Rice Mutant Research Articles

Natural mutation in Stay-Green (OsSGR) confers enhanced resistance to rice sheath blight through elevating cytokinin content.

Sheath blight (ShB), caused by Rhizoctonia solani, is a highly destructive disease in many crops worldwide and no major resistance genes are available. Here, we identified a sbr1 (sheath blight resistance 1) rice mutant, which shows enhanced ShB resistance and maintains wildtype agronomic traits including yield, but carries an undesired stay-green phenotype. Through map-based cloning and transgenic validation, we found that an insertion disrupting the Stay-Green (OsSGR) gene is responsible for sbr1 phenotypes. Mechanistically, the sbr1/Ossgr mutants reduce the expression of most OsCKX genes, which function in cytokinin (CK) degradation, to accumulate CK leading to ShB resistance. Importantly, knockout of OsCKX7, predominantly expressed in the leaf sheath and highly induced by R. solani, significantly enhances ShB resistance without stay-green phenotype nor yield penalty, showing high application potential. Thus, our study reveals novel insights that OsSGR and cytokinin play key roles in rice-R. solani interaction and generates a valuable ShB-resistant germplasm.

Distinct lignocelluloses of plant evolution are optimally selective for complete biomass saccharification and upgrading Cd2+/Pb2+ and dye adsorption via desired biosorbent assembly

In this study, 15 plant species representing plant evolution were selected, and distinct lignocellulose compositions for largely varied biomass enzymatic saccharification were detected. By comparison, the acid-pretreated lignocellulose of rice mutant was of the highest Congo-red adsorption (298 mg/g) accounting for cellulose accessibility, leading to complete cellulose hydrolysis and high bioethanol production. By conducting oxidative-catalysis with the acid-pretreated lignocellulose of moss plant, the optimal biosorbent was generated with maximum Cd/Pb adsorption (54/118 mg/g), mainly due to half-reduced cellulose polymerization degree and raised functional groups accountable for multiple physical and chemical interactions. Furthermore, the acid-pretreated lignocellulose of eucalyptus was of large and small pores for much higher adsorption capacities with direct-yellow and direct-blue than those of the previously-reported. Therefore, this study raises a mechanism model about how distinct lignocelluloses of plant evolution are selective for complete biomass saccharification and optimal biosorbents assembly, providing insights into lignocellulose biosynthesis and biomass conversion.

Disruption of the OsWRKY71 transcription factor gene results in early rice seed germination under normal and cold stress conditions

BackgroundEarly seed germination in crops can confer a competitive advantage against weeds and reduce the time to maturation and harvest. WRKY transcription factors regulate many aspects of plant development including seed dormancy and germination. Both positive and negative regulators of seed germination have been reported in many plants such as rice and Arabidopsis. Using a transient expression system, we previously demonstrated that OsWRKY71 is a negative regulator of gibberellin (GA) signaling in aleurone cells and likely forms a “repressosome” complex with other transcriptional repressors. Hence, it has the potential to impact seed germination properties.ResultsIn this study, we demonstrate that OsWRKY71, a Group IIa WRKY gene, appeared at the same time as seed-bearing plants. Rice mutants lacking OsWRKY71 have seeds and embryos that germinate earlier than wildtype controls. In oswrky71 aleurone layers, α-amylase activity was hypersensitive to stimulation by GA3 and hyposensitive to inhibition by abscisic acid (ABA). Early germination in oswrky71 intact seeds was also hyposensitive to ABA. Transcriptomic profiling during embryo germination and early post-germination growth demonstrates that OsWRKY71 influences the expression of 9–17% of genes in dry and imbibing embryos. Compared to wildtype embryos, the mutant transcriptomes have large temporal shifts at 4, 8 and 12 h after imbibition (HAI). Importantly, many genes involved in the ABA-dependent inhibition of seed germination were downregulated in oswrky71-1. This mutant also displayed altered expression of multiple ABA receptors (OsPYLs/RCARs) that control ABA signaling and the VP1-SDR4-DOG1L branch of ABA signaling that promotes seed dormancy. Association studies reveal an OsWRKY71-containing quantitative trait locus involved in low-temperature seed germinability, qLTG-2. Indeed, oswrky71 seeds germinated early at 15 °C.ConclusionsRice Group-IIa WRKY transcription factor OsWRKY71 is a master regulator of germination that influences the expression of 9–17% of genes in dry and imbibing embryos. It is also most likely the primary candidate of low-temperature seed germinability QTL, qLTG-2. We propose that knockouts of OsWRKY71 can generate rice varieties with improved germination properties under normal or low-temperature conditions.

