Rice Seedlings Research Articles

Effect of S-Allyl-L-Cysteine on Nitric Oxide and Cadmium Processes in Rice (Oryza sativa L. sp. Zhongzao35) Seedlings

Nitric oxide (NO) is an important signaling molecule involved in regulating plant processes to cope with abiotic stress. S-allyl-L-cysteine (SAC) is known to induce NO synthesis in animals. However, it is unknown whether SAC can trigger NO biosynthesis, regulate Cd transport, or alleviate Cd stress in plants. After being sprayed with 0.2 mM SAC, rice seedlings had a NO content that was 1.8 times higher than that of the control (ctrl) group at the ninth hour, which then gradually decreased. The expressions of Cd uptake and transport genes in the roots (including OsNRAMP5, OsNRAMP1, and OsHMA2) were markedly downregulated by 27.2%, 24.8%, and 49.1%, respectively, 72 h after SAC spraying treatment. The Cd content in seedling roots’ cell wall (CW) components significantly increased by 43.5% compared to that of the ctrl group. The Cd content in the shoots and roots decreased by 49.0% and 29.8%, respectively. Cd stress in the seedlings was also substantially alleviated. In conclusion, spraying rice seedlings with SAC triggered an increase in NO synthesis, regulated the expression of genes related to Cd transport, increased Cd fixation in the root CW components, and reduced Cd accumulation in the roots and shoots.

Hydroponic Culture of Rice Seedlings for Stress Response Assay.

The major environmental factors limiting rice growth and production are osmotic stresses such as drought and high salinity. High osmotic stresses directly disrupt cellular activities, leading to plant growth retardation or death. Plants have various response mechanisms to survive under such stresses. Understanding rice's stress response mechanisms is necessary to enhance the osmotic stress tolerance of rice. However, assessing specific physiological responses to osmotic stresses is difficult because multiple environmental factors affect rice growth. Here, we describe a simple method for analyzing the osmotic stress responses of rice plants using a hydroponic culture system. This method allows comprehensive gene expression and phenotypic analyses under osmotic stress conditions in rice. Various osmotic stress conditions and samples can be tested simultaneously because this method is small-scale. In addition, the procedure is easy, and highly reproductive results can be obtained.

An ABA biosynthesis enzyme gene OsNCED4 regulates NaCl and cold stress tolerance in rice

Rice (Oryza sativa L.) is susceptible to various abiotic stresses, such as salt, cold, and drought. Therefore, there is an urgent need to explore the relevant genes that enhance tolerance to these stresses. In this study, we identified a gene, OsNCED4 (9-cis-epoxycarotenoid dioxygenase 4), which regulates tolerance to multiple abiotic stresses. OsNCED4 encodes a chloroplast-localized abscisic acid (ABA) biosynthetic enzyme. The expression of OsNCED4 gene was significantly induced by 150 mM NaCl and cold stress. Disruption of OsNCED4 by CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)/Cas9-mediated mutagenesis resulted in significant sensitivity to NaCl and cold stress. The salt and cold sensitivity of osnced4 mutant was due to the reduction of ABA content and the excessive accumulation of reactive oxygen species (ROS) under stress. Moreover, OsNCED4 also regulates drought stress tolerance of rice seedlings. Taken together, these results indicate that OsNCED4 is a new regulator for multiple abiotic stress tolerance in rice, and provided a potential target gene for enhancing multiple stress tolerance in the future.

Toxicity and Glutathione S-Transferase-Catalyzed Metabolism of R-/S-Metolachlor in Rice.

