascorbate reductase (DHAR) and glutathione reductase (GR) in cucumber leaves exposed to excess Mn. As important antioxidants, ascorbate and glutathione contents in cucumber leaves exposed to excess Mn were significantly increased by SA treatment.     

Effects of 24-epibrassinolide on antioxidation defense and osmoregulation systems of young grapevines (V. vinifera L.) under chilling stress

The objective of this study was to investigate the influence of exogenous 24-epibrassinolide (EBR) on the substances involved in antioxidation and osmoregulation responses of young grape plants under chilling stress. The grapevine leaves were sprayed with 0 (control), 0.05, 0.10 or 0.15 mg/L of 24-epibrassinolide and then exposed to 4 and 0 °C for 24 h, respectively. The EBR treatment significantly enhanced the activities of antioxidative enzymes such as catalase, superoxide dismutase, peroxidase and ascorbate peroxidase in the plant leaves compared with the control. The contents of ascorbic acid and reduced glutathione increased after the EBR treatment, while reactive oxygen species (ROS) and lipid peroxidation were inhibited. In addition, the EBR treatment also greatly increased the contents of free proline, soluble protein, and soluble sugar. These results indicated that exogenous EBR treatment could enhance the antioxidation defense system and reduce oxidative damage caused by ROS and lipid oxidation in the young grapevine leaves. Meanwhile, it was found that the treatment could also increase the osmoregulation substance content in the grapevine leaves and improve their resistance against chilling stress.     

Heat and chilling induced disruption of redox homeostasis and its regulation by hydrogen peroxide in germinating rice seeds (Oryza sativa L., Cultivar Ratna)

Extremes of temperature (both heat and chilling) during early inbibitional phase of germination caused disruption of redox-homeostasis by increasing accumulation of reactive oxygen species (superoxide and hydrogen peroxide) and significant reduction of antioxidative defense (assessed in terms of total thiol content and activities of superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase) in germinating tissues of rice (Oryza sativa L., cultivar Ratna). Imbibitional heat and chilling stress also induced oxidative damage to newly assembled membrane system by aggravating membrane lipid peroxidation and protein oxidation [measured in terms of thiobarbituric acid reactive substances (TBARS), free carbonyl content (C = O groups) and membrane protein thiol level (MPTL)]. Treatment with standardized low titer hydrogen peroxide during early imbibitional phase of germination caused significant reversal in oxidative damages to the newly assembled membrane system imposed by heat and chilling stress [evident from the data of TBARS, C = O, MPTL, ROS accumulation, membrane permeability status, membrane injury index and oxidative stress index] in seedlings of experimental rice cultivar. Imbibitional H₂O₂ pretreatment also caused up-regulation of antioxidative defense (activities of superoxide dismutase, catalase, ascorbate peroxidase, glutathione reductase and total thiol content) in the heat and chilling stress-raised rice seedlings. When the parameters of early growth performances were assessed (in terms of relative growth index, biomass accumulation, relative germination performance, mean daily germination, T₅₀ value), it clearly exhibited significant improvement of early growth performances of the experimental rice cultivar. The result proposes that an ‘inductive pulse’ of H₂O₂ is required to switch on some stress acclimatory metabolism through which plant restores redox homeostasis and prevents or repairs oxidative damages to newly assembled membrane system caused by unfavorable environmental cues during early germination to the rice cultivar Ratna. The importance of mitigating oxidative damages to membrane lipid and protein necessary for post-germinative growth under extremes of temperature is also suggested.     

Studies on the superoxide anion production and peroxidase activity in potato leaf cell suspension exposed to salicylic acid

It is now widely accepted that salicylic acid (SA) signaling is mediated by reactive oxygen species (ROS) production. We have studied the effect of SA on peroxidase activity and superoxide anion production in potato leaf cell suspension. The results show that potato cells are insensitive to low concentrations of exogenous SA (< 1 mM) and the effect is observed at 1–5 mM SA. The cells exposed to SA exhibit higher peroxidase activity and show different peroxidase pattern when analyzed on native gels compared to the control. Superoxide anion production is enhanced after two hours of treatment and 2.5 mM SA gives the highest value. The results suggest peroxidase-mediated detoxification of ROS elicited by SA.     

