Al Tolerance In Rice Research Articles

Overexpression of OsGASR1 promotes Al tolerance in rice

Aluminum (Al) toxicity in acid soils poses a significant threat to rice, which exhibits highly complex genetic mechanisms for both external detoxification and internal tolerance among cereal crops. Although several genes involved Al tolerance have been identified, the molecular mechanisms underlying Al tolerance in rice remain to be fully explored. Here, we functionally characterized the gibberellin-stimulated transcription gene OsGASR1, which encodes a small cysteine-rich peptide localized to the nucleus and cytoplasm and plays a significant role in Al tolerance in rice. The expression of OsGASR1 is rapidly up-regulated by Al in rice root tips but not in the shoots. Its expression is not regulated by the central regulator Aluminum Resistance Transcription Factor 1 (ART1), indicating that OsGASR1 functions as a novel gene in rice Al resistance independent of ART1. Knockout of OsGASR1 reduced root length but did not affect Al tolerance in rice, whereas overexpression of OsGASR1 enhanced Al tolerance without affecting Al distribution and accumulation and promoted the accumulation of reactive oxygen species (ROS) in the root tips. RNA-seq analysis revealed that overexpression of OsGASR1 upregulated the expression of genes associated with cell wall modification, oxidative stress, and Al tolerance. Collectively, these findings suggest that OsGASR1 is involved in Al tolerance in rice independently of ART1, and the up-regulation of this gene is necessary for rice Al tolerance.

Overexpression of an ART1-Interacting Gene OsNAC016 Improves Al Tolerance in Rice

Rice (Oryza sativa) exhibits tremendous aluminum (Al)-tolerance. The C2H2-transcription factor (TF) ART1 critically regulates rice Al tolerance via modulation of specific gene expression. However, little is known about the posttranscriptional ART1 regulation. Here, we identified an ART1-interacted gene OsNAC016 via a yeast two-hybrid (Y2H) assay. OsNAC016 was primarily expressed in roots and weakly induced by Al. Immunostaining showed that OsNAC016 was a nuclear protein and localized in all root cells. Knockout of OsNAC016 did not alter Al sensitivity. Overexpression of OsNAC016 resulted in less Al aggregation within roots and enhanced Al tolerance in rice. Based on transcriptomic and qRT-PCR evaluations, certain cell-wall-related or ART-regulated gene expressions such as OsMYB30 and OsFRDL4 were altered in OsNAC016-overexpressing plants. These results indicated that OsNAC016 interacts with ART1 to cooperatively regulate some Al-tolerance genes and is a critical regulatory factor in rice Al tolerance.

Identification and expression pattern of aluminium-responsive genes in roots of rice genotype with reference to Al-sensitivity

Aluminium (Al) is the third most abundant element in the Earth's crust. Globally, acidic soil occupies 30–40% of ice-free land areas; Al toxicity is a major threat to crops. The first symptom of Al toxicity is the inhibition of root growth followed by poor root hair development, swollen root apices, necrosis of leaves and reduced yield. Although Rice (Oryza sativa) is an Al toxicity tolerant crop, it shows considerable variations among rice genotypes to Al exposure. Therefore, it is pertinent to understand Al toxicity and underlying mechanisms for Al tolerance in Rice. In the present study, 63 rice genotypes screened under Al stress showed significant variations of root growth. Expression stability of endogenous control genes (ECGs) revealed sulphite reductase (SR) as the most stable ECG that can be used as a reference gene for quantitative real-time PCR (qRT-PCR). Expression patterns of Al-responsive genes suggest genes associated with cytoskeletal dynamics, metabolism, and ion transporter could play significant roles in Al adaptation and tolerance in rice. The results showed Motodhan, Vietnam-1, Yimyu and N-861 as Al-toxicity tolerant, while Lespah, RCPL-13, VL-31329, and UPR2919-141-1 as most Al-sensitive genotypes among the studied rice lines cultivated in North-East India.

Transcriptomics and metabolomics reveal tolerance new mechanism of rice roots to Al stress.