Metolachlor, the chiral herbicide, inhibits the very-long-chain fatty acid (VLCFA) synthesis; elucidating the enantioselectivity between R- and S-metolachlor in the toxicological difference will facilitate the understanding of the site of action. We found that the endogenous accumulation of C22 VLCFAs decreased in both R-/S-metolachlor -treated plants by 6, 12, and 24 h after treatment, with more significant reduction in the S isomer group. Gene expression of glutathione S-transferase OsGST Tau members were obviously induced upon treatments with S or R isomer; both OsGSTU1 and OsGSTU4 can metabolize metolachlor effectively, with S isomer as the preference by directly catalyzing the conjugation between S-metolachlor and glutathione. In the current study, we provide the first evidence in rice seedlings that S-metolachlor showed herbicidal toxicity by blocking the synthesis of C22-type fatty acid, which eventually affects the whole elongation chain of (V)LCFA. Meanwhile, OsGSTU1 and 4 metabolize the metolachlor with the S isomer as preference. All of these discoveries broaden our knowledge about metolachlor toxicology and enantioselectivity.

CRISPR/Cas knockout of the NADPH oxidase gene OsRbohB reduces ROS overaccumulation and enhances heat stress tolerance in rice.

Heat stress (HS) has become a major factor limiting crop yields worldwide. HS inhibits plant growth by ROS accumulation, and NADPH oxidases (Rbohs) are major ROS producers in plants. Here, we show that CRISPR/Cas knockout of the OsRbohB (OsRbohB-KO) significantly increased rice tolerance to HS imposed at various different growth stages. We produced OsRbohB-KO and OsRbohB-overexpression (OsRbohB-OE) lines in a japonica cultivar, Nipponbare. Compared with nontransgenic wild-type (WT) plants, the OsRbohB-KO lines showed a significant increase in chlorophyll contents (5.2%-58.0%), plant growth (48.2%-65.6%) and grain yield (8.9%-20.5%), while reducing HS-induced ROS accumulation in seeds (21.3%-33.0%), seedlings (13.0%-30.4%), anthers (13.1%-20.3%) and grains (9.7%-22.1%), under HS conditions. Analysis of yield components revealed that the increased yield of OsRbohB-KO plants was due to increased starch synthetase activity, spikelets per panicle (2.0%-9.3%), filled spikelets (4.8%-15.5%), percentage of filled spikelets (2.4%-6.8%) and 1000-grain weight (2.9%-7.4%) under HS conditions during the reproductive stage. Grain milling and appearance quality, and starch content were also significantly increased in OsRbohB-KO plants under HS conditions during the mature stage. Furthermore, OsRbohB-KO significantly upregulated the expression levels of heat shock-related genes, OsHSP23.7, OsHSP17.7, OsHSF7 and OsHsfA2a, in rice seedlings and grains under long-term HS conditions. Conversely, OsRbohB-OE resulted in phenotypes that were opposite to OsRbohB-KO in most cases. Our results suggest that suppression of OsRbohB provides an effective approach for alleviating heat damage and improving grain yield and quality of rice under long-term HS conditions.

Magnesium-Doped Carbon Quantum Dot Nanomaterials Alleviate Salt Stress in Rice by Scavenging Reactive Oxygen Species to Increase Photosynthesis.

Salt stress has strongly impacted the long-term growth of eco-friendly farming worldwide. By targeting the oxidative stress induced by salt, the utilization of biomass-derived carbon dots (CDs) that possess high-efficiency antioxidant properties, are nontoxic, and have excellent biocompatibility represents a viable and effective approach for enhancing the salt tolerance of plants. In this study, we blended magnesium oxide nanoparticles with carbon sources derived from durian shells to construct Mg-doped carbon dots (Mg-CDs) through a hydrothermal reaction. We demonstrated that the foliar application of 150 μg/mL Mg-CDs to rice plants after treatment with 100 mM salt effectively increased the plant height (9.52%), fresh weight (22.41%), dry weight (33.33%), K+ content (21.46%), chlorophyll content (36.21%), and carotenoid content (16.21%); decreased the malondialdehyde (MDA) (9.43%), Na+ (25.75%), H2O2 (17.50%), and O2•- contents (37.99%); and promoted the photosynthetic system and antioxidant activity. Transcriptome analysis revealed that Mg-CD pretreatment triggered transcriptional reprogramming in rice seedlings. The enrichment analysis of the Kyoto Encyclopedia of Genes and Genomes pathways based on trend groups of gene expression patterns of Profile 8 and Profile 15 indicated that priming with Mg-CDs activated stress signaling- and defense-related pathways, such as metabolic pathways, biosynthesis of secondary metabolites, and photosynthesis pathways. These activations subsequently prompted the expression of genes related to the mitogen-activated protein kinase signaling pathway, hormone signal transduction, the oxidative stress response, and the photosynthetic system. This study demonstrated that the use of Mg-CDs represents a potential strategy to increase plant salt tolerance, creating the possibility for the regulation of crop salinity stress and offering valuable advancements in sustainable agriculture.

