Oxidative stress, protein carbonylation, proteolysis and antioxidative defense system as a model for depicting water deficit tolerance in Indica rice seedlings

Water deficit is an important constraint to rice (Oryza sativa L.) productivity. The present study was undertaken to investigate whether the level of oxidative stress, carbonylation of proteins, proteolysis and status of antioxidative defense could serve as a model to distinguish water deficit tolerant and sensitive rice cultivars. When 10-day-grown seedlings of two rice cultivars, Malviya-36 (drought-sensitive) and Brown Gora (drought-tolerant) were subjected to ?1.0 and ?2.1 MPa water deficit treatments for 24–72 h with polyethylene glycol 6000 in the medium, a greater decline in the growth of the seedlings and levels of leaf water potential, relative water content, Chl a, Chl b, carotenoids and greater increase in leaf water loss were observed in the sensitive cultivar than the tolerant. Under similar level of water deficit seedlings of sensitive cultivar showed higher level of superoxide anion generation, H₂O₂, lipid peroxidation and proteolysis in roots as well as shoots compared to the tolerant. Drought-tolerant cultivar had higher constitutive level of antioxidative enzymes superoxide dismutase and catalase and the activities of these two enzymes alongwith of guaiacol peroxidase showed greater increase in this cultivar under water deficit compared to the sensitive. A significant decline in the level of protein thiol and a higher increase in protein carbonyls content, also confirmed by protein gel blot analysis with an antibody against 2,4-dinitrophenylhydrazine was observed in the seedlings of drought sensitive cv. Malviya-36 compared to the tolerant cv. Brown Gora when subjected to similar level of water deficit. Seedlings of drought sensitive cultivar, under water deficit, showed higher proteolytic activity, higher number of in-gel activity stained proteolytic bands and higher expression of oxidized proteins in roots compared to the tolerant cultivar. Results suggest that poor capacity of antioxidative enzymes could be, at least partly, correlated with water deficit sensitivity of sensitive cultivar and that higher activity of antioxidative enzymes superoxide dismutase, catalase, guaiacol peroxidase, low proteolytic activity, lower level of protein carbonyls and protein thiolation could serve as a model to depict water deficit tolerance in Indica rice seedlings.     

Reactive oxygen species, ascorbate–glutathione pool, and enzymes of their metabolism in drought-sensitive and tolerant indica rice (Oryza sativa L.) seedlings subjected to progressing levels of water deficit

Water deficit for rice is a worldwide concern, and to produce drought-tolerant varieties, it is essential to elucidate molecular mechanisms associated with water deficit tolerance. In the present study, we investigated the differential responses of nonenzymatic antioxidants ascorbate (AsA), glutathione (GSH), and their redox pool as well as activity levels of enzymes of ascorbate–glutathione cycle in seedlings of drought-sensitive rice (Oryza sativa L.) cv. Malviya-36 and drought-tolerant cv. Brown Gora subjected to water deficit treatment of ?1.0 and ?2.1 MPa for 24–72 h using PEG-6000 in sand cultures. Water deficit caused increased production of reactive oxygen species such as O₂?^?, H₂O₂, and HO? in the tissues, and the level of production was higher in the sensitive than the tolerant cultivar. Water deficit caused reduction in AsA and GSH and decline in their redox ratios (AsA/DHA and GSH/GSSG) with lesser decline in tolerant than the sensitive seedlings. With progressive level of water deficit, the activities of monodehydroascorbate reductase, dehydroascorbate reductase, ascorbate peroxidase (APX), and glutathione transferase increased in the seedlings of both rice cultivars, but the increased activity levels were higher in the seedlings of drought-tolerant cv. Brown Gora compared to the sensitive cv. Malviya-36. Greater accumulation of proline was observed in stressed seedlings of tolerant than the sensitive cultivar. In-gel activity staining of APX revealed varying numbers of their isoforms and their differential expression in sensitive and tolerant seedlings under water deficit. Results suggest that an enhanced oxidative stress tolerance by a well-coordinated cellular redox state of ascorbate and glutathione in reduced forms and induction of antioxidant defense system by elevated activity levels of enzymes of ascorbate–glutathione cycle is associated with water deficit tolerance in rice.     