The prevalence of soluble aluminum (Al) ions is one of the major limitations to crop production worldwide on acid soils. Therefore, understanding the Al tolerance mechanism of rice and applying Al tolerance functional genes in sensitive plants can significantly improve Al stress resistance. In this study, transcriptomics and metabolomics analyses were performed to reveal the mechanism of Al tolerance differences between two rice landraces (Al-tolerant genotype Shibanzhan (KR) and Al-sensitive genotype Hekedanuo (MR) with different Al tolerance. The results showed that DEG related to phenylpropanoid biosynthesis was highly enriched in KR and MR after Al stress, indicating that phenylpropanoid biosynthesis may be closely related to Al tolerance. E1.11.1.7 (peroxidase) was the most significant enzyme of phenylpropanoid biosynthesis in KR and MR under Al stress and is regulated by multiple genes. We further identified that two candidate genes Os02g0770800 and Os06g0521900 may be involved in the regulation of Al tolerance in rice. Our results not only reveal the resistance mechanism of rice to Al stress to some extent, but also provide a useful reference for the molecular mechanism of different effects of Al poisoning on plants.

Using brefeldin A to disrupt cell wall polysaccharide components in rice and nitric oxide to modify cell wall structure to change aluminum tolerance.

The components and structure of cell wall are closely correlated with aluminum (Al) toxicity and tolerance for plants. However, the cell wall assembly and function construction in response to Al is not known. Brefeldin A (BFA), a macrolide, is used to disrupt cell wall polysaccharide components, and nitric oxide (NO), a signal molecule, is used to modify the cell wall structure. Pretreatment with BFA accelerated Al accumulation in root tips and Al-induced inhibition of root growth of two rice genotypes of Nipponbare and Zhefu 802, and significantly decreased the cell wall polysaccharide content including pectin, hemicellulose 1, and hemicellulose 2, indicating that BFA inhibits the biosynthesis of components in the cell wall and makes the root cell wall lose the ability to resist Al. The addition of NO donor (SNP) significantly alleviated the toxic effects of Al on root growth, Al accumulation, and oxidative damage, and decreased the content of pectin polysaccharide and functional groups of hydroxyl, carboxyl, and amino in the cell wall via FTIR analysis, while had no significant effect on hemicellulose 1 and hemicellulose 2 content compared with Al treatment. Furthermore, NO didn't change the inhibition effect of BFA-induced cell wall polysaccharide biosynthesis and root growth. Taken together, BFA disrupts the integrity of cell wall and NO modifies partial cell wall composition and their functional groups, which change the Al tolerance in rice.

Metabolome Analysis under Aluminum Toxicity between Aluminum-Tolerant and -Sensitive Rice (Oryza sativa L.).

Aluminum (Al) solubilizes into trivalent ions (Al3+) on acidic soils, inhibiting root growth. Since about 13% of global rice cultivation is grown on acidic soils, improving Al tolerance in rice may significantly increase yields. In the present study, metabolome analysis under Al toxicity between the Al-tolerant variety Nipponbare and the Al-sensitive variety H570 were performed. There were 45 and 83 differential metabolites which were specifically detected in Nipponbare and H570 under Al toxicity, respectively. Furthermore, the results showed that 16 lipids out of 45 total metabolites were down-regulated, and 7 phenolic acids as well as 4 alkaloids of 45 metabolites were up-regulated in Nipponbare, while 12 amino acids and their derivatives were specifically detected in H570, of which 11 amino acids increased, including L-homoserine and L-methionine, which are involved in cysteine synthesis, L-ornithine and L-proline, which are associated with putrescine synthesis, and 1-aminocyclopropane-1-carboxylate, which is associated with ethylene synthesis. The contents of cysteine and s-(methyl) glutathione, which were reported to be related to Al detoxification in rice, decreased significantly. Meanwhile, putrescine was accumulated in H570, while there was no significant change in Nipponbare, so we speculated that it might be an intermediate product of Al detoxification in rice. The differential metabolites detected between Al-tolerant and -sensitive rice variants in the present study might play important roles in Al tolerance. These results provide new insights in the mechanisms of Al tolerance in rice.

Distinct Patterns of Rhizosphere Microbiota Associated With Rice Genotypes Differing in Aluminum Tolerance in an Acid Sulfate Soil.

