Proteomic Analysis of Low Nitrogen Stress-Responsive Proteins in Roots of Rice

In recent years, the application of proteomic approaches as a tool for global expression analysis and protein identification has been highly efficient in the field of plant research. A solution culture experiment involving two nitrogen treatments, 0.14 mM NH₄NO₃ (low nitrogen (N)) and 1.07 mM NH₄NO₃ (control), was conducted to investigate the response of rice root to low N stress. Root system architecture changed markedly under low N stress, with more lateral roots occurring on the lower part of adventitious roots and longer lateral roots on the upper part, compared to the control. A proteomic approach was employed to further study the rice responses to low N stress. Proteins extracted from roots were profiled by two-dimensional gel electrophoresis, and differentially expressed proteins were analyzed by mass spectrometry. Twelve protein spots were successfully identified by mass spectrometry, 11 of which had known functions. Of these, four were involved with the tricarboxylic acid cycle, two with adenylate metabolism, two with phenylpropanoid metabolism, and two with protein degradation. These differentially expressed proteins play an important role in the responsive mechanisms of rice root to low N stress, and uncovering how the rice proteins respond to low N stress could contribute to improving the nitrogen use efficiency.     

Identification of Cu-binding proteins in embryos of germinating rice in response to Cu toxicity

Seed germination, an early and important process for the growth and development of plants, is hypersensitive to environmental changes. Copper (Cu) is a necessary micronutrient for plants; however, an excessive dose of Cu had an extremely negative effect at the cellular level as a result of inevitable binding to proteins. In contrast, some structural motifs of proteins can bind free Cu ions and relieve Cu toxicity. This study aimed to understand the expression characteristics of Cu-binding proteins induced by excess Cu during rice seed germination. We investigated Cu-binding proteins in germinating rice embryos treated with 200 µM Cu using a Sephadex G-50 column or immobilized Cu affinity chromatography combined with two-dimensional gel electrophoresis. Proteomics analysis indicated that 12 protein spots exhibited a?>?2.0-fold increase in intensity in response to Cu toxicity as compared with controls. Among nine proteins in ten spots identified as Cu-binding proteins, three proteins (from four spots) were involved in antioxidative defense: copper, zinc superoxide dismutase, glutathione S-transferase and protein disulfide isomerase. These results show that reactive oxygen species may be involved in the expression regulation of Cu-binding proteins in germinating rice in response to Cu stress.     

A Full-Length Dof1 Transcription Factor of Finger Millet and its Response to a Circadian Cycle

DOF1 (DNA binding with one finger) plays an important role in regulating C/N metabolism in cereals. In order to validate its role in the regulation of nitrogen use efficiency (NUE) and photosynthetic efficiency in finger millet, 5′–3′ RACE PCR was performed to obtain and characterize full-length Dof1 genes of high and low grain protein finger millet genotypes. The full-length DOF1 ORFs were both 1,284 nt long and were 98.8 % similar over 427 amino acids containing the characteristic Dof domain. Comparison of both the EcDof1 protein sequences with the Dof1 of other cereals revealed high sequence similarity to the Dof1 of rice. Southern hybridization carried out using the probe developed from the region encoding the highly variable C-terminal region of EcDof1 showed the presence of four copies of the DOF1 gene in finger millet, which might explain the high NUE and photosynthetic performance of finger millet. Since the genes involved in C/N metabolism are regulated diurnally and play crucial roles in determining grain protein content during grain filling, the diurnal expression of EcDOF1 was assessed in two finger millet genotypes (GE 3885 and GE 1437) with differing grain protein content (13.8 % and 6.15 % respectively). It was found that EcDOF1 exhibited diurnal regulation and peak differential pattern expression with early phasing in GE3885 and late phasing in GE1437. Differential expression of DOF1 might alter the regulation of genes involved in C/N metabolism affecting grain protein composition of finger millet genotypes.     

