Al Tolerance In Rice Research Articles

Identifying aluminum tolerance in rice with a molecular marker

Improving rice aluminum (Al) tolerance in acid soils is highly significant to rice production. So far, no molecular marker which can efficiently identify rice Al tolerance has been developed. In this study, Art1, a primer specific for the ART1 gene was designed to study whether some natural variations exist in ART1 between Al tolerant and sensitive varieties. The results showed that two Al tolerant varieties had identical nucleotides and amino acids, while three Al sensitive varieties had the same nucleotides and amino acids differing from the two Al tolerant varieties. The same results were discovered within 150 rice varieties of Ting’s core collection. Art1 could be used to judge rice Al tolerance.

ALUMINUM RESISTANCE TRANSCRIPTION FACTOR 1 (ART1) contributes to natural variation in aluminum resistance in diverse genetic backgrounds of rice (O. sativa).

Transcription factors (TFs) regulate the expression of other genes to indirectly mediate stress resistance mechanisms. Therefore, when studying TF‐mediated stress resistance, it is important to understand how TFs interact with genes in the genetic background. Here, we fine‐mapped the aluminum (Al) resistance QTL Alt12.1 to a 44‐kb region containing six genes. Among them is ART1, which encodes a C2H2‐type zinc finger TF required for Al resistance in rice. The mapping parents, Al‐resistant cv Azucena (tropical japonica) and Al‐sensitive cv IR64 (indica), have extensive sequence polymorphism within the ART1 coding region, but similar ART1 expression levels. Using reciprocal near‐isogenic lines (NILs) we examined how allele‐swapping the Alt12.1 locus would affect plant responses to Al. Analysis of global transcriptional responses to Al stress in roots of the NILs alongside their recurrent parents demonstrated that the presence of the Alt12.1 from Al‐resistant Azucena led to greater changes in gene expression in response to Al when compared to the Alt12.1 from IR64 in both genetic backgrounds. The presence of the ART1 allele from the opposite parent affected the expression of several genes not previously implicated in rice Al tolerance. We highlight examples where putatively functional variation in cis‐regulatory regions of ART1‐regulated genes interacts with ART1 to determine gene expression in response to Al. This ART1–promoter interaction may be associated with transgressive variation for Al resistance in the Azucena × IR64 population. These results illustrate how ART1 interacts with the genetic background to contribute to quantitative phenotypic variation in rice Al resistance.

Unbiased identification of decidual immune cell subsets in pregnant nonhuman primates using dimensionality reduction of flow cytometric data

The Natural Resistance Associated Macrophage Protein (Nramp) members play diverse roles in metal transport in plants. Recent studies have showed that OsNrat1 (OsNramp4) encodes an Al transporter, which is required for rice Al tolerance. In this study, we functionally characterized a Nramp member in sorghum, SbNrat1, which is homologous to OsNrat1 with 88% identity. SbNrat1 was expressed in both roots and shoots, and its expression was not induced by Al treatment. When expressed in yeast, SbNrat1 transports trivalent Al ion, but not Mn and Cd. Furthermore, introduction of SbNrat1 into the rice mutant osnrat1 can rescue its sensitivity to Al. However, no correlation between Al tolerance and the expression level of SbNrat1 was found in thirteen sorghum cultivars tested. These results indicate that SbNrat1 functions as an Al transporter that is possibly involved in basic Al tolerance in sorghum.

Functional characterization of the SbNrat1 gene in sorghum

The Natural Resistance Associated Macrophage Protein (Nramp) members play diverse roles in metal transport in plants. Recent studies have showed that OsNrat1 (OsNramp4) encodes an Al transporter, which is required for rice Al tolerance. In this study, we functionally characterized a Nramp member in sorghum, SbNrat1, which is homologous to OsNrat1 with 88% identity. SbNrat1 was expressed in both roots and shoots, and its expression was not induced by Al treatment. When expressed in yeast, SbNrat1 transports trivalent Al ion, but not Mn and Cd. Furthermore, introduction of SbNrat1 into the rice mutant osnrat1 can rescue its sensitivity to Al. However, no correlation between Al tolerance and the expression level of SbNrat1 was found in thirteen sorghum cultivars tested. These results indicate that SbNrat1 functions as an Al transporter that is possibly involved in basic Al tolerance in sorghum.