Rhizosphere microbes are important for plant tolerance to various soil stresses. Rice is the most aluminum (Al)-tolerant small grain cereal crop species, but the link between rice Al tolerance and rhizosphere microbiota remains unclear. This study aimed to investigate the microbial community structure of aluminum-sensitive and Al-tolerant rice varieties in acid sulfate soil under liming and non-liming conditions. We analyzed the rice biomass and mineral element contents of rice plants as well as the chemical properties and microbial (archaea, bacteria, and fungi) communities of rhizosphere and bulk soil samples. The results showed that the Al-tolerant rice genotype grew better and was able to take up more phosphorus from the acid sulfate soil than the Al-sensitive genotype. Liming was the main factor altering the microbial diversity and community structure, followed by rhizosphere effects. In the absence of liming effects, the rice genotypes shifted the community structure of bacteria and fungi, which accounted for the observed variation in the rice biomass. The Al-tolerant rice genotype recruited specific bacterial and fungal taxa (Bacillus, Pseudomonas, Aspergillus, and Rhizopus) associated with phosphorus solubilization and plant growth promotion. The soil microbial co-occurrence network of the Al-tolerant rice genotype was more complex than that of the Al-sensitive rice genotype. In conclusion, the bacterial and fungal community in the rhizosphere has genotype-dependent effects on rice Al tolerance. Aluminum-tolerant rice genotypes recruit specific microbial taxa, especially phosphorus-solubilizing microorganisms, and are associated with complex microbial co-occurrence networks, which may enhance rice growth in acid sulfate soil.

Expression Level of Transcription Factor ART1 Is Responsible for Differential Aluminum Tolerance in Indica Rice.

Rice is the most aluminum (Al)-tolerant species among the small grain cereals, but there are great variations in the Al tolerance between subspecies, with higher tolerance in japonica subspecies than indica subspecies. Here, we performed a screening of Al tolerance using 65 indica cultivars and found that there was also a large genotypic difference in Al tolerance among indica subspecies. Further characterization of two cultivars contrasting in Al tolerance showed that the expression level of ART1 (ALUMINUM RESISTANCE TRANSCRIPTION FACTOR 1) encoding a C2H2-type Zn-finger transcription factor, was higher in an Al-tolerant indica cultivar, Jinguoyin, than in an Al-sensitive indica cultivar, Kasalath. Furthermore, a dose-response experiment showed that ART1 expression was not induced by Al in both cultivars, but Jinguoyin always showed 5.9 to 11.4-fold higher expression compared with Kasalath, irrespectively of Al concentrations. Among genes regulated by ART1, 19 genes showed higher expression in Jinguoyin than in Kasalath. This is associated with less Al accumulation in the root tip cell wall in Jinguoyin. Sequence comparison of the 2-kb promoter region of ART1 revealed the extensive sequence polymorphism between two cultivars. Whole transcriptome analysis with RNA-seq revealed that more genes were up- and downregulated by Al in Kasalath than in Jinguoyin. Taken together, our results suggest that there is a large genotypic variation in Al tolerance in indica rice and that the different expression level of ART1 is responsible for the genotypic difference in the Al tolerance.

OsGERLP: A novel aluminum tolerance rice gene isolated from a local cultivar in Indonesia

There is a decrease in the land available for rice cultivation due to the rapid conversion to urban uses. Subsequently, acid soil could be an alternative land cultivating rice, but will require the use of aluminum (Al)-tolerant rice varieties. This Al tolerance trait is genetically controlled, and there is a need to discover more genes needed to develop Al-tolerant rice. Therefore, the objective of this study was to clone and characterize a novel Al tolerance gene isolated from a local cultivar of Indonesian rice. The gene cloning was conducted based on the rye/rice microsynteny relationship. In addition, the root growth and gene expression analyses were performed to verify the role of the gene on Al tolerance in gene-silenced rice and in overexpressed transgenic tobacco. The results showed an Al tolerance candidate gene, OsGERLP, was successfully cloned from rice cv. Hawara Bunar, with its gene encoding a protein similar to a bacterial ribosomal L32 protein. Additionally, the analysis showed that low gene expression caused the gene-silenced rice to be sensitive to Al, while high expression induced the Al tolerance in transgenic tobacco. Furthermore, it was discovered that the gene expression level in both plants was in line with the lower expression of the OsFRDL4 gene in the silenced rice and the high expression of the MATE gene in transgenic tobacco also with the higher citrate secretion from transgenic tobacco roots. In conclusion, the OsGERLP gene could act as a regulator for other Al tolerance genes, with the potential to develop Al-tolerant rice varieties.