Rice sucrose non-fermenting related protein kinase (SnRK1) has a limited role in defense against Fall armyworm (Spodoptera frugiperda)

Sucrose non-fermenting (SNF) related protein kinase 1 (SnRK1) is a master regulator of energy deprivation signaling and has also been implicated in abiotic and biotic stress regulation. SnRK1 promotes stress tolerance through metabolic and transcriptional changes and plays important roles in innate immunity against various pathogens. However, whether it plays a role against insect herbivory is not understood. To test this, using the wild type (with SnRK1) and snrk1 mutant lines in rice, we examined the potential role of SnRK1 in rice against the ruinous pest, Fall armyworm (FAW), Spodoptera frugiperda. We also investigated the response of FAW towards these lines at different time intervals after exogenous application of plant hormone, Jasmonic acid (JA), and a JA blocker (Ibuprofen). Additional experiments by feeding FAW with leaf infused diet, fresh leaves, and a short-term exposure of FAW to the lines were also carried out. FAW mass gain, growth and development, and host ecophysiological traits were observed. In addition, we also quantified the major surface defenses- trichomes, and wax before and after herbivory. Our results show that FAW response did not vary between mutants and wild type rice. Meanwhile, we found plant hormonal application influenced the ecophysiological traits regardless of mutants and wild type rice. Collectively, we show that while defense against FAW in rice is JA mediated, SnRK1 has a limited role as observed through manipulative experiments with the wild type and snrk1 mutant rice lines.

The MON1-CCZ1 complex plays dual roles in autophagic degradation and vacuolar protein transport in rice.

Autophagy is a highly conserved cellular program in eukaryotic cells which mediates the degradation of cytoplasmic components through the lysosome, also named the vacuole in plants. However, the molecular mechanisms underlying the fusion of autophagosomes with the vacuole remain unclear. Here, we report the functional characterization of a rice (Oryza sativa) mutant with defects in storage protein transport in endosperm cells and accumulation of numerous autophagosomes in root cells. Cytological and immunocytochemical experiments showed that this mutant exhibits a defect in the fusion between autophagosomes and vacuoles. The mutant harbors a loss-of-function mutation in the rice homolog of Arabidopsis thaliana MONENSIN SENSITIVITY1 (MON1). Biochemical and genetic evidence revealed a synergistic interaction between rice MON1 and AUTOPHAGY-RELATED 8a in maintaining normal growth and development. In addition, the rice mon1 mutant disrupted storage protein sorting to protein storage vacuoles. Furthermore, quantitative proteomics verified that the loss of MON1 function influenced diverse biological pathways including autophagy and vacuolar transport, thus decreasing the transport of autophagic and vacuolar cargoes to vacuoles. Together, our findings establish a molecular link between autophagy and vacuolar protein transport, and offer insights into the dual functions of the MON1-CCZ1 (CAFFEINE ZINC SENSITIVITY1) complex in plants.

The Effect of Gamma Irradiation 100 Gy on Stained Rice Seeds Msp-04 to on Vegetative and Generative Mutant Characters of M2 Generation

This study aims to obtain information on changes in the character of individual M2 generation mutant rice, similarities between individuals of the M2 generation mutant and its comparison, and correlation between variables. This research was conducted by observing the vegetative, generative, and potential yields of each individual rice mutant and its comparison. The data obtained were tested by cluster analysis and correlation analysis. The observations showed that almost all the characters of the M2 generation mutant individuals changed except for the qualitative vegetative characters. Cluster analysis based on quantitative vegetative character showed 1 different mutant individual (0% similarity), and 1 other individual with 38.83% similarity compared to other and non-mutant mutants, and commercial varieties. Based on quantitative and production generative characters, there are 2 distinct groups that differ from each other (0% similarity). The similarity range of each distinct group is 21.17-43.56%. The strong to very strong and very significant correlation that occurs in vegetative and generative characters with production is found between the number of tillers and panicles, total grain weight and dry grain weight of 98.7% each; 60.9% and 58.3%; the number of leaves is thus also between the number of panicles, total grain weight and dry grain weight respectively 91.3%; 53.3% and 53.4%; panicles with total grain weight and dry grain weight were 60.1% and 57.6%, respectively.