Willow seedlings from photooxidized seeds accelerate cotyledon death and anticipate first leaf emergence: a histological and biochemical study following germination

In willow seeds, photooxidative damage is mainly restricted to the outer cotyledonary tissues, significantly reducing normal germination. Here we analyzed the damage generated in cotyledonary tissues and investigated whether the increase in reactive oxygen species (ROS) generation in seedlings from photooxidized seeds can affect the morphogenetic capacity of the shoot apical meristem. Seeds were photooxidized under different light intensities and the evolution of the damage during seedling growth was studied by light and transmission electron microscopies. The level of lipid peroxidation and changes in antioxidant capacity were measured following the time course of superoxide dismutase, catalase, ascorbate peroxidase and guaiacol peroxidase enzyme activities, and the effect of photooxidative stress on the genesis of new leaf primordia and lateral roots was examined. Early and active endocytosis and autophagy, changes in chloroplast morphology, as well as the accumulation and diffusion of ROS all play important roles in the early cell death observed in cotyledonary tissues. Following germination, seedlings from photooxidized seeds anticipated the emergence of first leaves, which complemented the altered functionality of the damaged cotyledons.     

Waterlogging induced oxidative stress and antioxidant activity in pigeonpea genotypes

The objective of this study was to examine the role of antioxidant enzymes in waterlogging tolerance of pigeonpea (Cajanus cajan L. Halls) genotypes ICP 301 (tolerant) and Pusa 207 (susceptible). Waterlogging resulted in visible yellowing and senescence of leaves, decrease in leaf area, dry matter, relative water content and chlorophyll content in leaves, and membrane stability index in roots and leaves. The decline in all parameters was greater in Pusa 207 than ICP 301. Oxidative stress in the form of superoxide radical, hydrogen peroxide and thiobarbituric acid reactive substances (TBARS) contents initially decreased, however at 4 and 6 d of waterlogging it increased over control plants, probably due to activation of DPI-sensitive NADPH-oxidase. Antioxidant enzymes such as superoxide dismutase, ascorbate peroxidase, glutathione reductase and catalase also increased under waterlogging. The comparatively greater antioxidant enzyme activities resulting in less oxidative stress in ICP 301 could be one of the factor determining its higher tolerance to flooding as compared to Pusa 207. This study is the first to conclusively prove that waterlogging induced increase in ROS is via NADPH oxidase.     

An inductive pulse of hydrogen peroxide pretreatment restores redox-homeostasis and oxidative membrane damage under extremes of temperature in two rice cultivars

Imbibitional heat and chilling stress caused disruption of redox-homeostasis and oxidative damage to newly assembled membrane system by aggravating membrane lipid peroxidation and protein oxidation [measured in terms of thiobarbituric acid reactive substances (TBARS), free carbonyl content (C=O groups) and membrane protein thiol level (MPTL)] along with concomitant increase in accumulation of reactive oxygen species (superoxide and hydrogen peroxide) and significant reduction of antioxidative defense (assessed in terms of total thiol content and activities of superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase) in both the salt sensitive (Ratna) and resistant (SR 26B) germinating tissues of rice cultivars. When compared, salt resistant cultivar SR 26B found to suffer significantly less oxidative membrane damage as compared to salt sensitive cultivar Ratna. Treatment with low titer of hydrogen peroxide caused significant reversal in oxidative damages to the newly assembled membrane system imposed by imbibitional heat and chilling stress (evident from the data of TBARS, C=O, MPTL, ROS accumulation, membrane permeability status, membrane injury index and oxidative stress index) in seedlings of both the cultivars of rice (Ratna and SR 26B). Imbibitional H₂O₂ pretreatment also caused up-regulation of antioxidative defense (activities of superoxide dismutase, catalase, ascorbate peroxidase, glutathione reductase and total thiol content) in the heat and chilling stress-raised seedlings of experimental rice cultivars. When the parameters of early growth performances were assessed (in terms of relative growth index, biomass accumulation and vigor index), it clearly exhibited significant improvement of early growth performances of both the rice cultivars. Better response towards H₂O₂-mediated acclamatory performances and restoration of redox- homeostasis under extremes of temperature were noticed in salt sensitive rice cultivar Ratna compared to salt resistant SR 26B. Taken as a whole, the result suggests the significance of the role of ‘inductive pulse’ of H₂O₂ in acclimatizing adverse temperature stress by restoration of redox-homeostasis and mitigation of oxidative membrane protein and lipid damages during the recovery phase of post-germination event.     