Expression of OsFRDL1, a MATE gene family member, indicates its involvement in aluminum response in rice

In soils under acidic conditions, Aluminum (Al) is solubilized to its ionic form, which is toxic to plants. Al rapidly inhibits root elongation, water and nutrient uptake, resulting in crop yield reduction. Members of the MATE family are responsible for citrate transport and Al detoxification in different species. In rice, the OsFRDL1 gene (MATE family) is homologous to the HvAACT1 and SbMATE, which are involved in Al tolerance in barley and sorghum, respectively. Silencing OsFRDL1 showed that it is not involved in Al tolerance in rice. However, the OsFRDL1 expression was not accessed in rice genotypes contrasting for Al tolerance. Thus, in this study, four Brazilian rice genotypes were evaluated in response to Al treatment in different time of exposition and OsFRDL1 expression was analyzed. The analyzed cultivars displayed different responses to Al dose x time. Al affected root growth in all analyzed genotypes, however, a minor negative effect that only occurred after 72 and 48 hours of exposure was detected in Farroupilha and BRS Curinga cultivars, respectively. In contrast, BR-IRGA 410 and IAS 12-9 showed a negative effect in root growth from the first hours of exposure to Al. Two cultivars differing in Al tolerance were used for gene expression analysis. The expression of OsFRDL1 was highly increased in Al-tolerant cultivar Farroupilha than in the Al-sensitive cultivar BR IRGA 410. This results indicates that OsFRDL1 is regulated by Al. This finding suggests that OsFRDL1 is involved in Al stress response, however seems to be insufficient in controlling Al tolerance.

Nitric oxide reduces the aluminum-binding capacity in rice root tips by regulating the cell wall composition and enhancing antioxidant enzymes

Plant cell wall, the first interface or barrier for toxic ions entering into protoplast, suffers from risk. Nitric oxide (NO) plays an important role in plant growth and responses to abiotic stresses, however, it is not clear whether NO is connected with the response of cell wall to aluminum (Al) tolerance in rice (Oryza sativa L.). In this study, we found that the application of 50 µM Al induces nitrate reductase (NR) activity and endogenous NO production, but not nitric oxide synthase (NOS) activity in two rice genotypes. Pretreatment with 100 µM NO donor (sodium nitroprusside, SNP) reduced Al-induced inhibition of root elongation by 32.3% and 91.7%, and Al accumulation in root-tip by 38.4% and 44.3% in Nipponbare and Zhefu802, respectively. The addition of SNP significantly decreased Al-induced accumulation of pectin, hemicellulose 1 and hemicellulose 2 by 43.1%, 13.1% and 19.2% in Zhefu802 and by 16.9%, 13.4% and 14.0% in Nipponbare, compared with roots treated with Al alone, as well as pectin methylesterase (PME) activity. Therefore, the content of Al absorbed in cell walls was decreased, indicating that the Al-induced structure damage to cell walls was alleviated. Furthermore, the activities of peroxidase (POD), superoxide dismutase (SOD) and catalase (CAT) treated by Al were all increased by SNP pretreatment, and the lipid peroxidation and damage to plasma membrane of root tips detected with Schiff’s reagent and Evans blue reduced. In contrast, the effect was abolished when NO scavenger (cPTIO), and NR inhibitor (NaN3), were added. These results indicated that by regulating the Al-binding capacity to cell walls and lipid peroxidation, the structure of cell walls can be stabilized and that Al toxicity in rice can be alleviated with increased NO.