Heredity and fine mapping of a yellow leaf and less tillering mutant yllt10 in rice

Rice (Oryza sativa L.) is a major food crop and increasing rice yield is the primary objective of rice research. Photosynthesis and nitrogen utilization efficiency directly affect the tiller number of rice, which affects the yield of rice. In this study, a stable yellow leaf and less tillering rice mutant yllt10 (yellow leaf and less tillering 10) was obtained by heavy-ion beam mutagenesis of rice variety 'Ke-fu-geng 7'. Compared with the wild type, yllt10 showed reduced chlorophyll content, decreased photosynthesis rate, and abnormal chloroplast structure. The genetic analysis indicated that the phenotype of yllt10 was controlled by a single recessive nuclear gene. Map-based cloning localized YLLT10 between two molecular markers J4 and J5 on chromosome 10. The sequencing of candidate genes within this interval revealed that YLLT10 was an allelic mutation of CAO1/PGL with a single base deletion in the first exon resulting in the frame shift mutation of CAO1/PGL, and YLLT10 was a new allelic variation of CAO1/PGL. The mutant yllt10 was insensitive to changes in nitrogen concentration when being incubated with different nitrogen concentrations. YLLT10 controls leaf color and tiller number and affects photosynthesis and yield of rice. The study of this gene provides a theoretical basis for molecular breeding of rice.

A Cyclin Gene OsCYCB1;5 Regulates Seed Callus Induction in Rice Revealed by Genome Wide Association Study

Plant tissue culture is extensively employed in plant functional genomics research and crop genetic improvement breeding. The callus induction ability is critical for utilizing Agrobacterium-mediated genetic transformation. In this study, we conducted a genome-wide association study (GWAS) utilizing 368 rice accessions to identify traits associated with callus induction rate (CIR), resulting in the identification of a total of 104 significant SNP loci. Integrated with gene function annotation and transcriptome analysis, 13 high-confidence candidate genes involved in auxin-related, CYC cyclins, and histone H3K9-specific methyltransferase were identified in significant loci. Furthermore, we also verified a candidate gene, Os05g0493500 (OsCycB1;5), and employed the CRISPR/Cas9 system to generate OsCycB1;5 knockout mutants in rice (Oryza sativa L.). The OscycB1;5 mutant displays significantly reduced callus induction and proliferation capacity, this result indicating OsCycB1;5 is required for the callus formation in rice. Overall, this study provides several reliable loci and high-confidence candidate genes that may significantly affect callus formation in rice. This information will offer valuable insights into the mechanisms underlying callus formation not only in rice but also in other plants.

OsKANADI1 and OsYABBY5 regulate rice plant height by targeting GIBERELLIN 2-OXIDASE6.

Plant height is an important agronomic characteristic of rice (Oryza sativa L.). Map-based cloning analyses of a natural semi-dwarf rice mutant with inwardly curled leaves found in the field revealed that the defects were due to a mutation of a SHAQKYF-class MYB family transcription factor, OsKANADI1 (OsKAN1). OsKAN1 directly bound to the OsYABBY5 (OsYAB5) promoter to repress its expression and interacted with OsYAB5 to form a functional OsKAN1-OsYAB5 complex. GIBERELLIN 2-OXIDASE6 (OsGA2ox6), encoding an enzyme in the gibberellin (GA) catabolic pathway, was activated by OsYAB5. Furthermore, the OsKAN1-OsYAB5 complex suppressed the inhibitory effect of OsKAN1 toward OsYAB5 and inhibited OsYAB5-induced OsGA2ox6 expression. The proOsKAN1:OsYAB5 transgenic plants were taller than wild-type plants, whereas oskan1 proOsKAN1:OsYAB5 plants exhibited a severe dwarf phenotype due to the absence of the OsKAN1-OsYAB5 complex. The OsKAN1-OsYAB5 complex modulated OsGA2ox6 expression, thereby regulating the levels of bioactive gibberellins and, consequently, plant height. This study elucidated the mechanism underlying the effect of the OsKAN1-OsYAB5-OsGA2ox6 regulatory pathway on plant height at different positions in rice stems and provided insights on stem development and candidate genes for the aerial architecture improvement of crop plants.