Trivalent chromium stress trigger accumulation of secondary metabolites in rice plants: Integration of biochemical and transcriptomic analysis

Trivalent chromium [Cr (III)] is a frequently detected environmental contaminant that negatively affects plant metabolism, growth, and yield. Plant secondary metabolites are non-nutrient compounds involved in diverse physiological functions in response to abiotic stresses. This study performed biochemical and transcriptomic analyses to clarify the protective role and mechanism of secondary metabolites in rice seedlings under Cr(III) stress. The content of measured secondary metabolites i.e., total soluble phenolics, flavonoids, lignin, and anthocyanin in rice tissues was significantly enhanced under Cr(III) exposure. Additionally, the activities of several enzymes including chalcone synthase, phenylalanine ammonialyase, peroxidase, and anthocyanidin synthase, were positively activated. Transcriptomic analysis revealed a number of differentially expressed genes (DEGs) in Cr(III)-treated rice seedlings and their expression patterns are tissue-specific. Additionally, the DEGs involved in secondary metabolism pathways were evident after KEGG enrichment analysis. Cr(III) exposure resulted in distinct genetic regulation strategies in rice tissues, with different DEGs activating various sub-pathways in the secondary metabolism pathways to cope with Cr(III) stress. Furthermore, the integrated correlation analysis of the secondary metabolites and transcriptome data identified key genes involved in different steps of the sub-pathways of secondary metabolism pathway in rice plants under Cr(III) stress. This study indicates that the accumulation of secondary metabolites in rice plants is a survival and detoxification strategy to reduce the adverse effects of toxic compounds. Future research should explore how key genes in secondary metabolism pathways aid in Cr(III) detoxification and enhance crop stress tolerance.

RiGaD: An aerial dataset of rice seedlings for assessing germination rates and density

RiGaD: An aerial dataset of rice seedlings for assessing germination rates and density

Bacterial endophyte Pseudomonas mosselii PR5 improves growth, nutrient accumulation, and yield of rice (Oryza sativa L.) through various application methods

BackgroundPseudomonas spp. have drawn considerable attention due to their rhizospheric abundance and exceptional plant growth-promoting attributes. However, more research is needed on the optimal application methods of Pseudomonas mosselii for rice growth, nutrient accumulation, and yield improvement. This research explored the application of the endophytic bacterium P. mosselii PR5 on rice cultivar BRRI dhan29 with four treatments: control, seedling priming, root drenching, and bacterial cell-free culture (CFC) foliar application.ResultsPR5 led to better rice growth, improved nutrient acquisition, and higher yields compared to the control, regardless of the application method used. The highest results in fresh weight of root (146.93 g/pot), shoot (758.98 g/pot), and flag leaf (7.88 g/pot), dry weight of root (42.16 g/pot), shoot (97.32 g/pot), and flag leaf (2.69 g/pot), and grains/panicle (224.67), were obtained from seedling priming treatment, whereas root drenching resulted in maximum plant height (105.67 cm), root length (49.0 cm), tillers/pot (23.7), and panicles/pot (17.67). In all three application methods, rice grain yield per pot was higher in PR5 inoculated treatments, compared to the control. The amount of P, Mg and Zn in the shoot and N, P, Ca, Mg and Si content in the flag leaf was significantly increased along with effective suppression of naturally occurring blast disease in bacterial CFC foliar application, validated by multivariate analysis.ConclusionOur results indicated that rice seedlings priming with PR5 improved rice growth, yield and nutrient uptake, whereas CFC foliar application significantly increased the concentration of most nutrients in the rice plant and suppressed the naturally occurring rice blast disease. This research highlights the significant potential of P. mosselii PR5 in enhancing rice growth, yield, and nutrient uptake, particularly through seedling priming and CFC foliar application methods.