Improved Root Growth by Liming Aluminum-Sensitive Rice Cultivar or Cultivating an Aluminum-Tolerant One Does Not Enhance Fertilizer Nitrogen Recovery Efficiency in an Acid Paddy Soil.

The root is the main site of nitrogen (N) acquisition and aluminum (Al) toxicity. The objective of this study is to investigate whether liming and cultivation of an Al-tolerant rice (Oryza sativa L.) cultivar can improve root growth, thereby increasing N acquisition by rice plants in acid paddy soil. Two rice cultivars (‘B690’, Al-sensitive, and ‘Yugeng5’, Al-tolerant) were cultivated with 15N-labeled urea, and with or without lime in an acid paddy soil (pH 4.9) in pots. We examined root and shoot growth, soil pH, soil exchangeable Al, N uptake, 15N distribution in plant-soil system, and fertilizer N recovery efficiency. Results showed that liming improved the root growth of ‘B690’ by decreasing soil exchangeable Al concentrations, in both N-limited and N-fertilized soils. Liming enhanced the N uptake of ‘B690’ only in the absence of N fertilizer. The root weight of ‘Yugeng5’ was greater than that of ‘B690’ without lime, but the two cultivars showed similar N uptake. The fertilizer N recovery efficiency and N loss did not differ significantly between limed and non-limed conditions, or between the two rice cultivars. Thus, liming an Al-sensitive rice cultivar and cultivating an Al-tolerant one improves root growth, but does not enhance fertilizer N recovery efficiency in the present acid paddy soil.

Mining candidate gene for rice aluminum tolerance through genome wide association study and transcriptomic analysis

BackgroundThe genetic mechanism of aluminum (Al) tolerance in rice is great complicated. Uncovering genetic mechanism of Al tolerance in rice is the premise for Al tolerance improvement. Mining elite genes within rice landrace is of importance for improvement of Al tolerance in rice.ResultsGenome-wide association study (GWAS) performed in EMMAX for rice Al tolerance was carried out using 150 varieties of Ting’s core collection constructed from 2262 Ting’s collections with more than 3.8 million SNPs. Within Ting’s core collection of clear population structure and kinship relatedness as well as high rate of linkage disequilibrium (LD) decay, 17 genes relating to rice Al tolerance including cloned genes like NRAT1, ART1 and STAR1 were identified in this study. Moreover, 13 new candidate regions with high LD and 69 new candidate genes were detected. Furthermore, 20 of 69 new candidate genes were detected with significant difference between Al treatment and without Al toxicity by transcriptome sequencing. Interestingly, both qRT-PCR and sequence analysis in CDS region demonstrated that the candidate genes in present study might play important roles in rice Al tolerance.ConclusionsThe present study provided important information for further using these elite genes existing in Ting’s core collection for improvement of rice Al tolerance.

Method for initially selecting Al-tolerant rice varieties based on the charge characteristics of their roots

To explore the relationship between charge characteristics of rice roots and aluminum (Al) tolerance of rice, roots of 47 different rice genotypes were obtained by hydroponic experiment. The zeta potentials of roots were determined by streaming potential method, and the Al tolerance and the functional groups of rice were measured by relative root elongation and infrared spectroscopy (ATR-FTIR), respectively. The exchangeable, complexed and precipitated Al(III) sorbed on the root surface of rice was extracted with 1 mol L−1 KNO3, 0.05 mol L−1 EDTA-2Na and 0.01 mol L−1 HCl, respectively. There was a significant correlation between the zeta potentials and the relative elongation of rice roots, indicating that the zeta potentials of rice roots could be used to characterize rice tolerance to Al toxicity. Twelve Al-tolerant rice varieties, 25 medium Al-tolerant rice varieties, and 10 Al-sensitive rice varieties were obtained. The Al-tolerant rice varieties sorbed less complexed Al(III) and total Al(III) because there was lower negative charge on their roots compared to less tolerant genotypes. A correlation analysis showed that there were significant negative correlations between the zeta potential, relative root elongation, and the total Al(III) sorption capacity of the roots, which further confirmed the reliability of using the root zeta potential to characterize rice tolerance to Al toxicity. The results of this paper provide a new method for screening Al-tolerant rice varieties.