MECHANISM OF ALUMINUM TOXICITY AVOIDANCE IN TROPICAL RICE (Oryza sativa), MAIZE (Zea mays), AND SOYBEAN (Glycine max)

Planting Al tolerant crops is an economically justifiable approach in crop production on acid soils. Experiments were conducted to study the mechanisms of Al tolerance among species and varieties of tropical rice, maize, and soybean with previously known levels of Al tolerance. These varieties were hydroponically cultured in 0, 5, 10, and 30 mg l-1 Al with complete nutrient solution at pH 4. The results show that root/shoot ratio of dry weight at 10 mg l-1 Al treatment was an important parameter to indicate differential Al tolerance in maize. Oxalic acid exudation from roots cannot always explain the Al tolerance. Total organic acid concentration in roots at 10 mg l-1 Al treatment indicated a difference of Al tolerance in soybean and lowland rice. Aluminum translocation from roots to shoots was lower in tolerant varieties than in sensitive varieties of soybean. Increased Al concentration in shoots with increased Al level in the solution was larger in soybean and maize than in lowland or upland rice. Among varieties of soybean, the Al concentration in shoots increased drastically in Wilis (Al-sensitive variety) with increase Al level, while in Kitamusume (Al-tolerant variety) it did not.

MECHANISM OF ALUMINUM TOXICITY AVOIDANCE IN TROPICAL RICE (Oryza sativa), MAIZE (Zea mays), AND SOYBEAN (Glycine max)

Planting Al tolerant crops is an economically justifiable approach in crop production on acid soils. Experiments were conducted to study the mechanisms of Al tolerance among species and varieties of tropical rice, maize, and soybean with previously known levels of Al tolerance. These varieties were hydroponically cultured in 0, 5, 10, and 30 mg l-1 Al with complete nutrient solution at pH 4. The results show that root/shoot ratio of dry weight at 10 mg l-1 Al treatment was an important parameter to indicate differential Al tolerance in maize. Oxalic acid exudation from roots cannot always explain the Al tolerance. Total organic acid concentration in roots at 10 mg l-1 Al treatment indicated a difference of Al tolerance in soybean and lowland rice. Aluminum translocation from roots to shoots was lower in tolerant varieties than in sensitive varieties of soybean. Increased Al concentration in shoots with increased Al level in the solution was larger in soybean and maize than in lowland or upland rice. Among varieties of soybean, the Al concentration in shoots increased drastically in Wilis (Al-sensitive variety) with increase Al level, while in Kitamusume (Al-tolerant variety) it did not.

Association Mapping for Aluminum Tolerance in a Core Collection of Rice Landraces.

Trivalent aluminum (Al3+) has drastic effect on the rice production in acidic soils. Elite genes for aluminum (Al) tolerance might exist in rice landraces. Therefore, the purpose of this research is to mine the elite genes within rice landraces. Association mapping for Al tolerance traits [i.e., relative root elongation (RRE)] was performed by using a core collection of 150 accessions of rice landraces (i.e., Ting’s rice core collection). Our results showed that the Ting’s rice core collection possessed a wide-range of phenotypic variation for Al tolerance, and the index of Al tolerance (RRE) was ranged from 0.22 to 0.89. Moreover, the groups with different origins and compositions of indica and japonica rice showed different degrees of tolerance to varying levels of Al. These rice landraces were further screened with 274 simple sequence repeat markers, and association mapping was performed using a mixed linear model approach. The mapping results showed that a total of 23 significant (P < 0.05) trait–marker associations were detected for Al tolerance. Of these, three associations (13%) were identical to the quantitative trait loci reported previously, and other 20 associations were reported for the first time in this study. The proportion of phenotypic variance (R2) explained by 23 significant associations ranged from 5.03 to 20.03% for Al tolerance. We detected several elite alleles for Al tolerance based on multiple comparisons of allelic effects, which could be used to develop Al tolerant rice cultivars through marker-assisted breeding.

An Al-inducible expansin gene, OsEXPA10 is involved in root cell elongation of rice.