Azadirachta indica and Ocimum sanctum leaf extracts alleviate arsenic toxicity by reducing arsenic uptake and improving antioxidant system in rice seedlings

In the present study the potentials of aqueous extracts of the two plants, neem (Azadirachta indica) and Tulsi (Ocimum sanctum) were examined in alleviating arsenic toxicity in rice (Oryza sativa L.) plants grown in hydroponics. Seedlings of rice grown for 8 days in nutrient solution containing 50 μM sodium arsenite showed decline in growth, reduced biomass, altered membrane permeability and increased production of superoxide anion (O2·−), H2O2 and hydroxyl radicals (·OH). Increased lipid peroxidation marked by elevated TBARS (thiobarbituric acid reactive substances) level, increased protein carbonylation, alterated levels of ascorbate, glutathione and increased activities of enzymes SOD (superoxide dismutase), CAT (catalase), APX (ascorbate peroxidase) and GPX (glutathione peroxidase) were noted in the seedlings on As treatment. Exogenously added leaf aqueous extracts of Azadirachta indica (0.75 mg mL−1, w/v) and Ocimum sanctum (0.87 mg mL−1, w/v) in the growth medium considerably alleviated As toxicity effects in the seedlings, marked by reduced As uptake, restoration of membrane integrity, reduced production of ROS, lowering oxidative damage and restoring the levels of ascorbate, glutathione and activity levels of antioxidative enzymes. Arsenic uptake in the seedlings declined by 72.5% in roots and 72.8% in shoots, when A. indica extract was present in the As treatment medium whereas with O. sanctum extract, the uptake declined by 67.2% in roots and 70.01% in shoots. Results suggest that both A. indica and O. sanctum aqueous extracts have potentials to alleviate arsenic toxicity in rice plants and that A. indica can serve as better As toxicity alleviator compared to O. sanctum.     

Cadmium accumulation and oxidative burst in garlic (Allium sativum)

To investigate the temporal sequence of physiological reactions of garlic (Allium sativum) to cadmium (Cd) treatment, seedlings developed from cloves were grown in increasing concentrations of CdCl2, ranging from 1-10 mM, for up to 8 days in sand. Analysis of Cd uptake indicated that most Cd accumulated in roots, but some was also translocated and accumulated in leaves at longer exposure time (after 12h) and higher concentrations (5 and 10mM) of CdCl2. Changes in activities of antioxidative enzymes, including superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT), were characterized in leaves of garlic seedlings. Cd (5 and 10 mM) initially inhibited the activities of SOD and CAT but thereafter recovered or even increased compared with control plants. POD activities at 5 and 10 mM of Cd increased more than 3-4 times over control plants within 12 h and then dropped, but were still higher than controls at the end of the experiment. Otherwise lipid peroxidation enhanced with the increasing of incubation time and concentrations of external Cd. Leaves exposed to 1 mM CdCl2 showed a less pronounced response and only a small reduction in shoot growth. These results suggested that in leaves of garlic seedlings challenged by CdCl2 at higher concentrations, induction of these various enzymes is part of a general defense strategy to cope with overproduction of reactive oxygen. The possible mechanism of antioxidative enzymes changing before Cd accumulation in leaves of garlic seedlings is discussed.     

