Physiology,gene expression,and metabolome of two wheat cultivars with contrasting submergence tolerance

Responses of wheat (Triticum aestivum) to complete submergence are not well understood as research has focused on waterlogging (soil flooding). The aim of this study was to characterize the responses of 2 wheat cultivars differing vastly in submergence tolerance to test if submergence tolerance was linked to shoot carbohydrate consumption as seen in rice. Eighteen‐day‐old wheat cultivars Frument (intolerant) and Jackson (tolerant) grown in soil were completely submerged for up to 19 days while assessing responses in physiology, gene expression, and shoot metabolome. Results revealed 50% mortality after 9.3 and 15.9 days of submergence in intolerant Frument and tolerant Jackson, respectively, and significantly higher growth in Jackson during recovery. Frument displayed faster leaf degradation as evident from leaf tissue porosity, chlorophyll_a, and metabolomic fingerprinting. Surprisingly, shoot soluble carbohydrates, starch, and individual sugars declined to similarly low levels in both cultivars by day 5, showing that cultivar Jackson tolerated longer periods of low shoot carbohydrate levels than Frument. Moreover, intolerant Frument showed higher levels of phytol and the lipid peroxidation marker malondialdehyde relative to tolerant Jackson. Consequently, we propose to further investigate the role of ethylene sensitivity and deprivation of reactive O₂ species in submerged wheat.     

Examination of two lowland rice cultivars reveals that gibberellin-dependent early response to submergence is not necessarily mediated by ethylene

Using two lowland rice (Oryza sativa L.) cultivars we found that in both cases submerged-induced elongation early after germination depends on gibberellins (GAs). Submergence increases the content of the active GA₁ by enhancing the expression of GA biosynthesis genes, thus facilitating the seedlings to escape from the water and preventing asphyxiation. However, the two cultivars differ in their response to ethylene. The cultivar Senia (short), by contrast to cultivar Bomba (tall), does not elongate after ethylene application, and submerged-induced elongation is not negated by an inhibitor of ethylene perception. Also, while ethylene emanation in Senia is not altered by submergence, Bomba seedlings emanate more ethylene upon desubmergence, associated with enhanced expression of the ethylene biosynthesis gene OsACS5. The cultivar Senia thus allows the possibility of clarifying the role of ethylene and other factors as triggers of GA biosynthesis enhancement in rice seedlings under submergence.Key words: gibberellins, ethylene, submergence, lowland rice, Oryza sativa     

The key regulator of submergence tolerance,SUB1A,promotes photosynthetic and metabolic recovery from submergence damage in rice leaves

The submergence‐tolerance regulator, SUBMERGENCE1A (SUB1A), of rice (Oryza sativa L.) modulates gene regulation, metabolism and elongation growth during submergence. Its benefits continue during desubmergence through protection from reactive oxygen species and dehydration, but there is limited understanding of SUB1A's role in physiological recovery from the stress. Here, we investigated the contribution of SUB1A to desubmergence recovery using the two near‐isogenic lines, submergence‐sensitive M202 and tolerant M202(Sub1). No visible damage was detected in the two genotypes after 3 d of submergence, but the sublethal stress differentially altered photosynthetic parameters and accumulation of energy reserves. Submergence inhibited photosystem II photochemistry and stimulated breakdown of protein and accumulation of several amino acids in both genotypes at similar levels. Upon desubmergence, however, more rapid return to homeostasis of these factors was observed in M202(Sub1). Submergence considerably restrained non‐photochemical quenching (NPQ) in M202, whereas the value was unaltered in M202(Sub1) during the stress. Upon reaeration, submerged plants encounter sudden exposure to higher light. A greater capability for NPQ‐mediated photoprotection can benefit the rapid recovery of photosynthetic performance and energy reserve metabolism in M202(Sub1). Our findings illuminate the significant role of SUB1A in active physiological recovery upon desubmergence, a component of enhanced tolerance to submergence.     