Water deficit and aluminum tolerance are associated with a high antioxidative enzyme capacity in Indica rice seedlings

Plant growth and productivity are greatly affected due to changes in the environmental conditions. In the present investigation, the interactive effects of two important abiotic stresses, i.e., water deficit and Al toxicity, were examined in the seedlings of two rice (Oryza sativa L.) cvs. Malviya-36 (water deficit/Al sensitive) and Vandana (water deficit/Al tolerant). When 15 days grown seedlings were exposed to water deficit (created with 15 % polyethylene glycol 6000) or Al (1 mM AlCl₃) treatment or both the treatments together for 48 h, the lengths of root/shoot, relative water content, and chlorophyll greatly declined in the seedlings of the sensitive cultivar, whereas in the tolerant seedlings, either little or insignificant decline in these parameters was observed due to the treatments. Seedlings subjected to water deficit or Al treatment alone or in combination showed increased intensity of the isoenzyme activity bands of superoxide dismutase (SOD), guaiacol peroxidase (GPX), and ascorbate peroxidase (APX) in in-gel activity staining studies. Water deficit caused decrease in intensity of catalase (CAT) activity bands; however, when seedlings were exposed to AlCl₃ alone or in combination with water deficit, the intensity of the CAT isoforms increased in both the rice cultivars. The level of expression of the activity bands of SOD, CAT, GPX, and APX was always higher in the seedlings of tolerant cv. Vandana compared to the sensitive cv. Malviya-36 under both controls as well as stress treatments. Higher intensity of isozymes representing higher activity levels of antioxidative enzymes in the rice seedlings and their further increase under water deficit, Al exposure, or in combination of both the stresses appears to serve as useful marker for specifying a combination of water deficit and Al tolerance in rice.     

Ozone suppresses soil drying- and abscisic acid (ABA)-induced stomatal closure via an ethylene-dependent mechanism

Elevated atmospheric ozone concentrations (70 ppb) reduced the sensitivity of stomatal closure to abscisic acid (ABA) in Leontodon hispidus after at least 24 h exposure (1) when detached leaves were fed ABA, and (2) when intact plants were sprayed or injected with ABA. They also reduced the sensitivity of stomatal closure to soil drying around the roots. Such effects could already be occurring under current northern hemisphere peak ambient ozone concentrations. Leaves detached from plants which had been exposed to elevated ozone concentrations generated higher concentrations of ethylene, although leaf tissue ABA concentrations were unaffected. When intact plants were pretreated with the ethylene receptor binding antagonist 1-methylcyclopropene, the stomatal response to both applied ABA and soil drying was fully restored in the presence of elevated ozone. Implications of ethylene's antagonism of the stomatal response to ABA under oxidative stress are discussed. We suggest that this may be one mechanism whereby elevated ozone induces visible injury in sensitive species. We emphasize that drought linked to climate change and tropospheric ozone pollution, are both escalating problems. Ozone will exacerbate the deleterious effects of drought on the many plant species including valuable crops that respond to this pollutant by emitting more ethylene.     

Oxidative Stress Tolerance Mechanism in Rice under Salinity

The research was conducted to investigate comparative oxidative damage including probable protective roles of antioxidant and glyoxalase systems in rice (Oryza sativa L.) seedlings under salinity stress. Seedlings of two rice genotypes: Pokkali (tolerant) and BRRI dhan28 (sensitive) were subjected to 8 dSm⁻¹ salinity stress for seven days in a hydroponic system. We observed significant variation between Pokkali and BRRI dhan28 in phenotypic, biochemical and molecular level under salinity stress. Carotenoid content, ion homeostasis, antioxidant enzymes, ascorbate and glutathione redox system and proline accumulation may help Pokkali to develop defense system during salinity stress. However, the activity antioxidant enzymes particularly superoxide dismutase (SOD), catalase (CAT) and non-chloroplastic peroxidase (POD) were observed significantly higher in Pokkali compared to salt-sensitive BRRI dhan28. Higher glyoxalase (Gly-I) and glyoxalase (Gly-II) activity might have also accompanied Pokkali genotype to reduce potential cytotoxic MG through non-toxic hydroxy acids conversion. However, the efficient antioxidants and glyoxalase system together increased adaptability in Pokkali during salinity stress.     