Frost resistance and water status of winter rape leaves as affected by differential shoot/root temperature

Twenty day-old winter rape ( Brassica napus L. var. oleifera L. cv. Jantar) seedlings, grown in nutrient solution, were exposed to different shoot/root temperature (i. e. 20/20, 20/3, 3/20 and 3/3°C) for 2 or 4 weeks. Chilling treatments modified markedly the pattern of plant growth as indicated by changes in dry matter accumulation in individual plant parts (leaves, hypocotyls, roots) and decreased leaf specific area. Growth of roots was less sensitive to low temperature than that of shoots. This was reflected by a decrease in shoot/root biomass ratio. Chilling treatments increased freezing resistance, decreased water content and water potential and modified reducing sugar, soluble protein and phospholipid contents in the leaves. A biphasic character of tissue responses to chilling temperature was observed, the most remarkable changes being registered during the first 7 or 14 days of the treatment. Effects of root or shoot exposure to chilling temperature on ice nucleation temperature, LT₅₀, water potential, accumulation of sugars and phospholipids in leaves were additive. All the observations point to the important role of the root system in plant acclimation to cold. Its impact on water status of leaves is emphasized and some mechanisms of root involvement in acclimation processes are proposed.     

Variation in shoot mortality within crowns of severely defoliated <Emphasis Type="Italic">Betula maximowicziana</Emphasis> trees in Hokkaido,northern Japan

To clarify mortality patterns of current-year shoots within the crown of Betula maximowicziana Regel after severe insect herbivory in central Hokkaido, northern Japan, we investigated the degree of defoliation, pattern of shoot development, shoot mortality, and leaf tissue-water relations. One hundred current-year long shoots growing in a B. maximowicziana plantation were observed for defoliation and mortality in June 2002. An outbreak of herbivorous insects (Caligula japonica and Lymantria dispar praeterea) occurred in the stand in mid-to-late June, and the monitored shoots were defoliated to various degrees. Within 1 month of defoliation, some of the severely defoliated shoots had produced new leaves on short shoots that had emerged from axillary buds. Stepwise logistic regression revealed that the probability that current-year long shoots would put out axillary short shoots with leaves is closely related to the degree of defoliation. To evaluate the water relations of the leaves, we determined pressure–volume curves for the leaves that survived the herbivorous insect outbreak and the new leaves that emerged after defoliation. The water potential at turgor loss (Ψ_l,tlp) and the osmotic potential at full turgidity (Ψ_π,sat) were higher for the new leaves than for the surviving leaves, indicating a lower ability to maintain leaf cell turgor against leaf dehydration in the new leaves. Of the 100 shoots, 13 died after the emergence of new leaves. Stepwise logistic regression revealed that the probability that the long shoots would die generally increased with the emergence of new leaves, with increasing shoot height. This result suggests that the combined effect of the vulnerability of newly emerged leaves and low water availability, associated with higher shoot positions within the crown, caused shoot mortality. Based on our results, some possible mechanisms for mortality in severely defoliated B. maximowicziana are discussed.     

Effects of exogenous nitric oxide on cadmium toxicity, element contents and antioxidative system in perennial ryegrass

The effects of sodium nitroprusside (SNP, a donor of NO) on cadmium (Cd) toxicity in ryegrass seedlings (Lolium perenne L.) were studied by investigating the symptoms, plant growth, chlorophyll content, lipid peroxidation, H⁺-ATPase enzyme and antioxidative enzymes. Addition of 100???M CdCl₂ caused serious chlorosis and inhibited the growth of ryegrass seedlings, and dramatically increased accumulation of Cd in both shoots and roots, furthermore, the absorption of macro and micronutrients were inhibited. Addition of 50, 100, 200???M SNP significantly decreased the transport of Cd from roots to shoots, alleviated the inhibition of K, Ca, Mg and Fe, Cu, Zn absorption induced by Cd, reduced the toxicity symptoms and promoted the plant growth. The accumulation of reactive oxygen species (ROS) significantly increased in ryegrass seedlings exposed to Cd, and resulted in the lipid peroxidation, which was indicated by accumulated concentration of thiobarbituric acid-reactive substances. Addition of 50, 100, 200???M SNP significantly decreased the level of ROS and lipid peroxidation. Activities of antioxidant enzymes also showed the same changes. Addition of 50, 100, 200???M SNP increased activities of superoxide dismutase, peroxidase, catalase and ascorbate peroxidase in ryegrass seedlings exposed to Cd. Addition of 100???M SNP had the most significant alleviating effect against Cd toxicity while the addition of 400???M SNP had no significant effect with Cd treatment.     