Responses of the antioxidative enzymes in Malaysian rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) cultivars under submergence condition

The potential involvement of activated oxygen species by submergence stress was studied in two Malaysian rice cultivars, MR219-4 and MR219-9, and cultivar FR13A that is known to be tolerant to submergence. Seedlings of these three rice cultivars were subjected to different submergence periods (4, 8, and 12 days). Under 8 days of complete submergence, FR13A cultivar showed higher lipid peroxidation in terms of malondialdehyde level and activities of antioxidative enzymes, superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR) when compared to the MR219-4 and MR219-9 cultivars. MR219-9 showed higher SOD, APX, and GR activities after 12 days of submergence. The levels of SOD activity indicated that detoxification of O₂^·− to H₂O₂ was maintained at a stable level throughout the submergence stress until up to 8 days and increased rapidly at 12 days of submergence. The results indicated that tolerance to submergence in rice is associated until 8 days submergence for MR219-4 and FR13A cultivars. These findings suggested that tolerance to submergence stress in rice might be proven by increased the capacity of antioxidative system. In addition, CAT activity has much higher affinity for scavenges H₂O₂ than APX. Therefore, ascorbate glutathione cycle might be more efficient to scavenge H₂O₂.     

Relative proportion of antioxidative enzyme activity in locally grown Indian rice cultivars (Oryza sativa L.) under submergence condition

Abstract

Breeders are trying to develop true submergence-tolerant genotypes for lowland irrigated and/or rainfed areas, where rice is often affected by short-term submergence. This study was conducted to evaluate the submergence tolerance, response of antioxidative defense systems to hypoxic and re-aerated conditions, and growth and survival percentage among 19 rice genotypes. The rice seedlings submerged into concrete tanks for 10 days exhibited decreased activities of antioxidant enzymes. The antioxidant enzymes, superoxide dismutase, catalase, and ascorbate peroxidase increased, whereas polyphenol oxidase and glutathione reductase decreased in subsequent re-aerated situation in tolerant genotypes. On the other hand, the activities of enzymes decreased greatly in susceptible genotypes under hypoxic and re-aerated conditions. Complete submergence of the seedling increased the plant height of 12 genotypes. However, it decreased the dry matter production. Genotypes having better root and shoot growth maintained higher dry weight and exhibited better survival than did others. The results indicated that genotypes tolerant of flash flood maintained higher activities of antioxidant enzymes and dry matter accumulation to protect against postanoxic injury.     

Effect of nutrient application and water turbidity on submergence tolerance of rice (Oryza sativa)

Flooding imposes severe selection pressure on plants, principally because excess water in their surroundings deprives the plants from certain basic resources such as oxygen, carbon dioxide and light for photosynthesis. Here, the effects of nitrogen and phosphorus application and their application time, on four Indica rice cultivars and their tolerance to submergence under turbid and clear flood water was studied. Submergence tolerance on metabolic changes, photosynthetic (Pn) rate and ethylene accumulation were evaluated. Submergence substantially reduced survival, chlorophyll, soluble sugars, Pn rate across cultivars with drastic reduction in IR‐20. Sub1 cultivars showed greater Pn rate as compared to sensitive cultivar under submergence because of better protection of chlorophyll, more stomatal conductance leading to higher survival. Turbid water resulted in lower under‐water photosynthesis because of poor light transmission, chlorophyll retention and silt deposition and ultimately poor survival. Foliar spray of N after desubmergence along with basal P improved the chlorophyll, soluble sugars and Pn rate and extenuated ethylene accumulation and shoot elongation resulting in significantly higher survival. To the best of our knowledge, this is the first time that such a study has been performed. Crop establishment could therefore be enhanced in areas where untimely flooding is anticipated by applying basal P and foliar spray of post‐submergence N.     

Chlorophyll fluorescence parameters,CO2 photosynthetic rate and regeneration capacity as a result of complete submergence and subsequent re-emergence in rice (Oryza sativa L.)