Further enhancement of cold tolerance in rice seedlings by Piriformospora indica collaborating with plant growth-promoting bacteria: Evidence from the antioxidant defense, osmoregulation, photosynthesis, and related genes

As the global population continues to grow, so does the demand for food. However, cold stress due to global climate change greatly threatens the safe production of rice. Piriformospora indica plays a pivotal role as a functional microbiota in improving cold tolerance in rice. However, the addition of P. indica alone has a limited effect. In this study, an attempt was made to select plant growth-promoting bacteria (Agrobacterium rhizogenes and Bacillus subtilis) as helper bacteria to help P. indica function better and further enhance rice cold tolerance. Under cold stress, the co-addition of the three beneficial microorganisms significantly increased the biomass, photosynthetic performance indicators, osmoregulatory substance contents, and antioxidant enzyme activities of rice. P. indica and helper bacteria significantly reduced the cold stress-induced increases in malondialdehyde content and electrolyte leakage rate in rice leaves and triggered a substantial upregulation of cold-tolerant genes in rice. This suggests that P. indica and helper bacteria can synergistically improve the cold resistance of rice. The findings of the research offer a conceptual foundation and practical assistance for enhancing rice's resilience to cold temperatures.

PfGSTF2 endows resistance to quizalofop-p-ethyl in Polypogon fugax by GSH conjugation.

Populations of Polypogon fugax have developed resistance to many acetyl-CoA carboxylase (ACCase)-inhibiting herbicides. This resistance threats the effectiveness and sustainability of herbicide use. In our previous research, a field P. fugax population exhibited GST-based metabolic resistance to the widely used ACCase-inhibiting herbicide quizalofop-p-ethyl. Here, in this current study, we identified and characterized two GST genes (named as PfGSTF2 and PfGSTF58) that showed higher expression levels in the resistant than the susceptible population. Transgenic rice calli overexpressing PfGSTF2, but not PfGSTF58, became resistant to quizalofop-p-ethyl and haloxyfop-R-methyl. This reflects similar cross-resistance pattern to what was observed in the resistant P. fugax population. Transgenic rice seedlings overexpressing PfGSTF2 also exhibited resistance to quizalofop-p-ethyl. In contrast, CRISPR/Cas9 knockout of the orthologue gene in rice seedlings increased their sensitivity to quizalofop-p-ethyl. LC-MS analysis of in vitro herbicide metabolism by Escherichia coli-expressed recombinant PfGSTF2 revealed that quizalofop (but not haloxyfop) was detoxified at the ether bond, generating the GSH-quizalofop conjugate and a propanoic acid derivative with greatly reduced herbicidal activity. Equally, these two metabolites accumulated at higher levels in the resistant population than the susceptible population. In addition, both recombinant PfGSTF2 and PfGSTF58 can attenuate cytotoxicity by reactive oxygen species (ROS), suggesting a role in plant defence against ROS generated by herbicides. Furthermore, the GST inhibitor (NBD-Cl) reversed resistance in the resistant population, and PfGSTF2 (but not PfGSTF58) responded to NBD-Cl inhibition. All these suggest that PfGSTF2 plays a significant role in the evolution of quizalofop resistance through enhanced herbicide metabolism in P. fugax.