Manganese-excess induces oxidative stress,lowers the pool of antioxidants and elevates activities of key antioxidative enzymes in rice seedlings

Manganese (Mn) is an essential element for plant growth but in excess, specially in acidic soils, it can become phytotoxic. In order to investigate whether oxidative stress is associated with the expression of Mn toxicity during early seedling establishment of rice plants, we examined the changes in the level of reactive oxygen species (ROS), oxidative stress induced an alteration in the level of non-enzymic antioxidants and activities of antioxidative enzymes in rice seedlings grown in sand cultures containing 3 and 6 mM MnCl₂. Mn treatment inhibited growth of rice seedlings, the metal increasingly accumulated in roots and shoots and caused damage to membranes. Mn treated plants showed increased generation of superoxide anion (O₂ ^.−), elevated levels of H₂O₂ and thiobarbituric acid reactive substances (TBARS) and decline in protein thiol. The level of nonprotein thiol, however, increased due to Mn treatment. A decline in contents of reduced ascorbate (AsA) and glutathione (GSH) as well as decline in ratios of their reduced to oxidize forms was observed in Mn-treated seedlings. The activities of antioxidative enzymes superoxide dismutase (SOD) and its isoforms Mn SOD, Cu/Zn SOD, Fe SOD as well as guaiacol peroxidase (GPX) increased in the seedlings due to Mn treatment however, catalase (CAT) activity increased in 10 days old seedlings but it declined by 20 days under Mn treatment. The enzymes of Halliwell-Asada cycle, ascorbate peroxidase (APX) monodehydoascorbate reductase (MDHAR), dehyroascorbate reductase (DHAR) and glutathione reductase (GR) increased significantly in Mn treated seedlings over controls. Results suggest that in rice seedlings excess Mn induces oxidative stress, imbalances the levels of antioxidants and the antioxidative enzymes SOD, GPX, APX and GR appear to play an important role in scavenging ROS and withstanding oxidative stress induced by Mn.     

Effect of fungal fusaric acid on the root and leaf physiology of watermelon (Citrullus lanatus) seedlings

The impact of fusaric acid (FA) phytotoxin on the physiology of root leaf cells in watermelon seedlings was evaluated. Results revealed that the cell membrane potential treated with FA in 12 h was decreased by 61.9–81.8% compared with untreated controls. FA markedly accelerated the lipid peroxidase activity of watermelon leaves. Malondiadehyde in leaves treated with different concentrations of FA in 24 h was 5.2–11.0 fold as much as control. Phenylalanine ammonia-lyase activity (PAL) in leaves treated with FA was first increased and then decreased. The highest PAL activity was obtained after 6–12 h, which was 7.2–10.5 fold as much as control. Activities of catalase (CAT), superoxide dismutase (SOD), and peroxidase (POD) were first increased and then decreased when the seedlings were treated with FA. The highest SOD and POD activities were observed after 12 h and then both declined. The β-1,3-glucanase and chitinase activities in leaves treated with FA were first quickly increased and then declined, the highest activities were found after 12 and 3 h. We concluded that FA strongly inhibited root and leaf cells functions physiologically responsible for fusarium wilt of watermelon.     

<Emphasis Type="Italic">In vitro</Emphasis> organogenesis and antioxidant enzymes activity in <Emphasis Type="Italic">Acanthophyllum sordidum</Emphasis>

The effect of various hormonal combinations on callus formation and regeneration of shoot and root from leaf derived callus of Acanthophyllum sordidum Bunge ex Boiss. has been studied. Proteins and activity of antioxidant enzymes were also evaluated during shoot and root organogenesis from callus. Calli were induced from leaf explants excised from 30-d-old seedlings grown on Murashige and Skoog medium containing 4.52 μM 2,4-dichlorophenoxyacetic acid + 4.65 μM kinetin. Maximum growth of calli and the most efficient regeneration of shoots and roots occurred with 2.69 μM 1-naphthalene acetic acid (NAA), 2.69 μM NAA + 4.54 μM thidiazuron and 2.46 μM indole-3-butyric acid. Protein content decreased in calli and increased significantly during regeneration of shoots from callus. Superoxide dismutase activity decreased in calli comparing to that of seedlings, then increased in regenerated shoots and roots. High catalase activity was detected in seedlings and regenerated shoots, whereas high peroxidase activity was observed in calli and regenerated roots.     