Changes in the Distribution of Pectin in Root Border Cells Under Aluminum Stress.

Root border cells (RBCs) surround the root apices in most plant species and are involved in the production of root exudates. We tested the relationship between pectin content in root tips and aluminum (Al) tolerance by comparing these parameters in wild-type (WT) and sensitive-to-Al-rhizotoxicity (star1) mutant rice plants. Staining for demethylesterified pectin decreased after Al treatment in the WT. A high level of pectin was observed in RBCs of the root tips. The level of total pectin was increased by about 50% compared with the control. In the Al-sensitive star1 mutant, Al treatment decreased root elongation and pectin content, especially in RBCs. In addition, almost no Al accumulation was observed in the control, whereas more Al was accumulated in the RBCs of STAR1 roots. These results show that the amount of pectin influences Al tolerance; that Al accumulation in rice roots is reduced by the distribution of pectin in root-tip RBCs; and that these reactions occur in the field around the RBCs, including the surrounding mucilage. Al accumulation in rice roots is reduced by the distribution of pectin in root tips, and pectin in the root cell walls contributes to the acquisition of Al tolerance in rice by regulating its distribution. The release of Al-binding mucilage by RBCs could play a role in protecting root tips from Al-induced cellular damage.

Correlation among Snpb11 markers, root growth, and physiological characters of upland rice under aluminum stress

Abstract. Fendiyanto MH, Satrio RD, Suharsono, Tjahjoleksono A, Miftahudin. 2019. Correlation among Snpb11 markers, root growth, and physiological characters of upland rice under aluminum stress. Biodiversitas 20: 1243-1254. The cultivation of upland rice in acid soils faces aluminum (Al) toxicity. Development of Al-tolerant rice could be one of the solutions to overcome the problem. Marker-assisted breeding to develop Al-tolerant rice requires at least a molecular marker for foreground selection. Snpb11 is a molecular marker developed from the nucleotide differences in a specific allele between Al-tolerant and sensitive rice. Snpb11 has never been used as a molecular marker in rice. Therefore this study aimed to examine the correlation among Snpb11 marker, root growth, and physiological characters under Al stress in upland rice. We used physiological characters and the Snpb11 marker to justify the Al tolerance level in several upland rice varieties. We found that physiological characters, i.e.: primary root length, total root length, chlorophyll, and carotenoid content showed positive correlation to Snpb11. Conversely, root malondialdehyde content, which represents the level of lipid peroxidation showed a negative correlation to Snpb11. There is evidence that the Snpb11 highly correlated with primary and total root length characters, which are the Al tolerance parameters used in rice. Therefore, Snpb11 markers can be used to distinguish the Al tolerance level in upland rice.

Mining Beneficial Genes for Aluminum Tolerance Within a Core Collection of Rice Landraces Through Genome-Wide Association Mapping With High Density SNPs From Specific-Locus Amplified Fragment Sequencing.

Trivalent Aluminum (Al3+) in acidic soils is harmful to root growth and significantly reduce crop yields. Therefore, mining beneficial genes for Al tolerance is valuable for rice production. The objective of this research is to identify some beneficial genes for Al tolerance from rice landraces with high density SNP set from SLAF-seq (Specific-Locus Amplified Fragment sequencing). A total of 67,511 SNPs were obtained from SLAF-seq and used for genome-wide association study (GWAS) for Al tolerance with the 150 accessions of rice landraces in the Ting's rice core collection. The results showed that rice landraces in the Ting's rice core collection possessed a wide-range of variation for Al tolerance, measured by relative root elongation (RRE). With the mixed linear models, GWAS identified a total of 25 associations between SNPs and Al tolerant trait with p < 0.001 and false discovery rate (FDR) <10%. The explained percentage by quantitative trait locus (QTL) to phenotypic variation was from 7.27 to 13.31%. Five of twenty five QTLs identified in this study were co-localized with the previously cloned genes or previously identified QTLs related to Al tolerance or root growth/development. These results indicated that landraces are important sources for Al tolerance in rice and the mapping results could provide important information to breed Al tolerant rice cultivars through marker-assisted selection.

