Severe reduction in growth rate and grain filling of rice mutants lacking OsGS1;1, a cytosolic glutamine synthetase1;1

Rice (Oryza sativa L.) plants possess three homologous but distinct genes for cytosolic glutamine synthetase (GS1): these are OsGS1;1, OsGS1;2, and OsGS1;3. OsGS1;1 was expressed in all organs tested with higher expression in leaf blades, while OsGS1;2, and OsGS1;3 were expressed mainly in roots and spikelets, respectively. We characterized knockout mutants caused by insertion of endogenous retrotransposon Tos17 into the exon-8 (lines ND8037 and ND9801) or the exon-10 (line NC2327) of OsGS1;1. Mendelian segregation occurred in each progeny. Homozygously inserted mutants showed severe retardation in growth rate and grain filling when grown at normal nitrogen concentrations. Abnormal mRNA for GS1;1 was transcribed, and the GS1 protein and its activity in the leaf blades were barely detectable in these mutants. The glutamine pool in the roots and leaf blades of the mutants was lower than that of the wild type. Re-introduction of OsGS1;1 cDNA under the control of its own promoter into the mutants successfully complemented these phenotypes. Progeny where Tos17 was heterozygously inserted or deleted during segregation showed normal phenotypes. The results indicate that GS1;1 is important for normal growth and grain filling in rice; GS1;2 and GS1;3 were not able to compensate for GS1;1 function.     

不同耐旱性水稻幼苗对氧化胁迫的反应

干旱条件下水稻(Oryza sativa L.)耐旱性较强的“湘中籼2号”比耐旱性较弱的“湘早釉12号”O_2~-的产生速率低。百草枯(0.01mmol/L)和H_2O_2(10mmol/L)处理促进干旱对水稻幼苗的伤害,但耐旱性较强的品种伤害较轻。单独用百草枯处理后,耐旱性较强的品种比耐旱性较弱的品种能更大幅度地提高超氧物歧化酶和过氧化物酶的活性;而H_2O_2处理后耐旱性较强的品种能提高过氧化氢酶活性。这些结果表明,耐旱性较强的水稻品种有较强的抗氧化胁迫能力。     

Differential Drought Responses in F-box Gene Expression and Grain Yield Between Two Rice Groups with Contrasting Drought Tolerance

Understanding the molecular mechanisms regulating rice’s response to drought stress is important for cereal crop development. We investigated the physiological and gene expression responses of three drought-tolerant and two drought-sensitive unrelated rice cultivars at the reproductive stage. Leaf water loss and leaf rolling tests distinguished these two groups from each other. Both 7 and 14 days of drought stress affected most of the tested agronomic traits including grain yield, and the effects were stronger in the drought-sensitive cultivars. Fourteen days of drought stress severely reduced grain yield in the sensitive cultivars. Expression levels of 50 genes previously published were examined in panicles. Of these, 25 genes were expressed in panicles and could be classified into 6 groups. LOC_Os02g51350, a Kelch domain-containing F-box gene, was selected for further studies including expression analysis in panicles during the 14-day drought stress, domain analysis, and analysis of cis-acting elements in the promoters. All three drought-resistant cultivars possessed the F-box domain, which was absent in the two drought-sensitive cultivars. In addition, drought-related ABRE and DRE/CRT cis-acting elements were more abundant in Os02g51350 promoters of cultivars with good grain yield under drought stress than in promoters of cultivars with severe yield reduction. Our results suggest that the F-box version of Os02g51350 is important for maintenance of grain yield under drought.     

Overexpression of the glutamine synthetase gene modulates oxidative stress response in rice after exposure to cadmium stress

Key message

Overexpression of OsGS gene modulates oxidative stress response in rice after exposure to cadmium stress. Our results describe the features of transformants with enhanced tolerance to Cd and abiotic stresses.