Melatonin Mediates Trade–Off in Antioxidant Responses and Nitrogen Metabolism and Benefits the Growth of Fragrant Rice Seedlings Under Light–Nitrogen Levels

Melatonin Mediates Trade–Off in Antioxidant Responses and Nitrogen Metabolism and Benefits the Growth of Fragrant Rice Seedlings Under Light–Nitrogen Levels

Exogenous Brassinolide Ameliorates the Adverse Effects of Gamma Radiation Stress and Increases the Survival Rate of Rice Seedlings by Modulating Antioxidant Metabolism

Gamma irradiation-based mutant creation is one of the most important methods for rice plant mutagenesis breeding and molecular biology research. Although median lethal dose irradiation severely damages rice seedlings, applying brassinolide (BR) can increase the survival rate of irradiated seedlings. In this study, we investigated the effects of soaking seeds in solutions containing different BR concentrations (0.001, 0.01, 0.1, 1.0, and 5.0 μmol/L) and then spraying the resulting seedlings twice with 0.1 μmol/L BR. The combined BR treatments markedly decreased the superoxide anion (O2•−), hydrogen peroxide (H2O2), and malondialdehyde contents but increased the chlorophyll content. An appropriate BR treatment of gamma-irradiated samples substantially increased the activities of the antioxidant enzymes superoxide dismutase, peroxidase, and ascorbate peroxidase as well as the proline, ascorbic acid, and glutathione contents in rice seedling shoots. The BR treatment also promoted the growth of seedlings derived from irradiated seeds and increased the shoot and root fresh and dry weights. Most notably, soaking seeds in 0.01 or 0.1 μmol/L BR solutions and then spraying seedlings twice with 0.1 μmol/L BR significantly increased the final seedling survival rate and decreased mutant loss. The study results suggest that exogenous BR treatments can protect rice seedlings from gamma irradiation stress by enhancing antioxidant metabolism.

Seed Priming with Dynamically Transformed Selenium Nanoparticles to Enhance Salt Tolerance in Rice.

Seed priming with nanomaterials is an emerging approach for improving plant stress tolerance. Here, we demonstrated a mechanism for enhancing salt tolerance in rice under salt stress via priming with nonstimulatory nanoparticles such as selenium nanoparticles (SeNPs), distinct from stimulatory nanomaterials. Due to the dynamic transformation ability of SeNPs, SeNP priming could enhance rice salt tolerance by mediating the glutathione cycle to eliminate excess reactive oxygen species (ROS). During priming, SeNPs penetrated rice seeds and transitioned into a soluble form (99.9%) within the embryo endosperm. Subsequently, the soluble selenium (Se) was transported to rice roots and metabolized into various Se-related derivatives, including selenomethionine (SeMet), Na2SeO3 (Se IV), selenocysteine (SeCys2), and methylselenocysteine (MeSeCys). These derivatives significantly enhanced the root activities of key enzymes such as glutathione peroxidase (GSH-PX), glutathione reductase (GR), catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD) by 24.97%, 47.98%, 16.23%, 16.81%, and 14.82%, respectively, thus reinforcing the glutathione cycle and ROS scavenging pathways. Moreover, these alterations induced transcriptional changes in rice seedlings, with genes involved in signal transduction, transcription factors (TFs), ROS scavenging, and protein folding being upregulated, activating signal perception and self-repair mechanisms. These findings offer valuable insights for the agricultural application of nanomaterials.

Isolation and Characterization of Arsenic-Tolerable Bacteria from Groundwater and Their Implementation on Rice Seedling's Shoot and Root Enhancement.

Arsenic exerts detrimental impacts on primary metabolism in plants, leading to reduced crop yield. Some arsenic-resistant plant growth-promoting bacteria (PGPB) help plants by providing some plant hormones to sustain their growth and development under arsenic stress. Here, seven different species of Bacillus were isolated from arsenic-contaminated groundwater of West Bengal, India. Those species were capable of growing in the presence of > 3.12g/L arsenate (AsV) and > 0.65g/L arsenite (AsIII) salts and also resist different heavy metals like Cu2+, Fe2+, Co2+, Zn2+, Pb2+, etc. They were susceptible to multiple groups of antibiotics like beta-lactam, aminoglycosides, etc. All species were capable of detoxifying arsenite and influenced rice seedlings' growth in the presence of arsenic salts by their capabilities likenitrogen-fixing ability, phosphate solubilization, indole 3-acetic acid (IAA), gibberellic acid (GA), proline production, etc. Most species helped enhance root and shoot lengths under arsenic stress. These primary findings suggest that those Bacillus spp. could be used as potential bio-fertilizers in arsenic-contaminated agricultural fields.