The alleviation of zinc toxicity by silicon is related to zinc transport and antioxidative reactions in rice

The objective of this study is to elucidate the roles of silicon (Si) in enhancing tolerance to excess zinc (Zn) in two contrasting rice (Oryza sativa L.) cultivars: i.e. cv. TY-167 (Zn-resistant) and cv. FYY-326 (Zn-sensitive). Root morphology, antioxidant defense reactions and lipid peroxidation, and histochemical staining were examined in rice plants grown in the nutrient solutions with normal (0.15 μM) and high (2 mM) Zn supply, without or with 1.5 mM Si. Significant inhibitory effects of high Zn treatment on plant growth were observed. Total root length (TRL), total root surface area (TRSA) and total root tip amount (TRTA) of both cultivars were decreased significantly in plants treated with high Zn, whereas these root parameters were significantly increased when Zn-stressed plants were supplied with 1.5 mM Si. Supply of Si also significantly decreased Zn concentration in shoots of both cultivars, indicating lower root-to-shoot translocation of Zn. Moreover, superoxide dismutase (SOD), catalase (CAT), and asorbate peroxidase (APX) activities were increased, whereas malondialdehyde (MDA) and hydrogen peroxide (H₂O₂) concentrations were decreased in Si-supplied plants of both Zn-sensitive and Zn-resistant rice cultivars exposed to Zn stress. These alleviative effects of Si, further confirmed by the histochemical staining methods, were more prominent in the Zn-resistant cultivar than in the Zn-sensitive one. Taken together, all these results suggest that Si-mediated alleviation of Zn toxicity is mainly attributed to Si-mediated antioxidant defense capacity and membrane integrity. The possible role of Si in reduction of root-to-shoot translocation of Zn can also be considered.     

Nitrogen Use Efficiency of Rice Cultivars (Oryza sativa L.) Under Salt Stress and Low Nitrogen Conditions

Improving nitrogen use efficiency (NUE) under salt stress has become crucial for rice as it is increasingly facing two major environmental constraints: excessive nitrogen fertilization and soil salinization. However, the interaction between salinity and N levels is very complex and has not yet been considered from the perspective of reduced nitrogen input. We conducted a hydroponic experiment at the early tillering stage on the Yoshida solution to evaluate the impact of rising NaCl and decreasing N application on NUE of four rice cultivars cultivated under three NaCl (0, 56, and 113 mM) and four N (2.86, 1.43, 0.72, and 0.36 mM) concentrations. After 4 weeks, physiological NUE (pNUE), absorption NUE (aNUE), agronomical NUE (agNUE), N transport efficiency (NTE), and physiological traits were evaluated. Significant interactions between N and NaCl-applied concentrations were found in all measured parameters. In all cultivars, increasing the NaCl-applied concentration markedly decreased aNUE and agNUE. For each NaCl treatment, lowering the N applied sharply increased aNUE and agNUE, and this effect was stronger when the NaCl applied was higher. The effect of N lowering on pNUE depended on the NaCl treatment: it enhanced pNUE in the absence of NaCl but had no influence under the highest NaCl-applied concentration. Cultivars largely differed in response to NaCl. The aNUE—but not pNUE—differed between salt-tolerant and salt-sensitive cultivars: aNUE markedly decreased with NaCl concentration in the most salt-sensitive cultivar, whereas it was the highest at the intermediate NaCl concentration in the most salt-tolerant cultivar, especially under low N levels. This finding suggests that under salt conditions, the use of salt-tolerant rice genotypes combined with reducing N level application is necessary to improve NUE. The study of NUE in rice should be focused on the improvement of aNUE with a strong emphasis on the salt tolerance of cultivars.

     

Physiological and proteomic analysis of salinity tolerance of the halotolerant cyanobacterium <Emphasis Type="Italic">Anabaena</Emphasis> sp

The halotolerant cyanobacterium Anabaena sp was grown under NaCl concentration of 0, 170 and 515 mM and physiological and proteomic analysis was performed. At 515 mM NaCl the cyanobacterium showed reduced photosynthetic activities and significant increase in soluble sugar content, proline and SOD activity. On the other hand Anabaena sp grown at 170 mM NaCl showed optimal growth, photosynthetic activities and comparatively low soluble sugar content, proline accumulation and SOD activity. The intracellular Na⁺ content of the cells increased both at 170 and 515 mM NaCl. In contrast, the K⁺ content of the cyanobacterium Anabaena sp remained stable in response to growth at identical concentration of NaCl. While cells grown at 170 mM NaCl showed highest intracellular K⁺/Na⁺ ratio, salinity level of 515 mM NaCl resulted in reduced ratio of K⁺/Na⁺. Proteomic analysis revealed 50 salt-responsive proteins in the cyanobacterium Anabaena sp under salt treatment compared with control. Ten protein spots were subjected to MALDI-TOF–MS/MS analysis and the identified proteins are involved in photosynthesis, protein folding, cell organization and energy metabolism. Differential expression of proteins related to photosynthesis, energy metabolism was observed in Anabaena sp grown at 170 mM NaCl. At 170 mM NaCl increased expression of photosynthesis related proteins and effective osmotic adjustment through increased antioxidant enzymes and modulation of intracellular ions contributed to better salinity tolerance and optimal growth. On the contrary, increased intracellular Na⁺ content coupled with down regulation of photosynthetic and energy related proteins resulted in reduced growth at 515 mM NaCl. Therefore reduced growth at 515 mM NaCl could be due to accumulation of Na⁺ ions and requirement to maintain higher organic osmolytes and antioxidants which is energy intensive. The results thus show that the basis of salt tolerance is different when the halotolerant cyanobacterium Anabaena sp is grown under low and high salinity levels.     