Expansins are cell wall loosening proteins, which are encoded by multigene families. However, the physiological role of most expansin genes is still poorly understood. Here, we functionally characterized an Al-inducible expansin gene, OsEXPA10, which is regulated by a C2H2-type zinc-finger transcription factor, ART1 in rice. A detailed expression analysis showed that OsEXPA10 was expressed in both the roots and shoots at a similar level, but only the expression in the roots was rapidly upregulated in response to Al. Furthermore, spatial expression analysis showed that the Al-induced expression was only found in the root tips (0-3mm), but not in the mature root zones. The expression was neither induced by other metals including Cd and La nor by low pH. Immunostaining showed that OsEXPA10 was localized at all cells of the root tips. Knockout of OsEXPA10 resulted in a significant decrease in the cell elongation of the roots in the absence of Al. In the presence of Al, knockout of OsEXPA10 did not alter the Al sensitivity evaluated by relative root elongation, but the root cell wall of knockout lines accumulated less Al compared to those of the wild-type rice. Collectively, our results indicate that OsEXPA10 expressed in the root tips is required for the root cell elongation, but that the contribution of this gene to high Al tolerance in rice is small.

Construction of RNA Interference Vector to Silence Aluminum Tolerance Gene Candidate in Rice cv Hawara Bunar

One of the aluminum (Al) tolerance gene candidates, namely B11 gene, has been successfully isolated from Al-tolerant rice cv Hawara Bunar. However, the role of the gene in Al tolerance in rice has not been known. RNA interference (RNAi) technique is an effective tool to examine the biological function of the target gene in plant. The objective of the research was to construct RNAi recombinant vector carrying untranslated region of the B11 gene. RNAi recombinant vector carrying 195 bp sized 3′UTR_B11 fragment as a double-stranded RNA (dsRNA) trigger has been successfully constructed using GATEWAY™ cloning technology, pENTR™/D-TOPO® as a shuttle vector, and pANDA vector as a destination vector. RNAi construct was successfully introduced into Agrobacterium tumefaciens AgL0, and has been infected to rice cv Hawara Bunar. Analysis of putative transgenic rice showed eight of 20 plants were transgenic carrying the B11-RNAi construct.

Functional Analysis of a MATE Gene OsFRDL2 Revealed its Involvement in Al-Induced Secretion of Citrate, but a Lower Contribution to Al Tolerance in Rice.

The multidrug and toxic compound extrusion (MATE) transporters represent a large transporter family in plants, but the role of most genes in this family has not been examined. We functionally characterized a MATE family member, OsFRDL2, in rice (Oryza sativa). OsFRDL2 showed an efflux transport activity for citrate when it was expressed in both Xenopus oocytes and cultured tobacco cells. OsFRDL2 was mainly expressed in the roots and its expression was not induced by iron (Fe) deficiency, but it was rapidly up-regulated by aluminum (Al). Furthermore, the expression of OsFRDL2 was regulated by ART1, a C2H2-type zinc-finger transcription factor for Al tolerance. OsFRDL2 protein was localized at unidentified vesicles in the cytosol, but not co-localized with either mitochondria or peroxisomes when expressed in both onion epidermal cells and cultured tobacco cells. Knockout of OsFRDL2 decreased Al-induced secretion of citrate from the roots, but did not affect the internal citrate concentration. The Al-induced inhibition of root elongation was similar between the OsFRDL2 knockout line and its wild-type rice. Knockout of OsFRDL2 did not affect the translocation of Fe from the roots to the shoots. A double mutant between osfrdl2 and osfrdl4 or osfrdl1 did not further decrease the Al-induced citrate secretion and Fe translocation compared with the single mutant. Collectively, our results indicate that although OsFRDL2 is involved in the Al-induced secretion of citrate, its contribution to high Al tolerance is relatively small in rice.

Overexpressing OsPIN2 enhances aluminium internalization by elevating vesicular trafficking in rice root apex.

Aluminium (Al) sequestration is required for internal detoxification of Al in plant cells. In this study, it was found that the rice OsPIN2 overexpression line (OX1) had significantly reduced Al content in its cell wall and increased Al concentration in cell sap only in rice root tips relative to the wild-type (WT). In comparison with WT, OX1 reduced morin staining of cytosolic Al, enhanced FM 4-64 staining of membrane vesicular trafficking in root tip sections (0-1mm), and showed morin-FM 4-64 fluorescence overlap. Recovery treatment showed that cell-wall-bound Al was internalized into vacuoles via endocytic vesicular trafficking after removal of external Al. In this process, OX1 showed a higher rate of Al internalization than WT. Brefeldin A (BFA) interfered with vesicular trafficking and resulted in inhibition of Al internalization. This inhibitory effect could be alleviated when BFA was washed out, and the process of alleviation was slower in the cells of WT than in those of OX1. Microscopic observations revealed that, upon Al exposure, numerous multilamellar endosomes were detected between the cell wall and plasma membrane in the cells of OX1. Moreover, more vesicles enriched with Al complexes accumulated in the cells of OX1 than in those of WT, and these vesicles transformed into larger structures in the cells of OX1. Taken together, the data indicate that endocytic vesicular trafficking might contribute to Al internalization, and that overexpressing OsPIN2 enhances rice Al tolerance via elevated endocytic vesicular trafficking and Al internalization.