Regeneration capacity of submerged rice (Oryza sativa) seedlings in terms of CO₂ photosynthetic rate, chlorophyll a fluorescence and chlorophyll and carbohydrate content were investigated in three Indica rice cultivars namely FR 13A, Kalaputia and IR 42 that differed in submergence tolerance. Twenty-one day old plants were completely submerged under water for 8 days. Subsequently, plants were kept under normal conditions with 5–10 cm of stagnant water above soil surface for a further period of 15 days. After complete submergence, all genotypes showed inhibition of CO₂ photosynthetic rate. Submergence treatment resulted in a significant reduction of Rubisco activity. Maximal photochemical efficiency (F_v/F_m) of PS II and area above the fluorescence curve between F_o and F_m decreased more under submergence especially in susceptible cultivar IR 42. When re-aerated, the plants recovered to various degrees. The carbohydrate content of plants was found to be significantly and positively associated with submergence tolerance and regeneration growth. The tolerant cultivar (FR 13A) could survive submergence apparently because it possessed 1.9–2.0 and 3.2–3.7-fold more non-structural carbohydrate content before and after submergence compared to the susceptible cultivar (IR 42) and it had a better capability to restore its photosynthetic capacity during post-submergence periods.     

Mechanism associated with nonstructural carbohydrate accumulation in submergence tolerant rice (Oryza sativa L.) cultivars

Nonstructural carbohydrate (NSC) accumulation in submergence tolerant rice cultivars (cv) was studied in six Indica rice [Oryza sativa (L.)] cv under control and simulated submerged conditions. Tolerant cultivars accumulated greater contents of NSC compared to the susceptible cultivars. Starch and total NSC content showed significant positive association with survival percentage. On the other hand, elongation due to submergence was significantly a negative association with survival. The CO₂ photosynthetic rate, chlorophyll content, maximum photochemical efficiency of PS II (Fv/Fm), and activities of Rubisco were not significantly different between tolerant and susceptible cv under control condition. The ADP glucose pyrophosphorylase (AGPPase) activity was significantly higher in the tolerant cv and was a positive association with starch/NSC, whereas Fructose 1,6-diphosphatase (FDPase) activity was significantly higher in susceptible cv compared to tolerant cv and was a negative association with starch/NSC. Greater activities of AGPPase along with lower activities of FDPase might facilitate greater accumulation of NSC in tolerant rice cultivars.     

Submergence-tolerant rice withstands complete submergence even in saline water: Probing through chlorophyll a fluorescence induction O-J-I-P transients

Plants experience multiple abiotic stresses during the same growing season. The implications of submergence with and without saline water on growth and survival were investigated using four contrasting rice cultivars, FR13A (submergence-tolerant, salinity-susceptible), IR42 (susceptible to salinity and submergence), and Rashpanjor and AC39416 (salinity-tolerant, submergence-susceptible). Though both FR13A and IR42 showed sensitivity to salinity, FR13A exhibited higher initial biomass as well as maintained greater dry mass under saline condition. Greater reduction of chlorophyll (Chl) contents due to salinity was observed in the susceptible cultivars, including FR13A, compared to the salinity-tolerant cultivars. Exposure of plants to salinity before submergence decreased the survival chance under submergence. Yet, survival percentage under submergence was greater in FR13A compared to other cultivars. Generally, the reduction in the Chl content and damage to PSII were higher under the submergence compared to salinity conditions. The submergence-tolerant cultivar, FR13A, maintained greater quantities of Chl during submergence compared to other cultivars. Quantification of the Chl a fluorescence transients (JIP-test) revealed large cultivar differences in the response of PSII to submergence in saline and nonsaline water. The submergence-tolerant cultivar maintained greater chloroplast structural integrity and functional ability irrespective of the quality of flooding water.     

Submergence effects on rice genotypes during seedling stage: Probing of submergence driven changes of photosystem 2 by chlorophyll a fluorescence induction O-J-I-P transients