Evaluation of nitrogen efficiency associated traits of starch potato cultivars under in vitro conditions

Genotype dependent responses to N-deficiency were investigated in vitro as well as in pots grown in a rain-out shelter with a set of 17 potato cultivars with the aim to identify genotypic differences in traits associated with nitrogen uptake and utilization and to compare results obtained in the two test environments. In vitro plantlets were grown under four nitrogen (N) levels (60, 30, 15 and 7.5 mmol/l) for 18 days and their respective traits were assessed after 7, 11, 14 and 18 days of culture (DoC). Considerable differences between genotypes were identified as early as seven DoC regarding N uptake, biomass production, root percentages of the total plant fresh and dry matter as well as chlorophyll (SPAD values) and crude protein concentrations. Significant differences were obtained for total biomass production between the 60, 30 and 15 mmol/l N-level as well as genotypic differences. Furthermore, significant variation between cultivars was identified for traits associated with efficient nitrogen use, e.g. the ability to maintain photosynthesis and N metabolization under N limitation as well as increased root development. No direct correlation was found between biomass production under in vitro conditions and tuber yields determined in pot trials. However, the stability of the plant performance under N-deficiency in the in vitro system as expressed by the membership function value of stress tolerance calculated from 11 traits correlated to the stability of the tuber yield of cultivars grown under limited N supply in pot trials (r?=?0.70). The experimental system is thought to be useful for pre-screening of germplasm and investigations of physiological processes associated with nitrogen use efficiency.     

Differential protein expression assessed by two-dimensional gel electrophoresis for two wheat varieties grown at four nitrogen levels

To limit N-fertilizer applied on wheat, cultivars that use N more efficiently are needed. Our objective was to investigate differences of nitrogen utilization in varieties by studying qualitative and quantitative proteins expression. Two wheat varieties, 'Arche' and 'Récital', were grown under controlled conditions at four N levels (0, 2, 8, and 20 mg N/plant/day) with two replicates. The number of tillers/plant, aerial dry weight/plant and total N content were measured after two months. Two-dimensional gel electrophoresis was also performed on leaf protein extracts. Analyses of variance showed that the N level effect was highly significant for the number of tillers/plant, aerial dry weight and N content. The variety x N level interaction was significant for N content. Analyses of variance on % volume carried out for 524 spots showed a significant variety effect for 55 spots and a significant N treatment effect for 76 spots. Twenty spots showed a significant variety x N treatment interaction. Fourteen proteins were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The possible role of these proteins, eight of which belong to the carbon metabolism, is discussed.     

Discovery of QTLs for water mining and water use efficiency traits in rice under water-limited condition through association mapping

Developing trait introgressed rice cultivars is essential to sustain yield under aerobic conditions. Here, we report DNA markers governing variability in root traits, water use efficiency (WUE) and other biometric traits like total leaf area by association mapping. A set of 173 diverse rice germplasm accessions were phenotyped for root traits in specially designed root structures and WUE using carbon isotope discrimination (Δ¹³C) during the monsoon season (July to October) of two consecutive years (2007 and 2008). The panel was genotyped using 291 SSR markers spanning the entire genome of rice. Root biomass varied between 1.8 and 16.3 g plant^?1 while root length between 22 and 78 cm representing significant genetic variability. Similarly, Δ¹³C varied from 18 to 23 ‰. The SSR markers showed extensive polymorphism with around 73 % of all the markers revealing polymorphism information content values more than 0.5. Model-based structure analysis using the squared-allele frequency correlations revealed six subgroups among the panel with an average LD decay of about 10–20 cM. The Benjamini–Hochberg analysis was carried out to compute the false discovery rate combined with the analysis of effective LD. A total of 82 markers were involved in 175 significant (corrected P values and Q values <0.05) marker–trait associations (MTAs) across experiment 1 and experiment 2 and for the pooled data. Out of these, 22 markers were found to be associated with more than one trait. Common markers with significant associations were discovered for root biomass, total leaf area and total biomass suggesting the interdependency of these traits. Finally, 12 markers showed significant and stable MTAs across the experiments for different traits. An in silico analysis indicated that 45 % of the MTAs overlapped with previously reported QTLs and can be used for QTL introgression through breeding.     