The Indica Hybrid Rice Containing an OsNRAMP5 Knockout Exhibits Better Adaptability Compared to Its Paternal Parent in the Middle Reaches of the Yangtze River

OsNRAMP5 is a pivotal gene involved in the uptake and transport of cadmium and manganese in rice plants. While the knockout of OsNRAMP5 has significantly decreased cadmium accumulation in rice grains, its impact on the adaptability of indica hybrid rice and its paternal parent still needs to be more adequately explored. This study conducted a three-year field experiment with two of the OsNRAMP5 knockout lines (indica hybrid rice LYDG-1 and its paternal parent DG-1) and their wild-type at four distinct locations in the middle reaches of the Yangtze River. Their adaptability was evaluated by cadmium and manganese accumulation, yield traits, and grain quality. The findings reveal that OsNRAMP5 knockout lines, when grown in mildly and moderately Cd-contaminated paddies, exhibit significantly and stably lower cadmium and manganese accumulation compared with the wild-type. However, there were no significant differences in cadmium and manganese content between LYDG-1 and DG-1. Regarding yield traits, OsNRAMP5 knockout lines did not show a significant reduction compared with the wild-type, except for DG-1 in soil with lower manganese content. For grain quality, the quality of the appearance of OsNRAMP5 knockout lines significantly improved compared with the wild-type. However, the milling quality of DG-1 significantly reduced compared with the wild-type, while this negative impact for LYDG-1 was only observed in soils with lower manganese content. Overall, the indica hybrid rice with an OsNRAMP5 knockout exhibits better adaptability than its paternal parent in the middle reaches of the Yangtze River. These findings offer valuable insights into the potential application and promotion of OsNRAMP5 mutant rice varieties in the middle reaches of the Yangtze River.

Loss of OsMATE6 Function Enhances Drought Resistance Without Yield Penalty by Regulating Stomatal Closure in Rice.

Drought is one of the most severe environmental factors limiting plant growth and crop yield, necessitating the identification of genes that enhance drought resistance for crop improvement. Through screening an ethyl methyl sulfonate-mutagenized rice mutant library, we isolated the PEG tolerance mutant 97-1 (ptm97-1), which displays enhanced resistance to osmotic and drought stress, and increased yield under drought conditions. A point mutation in OsMATE6 was identified as being associated with the drought-resistant phenotype of ptm97-1. The role of OsMATE6 in conferring drought resistance was confirmed by additional OsMATE6 knockout mutants. OsMATE6 is expressed in guard cells, shoots and roots and the OsMATE6-GFP fusion protein predominantly localizes to the plasma membrane. Our ABA efflux assays suggest that OsMATE6 functions as an ABA efflux transporter; mutant protoplasts exhibited a slower ABA release rate compared to the wild type. We hypothesize that OsMATE6 regulates ABA levels in guard cells, influencing stomatal closure and enhancing drought resistance. Notably, OsMATE6 knockout mutants demonstrated greater yields under field drought conditions compared to wild-type plants, highlighting OsMATE6 as a promising candidate for improving crop drought resistance.

Application of CRISPR/Cas9 system to knock out GluB gene for developing low glutelin rice mutant