The effects of submergence on chlorophyll (Chl) a fluorescence were compared in seven Oryza sativa (L.) cultivars, namely FR 13A, Khoda, Khadara, Kalaputia (tolerant), Sabita, and Hatipanjari (avoiding type), and IR 42 (susceptible). Seedlings were submerged for 4 d under complete darkness. Oxygen concentration of flood water decreased with the period of submergence with concomitant increase in concentration of carbon dioxide. Submergence caused diminution in the amount of total Chl. Genotypic differences were observed for Chl content and survival percentage. Quantification of the Chl a fluorescence transients (JIP-test) revealed large cultivar differences in the response of photosystem 2 (PS2) to submergence. The kinetics of Chl a fluorescence rise showed complex changes in the magnitudes and rise of O-J, J-I, and I-P phases caused by submergence. The selective suppression of the J-I phase of fluorescence especially after 2 d of submergence provided evidence for weakened electron donation from the oxygen evolving complex whereas under severe submergence stress (4 d) both O-J and J-I steps were suppressed greatly with highly suppressed P-step, which resulted in lowering of variable fluorescence. Grouping probability or energetic connectivity between PS2 obtained through JIP-test from the data after 2 d of submergence showed a direct relation with survival percentage, i.e. fluorescence measurements contained the information of the survival chance of a plant under submerged conditions. The information could be used in identifying the submergence tolerant cultivars when the damage is not very severe.     

The role of ethylene in the growth response of submerged deep water rice

We investigated the effect of partial submergence on internode elongation in a Bangladesh variety of floating or deep water rice (Oryza sativa L., cv. Habiganj Aman II). In plants which were at least 21 days old, 7 days of submergence led to a 3- to 5-fold increase in internodal length. During submergence, the ethylene concentration in the internodes increased from about 0.02 to 1 microliters per liter. Treatment of nonsubmerged plants with ethylene also stimulated internode elongation. When ethylene synthesis in partially submerged plants was blocked with aminooxyacetic acid and aminoethoxyvinylglycine, internode elongation was inhibited. This growth inhibition was reversed when ethylene biosynthesis was restored with 1-aminocyclopropane-1-carboxylic acid (ACC). Radio-labeling studies showed that ethylene in floating rice was synthesized from methionine via ACC. Internodal tissue from submerged plants had a much higher capacity to form ethylene than did internodal tissue from nonsubmerged plants. This increase in ethylene synthesis appeared to be due to enhanced ACC formation rather than to increased conversion of ACC to ethylene. Our results indicate that ethylene produced during submergence is required for the stimulation of growth in submerged floating rice plants.     

Salinity induced oxidative stress and antioxidant system in salt-tolerant and salt-sensitive cultivars of rice (Oryza sativa L.)

Indices of oxidative stress viz., superoxide radical and H₂O₂ content increased in leaves of all the cultivars with the rise in salinity level, the increase was more pronounced and significant in salt-sensitive varieties and non-significant in resistant cultivars. Except for glutathione reductase (GR), basal activities of all other antioxidative enzymes viz. superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), ascorbate peroxidase (APX) and glutathione reductase (GR) were significantly higher in leaves of all the resistant cultivars as compared to the sensitive ones. A differential response of salinity was observed on various enzymatic and non-enzymatic components of antioxidant system in leaves of salt-tolerant and salt-sensitive cultivars of rice (Oryza sativa L.). Activities of superoxide dismutase and glutathione reductase enhanced in all the tolerant cultivar while declined in the sensitive cultivars with increasing salinity from 0 to 100 mM. Salt-stress induced the activities of catalase and peroxidase in all the cultivars but the magnitude of increase was more pronounced in the sensitive cultivars than in the tolerant cultivars. Contrarily, APX activity increased in the salt-sensitive cultivars but showed no significant change in the salt-tolerant cultivars. The amount of ascorbic acid content, reduced glutathione (GSH), reduced/oxidized glutathione (GSSG) ratio was higher in leaves of the tolerant cultivars than that of the sensitive cultivars under saline conditions. It is inferred that leaves of salt-tolerant cultivars tend to attain greater capacity to perform reactions of antioxidative pathway under saline conditions to combat salinity-induced oxidative stress.     