Effects of Decabromodiphenyl Ethane and Cadmium Coexposure on Their Bioaccumulation, Oxidative Stress, Root Metabolism, and Rhizosphere Soil Microorganisms in a Soil-Rice System.

Decabromodiphenyl ethane (DBDPE) and cadmium (Cd) are typical pollutants in e-waste, seriously threatening crop growth. This study investigated the bioaccumulation and toxicity mechanisms of DBDPE and Cd in a soil-rice system. The results showed that 50 mg/kg DBDPE could reduce the level of accumulation of Cd in rice roots. DBDPE and Cd induced the antioxidant system (SOD, POD, and MDA) in rice seedlings. The combined exposure reduced the contents of carbohydrates, lipids, amino acids, and organic acids. Phenylalanine and phenylpropanoid metabolisms were identified as the key detoxification metabolic pathways under combined exposure. DBDPE and Cd disrupted the functional cycling of carbon and nitrogen in rhizosphere soil, while Gemmatimonadetes, Actinobacteria, and Bacteroidetes were the key bacterial groups responding to DBDPE and Cd stress. This work provides data for the toxicity risk evaluation of DBDPE and Cd combined exposure to food crops.

Influence of Foliar Application of Nanoparticles on Low Temperature Resistance of Rice Seedlings

Chilling stress, a common abiotic factor, adversely affects the growth and biomass of rice seedlings during the early stages, ultimately reducing the yield. Effective strategies to mitigate these negative impacts are essential for improving rice productivity. The application of nanotechnology in agriculture, particularly nanoparticles (NPs), has shown a promising effect in alleviating chilling stress in plants. This study evaluates the effects of various nanoparticles, ZnO (0, 50, 100, and 200 mg/L), Fe2O3 (0, 50, 75, and 100 mg/L), TiO2 (0, 50, 75, and 100 mg/L), and CeO2 (0, 50, 75, and 100 mg/L) on the chilling resistance with one control (a water spray) under a normal temperature. Four rice cultivars: LLY-7108 and XZX-6 (Low-temperature-tolerant), and LLY-32 and ZJZ-17 (Low-temperature-susceptible) were tested in this experiment. Rice seedlings were subjected to low temperature conditions (12 h light 14 °C/12 h dark, at 10 °C) for five days, followed by seven days of recovery. The results of this study demonstrate that NPs significantly enhanced seedling height fresh/dry weight and root length compared to untreated controls under chilling stress. NP treatment also reduced the reactive oxygen species (ROS), malondialdehyde (MDA), and proline content, while enhancing superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities, thereby mitigating oxidative damage. The four rice varieties exhibited clear signs of rapid growth recovery and positive physiological changes due to NPs’ application. Among the tested cultivars, LLY-7108 showed the most substantial recovery and physiological responses, while ZJZ-17 exhibited the least. The findings of this study indicate that the foliar application of ZnO (100 mg/L), Fe2O3 (50 mg/L), TiO2 (50 mg/L), and CeO2 (75 mg/L) NPs effectively mitigates chilling stress in rice seedlings, likely by enhancing the antioxidant enzymatic activity while reducing the oxidative damage. This study highlights the potential of NPs as effective agents in reducing the adverse effects of chilling stress on rice.

Elevated CO2 enhances growth and cyanide assimilation in nitrogen-deficient rice: A transcriptome and metabolomic perspective