The nutritional quality improvement is among the most integral objective for any rice molecular breeding programs. The seed storage proteins (SSPs) have greater role to determine the nutritional quality of any cereal grains. Rice contains relatively balanced amino acid composition and the SSPs are fractioned into albumins (ALB), globulins (GLO), prolamins (PRO) and glutelins (GLU) according to differences in solubility. GLUs are further divided into subfamilies: GluA, GluB, GluC, and GluD depending on resemblance in amino acid. The GLU protein accounts for 60–80% of total protein contents, encoded by 15 genes located on different chromosomes of rice genome. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system was employed to knockout Glu-B (LOC-Os02g15070) gene in non-basmati rice PK386 cultivar. The mutant displayed two base pair and three base pair mutation in the targeted regions. The homozygous mutant plant displayed reduction for both in total protein contents and GLU contents whereas, elevation in GLO, ALB and PRO. Moreover, the mutant plant also displayed reduction in physio-chemical properties e.g., total starch, amylose and gel consistency. The agronomic characteristics of both mutant and wild type displayed non-significant differences along with increase in higher percentage of chalkiness in mutant plants. The results obtained from scanning electron microscopy showed the loosely packed starch granules compared to wild type. The gene expression analysis displayed the lower expression of gene at 5 days after flowering (DAF), 10 DAF, 15 DAF and 20 DAF compared to wild type. GUS sub-cellular localization showed the staining in seed which further validated the results obtained from gene expression. Based on these findings it can be concluded Glu-B gene have significant role in controlling GLU contents and can be utilized in breeding programs to enhance the nutritional quality of rice, and may serve as healthy diet for patient allergic with high GLU contents.

Growth and Production Character Response of Kuantan Singingi Local Rice Mutants Resulting from Gamma Irradiation

Rice (Oryza sativa L.) is a crucial crop with significant importance in various regions. In Riau, specifically in Kuantan Singingi Regency, it ranks as the fourth largest area for rice production, showcasing numerous local rice varieties with substantial development potential. This study aimed to generate mutants exhibiting early maturity and reduced plant height through gamma-ray induced mutations. A non-factorial Randomized Block Design (RAK) was employed, with the primary factor being the irradiation dose: R0 served as the control group without radiation, while the experimental group received an irradiation dose of 300 Gy, determined to be effective in prior research. Each treatment was replicated five times, utilizing a total of 600 plants per treatment. The findings indicated that rice plants subjected to 300 Gy irradiation exhibited shorter heights compared to those in the control group. Additionally, the stem circumference of plants irradiated with 300 Gy was significantly greater than that of the non-irradiated plants. Furthermore, the number of productive tillers was higher in the 300 Gy group compared to the control. However, the harvest age for rice plants exposed to 300 Gy was extended relative to those without radiation. Conversely, the length of the flag leaf in rice plants irradiated at 300 Gy was shorter than that of the control group. There remains a considerable need for data on the desired characteristics of rice mutants to facilitate advanced breeding efforts.

Mitigating nitrogen loss in paddy field microcosms through indigenous arbuscular mycorrhizal fungi assemblage

AbstractWhether farmers should consider the role of arbuscular mycorrhizal fungi (AMF) in agriculture is a hotly debated topic. We aimed to investigate the role of indigenous AMF in reducing nitrogen (N) loss from paddy fields via runoff, leaching, NH3 volatilization, and N2O emission. We conducted a pot experiment employing a mycorrhiza‐defective rice mutant (non‐mycorrhizal) as the control, grown in soil containing indigenous AMF. The corresponding AMF treatment used the progenitor of this mutant with the same soil. The plants were fertilized with nitrogen, phosphorus and potassium 6 weeks after sowing. The root colonization was 23% in mycorrhizal rice, and no typical AMF structures were observed in the roots of non‐mycorrhizal rice. Our findings indicated that the mycorrhizal system exhibited lower N concentrations of runoff and leachate further compounded by reduced fluxes of N2O and NH3. This led to 14% decrease (mycorrhizal rice 111 kg N ha−1; the non‐mycorrhizal rice: 129 kg N ha−1) in cumulative N loss within 3 days post‐fertilization. While this AMF effect was consistent across the four tested N loss pathways, differences were observed between NH4+‐N and NO3−‐N in the runoff pathway. Notably, our results revealed no evidence of trade‐offs in AMF effect on N loss among the tested pathways. Additionally, mycorrhizal rice had larger shoots and roots than their non‐mycorrhizal counterparts. Our study underscores the potential benefits of indigenous AMF in paddy fields for mitigating water pollution and reducing greenhouse gas emission.

Mutation of KAP, which encodes a keratin-associated protein, affects grain size and yield production in rice.