Relationship between Coleoptile Elongation and Alcoholic Fermentation in Rice Exposed to Anoxia. II. Cultivar Differences

The relationship between coleoptile elongation and survivalvs. alcoholic fermentation of rice under anoxia is examinedusing eight cultivars differing in submergence tolerance. Anoxiawas imposed on either 1 or 4 d aerated seeds either by N₂ flushingsubmerged tissues or by incubating tissues in stagnant deoxygenatedagar at 0·1% w/v; the latter simulated the stagnant conditionsof waterlogged soil. Two cultivars that were most tolerant tosubmergence also had the greatest tolerance to anoxia, whilea submergence intolerant cultivar was also intolerant to anoxia. Coleoptile growth under anoxia was related to rates of ethanolsynthesis (R_E), however differences between growth during anoxiaand survival after anoxia indicated that post-anoxic injurymay also be important in rice seeds exposed to anoxia. The correlationbetween coleoptile growth and R_E measured on a tissue basisusing intact seeds was r² = 0·67 among six varietiesover 0-3 d anoxia. This correlation improved to about r² = 0·85using R_E of (embryos plus coleoptiles) over 0-3 d, or coleoptilesat 3 d after anoxia. Coleoptile growth of individual seeds wasusually poorly correlated to R_E in these cultivars at 2-3 dafter anoxia. When coleoptiles of similar lengths were obtainedfrom different cultivars using 4 d aerated seeds, there weredifferences in R_E and coleoptile growth which were related tocoleoptile growth during 3 or 5 d anoxia, either on a tissue(r² = 0·85) or a fresh weight basis (r² = 0·70-0·97respectively). Results are discussed in relation to factorswhich may limit ethanol synthesis in rice exposed to anoxiaand the importance of growth to the survival of seeds and matureplants during submergence in the natural environment.Copyright1994, 1999 Academic Press Anoxia, ethanol, alcoholic fermentation, Oryza sativa L., rice, submergence     

Starch Phosphorylase Enzyme is Over-expressed in Submerged Rice (Oryza sativa L) Plant

Activity of starch phosphorylase enzyme and its isozymes were studied in one-week and two-week-old seedlings of four cultivars of rice, viz. FR13A, Gangasiuli (submergence tolerant), Sarala and IR42 (submergence susceptible) under submerged as well as control conditions. In one-week-old seedlings there was no apparent increase in enzyme activity, whereas in the two-week-old seedlings the activity rose steadily and reached to a maximum after 144 h of submergence and then declined. The increase in activity was also reflected in the isozyme profiles of this enzyme wherein both qualitative as well as quantitative (suppression and over-expression of certain bands) differences were observed. It was concluded that, in addition to other enzymes of starch degradation, starch phosphorylase too may be involved in starch degradation during submergence stress in rice plants.     

Responses of antioxidative system to chilling stress in two rice cultivars differing in sensitivity

The responses of antioxidative system of rice to chilling were investigated in a tolerant cultivar, Xiangnuo-1, and a susceptible cultivar, IR-50. The electrolyte leakage and malondialdehyde content of Xiangnuo-1 were little affected by chilling treatment but those of IR-50 increased. Activities of suoperoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase, and ascorbic acid content of Xiangnuo-1 were remained high, while those of IR-50 decreased under chilling. The results indicated that higher activities of defense enzymes and higher content of antioxidant under stress were associated with tolerance to chilling.     

Morphological and physiological responses of rice seedlings to complete submergence (flash flooding)

Background and Aims

Reducing damage to rice seedlings caused by flash flooding will improve the productivity of rainfed lowland rice in West Africa. Accordingly, the morphological and physiological responses of different forms of rice to complete submergence were examined in field and pot experiments to identify primary causes of damage.

Methods

To characterize the physiological responses, seedlings from a wide genetic base including Oryza sativa, O. glaberrima and interspecific hybrids were compared using principle component analysis.

Key Results

Important factors linked to flash-flood tolerance included minimal shoot elongation underwater, increase in dry matter weight during submergence and post-submergence resistance to lodging. In particular, fast shoot elongation during submergence negatively affected plant growth after de-submergence. Also shoot-elongating cultivars showed a strong negative correlation between dry matter weight of the leaves that developed before submergence and leaves developing during submergence.