The role of active oxygen in iron tolerance of rice (Oryza sauva L.)

Summary Iron tolerance of rice (Oryza sativa L.) was investigated using an oxygen depleted hydroculture system. Treatment with high concentrations of Fe²⁺ induced yellowing and bronzing symptoms as well as iron coatings at the root surface. Root and shoot growth were inhibited by increasing iron concentration in the medium. All symptoms were more pronounced in an iron sensitive cultivar (IR 64) compared to an iron tolerant one (IR 9764-45-2). Superoxide dismutase and peroxidase activity of root extracts of IR 97 were about twice that of IR 64 in untreated control plants. No significant increase of peroxidase activity was detected with increasing iron concentration in the medium. Catalase activity of IR 64 was slightly higher than that of IR 97, independent of iron concentration.Abbreviations SOD Superoxide dismutase (EC 1.15.1.1) - POD peroxidase (EC 1.11.1.7) - EDTA ethylenediamintetraacetic acid - fwt fresh weight - Hepes (N-[2-hydroxyethyl]piperazine-N-[2-ethanesulfonic acid]) - BSA bovine serum albumin - IR 97 IR 9764-45-2 an iron tolerant rice cultivar - IR 64 iron sensitive rice cultivar - PM plasma membrane     

Stomatal Closure During Tobacco Leaf Desiccation as Affected by Ozone

The effect of acute ozone exposure on the stomatal conductance and leaf water content during rapid desiccation was examined in leaves of two tobacco cultivars, ozone sensitive cv. BelW3 and ozone tolerant cv. Samsun. The relative rate of stomatal closure was constant during leaf desiccation in cv. Samsun but decreased in cv. BelW3 in both ozonated and control plants. Ozone exposure increased the relative rate of stomatal closure and transpiration rate (measured on the following day) in cv. Samsun, but reduced the respective parameters in cv. BelW3. As a result, the plants of ozone-sensitive cultivar, treated with ozone, lost more water during desiccation than control plants. This revised version was published online in July 2006 with corrections to the Cover Date.     

Evaluation of oxidative stress tolerance in maize (Zea mays L.) seedlings in response to drought

Seedlings of selected six genotypes of maize (Zea mays L.) differing in their drought sensitivity (LM5 and Parkash drought-tolerant and PMH2, JH3459, Paras and LM14 as drought-sensitive) were exposed to 72 h drought stress at two leaf stage. Alterations in their antioxidant pools combined with activities of enzymes involved in defense against oxidative stress were investigated in leaves. Activities of some reactive oxygen species (ROS)-scavenging enzymes, catalase (CAT) and ascorbate peroxidase (APX) were enhanced in tolerant genotypes in response to drought stress. Superoxide dismutase (SOD) activity was significantly decreased in sensitive genotypes, but remained unchanged in tolerant genotypes under stress. Peroxidase (POX) activity was significantly induced in tolerant, as well as sensitive genotypes. Imposition of stress led to increase in H2O2 and malondialdehyde (MDA, a marker for lipid peroxidation) content in sensitive genotypes, while in tolerant genotypes no change was observed. Significant increase in glutathione content was observed in sensitive genotypes. Ascorbic acid pool was induced in both tolerant and sensitive genotypes, but induction was more pronounced in tolerant genotypes. Significant activation of antioxidative defence mechanisms correlated with drought-induced oxidative stress tolerance was the characteristic of the drought tolerant genotypes. These studies provide a mechanism for drought tolerance in maize seedlings.     