Brassinosteroids alleviate chilling-induced oxidative damage by enhancing antioxidant defense system in suspension cultured cells of <Emphasis Type="Italic">Chorispora bungeana</Emphasis>

Brassinosteroids (BRs), a class of plant steroid hormones, play a significant role in the amelioration of various biotic and abiotic stresses. In order to further explore and elaborate their roles in plants subjected to chilling stress, suspension cultured cells of Chorispora bungeana with or without 24-epibrassinolide (EBR) application were exposed to 4 and 0°C for 5 days. The EBR treated cells exhibited higher viability after exposure to low temperatures compared with the control. Under chilling stress, reactive oxygen species (ROS) levels and lipid peroxidation were increased in the cultured cells, which were significantly inhibited by EBR application. The activities of antioxidative enzymes such as ascorbate peroxidase (APX), catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD) were increased during chilling treatments, and these increases were more significant in the EBR applied suspension cells. The EBR treatment also greatly enhanced contents of ascorbic acid (AsA) and reduced glutathione (GSH) under chilling stress. From these results, it can be concluded that EBR could play the positive roles in the alleviation of oxidative damage caused by ROS overproduction through enhancing antioxidant defense system, resulting in improving the tolerance of C. bungeana suspension cultures to chilling stress.     

Effect of salicylic acid potentiates cadmium-induced oxidative damage in <Emphasis Type="Italic">Oryza sativa</Emphasis> L. leaves

In the present investigation, we studied the possible potentiating effect of salicylic acid (SA) under Cd toxicity in Oryza sativa L. leaves. Cd treatments for 24 h reduced the shoot length, dry biomass and total chlorophyll content followed by high Cd accumulation in shoots. About 16 h presoaking with SA resulted in partial protection against Cd, as observed by minor changes in length, biomass and total chlorophyll. SA priming resulted in low Cd accumulation. Enhanced thiobarbituric acid reactive substances (TBARS), hydrogen peroxide (H₂O₂) and superoxide anion (O₂ ⁻) content were seen when Cd was applied alone, while under SA priming the extent of TBARS, H₂O₂ and O₂ ⁻ were significantly low, suggesting SA-regulated protection against oxidative stress. The antioxidant enzymes like Catalase (CAT), guaiacol peroxidase (GPx), glutathione reductase (GR) and superoxide dismutase (SOD) showed varied activities under Cd alone. CAT activity increased after Cd treatment, followed by a decline in GPX and GR activity. SOD also declined at the highest concentrations with an initial increase. Under SA-priming conditions, the efficiency of the antioxidant enzymes was significantly elevated. GPx and SOD activity showed significant increase in activity. The ascorbate activity increased after Cd treatment, followed by a decline in glutathione under SA-free condition. SA priming showed gradual increase in these non-enzymic antioxidants. Our results indicate that Cd-induced oxidative stress can be regulated by SA.     

Modelling foliage characteristics in 3D tree crowns: influence on light interception and leaf irradiance