Grain size and shape are critical agronomic traits that directly impact rice grain yield. Identifying genes that control these traits can provide new strategies for yield improvement. In this study, we characterized a rice mutant, reduced grain length (rgl), which exhibited decreased grain length due to reduced cell proliferation. Map-based cloning identified a base deletion in OsRGL2, a gene encoding a keratin-associated protein (KAP), as the cause of the mutant phenotype. CRISPR-Cas9-generated OsRGL2 knockout mutants also displayed reduced grain length, confirming its role. OsRGL2 transcripts were detected in various tissues, with relative higher gene expression in young panicles, and OsRGL2 was localized to the plasma membrane. Overexpression of OsRGL2 increased grain size by promoting cell proliferation in the spikelet hull and significantly enhanced grain yield per plant. Importantly, OsRGL2 was found to interact with RGB1, indicating that OsRGL2 positively regulates grain size and yield through its interaction with RGB1. Additionally, OsRGL2 regulated the expression of cell cycle-related genes, further elucidating its role in grain development. These findings demonstrate that OsRGL2 is a positive regulator of grain size in rice, and manipulating its expression may offer a novel strategy for enhancing rice grain yield.

Application of Jarwo Planting System and Fertiphos Fertilizer on The Growth and Yield of Black Rice Mutant M4

This study aims to examine the interaction between the dose of Fertiphos fertilizer and the JARWO (JARWO) planting pattern on the growth and yield of black rice mutant rice. The experiment was conducted in Jorong Labuah, Agam Regency, and the Seed Science and Technology Laboratory, Andalas University, Padang, from February to June 2024. A Completely Randomized Design (CRD) with two factors was used, namely the JARWO planting system (2:1, 3:1, 4:1) and the dose of Fertiphos fertilizer (0 kg/ha, 100 kg/ha, 200 kg/ha, 300 kg/ha). Data were analyzed using the F test at the 5% level, followed by Duncan's New Multiple Range Test (DNMRT) if significant. The results showed a significant interaction between the dose of Fertiphos fertilizer and the JARWO planting pattern on the growth and yield of black rice mutant rice. Fertiphos fertilizer dose of 200 kg/ha gave the best results on the total number of grains per panicle, the number of filled grains per panicle, the total weight of grains per panicle, and the weight of filled grains per clump. The JARWO 2:1 planting pattern gave the best results on the total number of tillers, the total number of grains per panicle, the number of filled grains per panicle, and the total weight of grains per panicle.

OsSNDP4, a Sec14-nodulin Domain Protein, is Required for Pollen Development in Rice

Pollen is encased in a robust wall that shields the male gametophyte from various stresses and aids in pollination. The pollen wall consists of gametophyte-derived intine and sporophyte-derived exine. The exine is mainly composed of sporopollenin, which is biopolymers of aliphatic lipids and phenolics. The process of exine formation has been the subject of extensive research, yet the underlying molecular mechanisms remain elusive. In this study, we identified a rice mutant of the OsSNDP4 gene that is impaired in pollen development. We demonstrated that OsSNDP4, a putative Sec14-nodulin domain protein, exhibits a preference for binding to phosphatidylinositol (3)-phosphate [PI(3)P], a lipid primarily found in endosomal and vacuolar membranes. The OsSNDP4 protein was detected in association with the endoplasmic reticulum (ER), vacuolar membranes, and the nucleus. OsSNDP4 expression was detected in all tested organs but was notably higher in anthers during exine development. Loss of OsSNDP4 function led to abnormal vacuole dynamics, inhibition in Ubisch body development, and premature degradation of cellular contents and organelles in the tapetal cells. Microspores from the ossndp4 mutant plant displayed abnormal exine formation, abnormal vacuole enlargement, and ultimately, pollen abortion. RNA-seq assay revealed that genes involved in the biosynthesis of fatty acid and secondary metabolites, the biosynthesis of lipid polymers, and exosome formation were enriched among the down-regulated genes in the mutant anthers, which correlated with the morphological defects observed in the mutant anthers. Base on these findings, we propose that OsSNDP4 regulates pollen development by binding to PI(3)P and influencing the dynamics of membrane systems. The involvement of membrane systems in the regulation of sporopollenin biosynthesis, Ubisch body formation, and exine formation provides a novel mechanism regulating pollen wall development.

Leaf Gas Film 1 promotes glycerol ester accumulation and formation of a tight root barrier to radial O2 loss in rice.