Natural variation underlies alterations in Nramp aluminum transporter (NRAT1) expression and function that play a key role in rice aluminum tolerance.

Aluminum (Al) toxicity is a major constraint for crop production on acid soils which compose ∼ 40% of arable land in the tropics and subtropics. Rice is the most Al-tolerant cereal crop and offers a good model for identifying Al tolerance genes and mechanisms. Here we investigated natural variation in the rice Nramp aluminum transporter (NRAT1) gene encoding a root plasma membrane Al uptake transporter previously hypothesized to underlie a unique Al tolerance mechanism. DNA sequence variation in the NRAT1 coding and regulatory regions was associated with changes in NRAT1 expression and NRAT1 Al transport properties. These sequence changes resulted in significant differences in Al tolerance that were found to be associated with changes in the Al content of root cell wall and cell sap in 24 representative rice lines from a rice association panel. Expression of the tolerant OsNRAT1 allele in yeast resulted in higher Al uptake than did the sensitive allele and conferred greater Al tolerance when expressed in transgenic Arabidopsis. These findings indicate that NRAT1 plays an important role in rice Al tolerance by reducing the level of toxic Al in the root cell wall and transporting Al into the root cell, where it is ultimately sequestered in the vacuole. Given its ability to enhance Al tolerance in rice and Arabidopsis, this work suggests that the NRAT1 gene or its orthologs may be useful tools for enhancing Al tolerance in a wide range of plant species.

New Insights into Aluminum Tolerance in Rice: The ASR5 Protein Binds the STAR1 Promoter and Other Aluminum-Responsive Genes

Aluminum (Al) toxicity in plants is one of the primary constraints in crop production. Al³⁺, the most toxic form of Al, is released into soil under acidic conditions and causes extensive damage to plants, especially in the roots. In rice, Al tolerance requires the ASR5 gene, but the molecular function of ASR5 has remained unknown. Here, we perform genome-wide analyses to identify ASR5-dependent Al-responsive genes in rice. Based on ASR5_RNAi silencing in plants, a global transcriptome analysis identified a total of 961 genes that were responsive to Al treatment in wild-type rice roots. Of these genes, 909 did not respond to Al in the ASR5_RNAi plants, indicating a central role for ASR5 in Al-responsive gene expression. Under normal conditions, without Al treatment, the ASR5_RNAi plants expressed 1.756 genes differentially compared to the wild-type plants, and 446 of these genes responded to Al treatment in the wild-type plants. Chromatin immunoprecipitation followed by deep sequencing identified 104 putative target genes that were directly regulated by ASR5 binding to their promoters, including the STAR1 gene, which encodes an ABC transporter required for Al tolerance. Motif analysis of the binding peak sequences revealed the binding motif for ASR5, which was confirmed via in vitro DNA-binding assays using the STAR1 promoter. These results demonstrate that ASR5 acts as a key transcription factor that is essential for Al-responsive gene expression and Al tolerance in rice.

Effect of aluminum on protein oxidation, non-protein thiols and protease activity in seedlings of rice cultivars differing in aluminum tolerance

The effect of toxic concentrations of aluminum (Al) was investigated on contents of protein-thiols, non-protein and total thiols, protein carbonylation and protease activity in the seedlings of Al-sensitive (Al-S) Indica rice cv. HUR-105 and Al-tolerant (Al-T) cv. Vandana grown in sand cultures. Al treatment of 178 μM and 421 μM for 3-12 days caused a significant decline in the level of protein thiols, rise in non-protein thiols (NPTs) as well as protein carbonyl content and an insignificant alteration in the level of total thiols in cv. HUR-105 seedlings. However, in the seedlings of Al-T cv. Vandana, no significant alteration could be observed on any of these parameters with Al treatment. Al treatment inhibited protease activity in roots, whereas the opposite trend was seen in shoots. New isozymes of protease appeared in shoots of cv. Vandana with increased level of Al treatment. Our results show a link between protein thiols and NPTs and suggest the role of NPTs in the repair and protection of protein thiols. Inhibitory effect of Al on protease activity in roots could be a major reason for Al rhizotoxic effects. Al tolerance in rice appears to be associated with lesser content of protein thiols in roots, smaller amount of carbonylated proteins in roots as well as shoots and higher protease activity in shoots.