Because of the difficulty and time involved in making exhaustive measurements of the geometric parameters of large tree crowns, simplifying hypotheses are often used in 3D virtual plant modelling, but the effects on the radiation balance of each approximation are rarely assessed. Three hybrid walnut trees aged 7–9 years were digitized to analyse the effect of the crown geometric variables on light capture. The six studied variables were: (1) leaf area, (2) number of leaves per annual shoot, (3) position of leaves, (4) orientation of leaves, (5) leaflet inclination, and (6) lamina shape. For each variable, a sensitivity analysis compared a reference, based on observed values, with scenarios consisting of simplifying hypotheses. The total incident light intercepted during a bright day and the distributions of leaf irradiance were calculated using the Archimed radiative transfer model. Since some of the crown parameters were generated stochastically, the radiation simulations were repeated until results stabilised. Simplified models can be used to calculate with satisfactory results individual leaf area and number of leaves per shoot. Conversely, differentiating statistical distributions of individual leaf area between short and long shoots is more difficult and may generate errors up to 30%. Leaf clumping is a determining factor and requires correct grouping of leaves around the annual shoots bearing them. The effect of position of leaves along the shoot is less than 2%. Simple statistical distributions are adequate for representing leaf angle. Finally, the effect of specific leaf geometry is very important, but it can be approached using a limited number of representative leaf shapes.     

Transgenic cotton (<Emphasis Type="Italic">Gossypium hirsutum</Emphasis> L.) seedlings expressing a tobacco glutathione <Emphasis Type="Italic">S</Emphasis>-transferase fail to provide improved stress tolerance

Transgenic cotton (Gossypium hirsutum L.) lines expressing the tobacco glutathione S-transferase (GST) Nt107 were evaluated for tolerance to chilling, salinity, and herbicides, antioxidant enzyme activity, antioxidant compound levels, and lipid peroxidation. Although transgenic seedlings exhibited ten-fold and five-fold higher GST activity under normal and salt-stress conditions, respectively, germinating seedlings did not show improved tolerance to salinity, chilling conditions, or herbicides. Glutathione peroxidase (GPX) activity in transgenic seedlings was 30% to 60% higher under normal conditions, but was not different than GPX activity in wild-type seedlings under salt-stress conditions. Glutathione reductase, superoxide dismutase, ascorbate peroxidase, and monodehydroascorbate reductase activities were not increased in transgenic seedlings under salt-stress conditions, while dehydroascorbate reductase activity was decreased in transgenic seedlings under salt-stress conditions. Transgenic seedlings had 50% more oxidized glutathione when exposed to salt stress. Ascorbate levels were not increased in transgenic seedlings under salt-stress conditions. Malondialdehyde content in transgenic seedlings was nearly double that of wild-type seedlings under normal conditions and did not increase under salt-stress conditions. These results show that expression of Nt107 in cotton does not provide adequate protection against oxidative stress and suggests that the endogenous antioxidant system in cotton may be disrupted by the expression of the tobacco GST.     

Effects of root and foliar applications of exogenous NO on alleviating cadmium toxicity in lettuce seedlings

The effects of sodium nitroprusside (SNP, a donor of NO) on cadmium (Cd) toxicity in lettuce seedlings were studied. SNP was added into hydroponic systems or sprayed directly on the leaves of plants grown with and without Cd. Excess supply of Cd (100 μM) caused growth inhibition, dramatically increased Cd accumulation in both leaves and roots, and inhibited the absorption of Ca, Mg, Fe and Cu. Excess Cd also decreased activities of superoxide dismutase peroxidase and catalase in leaves and roots, and increased the accumulation of superoxide anion (O ₂ ^·? ), hydrogen peroxide (H₂O₂) and malondialdehyde (MDA). Root or foliar applications of exogenous NO alleviated Cd-induced growth suppression, especially root application of 250 μM SNP and foliar addition of 500 μM SNP. Addition of SNP promoted the chlorophyll synthesis suggesting that the photosynthesis was up-regulated. Exogenous NO increased Cd-decreased activities of antioxidant enzymes and markedly diminished Cd-induced reactive oxygen species (ROS) and MDA accumulation. Moreover, the absorption of Ca, Mg, Fe and Cu was increased, indicating that exogenous NO stimulated H⁺-ATPase activity to promote sequestration or uptake of ions. In addition, exogenous NO also inhibited Cd transfer from roots to shoots, which may indicate that Cd retention in roots induced by NO plays a significant role in Cd tolerance in lettuce seedlings. These data suggest that under Cd stress, exogenous NO improves photosynthesis by increasing chlorophyll synthesis, protects lettuce seedlings against oxidative damage by scavenging ROS, helps to maintain the uptake of nutrient elements, and inhibits Cd transferred to shoots effectively.