Abstract

Glutamine synthetase (GS) (EC 6.3.1.2) is an enzyme that plays an essential role in the metabolism of nitrogen by catalyzing the condensation of glutamate and ammonia to form glutamine. Exposure of plants to cadmium (Cd) has been reported to decrease GS activity in maize, pea, bean, and rice. To better understand the function of the GS gene under Cd stress in rice, we constructed a recombinant pART vector carrying the GS gene under the control of the CaMV 35S promoter and OCS terminator and transformed using Agrobacterium tumefaciens. We then investigated GS overexpressing rice lines at the physiological and molecular levels under Cd toxicity and abiotic stress conditions. We observed a decrease in GS enzyme activity and mRNA expression among transgenic and wild-type plants subjected to Cd stress. The decrease, however, was significantly lower in the wild type than in the transgenic plants. This was further validated by the high GS mRNA expression and enzyme activity in most of the transgenic lines. Moreover, after 10 days of exposure to Cd stress, increase in the glutamine reductase activity and low or no malondialdehyde contents were observed. These results showed that overexpression of the GS gene in rice modulated the expression of enzymes responsible for membrane peroxidation that may result in plant death.     

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.     

High content of cytosolic glutamine synthetase does not accompany a high activity of the enzyme in rice (Oryza sativa) leaves of indica cultivars

Reproductive stages of 5 japonica, 9 indica, and 2 javanica cultivars of rice ( Oryza sativa L.) were provided to compare the contents of protein for cytosolic glutamine synthetase (GS1; EC 6.3.1.2) in the lowest position of the attached leaf blade (position 6 from the primary leaf) and those for NADH-glutamate synthase (NADH-GOGAT; EC 1.4.1.14) in non-green portion of the expanding 10th leaf blade. Some of the indica cultivars, including Kasalath, contained GS1 protein twice as high as other japonica and javanica cultivars based on total leaf nitrogen. Most of the indica cultivars, on the other hand, contained less NADH-GOGAT protein than japonica and javanica cultivars. Immunostaining proved that GS1 protein was located in vascular tissues of the leaf blades of Kasalath, which was identical to our previous results with a japonica cultivar [Sakurai et al. (1996) Planta 200: 306–311]. Although relative contents of GS1 protein in the leaf blade of Kasalath increased as a function of leaf age, GS1 activity remained relatively constant. In addition, Kasalath showed lower activity than other japonica and javanica cultivars, especially during leaf expansion. GS1 activity, based on GS1 protein amount, changed during the life span of the leaf blade and we thus assume that GS1 activity was modulated post-translationally in rice leaves.     

水稻幼苗叶绿体保护系统对干旱的反应

干旱条件下水稻耐旱品种湘中籼２号比不耐旱品种湘早籼１２号具有较低的质膜相对透性、叶绿素含量降低较少,表明耐旱品种受伤害较轻,两个品种的抗坏血酸。过氧化物酶（ＡｓＡ－ＰＯＤ）均降低,但耐旱品种降低较少。耐旱品种的谷胱甘肽还原酶（ＧＲ）活性在干旱初期提高,不耐旱品种的则变化不大。０．０１ｍｍｏｌ／Ｌ的百草枯和０．１ｍｍｏｌ／Ｌ的Ｈ２Ｏ２亦能诱导耐旱品种的ＧＲ活性,但对不耐旱品种的ＧＲ活性不受影响,说明可能是干旱诱导的氧化胁迫诱导了ＧＲ活性。干旱条件下两品种的抗坏血酸（ＡｓＡ）和还原型谷胱甘肽（ＧＳＨ）含量均是先升后降,但耐旱品种提高幅度较大。结果表明,耐旱品种的叶绿体保护系统的活性比不耐旱品种的高。     

Activity of the tetramer and octamer of glutamine synthetase isoforms during primary leaf ontogeny of sugar beet (Beta vulgaris L.)