Rice (Oryza sativa L.) and many other wetland plants form an apoplastic barrier in the outer parts of the roots to restrict radial O2 loss to the rhizosphere during soil flooding. This barrier facilitates longitudinal internal O2 diffusion via gas-filled tissues from shoot to root apices, enabling root growth in anoxic soils. We tested the hypothesis that Leaf Gas Film 1 (LGF1), which influences leaf hydrophobicity in rice, plays a crucial role in tight outer apoplastic barriers formation in rice roots. We examined the roots of a rice mutant (dripping wet leaf 7, drp7) lacking functional LGF1, its wild type, and an LGF1 overexpression line for their capacity to develop outer apoplastic barriers that restrict radial O2 loss. We quantified the chemical composition of the outer part of the root and measured radial O2 diffusion from intact roots. The drp7 mutant exhibited a weak barrier to radial O2 loss compared to the wild type. However, introducing functional LGF1 into the mutant fully restored tight barrier function. The formation of a tight barrier to radial O2 loss was associated with increased glycerol ester levels in exodermal cells, rather than differences in total root suberization or lignification. These results demonstrate that, in addition to its role in leaf hydrophobicity regulation, LGF1 plays an important role in controlling the function of the outer apoplastic barriers in roots. Our study suggests that increased deposition of glycerol esters in the suberized root exodermis establishes a tight barrier to radial O2 loss in rice roots.

The tonoplast-localized OsTIP2;1 is involved in aluminum detoxification in rice

Aluminum (Al) stress is a significant issue in acidic soils, severely affecting crop growth and yield. Rice is notably resilient to Al toxicity, yet the internal tolerance mechanisms remain inadequately addressed. Here, we examined the role of OsTIP2;1, a tonoplast-bound intrinsic protein (TIP), in rice's internal Al detoxification. Our findings reveal that OsTIP2;1 expression was quickly and explicitly activated by Al ions in roots but not in shoots. The OsTIP2;1-GFP protein localizes to the tonoplast in plant and yeast cells. Non-functional ostip2;1 rice mutants were more vulnerable to Al toxicity. In the roots, the ostip2;1 mutants exhibited considerably lower levels of Al in the cell sap, primarily the vacuolar contents, than in the wild-type plant. Moreover, the ostip2;1 mutants showed reduced Al accumulation in the roots but increased translocation to the shoots. Heterologous expression of tonoplast-localized OsTIP2;1 in yeast led to enhanced Al tolerance, suggesting that OsTIP2;1 facilitates Al sequestration to the vacuole. These findings indicate that OsTIP2;1 mediates internal detoxification by transporting Al into the vacuole in the root and restricting its transport to above-ground tissues, thus contributing to Al resistance in rice.

Mitogen-Activated Protein Kinase Kinase OsMEK2 Positively Regulates Ca2+ Influx and Ferroptotic Cell Death during Rice Immune Responses.

Mitogen-activated protein (MAP) kinase (MAPK) signaling pathway is important in plant immune responses, involved in iron- and reactive oxygen species (ROS)-dependent ferroptotic cell death mediated by Ca2+. High Ca2+ influx triggered iron-dependent ROS accumulation, lipid peroxidation, and subsequent hypersensitive response (HR) cell death in rice (Oryza sativa). Apoplastic Ca2+ chelation by EGTA during avirulent Magnaporthe oryzae infection altered Ca2+, ROS, and Fe2+ accumulation, increasing rice susceptibility to infection. By contrast, acibenzolar-S-methyl (ASM), a plant defense activator, significantly enhanced Ca2+ influx, and H2O2 accumulation, triggering rice ferroptotic cell death during virulent Magnaporthe oryzae infection. Here, we report a novel role of the MAPK signaling pathway in regulating cytoplasmic Ca2+ increase during ferroptotic cell death in rice immunity, using the ΔOsmek2 knockout mutant rice. The knockout of rice OsMEK2 impaired the ROS accumulation, lipid peroxidation, and iron accumulation during avirulent M. oryzae infection. This study has shown that OsMEK2 could positively regulate iron- and ROS-dependent ferroptotic cell death in rice by modulating the expression of OsNADP-ME, OsRBOHB, OsPLC, and OsCNGC. This modulation indicates a possible mechanism for how OsMEK2 participates in Ca2+ regulation in rice ferroptotic cell death, suggesting its broader role in plant immune responses in response to M. oryzae infection.