Genetic dissection of nitrogen nutrition in pea through a QTL approach of root, nodule, and shoot variability

Pea (Pisum sativum L.) is the third most important grain legume worldwide, and the increasing demand for protein-rich raw material has led to a great interest in this crop as a protein source. Seed yield and protein content in crops are strongly determined by nitrogen (N) nutrition, which in legumes relies on two complementary pathways: absorption by roots of soil mineral nitrogen, and fixation in nodules of atmospheric dinitrogen through the plant–Rhizobium symbiosis. This study assessed the potential of naturally occurring genetic variability of nodulated root structure and functioning traits to improve N nutrition in pea. Glasshouse and field experiments were performed on seven pea genotypes and on the ‘Cameor’ × ‘Ballet’ population of recombinant inbred lines selected on the basis of parental contrast for root and nodule traits. Significant variation was observed for most traits, which were obtained from non-destructive kinetic measurements of nodulated root and shoot in pouches, root and shoot image analysis, ¹⁵N quantification, or seed yield and protein content determination. A significant positive relationship was found between nodule establishment and root system growth, both among the seven genotypes and the RIL population. Moreover, several quantitative trait loci for root or nodule traits and seed N accumulation were mapped in similar locations, highlighting the possibility of breeding new pea cultivars with increased root system size, sustained nodule number, and improved N nutrition. The impact on both root or nodule traits and N nutrition of the genomic regions of the major developmental genes Le and Af was also underlined.     

Proteomic analysis of salt stress-responsive proteins in rice root

Salt stress is one of the major abiotic stresses in agriculture worldwide. We report here a systematic proteomic approach to investigate the salt stress-responsive proteins in rice (Oryza sativa L. cv. Nipponbare). Three-week-old seedlings were treated with 150 mM NaCl for 24, 48 and 72 h. Total proteins of roots were extracted and separated by two-dimensional gel electrophoresis. More than 1100 protein spots were reproducibly detected, including 34 that were up-regulated and 20 down-regulated. Mass spectrometry analysis and database searching helped us to identify 12 spots representing 10 different proteins. Three spots were identified as the same protein, enolase. While four of them were previously confirmed as salt stress-responsive proteins, six are novel ones, i.e. UDP-glucose pyrophosphorylase, cytochrome c oxidase subunit 6b-1, glutamine synthetase root isozyme, putative nascent polypeptide associated complex alpha chain, putative splicing factor-like protein and putative actin-binding protein. These proteins are involved in regulation of carbohydrate, nitrogen and energy metabolism, reactive oxygen species scavenging, mRNA and protein processing, and cytoskeleton stability. This study gives new insights into salt stress response in rice roots and demonstrates the power of the proteomic approach in plant biology studies.     

Protein changes in the shoot-tips of vanilla (<Emphasis Type="Italic">Vanilla planifolia</Emphasis>) in response to osmoprotective treatments

Cryogenic storage of vanilla shoot-tips represents the safest biotechnological strategy for the long-term conservation of the vanilla germplasm, but successful cryopreservation depends on its tolerance to both dehydration stress imposed by cryoprotective treatments and thermal stress produced by immersion in liquid nitrogen. In this work, we evaluated the impact of various osmoprotective treatments on protein expression patterns in vanilla (Vanilla planifolia) shoot-tips subjected to successive dehydration steps prior to cryopreservation. Two-dimensional electrophoretic protein profiles of shoot-tips dissected from in vitro grown plants and preconditioned on semisolid media with 0.3 M sucrose for one day, and shoot-tips preconditioned, loaded with a solution of 0.4 M sucrose and 2 M glycerol, and subsequently exposed to plant vitrification solution 3 (50% (w/v) sucrose and 50% (w/v) glycerol), were compared with non-treated dissected shoot-tips. We observed an increase in the expression level of six protein spots (fold change exceeding 1.5) and a decrease (fold change not exceeding 0.6) of ten protein spots after preconditionig treatment, whereas the profiles after preconditioning, loading and exposure to vitrification solution showed an increase in the expression level of 21 protein spots and a decrease in the expression level of 13. Most proteins identified were down-regulated and belonged to groups of biosynthesis, folding, and protein degradation. Many others were related to energetic metabolism, defense, and cell structure. These preliminary results contribute to knowledge of the proteome of this species and partially clarify its sensitivity to osmotic dehydration treatments.     