In extracts from the primary leaf blade of sugar beet (Beta vulgaris L.) we separated a chloroplastic isoform (GS 2) of glutamine synthetase (GS, EC 6.3.1.2) and one or two (depending on leaf age) cytosolic isoforms (GS 1a and GS 1b). The latter were prominent in the early (GS 1a) and late stages of leaf ontogeny (GS 1a and GS 1b), whereas during leaf maturation GS 2 was the predominantly active GS isoform. The GS 1 isoforms were active exclusively in the octameric state although tetrameric GS 1 protein was detected immunologically. Their activity stayed at a relatively constant level during leaf ontogeny; an increase was observed only in the senescent leaf. The activity of GS 2, however, changed drastically during primary leaf ontogeny and was modulated by changes in the oligomeric state of the active enzyme. In the early and late stages of leaf ontogeny when GS 2 activity was low (lower than that of the GS 1 isoforms), GS 2 was active only in the octameric state. In the maturing leaf, when GS 2 activity had reached its maximum level (much higher than that of the GS 1 isoforms), 80 of total GS 2 activity was due the activity of the tetrameric form of the enzyme and 20 was due to octameric GS 2. Tetrameric GS 2 was a hetero-tetramer and thus not the unspecific dissociation product of homo-octameric GS 2. In addition, GS 2 activity was modulated by an activation/inactivation of the tetrameric GS 2 protein. Due to an activation of the GS 2 tetramer, the activity of tetrameric GS 2 increased during leaf maturation from zero level 23-fold compared with that of GS 1a and 18-fold compared with that of GS 1b. Possible activators of tetrameric GS 2 are thiol-reactive substances. During leaf senescence, GS 2 activity decreased to zero; this decrease was due to an inactivation of the tetrameric GS 2 protein probably caused by oxidation.Abbreviations FLL final lamina length - FPLC fast protein liquid chromatography - GS glutamine synthetase - GHA -glutamyl hydroxamate - Rubisco ribulose-1,5-bisphosphate carboxylase/oxygenase Dr. Roger Wallsgrove's (Rothamsted Experimental Station, Harpenden, UK) generous gift of GS antiserum is greatly appreciated.     

Biochar coupling with phosphorus fertilization modifies antioxidant activity,osmolyte accumulation and reactive oxygen species synthesis in the leaves and xylem sap of rice cultivars under high-temperature stress

Increasing temperature poses a serious threat to rice productivity. This study investigated the impact of various biochar treatments and phosphorous (P) fertilization on osmolyte accumulation, ROS development, and antioxidant activity in two rice cultivars (IR-64 and Huanghuazhan) under high-temperature stress. All plants of both cultivars were grown in a controlled environment under ambient temperatures (AT), high day temperatures (HDT) or high night temperatures (HNT). The different fertilization treatments were biochar alone, P alone and biochar + P with control. In the leaves and xylem sap of both rice cultivars, particularly in the susceptible cv. IR-64, high-temperature stress increased the production of MDA and H₂O₂. HDT and HNT decreased total soluble sugars, protein, and proline levels in both rice cultivars. HNT was observed as more harmful compared to HDT during most of the studied characteristics. The response of antioxidant enzyme activities, viz, SOD, POD, CAT, APX, ASC, GSH, GR, and GSSC activities, to the temperature treatments varied between the two cultivars. Antioxidant activities decreased in the leaves and xylem sap of IR-64 but increased in those of Huanghuazhan upon exposure to high-temperature stress. Huanghuazhan exhibited better heat tolerance compared to IR-64, which was linked to its increased antioxidant enzyme activation and metabolite synthesis. As compared to the control, all soil fertilization treatments considerably reduced the adverse impacts of high temperature on the rice cultivars. The combination of biochar and P resulted in better performance compared to the other treatments in terms of all studied attributes.     

Utility of phyllosphere N2-fixing micro-organisms in the improvement of crop growth

Summary The beneficial effect of spraying some highly active phyllosphere N₂-fixing microorganisms on high and low yielding cultivars of rice plants as compared with that of urea applied at different doses are described. The dry weight, N-content, 1000 grain weight, and yield were remarkably increased in all cases with the application of phyllosphere microorganisms. The performance of two isolates KUP₄ and KUPBR₂ with IR-8 and IR-26 rice, was better than that of 52 kg urea-N per hectare. IR-579 rice leaves in association with some phyllosphere bacteria reduced acetylene at the rate of 664–816 nmoles/g leaf/h. In IR-26 rice the effect of application of bacterial suspension at three phases of plant growth corresponded very well with that of urea application in three split doses under identical conditions. Recommended fertilizer rates produced the same yield as the half dose plus bacterial spray in the cultivars Pankaj and Rupsail. Fertilizer application in Pankaj and Rupsail rice reduced nitrogenase activity and the beneficial effects of phyllosphere N₂-fixation was reduced by 40–55%.     