Plants face multiple challenges from environmental pollutants and higher emissions of atmospheric CO2. Therefore, a hydroponic-based experiment was used to explore the combined effects of elevated [CO2] (700 ppm) and exogenous cyanide (CN−) (3.0 mg CN/L) on rice seedlings under nitrogen deficiency, utilizing metabonomic and transcriptomic analysis. Elevated [CO2] significantly improved the growth of CN−-treated rice seedlings compared to those with ambient [CO2] (350 ppm), and it also significantly affected CN− assimilation. Transcriptome analysis revealed distinct impacts on differentially expressed genes (DEGs) across treatments and tissues. KEGG analysis showed variability in DEGs enriched in amino acid (AA) and energy metabolism pathways due to elevated [CO2] and CN−. Metabonomic indicated that higher input of [CO2] and exogenous CN− more severely impacted energy metabolism elements than the individual species of AAs. Positive synergistic effects of elevated [CO2] and CN− were observed for glutamine and asparagine in shoots, and methionine in roots, wherein negative effects were noted for phenylalanine in shoots, and phenylalanine, valine, and alanine in roots. Meanwhile, positive effects on fumarate in shoots and α-ketoglutarate and succinate in roots were also found. Overall, elevated [CO2] enhanced growth in CN−-treated rice seedlings under nitrogen deficiency by altering AA and energy metabolism. This is the first attempt to provide new evidence of [CO2]-based gaseous fertilization as an energy-saving strategy for rice plants fed with biodegradable N-containing pollutants as a supporting N source under N deficient conditions.

OsAAH confers salt tolerance in rice seedlings.

Soil salinization is becoming a great threat that reduces crop productivity worldwide. In this study, we found that rice allantoate amidohydrolase (OsAAH) expression was significantly upregulated by salt stress, and its overexpression conferred salt tolerance at the seedling stage. Compared to wild type (WT), the contents of ureides (allantoin and allantoate) were significantly increased in Osaah mutants and reduced in OsAAH overexpression lines both before and after salt treatments. Exogenous allantoin significantly promoted salt tolerance in OsAAH overexpression, but not in Osaah mutants. Subcellular localization showed that OsAAH was also localized to the peroxisomes in addition to the previously reported endoplasmic reticulum (ER). The differential expression of peroxisome-related genes was identified between Osaah mutants and WT. Furthermore, the contents of H2O2 and malondialdehyde (MDA) were significantly accumulated in Osaah mutants and reduced in OsAAH overexpression lines. The activities of antioxidant enzymes were significantly reduced in Osaah mutants and enhanced in OsAAH overexpression under NaCl treatment. The transcription factor OsABI5 could directly bind to OsAAH promoter and activate OsAAH expression. Our findings reveal that OsAAH could be induced by salt stress through the activation of OsABI5 and then confer salt tolerance by enhancing the scavenging capacity of reactive oxygen species (ROS), which contributes to rice breeding in salt tolerance.

Overexpression of the RAV Transcription Factor OsAAT1 Confers Enhanced Arsenic Tolerance by Modulating Auxin Hemostasis in Rice.

Characterization of arsenic (As)-responsive genes is fundamental to solving the issue of As contamination in rice. Herein, we establish the involvement of an RAV transcription factor OsAAT1 (Arsenic Accumulation and Tolerance 1) in regulating As response in rice. The expression of OsAAT1 is significantly higher in roots and stems of rice seedlings and is clearly upregulated by higher concentrations of arsenite [As(III)]. Compared with wild-type (WT) plants, OsAAT1-overexpressed transgenic lines (OE-OsAAT1) exhibit tolerance, while OsAAT1-knockout mutants (Osaat1) are sensitive to As(III) stress. Notably, the application of exogenous 1-naphthylacetic acid (NAA) greatly enhances the As tolerance of WT and transgenic lines, with stronger effects on OE-OsAAT1. The change in OsAAT1 expression leads to the alteration of As and auxin accumulation in transgenic plants by regulating the expression of OsLsi1, OsLsi2, OsCRL4, and OsAUX1 genes. Moreover, overexpression of OsAAT1 accelerates ROS scavenging and phytochelatins (PCs) synthesis, especially with the addition of exogenous NAA. OsAAT1 localizes in the nucleus and works as a transcriptional suppressor. OsGH3-12, belonging to the auxin-responsive GH3 gene family, is the downstream target gene of OsAAT1, whose expression is extensively downregulated by As(III). These findings provide new insights into As response via auxin signaling pathway in rice.