Genetic dissection of Al tolerance QTLs in the maize genome by high density SNP scan.

BackgroundAluminum (Al) toxicity is an important limitation to food security in tropical and subtropical regions. High Al saturation on acid soils limits root development, reducing water and nutrient uptake. In addition to naturally occurring acid soils, agricultural practices may decrease soil pH, leading to yield losses due to Al toxicity. Elucidating the genetic and molecular mechanisms underlying maize Al tolerance is expected to accelerate the development of Al-tolerant cultivars.ResultsFive genomic regions were significantly associated with Al tolerance, using 54,455 SNP markers in a recombinant inbred line population derived from Cateto Al237. Candidate genes co-localized with Al tolerance QTLs were further investigated. Near-isogenic lines (NILs) developed for ZmMATE2 were as Al-sensitive as the recurrent line, indicating that this candidate gene was not responsible for the Al tolerance QTL on chromosome 5, qALT5. However, ZmNrat1, a maize homolog to OsNrat1, which encodes an Al3+ specific transporter previously implicated in rice Al tolerance, was mapped at ~40 Mbp from qALT5. We demonstrate for the first time that ZmNrat1 is preferentially expressed in maize root tips and is up-regulated by Al, similarly to OsNrat1 in rice, suggesting a role of this gene in maize Al tolerance. The strongest-effect QTL was mapped on chromosome 6 (qALT6), within a 0.5 Mbp region where three copies of the Al tolerance gene, ZmMATE1, were found in tandem configuration. qALT6 was shown to increase Al tolerance in maize; the qALT6-NILs carrying three copies of ZmMATE1 exhibited a two-fold increase in Al tolerance, and higher expression of ZmMATE1 compared to the Al sensitive recurrent parent. Interestingly, a new source of Al tolerance via ZmMATE1 was identified in a Brazilian elite line that showed high expression of ZmMATE1 but carries a single copy of ZmMATE1.ConclusionsHigh ZmMATE1 expression, controlled either by three copies of the target gene or by an unknown molecular mechanism, is responsible for Al tolerance mediated by qALT6. As Al tolerant alleles at qALT6 are rare in maize, marker-assisted introgression of this QTL is an important strategy to improve maize adaptation to acid soils worldwide.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2164-15-153) contains supplementary material, which is available to authorized users.

A plasma membrane‐localized small peptide is involved in rice aluminum tolerance

A transcription factor for Al tolerance, ART1, regulates the expression of at least 30 genes in rice. Here we functionally characterized one of the downstream genes, OsCDT3, which encodes a predicted peptide of only 53 amino acid residues rich in cysteine. Knockdown of this gene resulted in decreased tolerance to Al, but did not affect the tolerance to Cd. The aluminum (Al) content in the root residues including cell wall and the plasma membrane of knockdown lines decreased, but the Al concentration in the root cell sap increased compared with those of the wild-type rice. OsCDT3 was mainly expressed in the roots and its expression was specifically induced by Al exposure, not by low pH and other metals. There was a small genotypic variation in OsCDT3 expression level, but no correlation between Al tolerance and the OsCDT3 variation was found among 17 rice cultivars. Analysis of pOsCDT3::GFP transgenic rice showed that OsCDT3 was expressed at all cells in the root tips. Transient expression of OsCDT3 fused with GFP at both N- and C-termini showed that OsCDT3 was anchored to the plasma membrane. Expression of OsCDT3 in yeast conferred tolerance to Al, but not to Cd. Furthermore, OsCDT3 did not show transport activity for Al in yeast, but was able to directly bind Al in vitro. Taken together, our results indicate that OsCDT3 anchoring to the plasma membrane may play a role in stopping entry of Al into the root cells by binding Al, therefore, contributing to high Al 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.