Differential ozone sensitivity of rice cultivars as indicated by visible injury and grain yield

Surface ozone pollution may cause reductions in rice ( Oryza sativa L.) yield. Ozone sensitivity in rice cultivars is often evaluated based on visible leaf injury at an early growth stage. However, it is not clear whether reduction in grain yield is related to visible injury. Therefore, visible damage and grain yield reduction were examined in Japanese and Asian rice cultivars exposed to ozone. In experiment 1, 3-week-old rice seedlings were exposed to ozone (min.: 20 nl·l⁻¹, max.:120 nl·l⁻¹) for 12 h in open-top chambers (OTCs). Visible leaf injury was quantified according to a leaf bronzing score. In experiment 2, rice plants were exposed to ozone in OTCs throughout the cropping season until grain harvest. Daily mean ozone concentrations were maintained at 2, 23, 28, 42, and 57 nl·l⁻¹ with a regular diurnal pattern of exposure. After harvest, grain yield was determined. Based on visible injury to the uppermost fully expanded leaf, the indica cultivar 'Kasalath' was most tolerant, and the japonica cultivar 'Kirara 397' was most sensitive to ozone. However, grain yields for both 'Kasalath' and 'Kirara 397' were significantly decreased after ozone exposure. The indica cultivar 'Jothi' suffered severe injury after ozone exposure but had no reduction in grain yield. Therefore, ozone sensitivity of rice cultivars evaluated by visible injury did not coincide with that evaluated by the reductions in grain yield. These results suggest that mechanisms that induce acute leaf injury do not relate to chronic ozone toxicity that reduces yield.     

Elucidation of salt-tolerance metabolic pathways in contrasting rice genotypes and their segregating progenies

Key message

Differentially expressed antioxidant enzymes, amino acids and proteins in contrasting rice genotypes, and co-location of their genes in the QTLs mapped using bi-parental population, indicated their role in salt tolerance.

Abstract

Soil salinity is a major environmental constraint limiting rice productivity. Salt-tolerant ‘CSR27’, salt-sensitive ‘MI48’and their extreme tolerant and sensitive recombinant inbred line (RIL) progenies were used for the elucidation of salt stress tolerance metabolic pathways. Salt stress-mediated biochemical and molecular changes were analyzed in the two parents along with bulked-tolerant (BT) and bulked-sensitive (BS) extreme RILs. The tolerant parent and BT RILs suffered much lower reduction in the chlorophyll as compared to their sensitive counterparts. Activities of antioxidant enzymes superoxide dismutase (SOD) and peroxidase (POD) and non-enzymatic antioxidant ascorbic acid were much higher in salt-stressed CSR27 and BT RILs than MI48 and BS RILs. Further, the tolerant lines showed significant enhancement in the levels of amino acids methionine and proline in response to salt stress in comparison to the sensitive lines. Similarly, the tolerant genotypes showed minimal reduction in cysteine content whereas sensitive genotypes showed a sharp reduction. Real time PCR analysis confirmed the induction of methionine biosynthetic pathway (MBP) enzymes cystathionine-β synthase (CbS), S-adenosyl methionine synthase (SAMS), S-adenosyl methionine decarboxylase (SAMDC) and serine hydroxymethyl transferase (SHMT) genes in tolerant lines, suggesting potential role of the MBP in conferring salt tolerance in rice variety CSR27. Proteome profiling also confirmed higher expression of SOD, POD and plastidic CbS and other proteins in the tolerant lines, whose genes were co-located in the QTL intervals for salt tolerance mapped in the RIL population. The study signifies integrated biochemical-molecular approach for identifying salt tolerance genes for genetic improvement for stress tolerant rice varieties.

     

Role of abscisic acid in cadmium tolerance of rice (Oryza sativa L.) seedlings

Changes in abscisic acid (ABA) contents in Cd-treated rice (Oryza sativa L.) seedlings of two cultivars were investigated. On treatment with CdCl2, the ABA content rapidly increased in the leaves and roots of Cd-tolerant cultivar (cv. Tainung 67, TNG67) but not in the Cd-sensitive cultivar (cv. Taichung Native 1, TN1). The reduction of transpiration rate of TN1 caused by Cd was less than that of TNG67. Exogenous application of ABA reduced transpiration rate, decreased Cd content, and enhanced Cd tolerance of TN1 seedlings. Exogenous application of the ABA biosynthesis inhibitor, fluridone, reduced ABA accumulation, increased transpiration rate and Cd content, and decreased Cd tolerance of TNG67 seedlings. Fluridone effect on Cd toxicity of TNG67 seedlings was reversed by the application of ABA. The roles of endogenous ABA in Cd tolerance of rice seedlings are discussed and suggested.     