Physiological and proteomic analysis of young rice leaves grown under nitrogen-starvation conditions

Rice grown in anaerobic waterlogged soil accumulates ammonium as a major source of nitrogen (N). We have compared the physiological symptoms of rice seedlings subjected to N-starvation stress with those receiving sufficient N, based on measurements of shoot/root length and weight and an analysis of protein expression patterns. N starvation marginally increased root growth but notably decreased shoot biomass. N uptake was reduced by >50% in the roots and shoots of N-starved seedlings. To better understand the mechanism of N starvation in rice, we performed a comparative proteome analysis of proteins isolated from rice leaves. Twenty-five differentially expressed proteins were analyzed by matrix-assisted laser desorption/ionization time-of-flight (TOF) mass spectrometry and electron spray ionization quadrupole TOF. Functional analysis of the N-starvation response proteins suggested their involvement in protein synthesis and fate, metabolism, and defense. These results indicate that these proteins may play important roles in regulating the plant’s complex adaptation responses for N use during N starvation. The proteins may be useful for further characterization of protein function in plant N nutrition.     

Identification of flooding stress responsible cascades in root and hypocotyl of soybean using proteome analysis

Flooding inducible proteins were analyzed using a proteomic technique to understand the mechanism of soybean response to immersion in water. Soybeans were germinated for 2 days, and then subjected to flooding for 2 days. Proteins were extracted from root and hypocotyl, separated by two-dimensional polyacrylamide gel electrophoresis, stained by Coomassie brilliant blue, and analyzed by protein sequencing and mass spectrometry. Out of 803 proteins, 21 proteins were significantly up-regulated, and seven proteins were down-regulated by flooding stress. Of the total, 11 up-regulated proteins were classified as related to protein destination/storage and three proteins to energy, while four down-regulated proteins were related to protein destination/storage and three proteins to disease/defense. The expression of 22 proteins significantly changed within 1 day after flooding stress. The effects of flooding, nitrogen substitution without flooding, or flooding with aeration were analyzed for 1–4 days. The expression of alcohol dehydrogenase increased remarkably by nitrogen substitution compared to flooding. The expression of many proteins that changed due to flooding showed the same tendencies observed for nitrogen substitution; however, the expression of proteins classified into protein destination/storage did not.     

Effect of foliar spray of zinc on chloroplast β-carbonic anhydrase expression and enzyme activity in rice (Oryza sativa L.) leaves

The β-carbonic anhydrase (β-CA) is regarded as a zinc-containing enzyme involved in photosynthesis. Here, the rice plants of cv. N22 were treated with foliar spray at a Zn²⁺ concentration range from 0–35.0 mM during the tillering stage. The β-CA expression in the treated leaves was quantitatively determined by RT-qPCR and gel-based immunoblotting techniques, and its enzyme activity and relative chlorophyll concentration were measured. Results indicated that exogenous zinc could benefit rice plants at the tillering stage, particularly chloroplast β-CA with a fourfold enhancement in gene expression and a 14.6 % increase in its activity by treating the rice leaves with the 7.0-mM Zn²⁺ concentration, thereby promoting photosynthesis by a 19.4 % increase in relative chlorophyll concentration per unit leaf area. Results also showed that the application of Zn²⁺ at a concentration exceeding 7.0 mM could result in leaf senescence, and in some cases leaf hurts with significant inactivation (decreasing by approximately 70 %) of β-CA enzyme. It could be concluded that the application of 7.0-mM Zn²⁺ benefits rice plants at the tillering stage. The β-CA activity was associated with the catalytic microenvironment, thus providing an indicator for physiological response to exogenous zinc in rice.     

Proteomic analysis of copper stress responses in the roots of two rice (Oryza sativa L.) varieties differing in Cu tolerance

Background and aims

Copper (Cu) is an essential micronutrient required for growth and development of plants. However, excess Cu is toxic to plants. To understand the mechanisms involved in copper stress response, a proteomic approach was used to investigate the differences in Cu stress-induced protein expression between a Cu-tolerant variety (B1139) and a Cu-sensitive one (B1195) of rice.