Interactive regulation of nitrogen and aluminum in rice

Despite many studies on the high aluminum (Al) tolerance of rice (Oryza sativa), its exact mechanisms remain largely unknown. It is also unclear why Al improves growth of some plants. Our research on interactions between nitrogen (N) and Al may help to understand these phenomena. Previously, we found that ammonium-supplemented rice was more Al tolerant than nitrate-supplemented rice. Furthermore, Al-tolerant rice varieties preferred ammonium, while Al-sensitive ones preferred nitrate; in fact, Al tolerance was significantly correlated with the ammonium/nitrate preference among rice varieties. Al even enhanced growth of ammonium-supplemented rice, while it inhibited growth of nitrate-supplemented rice. Based on our own and other reports on N-Al interactions, we propose that intermediate products of N metabolism may play a role in rice Al tolerance. Al-enhanced ammonium utilization may explain why Al promotes growth of some plants, since Al often coexists with higher levels of ammonium than nitrate in acid soils.

Differential responses of antioxidant enzymes to aluminum toxicity in two rice (Oryza sativa L.) cultivars with marked presence and elevated activity of Fe SOD and enhanced activities of Mn SOD and catalase in aluminum tolerant cultivar

Seedlings of two Indica rice (Oryza sativa L.) cvs. HUR-105 and Vandana, differing in Al-tolerance were used to identify the key mechanisms involved in their differential behaviour towards Al toxicity. Cv. HUR-105 appeared to be Al sensitive by showing significant reduction (p ≤ 0.01) in root/shoot length, fresh weight, dry weight and water content in presence of 421 μM Al3+ in growth medium whereas cv. Vandana appeared to be fairly Al3+ tolerant. A conspicuous and significant reduction in dry weight of root and shoot was observed in Al sensitive cv. HUR-105 with 178 μM Al3+ treatment for 3 days. Al was readily taken up by the roots and transported to shoots in both the rice cultivars. Localization of absorbed Al was always greater in roots than in shoots. Our results of the production of reactive oxygen species (ROS) H2O2 and O2.− and activities of major antioxidant enzymes such as total superoxide dismutase (SOD), Cu/Zn SOD, Mn SOD, Fe SOD, catalase (CAT) and guaiacol peroxidase revealed Al induced higher oxidative stress, greater production of ROS and lesser capacity to scavenge ROS in cv. HUR-105 than Vandana. With Al treatment, higher oxidative stress was noted in shoots than in roots. Greatly enhanced activities of SOD (especially Fe and Mn SOD) and CAT in Al treated seedlings of cv. Vandana suggest the role of these enzymes in Al tolerance. Furthermore, a marked presence of Fe SOD in roots and shoots of the seedlings of Al tolerant cv. Vandana and its significant (p ≤ 0.01) increase in activity due to Al-treatment, appears to be the unique feature of this cultivar and indicates a vital role of Fe SOD in Al-tolerance in rice.

Comparative Genome-Wide Transcriptional Analysis of Al-Responsive Genes Reveals Novel Al Tolerance Mechanisms in Rice

Rice (Oryza sativa) is the most aluminum (Al)-tolerant crop among small-grain cereals, but the mechanism underlying its high Al resistance is still not well understood. To understand the mechanisms underlying high Al-tolerance, we performed a comparative genome-wide transcriptional analysis by comparing expression profiling between the Al-tolerance cultivar (Koshihikari) and an Al-sensitive mutant star1 (SENSITIVE TO AL RHIZOTOXICITY 1) in both the root tips and the basal roots. Exposure to 20 µM AlCl3 for 6 h resulted in up-regulation (higher than 3-fold) of 213 and 2015 genes including 185 common genes in the root tips of wild-type and the mutant, respectively. On the other hand, in the basal root, genes up-regulated by Al were 126 and 2419 including 76 common genes in the wild-type and the mutant, respectively. These results indicate that Al-response genes are not only restricted to the root tips, but also in the basal root region. Analysis with genes up- or down-regulated only in the wild-type reveals that there are other mechanisms for Al-tolerance except for a known transcription factor ART1-regulated one in rice. These mechanisms are related to nitrogen assimilation, secondary metabolite synthesis, cell-wall synthesis and ethylene synthesis. Although the exact roles of these putative tolerance genes remain to be examined, our data provide a platform for further work on Al-tolerance in rice.