Metabolomics data reveal a crucial role of cytosolic glutamine synthetase 1;1 in coordinating metabolic balance in rice

Rice plants grown in paddy fields preferentially use ammonium as a source of inorganic nitrogen. Glutamine synthetase (GS) catalyses the conversion of ammonium to glutamine. Of the three genes encoding cytosolic GS in rice, OsGS1;1 is critical for normal growth and grain filling. However, the basis of its physiological function that may alter the rate of nitrogen assimilation and carbon metabolism within the context of metabolic networks remains unclear. To address this issue, we carried out quantitative comparative analyses between the metabolite profiles of a rice mutant lacking OsGS1;1 and its background wild type (WT). The mutant plants exhibited severe retardation of shoot growth in the presence of ammonium compared with the WT. Overaccumulation of free ammonium in the leaf sheath and roots of the mutant indicated the importance of OsGS1;1 for ammonium assimilation in both organs. The metabolite profiles of the mutant line revealed: (i) an imbalance in levels of sugars, amino acids and metabolites in the tricarboxylic acid cycle, and (ii) overaccumulation of secondary metabolites, particularly in the roots under a continuous supply of ammonium. Metabolite-to-metabolite correlation analysis revealed the presence of mutant-specific networks between tryptamine and other primary metabolites in the roots. These results demonstrated a crucial function of OsGS1;1 in coordinating the global metabolic network in rice plants grown using ammonium as the nitrogen source.     

Ammonia emission from rice leaves in relation to photorespiration and genotypic differences in glutamine synthetase activity

Background and Aims

Rice (Oryza sativa) plants lose significant amounts of volatile NH₃ from their leaves, but it has not been shown that this is a consequence of photorespiration. Involvement of photorespiration in NH₃ emission and the role of glutamine synthetase (GS) on NH₃ recycling were investigated using two rice cultivars with different GS activities.

Methods

NH₃ emission (AER), and gross photosynthesis (P_G), transpiration (Tr) and stomatal conductance (g_S) were measured on leaves of ‘Akenohoshi’, a cultivar with high GS activity, and ‘Kasalath’, a cultivar with low GS activity, under different light intensities (200, 500 and 1000 µmol m⁻² s⁻¹), leaf temperatures (27·5, 32·5 and 37·5 °C) and atmospheric O₂ concentrations ([O₂]: 2, 21 and 40 %, corresponding to 20, 210 and 400 mmol mol⁻¹).

Key Results

An increase in [O₂] increased AER in the two cultivars, accompanied by a decrease in P_G due to enhanced photorespiration, but did not greatly influence Tr and g_S. There were significant positive correlations between AER and photorespiration in both cultivars. Increasing light intensity increased AER, P_G, Tr and g_S in both cultivars, whereas increasing leaf temperature increased AER and Tr but slightly decreased P_G and g_S. ‘Kasalath’ (low GS activity) showed higher AER than ‘Akenohoshi’ (high GS activity) at high light intensity, leaf temperature and [O₂].

Conclusions

Our results demonstrate that photorespiration is strongly involved in NH₃ emission by rice leaves and suggest that differences in AER between cultivars result from their different GS activities, which would result in different capacities for reassimilation of photorespiratory NH₃. The results also suggest that NH₃ emission in rice leaves is not directly controlled by transpiration and stomatal conductance.     

Occluded C in rice phytoliths: implications to biogeochemical carbon sequestration

Aims

Carbon (C) bio-sequestration within the phytoliths of plants, a mechanism of long-term biogeochemical C sequestration, may play a major role in the global C cycle and climate change. In this study, we explored the potential of C bio-sequestration within phytoliths produced in cultivated rice (Oryza sativa), a well known silicon accumulator.