Abscisic acid-induced hydrogen peroxide is required for anthocyanin accumulation in leaves of rice seedlings

The role of hydrogen peroxide (H(2)O(2)) in abscisic acid (ABA)-induced anthocyanin accumulation in detached and intact leaves of rice seedlings was investigated. Treatment with ABA resulted in an accumulation of anthocyanins in detached rice leaves. Dimethylthiourea, a chemical trap for H(2)O(2), was observed to be effective in inhibiting ABA-induced accumulation of anthocyanins. Inhibitors of NADPH oxidase (diphenyleneiodonium chloride and imidazole), phosphatidylinositol 3-kinase (wortmannin and LY 294002), and a donor of nitric oxide (N-tert-butyl-alpha-phenylnitrone), which have previously been shown to prevent ABA-induced H(2)O(2) accumulation in detached rice leaves, inhibited ABA-induced anthocyanin increase. Exogenous application of H(2)O(2), however, was found to increase the anthocyanin content of detached rice leaves. In terms of H(2)O(2) accumulation, intact (attached) leaves of rice seedlings of cultivar Taichung Native 1 (TN1) are ABA sensitive and those of cultivar Tainung 67 (TNG67) are ABA insensitive. Upon treatment with ABA, H(2)O(2) and anthocyanins accumulated in leaves of TN1 seedlings but not in leaves of TNG67. Our results, obtained from detached and intact leaves of rice seedlings, suggest that H(2)O(2) is involved in ABA-induced anthocyanin accumulation in this species.     

Role of abscisic acid in chilling tolerance of rice (Oryza sativa L.) seedlings. I. Endogenous abscisic acid levels

Changes of ABA levels in chilled rice (Oryza sativa L.) seedlings of two varieties were determined. On exposure to chilling, ABA concentration rapidly increased in the chilling-tolerant cultivar (cv. Tainung 67, TNG.67) but not in the chilling-sensitive cultivar (cv. Taichung Native 1, TN.1). Both detached shoots and roots of TNG.67 seedlings showed a significant ABA increase after exposure to chilling. TN.1 seedlings could not accumulate ABA under low temperature but well-watered status. Exogenous application of the ABA biosynthetic inhibitor, fluridone, reduced ABA accumulation, as well as survival ratio of chilled TNG.67 seedlings. Electrolyte leakage and leaf conductance were also increased by the inhibitor and the effects could be reversed by exogenously applied ABA. ABA concentrations in xylem sap of TNG.67 seedlings increased within 4 h after chilling, and this was temporally coincident with the reduction of leaf conductance. The roles of endogenous ABA in the tolerance of rice seedlings to chilling on a whole plant basis are discussed and suggested.     

Antioxidant metabolite levels in ozone-sensitive and tolerant genotypes of snap bean

Ozone-sensitive and tolerant genotypes of snap bean ( Phaseolus vulgaris L.) were compared for differences in leaf ascorbic acid (vitamin C), glutathione and α -tocopherol (vitamin E) content to determine whether antioxidant levels were related to ozone tolerance. Seven genotypes were grown in pots under field conditions during the months of June and July. Open top chambers were used to establish either a charcoal filtered (CF) air control (36 nmol mol⁻¹ ozone) or a treatment where CF air was supplemented with ozone from 8:00 to 20:00 h with a daily 12 h mean of 77 nmol mol⁻¹. Fully expanded leaves were analyzed for ascorbic acid, chlorophyll, glutathione, guaiacol peroxidase (EC 1.11.1.7) and α -tocopherol. Leaf ascorbic acid was the only variable identified as a potential factor in ozone tolerance. Tolerant genotypes contained more ascorbic acid than sensitive lines, but the differences were not always statistically significant. Genetic differences in glutathione and α -tocopherol were also observed, but no relationship with ozone tolerance was found. Guaiacol peroxidase activity and leaf α -tocopherol content increased in all genotypes following a one week ozone exposure, indicative of a general ozone stress response. Ozone had little effect on the other variables tested. Overall, ozone sensitive and tolerant plants were not clearly distinguished by differences in leaf antioxidant content. The evidence suggests that screening for ozone tolerance based on antioxidant content is not a reliable approach.     