Conclusions

Enhancement of shoot elongation during submergence in water that is too deep to permit re-emergence by small seedlings represents a futile escape strategy that takes place at the expense of existing dry matter in circumstances where underwater photosynthetic carbon fixation is negligible. Consequently, it compromises survival or recovery growth once flood water levels recede and plants are re-exposed to the aerial environment. Tolerance is greater in cultivars where acceleration of elongation caused by submergence is minimal.Key words: Africa, flash floods, Oryza glaberrima, rainfed lowland, rice, shoot elongation, stress tolerance, submergence     

Injury to Rice Plants Caused by Complete Submergence; A Contribution by Ethylerie (Ethene)

Complete submergence of rice plants (Oryza sativa L. cv. ‘IR42’)in dilute nutrient solution for 3–6 d almost stopped theaccumulation of dry matter, depressed soluble carbohydrate concentrationby over 75% and promoted chlorosis in fully expanded leaves.Increase in fresh weight by the shoots was not impaired. Extensionby the youngest visible leaf was stimulated. Extension by thenext leaf to appear was retarded by submergence. These growthresponses to submergence were associated with a 1-5-fold increasein the partial pressure of endogenous ethylene (ethene). Applying ethylene (0.3–0.35 Pa) in the gas-phase to non-submergedplants reproduced some, but not all, of these effects of submergence.Thus, greater leaf extension and chlorosis of submerged plantscould be attributable to accumulated ethylene but neither theslow relative growth rate nor the decreased extension of leavesemerging after the start of submergence could be so attributed. Two cultivars (‘FR13A’ and ‘Kurkaruppan’)already known to tolerate submergence, differed little fromsubmergence-intolerant ‘IR42’ in their relativegrowth rate and soluble carbohydrate concentration during submergence.However, their underwater leaf extension was less than in ‘IR42’and chlorosis was much less prevalent, especially in ‘FR13A’.Similarly, ethylene supplied to non-submerged plants was a lesseffective promotor of leaf extension and chlorosis in the twosubmergence tolerant cultivars. Application of 1.0 kPa carbondioxide in the gas-phase prevented the chlorosis response toethylene. The results indicate that accumulated ethylene is a likely causeof fast leaf extension and chlorosis in submergence intolerantforms of rice, particularly when amounts of dissolved carbondioxide are minimal. Key words: Oryza sativa L., aeration, ethylene (ethene), stress-tolerance     

Distinct mechanisms for aerenchyma formation in leaf sheaths of rice genotypes displaying a quiescence or escape strategy for flooding tolerance

Background and Aims

Rice is one of the few crops able to withstand periods of partial or even complete submergence. One of the adaptive traits of rice is the constitutive presence and further development of aerenchyma which enables oxygen to be transported to submerged organs. The development of lysigenous aerenchyma is promoted by ethylene accumulating within the submerged plant tissues, although other signalling mechanisms may also co-exist. In this study, aerenchyma development was analysed in two rice (Oryza sativa) varieties, ‘FR13A’ and ‘Arborio Precoce’, which show opposite traits in flooding response in terms of internode elongation and survival.

Methods

The growth and survival of rice varieties under submergence was investigated in the leaf sheath of ‘FR13A’ and ‘Arborio Precoce’. The possible involvement of ethylene and reactive oxygen species (ROS) was evaluated in relation to aerenchyma formation. Cell viability and DNA fragmentation were determined by FDA/FM4-64 staining and TUNEL assay, respectively. Ethylene production was monitored by gas chromatography and by analysing ACO gene expression. ROS production was measured by using Amplex Red assay kit and the fluorescent dye DCFH₂-DA. The expression of APX1 was also evaluated. AVG and DPI solutions were used to test the effect of inhibiting ethylene biosynthesis and ROS production, respectively.

Key Results

Both the varieties displayed constitutive lysigenous aerenchyma formation, which was further enhanced when submerged. ‘Arborio Precoce’, which is characterized by fast elongation when submerged, showed active ethylene biosynthetic machinery associated with increased aerenchymatous areas. ‘FR13A’, which harbours the Sub1A gene that limits growth during oxygen deprivation, did not show any increase in ethylene production after submersion but still displayed increased aerenchyma. Hydrogen peroxide levels increased in ‘FR13A’ but not in ‘Arborio Precoce’.

Conclusions

While ethylene controls aerenchyma formation in the fast-elongating ‘Arborio Precoce’ variety, in ‘FR13A’ ROS accumulation plays an important role.     

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.