Methods

The rice phytolith extraction was undertaken with microwave digestion procedures and the determination of occluded C in phytoliths was based on dissolution methods of phytolith-Si.

Results

Chemical analysis indicates that the phytolith-occluded C (PhytOC) contents of the different organs (leaf, stem, sheath and grains) on a dry weight basis in 5 rice cultivars range from 0.4 mg?g^?1 to 2.8 mg?g^?1, and the C content of phytoliths from grains is much lower than that of leaf, stem and sheath. The data also show that the PhytOC content of rice depends on both the content of phytoliths and the efficiency of C occlusion within phytoliths during rice growth. The biogeochemical C sequestration flux of phytoliths in 5 rice cultivars is approximately 0.03–0.13 Mg of carbon dioxide (CO₂) equivalents (Mg-e-CO₂) ha^?1?year^?1. From 1950 to 2010, about 2.37?×?10⁸?Mg of CO₂ equivalents might have been sequestrated within the rice phytoliths in China. Assuming a maximum phytoliths C bio-sequestration flux of 0.13 Mg-e-CO₂ ha^?1?year^?1, the global annual potential rate of CO₂ sequestrated in rice phytoliths would approximately be 1.94?×?10⁷?Mg.

Conclusions

Therefore rice crops may play a significant role in long-term C sequestration through the formation of PhytOC.     

Arsenic Toxicity is Counteracted by Exogenous Application of Melatonin to Different Extents in Arsenic-susceptible and Arsenic-tolerant Rice Cultivars

The current study established the protective effects of exogenous melatonin in ameliorating arsenic toxicity in Khitish (arsenic-sensitive) and Muktashri (arsenic-tolerant) rice cultivars. Melatonin highly improved the overall growth performance of arsenic-treated seedlings, more prominently in the sensitive variety, Khitish. Although the level of arsenic increased in both the cultivars, Khitish accumulated comparatively higher arsenic level. However, melatonin supplementation reduced arsenic bioaccumulation and restored physiological growth attributes, as supported by lowering of electrolyte leakage, chlorophyll loss (by inducing RuBisCo), protein carbonylation, malondialdehyde accumulation, lipoxygenase (LOX), NADPH oxidase (NOX) and protease activity, and improvement of membrane stability index. Isoforms of LOX and NOX showed varietal differences during arsenic stress, both in the presence and absence of melatonin. Melatonin reduced methylglyoxal content during arsenic stress, concomitant with down-regulated gene expression and enzyme activity for glyoxalases. The nitrogen assimilation was improved via induced nitrate reductase (NR) activity and NR expression. The variable accumulation of osmolytes like proline, glycine betaine and total amino acids, concomitant with suppressed P5CS and BADH1 expression, and induced PDH was noteworthy. Antioxidant metabolites like anthocyanins, flavonoids, carotenes, xanthophylls and total phenolics were accumulated upon supplementation of melatonin in arsenic-stressed Khitish, supported by the activation of ANS and PSY genes. Melatonin lowered the ascorbic-acid oxidase activity and restored the ascorbate sink in arsenic-affected seedlings. Overall, the study revealed the potential role of exogenous melatonin in mitigating arsenic-induced injuries by strengthening osmolytes and antioxidative machinery, leading to the restoration of growth and metabolism in rice, especially in the susceptible cultivar.

     

Differential changes in cell wall matrix polysaccharides and glycoside-hydrolyzing enzymes in developing wheat seedlings differing in drought tolerance