Methods

Rice seedlings were exposed to 8 μM Cu for 3 days, with plants grown in the normal nutrient solution containing 0.32 μM Cu serving as the control. Proteins were extracted from the roots and separated by two-dimensional PAGE. Thirty four proteins were identified using MALDI-TOF mass spectrometry.

Results

Thirty-four protein spots were found to be differently expressed in the Cu-stressed roots in at least one variety of rice, including those involved in antioxidative defense, redox regulation, stress response, sulfur and glutathione (GSH) metabolism, carbohydrate metabolism, signal transduction, and some other proteins with various functions. Nine proteins, including putative cysteine synthase, probable serine acetyltransferase 3, L-ascorbate peroxidase 1, putative glutathione S-transferase 2, and thioredoxin-like 3-3, exhibited a greater increase in response to Cu stress in the Cu-tolerant variety B1139 compared with the Cu–sensitive variety B1195.

Conclusion

The majority of the proteins showing differential expression in response to Cu exposure are involved in the redox regulation, and sulfur and GSH metabolism, suggesting that these proteins, together with antioxidant enzymes, play an important role in the detoxification of excess Cu and maintaining cellular homeostasis.     

Alleviation of NaCl toxicity in the cyanobacterium <Emphasis Type="Italic">Synechococcus</Emphasis> sp. PCC 7942 by exogenous calcium supplementation

Salinity (NaCl) is one of the major problems associated with irrigated agricultural lands, especially rice fields. Being the common inhabitants of rice fields, cyanobacteria frequently experience high concentration of NaCl which in turn causes cellular damage. Therefore, mitigation of NaCl stress in cyanobacteria, plant growth-promoting microorganisms, is of utmost importance. The present study was designed to investigate the role of calcium in the alleviation of NaCl stress-induced cellular in Synechococcus sp. PCC 7942. The cyanobacterium was subjected to sub-lethal concentration of NaCl (800 mM) with and without the supplementation of calcium (1 mM CaCl₂) for 8 days. The results showed a drastic reduction in growth due to excess NaCl, but supplementation of CaCl₂ reduced the salt stress damage and partially restored growth. Application of calcium increased pigment contents, photosynthetic efficiency, antioxidative enzyme activity, osmolyte contents and reduced the intracellular sodium ion concentration, MDA content, electrolyte leakage and free oxygen radical generation. Furthermore, proteins involved in photosynthesis, respiration, ATP synthesis and protein synthesis along with two hypothetical proteins were also observed to be upregulated in the cyanobacterium in presence of calcium. Furthermore, proteins related to oxidative stress defence, nitrogen metabolism, carbohydrate metabolism, fatty acid metabolism and secondary metabolism were found to be upregulated by several fold. Therefore, our study suggests that calcium suppresses salt toxicity in Synechococcus sp. PCC 7942 by restricting the entry of Na⁺ into the cell, increasing osmolyte production and upregulating defence-related proteins.     

Nitrogen application enhanced the expression of developmental plasticity of root systems triggered by mild drought stress in rice

Background

The promoted root growth under developmental plasticity triggered specifically by mild drought stress (MDS) is known to contribute to maintained water uptake and dry matter production (DMP).

Aims

To examine whether the expression of developmental plasticity of root systems and its contribution to DMP would be affected by the levels of nitrogen (N) application.

Methods

Two genotypes (CSSL50 derived from Nipponbare/Kasalath cross and Nipponbare) were grown under soil moisture gradients with a line source sprinkler system. Three N fertilizer treatments were used; 25 (low), 75 (standard) and 150 kg N ha^?1 (high) in 2009 and 60 (low), 120 (standard) and 180 kg N ha^?1 (high) in 2011.

Results

Across varying N level treatments, there were no significant differences in any of the traits examined between the two genotypes under well-watered and severe drought stress conditions. In contrast, under MDS conditions (15–25 % w/w of soil moisture content (SMC) in 2009 and 17–25 % w/w of SMC in 2011), CSSL50 showed greater DMP than Nipponbare. The difference, however, varied with N level treatments since CSSL50’s greater root system development under MDS, was more pronounced at standard and high N levels than at low N level than it was for Nipponbare.

Conclusions

N application enhanced the expression of plasticity in root system development at standard and high N levels as compared with low N level under MDS conditions, which contributed to the maintenance of DMP.