Effects of abscisic acid and its related compounds on rice seedling growth

Rice seedlings with the mesocotyl and coleoptile (the undeveloped leaves enclosed in the coleoptile) are here referred to as MC type seedlings and are considered to be suitable for deep sowing. We investigated the effects of abscisic acid (ABA) and several of its related compounds on the occurrence of MC type seedlings and on rice mesocotyl growth. Rice (Oryza sativa L. cv. JC 91) seedlings were grown on 0.8% agar medium in the presence or absence of various kinds of ABAs under aseptic conditions at 30 °C in the dark for 14 days. The activity of the R isomer of ABA (R-ABA) was slightly less than that of the naturally occurring S form (S-ABA) concerning the occurrence of MC type rice seedlings and the growth of the rice mesocotyl. In addition, the racemate of R-and S-ABA (RS-ABA) is less effective than R-ABA and S-ABA alone.Trans-ABA had no activity in relation to both percent occurrence of MC type seedlings and mesocotyl growth. The results of the present study suggest that the occurrence of MC type rice seedlings and the growth of rice mesocotyls were closely related to structure-activity relationships with analogs of ABA.     

Chronic exposure to increasing background ozone impairs stomatal functioning in grassland species

Two species found in temperate calcareous and mesotrophic grasslands (Dactylis glomerata and Leontodon hispidus) were exposed to eight ozone treatments spanning preindustrial to post‐2100 regimes, and late‐season effects on stomatal functioning were investigated. The plants were grown as a mixed community in 14 L containers and were exposed to ozone in ventilated solardomes (dome‐shaped greenhouses) for 20 weeks from early May to late September 2007. Ozone exposures were based on O₃ concentrations from a nearby upland area, and provided the following seasonal 24 h means: 21.4, 39.9 (simulated ambient), 50.2, 59.4, 74.9, 83.3, 101.3 and 102.5 ppb. In both species, stomatal conductance of undamaged inner canopy leaves developing since a midseason cutback increased linearly with increasing background ozone concentration. Imposition of severe water stress by leaf excision indicated that increasing background ozone concentration decreased the ability of leaves to limit water loss, implying impaired stomatal control. The threshold ozone concentrations for these effects were 15–40 ppb above current ambient in upland UK, and were within the range of ozone concentrations anticipated for much of Europe by the latter part of this century. The potential mechanism behind the impaired stomatal functioning was investigated using a transpiration assay. Unlike for lower ozone treatments, apparently healthy green leaves of L. hispidus that had developed in the 101.3 ppb treatment did not close their stomata in response to 1.5 μm abscisic acid (ABA); indeed stomatal opening initially occurred in this treatment. Thus, ozone appears to be disrupting the ABA‐induced signal transduction pathway for stomatal control thereby reducing the ability of plants to respond to drought. These results have potentially wide‐reaching implications for the functioning of communities under global warming where periods of soil drying and episodes of high vapour pressure deficit are likely to be more severe.     

ABA application to maize hybrids contrasting for drought tolerance: changes in water parameters and in antioxidant enzyme activity