The auxin influx carrier, OsAUX3, regulates rice root development and responses to aluminium stress

In rice, there are five members of the auxin carrier AUXIN1/LIKE AUX1 family; however, the biological functions of the other four members besides OsAUX1 remain unknown. Here, by using CRISPR/Cas9, we constructed two independent OsAUX3 knock-down lines, osaux3-1 and osaux3-2, in wild-type rice, Hwayoung (WT/HY) and Dongjin (WT/DJ). osaux3-1 and osaux3-2 have shorter primary roots (PRs), decreased lateral root (LR) density, and longer root hairs (RHs) compared with their WT. OsAUX3 expression in PRs, LRs, and RHs further supports that OsAUX3 plays a critical role in the regulation of root development. OsAUX3 locates at the plasma membrane and functions as an auxin influx carrier affecting acropetal auxin transport. OsAUX3 is up-regulated in the root apex under aluminium (Al) stress, and osaux3-2 is insensitive to Al treatments. Furthermore, 1-naphthylacetic acid accented the sensitivity of WT/DJ and osaux3-2 to respond to Al stress. Auxin concentrations, Al contents, and Al-induced reactive oxygen species-mediated damage in osaux3-2 under Al stress are lower than in WT, indicating that OsAUX3 is involved in Al-induced inhibition of root growth. This study uncovers a novel pathway alleviating Al-induced oxidative damage by inhibition of acropetal auxin transport and provides a new option for engineering Al-tolerant rice species.

Functional characterization of an aluminum (Al)-inducible transcription factor, ART2, revealed a different pathway for Al tolerance in rice.

High aluminum (Al) tolerance in rice (Oryza sativa) is controlled by a Cys2His2-type zinc finger transcription factor ART1 (Al resistance transcription factor1). There are five close homologs of ART1 in the rice genome, but the role of these homologs is unknown. We functionally characterized one of the ART1 homologs, ART2, in terms of tissue and spatial expression, subcellular localization, transcriptional activation activity, and phenotypic analysis of the knockout lines. ART2 was localized to the nucleus and showed a transcriptional activation potential in yeast. ART2 was mainly expressed in the roots, but the expression level was much lower than that of ART1. The ART2 expression was rapidly induced by Al in the roots of the wild-type rice, but not in art1 mutant. Knockout of ART2 resulted in increased sensitivity to Al toxicity, but did not alter sensitivity to different pH values. Expression profile analysis by RNA-sequencing showed that ART2 was not involved in activation of genes regulated by ART1; however, four genes seems to be regulated by ART2, which are implicated in Al tolerance. These results indicate that ART1 and ART2 regulate different pathways leading to Al tolerance, and ATR2 plays a supplementary role in Al tolerance in rice.

Transcription factor WRKY22 promotes aluminum tolerance via activation of OsFRDL4 expression and enhancement of citrate secretion in rice (Oryza sativa).

Whilst WRKY transcription factors are known to be involved in diverse plant responses to biotic stresses, their involvement in abiotic stress tolerance is poorly understood. OsFRDL4, encoding a citrate transporter, has been reported to be regulated by ALUMINUM (Al) RESISTANCE TRANSCRIPTION FACTOR 1 (ART1) in rice, but whether it is also regulated by other transcription factors is unknown. We define the role of OsWRKY22 in response to Al stress in rice by using mutation and transgenic complementation assays, and characterize the regulation of OsFRDL4 by OsWRKY22 via yeas one-hybrid, electrophoretic mobility shift assay and ChIP-quantitative PCR. We demonstrate that loss of OsWRKY22 function conferred by the oswrky22 T-DNA insertion allele causes enhanced sensitivity to Al stress, and a reduction in Al-induced citrate secretion. We next show that OsWRKY22 is localized in the nucleus, functions as a transcriptional activator and is able to bind to the promoter of OsFRDL4 via W-box elements. Finally, we find that both OsFRDL4 expression and Al-induced citrate secretion are significantly lower in art1 oswrky22 double mutants than in the respective single mutants. We conclude that OsWRKY22 promotes Al-induced increases in OsFRDL4 expression, thus enhancing Al-induced citrate secretion and Al tolerance in rice.