The growth kinetics and variations in cell wall matrix polysaccharides and glycoside hydrolases during seedling development of the drought-tolerant wheat cultivar (cv. Hong Mang Mai) were compared with the drought-sensitive cultivar (cv. Shirasagikomugi). After 15d of culture in water at 22 degrees C under constant irradiance of 98mumolm(-2)s(-1), the length of the coleoptile and leaf sheath of Hong Mang Mai seedlings was 1.7 times longer than those of Shirasagikomugi seedlings. In the cell walls isolated from coleoptiles and leaf sheaths of the seedling of the two cultivars, the contents of arabinose, xylose, and glucose changed during development. The cell walls were fractionated progressively with 50mM CDTA, 50mM Na(2)CO(3), 1M KOH and 4M KOH, and sugar composition was determined. The amount of CDTA-soluble fraction from the Hong Mang Mai cell walls was 2.4-fold higher than that from the Shirasagikomugi cell walls at 6d of culture, and a considerable decrease was observed during development. The ratio of arabinose to xylose in 1M KOH-soluble fraction from the two cultivars decreased. The amount of 4M KOH-soluble fraction from the Shirasagikomugi cell walls was affected much more than those of the Hong Mang Mai cell walls. Many glycoside hydrolase activities were detected in the protein fractions from coleoptiles and leaf sheaths of the two cultivars, and the activities of licheninase, 1,3-1,4-beta-glucanase, and 1,3-beta-glucanase in the LiCl-soluble protein fraction increased drastically during development of the Shirasagikomugi seedlings. These findings suggest that the metabolism of the cell wall matrix polysaccharides of the drought-tolerant wheat cultivar is far different from that of the drought-sensitive wheat cultivar during seedling development.     

Regional-scale patterns of δ<Superscript>13</Superscript>C and δ<Superscript>15</Superscript>N associated with multiple ecosystem functions along an aridity gradient in grassland ecosystems

Background and aims

Nitrogen (N) deficiency and drought are two key limiting factors for rice production worldwide, but the relationship of drought stress with N homeostasis in rice is rarely advanced. The aim of this study was to dissect the physiological effects of drought stress on rice growth that coupled unbalanced N metabolism.

Results

Water-deficient stress (WD) limited stomatal aperture function and activity of Rubisco carboxylase to photosynthesis. The rate of total electron transport (J_t) and the electron to carboxylation (J_c) were considerably decreased, whereas the proportion of e^? flow to photorespiration was stimulated by WD, especially at 1600 μmol m^?2 s^?1 PPFD. Concurrently, the expressions of glycolate oxidase genes (GOX1, GOX5) and glycine decarboxylase complex (GDCH, GDCP and GDCT) were significantly induced in leaves of WD treatment, which led to the accumulation of reactive oxygen species in leaves. With the photosynthetic change, nitrate uptake and reduction were suppressed. Moreover, the enhanced photorespiration generated excess NH₃ accumulation in leaves and stimulated the expressions of GS1;1, GS1;2 and GS2, which were tightly coupled with the expressions of PEPC1 and PEPC2 under WD stress.

Conclusions

Our results suggest that the inhibited nitrate reduction associated with diminished electron transport rate, and the photorespiration-associated accumulation of hydrogen peroxide and NH₃ were critical in the drought-induced rice growth inhibition.

     

Expansin-regulated cell elongation is involved in the drought tolerance in wheat

Water stress restrains plant growth. Expansin is a cell wall protein that is generally accepted to be the key regulator of cell wall extension during plant growth. In this study, we used two different wheat cultivars to study the involvement of expansin in drought tolerance. Wheat coleoptile was used as the material in experiment. Our results indicated that water stress induced an increase in acidic pH-dependant cell wall extension, which is related to expansin activity; however, water stress inhibited coleoptile elongation growth. The increased expansin activity was mainly due to increased expression of expansin protein that was upregulated by water stress, but water stress also resulted in a decrease in cell wall acidity, a negative factor for cell wall extension. Decreased plasma membrane H⁺-ATPase activity was involved in the alkalinization of the cell wall under water stress. The activity of expansin in HF9703 (a drought-tolerant wheat cultivar) was always higher than that in 921842 (a drought-sensitive wheat cultivar) under both normal and water stress conditions, which may be correlated with the higher expansin protein expression and plasma membrane H⁺-ATPase activity observed in HF9703 versus 921842. However, water stress did not change the susceptibility of the wheat cell wall to expansin, and no difference in this susceptibility was observed between the drought-tolerant and drought-sensitive wheat cultivars. These results suggest the involvement of expansin in cell elongation and the drought resistance of wheat.