The objective of this study was to evaluate the effects of abscisic acid (ABA) related to the increase of water-stress tolerance in two drought contrasting maize hybrids: DKB 390 (tolerant) and BRS 1030 (sensitive). The characterization of water status (pre-dawn leaf water potential, Ψ_pd; midday leaf water potential, Ψ_md and stem water potential, Ψ_st) and antioxidant enzyme activity was conducted on greenhouse grown plants. The ABA, hydrogen peroxide (H₂O₂), and malondialdehyde (MDA) contents were also analyzed. Water deficit was imposed for 10 days at the flowering stage and a dosage of 100 μM ABA was applied to plant canopy. Measurements were taken during 10 days after the water recovery. With 5 days of stress, the tolerant hybrid showed lower MDA content, decrease in the water status, and higher activity of the enzymes superoxide dismutase, catalase, ascorbate peroxidase, as well as guaiacol, glutathione reductase, dehydroascorbate reductase, polyphenol oxidase, and l-phenylalanine ammonia-lyase, as compared to the sensitive hybrid. With 10 days of stress, DKB 390 had a decrease in the activity of enzymes whereas BRS 1030 showed a higher activity. In addition, the latter showed greater amounts of H₂O₂ and MDA. ABA application led to a higher tolerance only in DKB 390, due to the increase of water status and the enzymatic activity, mainly the catalase.     

Effect of water deficit on carbohydrate status and enzymes of carbohydrate metabolism in seedlings of wheat cultivars

The effect of water deficit on carbohydrate status and enzymes of carbohydrate metabolism (alpha and beta amylases, sucrose phosphate synthase, sucrose synthase, acid and alkaline invertases) in wheat (Triticum aestivum L.) was investigated in the seedlings of drought-sensitive (PBW 343) and drought-tolerant (C 306) cultivars. The water deficit was induced by adding 6% mannitol (water potential -0.815 Mpa) in the growth medium. The water deficit reduced starch content in the shoots of tolerant seedlings as compared to the sensitive ones, but increased sucrose content in the shoots and roots of tolerant seedlings, indicating their protective role during stress conditions. It also decreased the alpha-amylase activity in the endosperm of seedlings of both the cultivars, but increased alpha and beta amylase activities in the shoots of tolerant ones. Sucrose phosphate synthase (SPS) activity showed a significant increase at 6 days of seedling growth (DSG) in the shoots of stressed seedlings of tolerant cultivar. However, SPS activity in the roots of stressed seedlings of sensitive cultivar was very low at 4 DSG and appeared significantly only at day 6. Sucrose synthase (SS) activity was lower in the shoots and roots of stressed seedlings of tolerant cultivar than sensitive ones at early stage of seedling growth. Higher acid invertase activity in the shoots of seedlings of tolerant cultivar appeared to be a unique characteristic of this cultivar for stress tolerance. Alkaline invertase activity, although affected under water deficit conditions, but was too low as compared to acid invertase activity to cause any significant affect on sucrose hydrolysis. In conclusion, higher sucrose content with high SPS and low acid invertase and SS activities in the roots under water deficit conditions could be responsible for drought tolerance of C 306.     

Shade light interaction with salicylic acid in regulating growth of sun (alpine) and shade (prairie) ecotypes of Stellaria longipes

Magnesium (Mg) deficiency in plants is a widespread problem, affecting productivity and quality in agriculture. The mechanism of Mg deficiency inducing antioxidant enzyme activities has not been elucidated in rice. We examined the relationship among abscisic acid (ABA), H₂O₂, and antioxidant enzymes in the leaves of rice seedlings grown under conditions of Mg deficiency. The expression of OsRab16A, an ABA responsive gene, was used to determine the content of ABA. Mg deficiency resulted in increased ABA content in leaves of rice seedlings. The production of H₂O₂ was examined by 3,3-diaminobenzidine staining and a colorimetric method. Mg deficiency also induced H₂O₂ production in leaves, which was blocked by dipehnyleneiodonium chloride (DPI), an NADPH oxidase inhibitor. Tungstate (Tu), an ABA biosynthesis inhibitor, was effective in reducing Mg deficiency-increased ABA content, as well as Mg deficiency-induced H₂O₂ production. Both Tu and DPI were effective in reducing Mg deficiency-induced activities of superoxide dismutase, ascorbate peroxidase, glutathione reductase, and catalase in the leaves. Mg deficiency-induced ABA accumulation may trigger increased production of H₂O₂, which may involve plasma-membrane NADPH oxidase, and, in turn, up-regulates the activities of antioxidant enzymes in leaves of rice seedlings.