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Cold tolerance at the booting (CTB) stage is a major factor limiting rice (Oryza sativa L.) productivity and geographical distribution. A few cold-tolerance genes have been identified, but they either need to be overexpressed to result in CTB or cause yield penalties, limiting their utility for breeding. Here, we characterize the function of the cold-induced transcription factor WRKY53 in rice. The wrky53 mutant displays increased CTB, as determined by higher seed setting. Low temperature is associated with lower gibberellin (GA) contents in anthers in the wild type but not in the wrky53 mutant, which accumulates slightly more GA in its anthers. WRKY53 directly binds to the promoters of GA biosynthesis genes and transcriptionally represses them in anthers. In addition, we uncover a possible mechanism by which GA regulates male fertility: SLENDER RICE1 (SLR1) interacts with and sequesters two critical transcription factors for tapetum development, UNDEVELOPED TAPETUM1 (UDT1), and TAPETUM DEGENERATION RETARDATION (TDR), and GA alleviates the sequestration by SLR1, thus allowing UDT1 and TDR to activate transcription. Finally, knocking out WRKY53 in diverse varieties increases cold tolerance without a yield penalty, leading to a higher yield in rice subjected to cold stress. Together, these findings provide a target for improving CTB in rice.
Assuntos
Oryza , Oryza/metabolismo , Giberelinas/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Genes de Plantas , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
MAIN CONCLUSION: The rice SnRK2 members SAPK4, SAPK5, SAPK7 and SAPK10 are positive regulators involved in the regulation of rice flowering, while other single mutants exhibited no effect on rice flowering. The rice SnRK2 family, comprising 10 members known as SAPK (SnRK2-Associated Protein Kinase), is pivotal in the abscisic acid (ABA) pathway and crucial for various biological processes, such as drought resistance and salt tolerance. Additionally, these members have been implicated in the regulation of rice heading date, a key trait influencing planting area and yield. In this study, we utilized gene editing technology to create mutants in the Songjing 2 (SJ2) background, enabling a comprehensive analyze the role of each SAPK member in rice flowering. We found that SAPK1, SAPK2, and SAPK3 may not directly participate in the regulatory network of rice heading date, while SAPK4, SAPK5, and SAPK7 play positive roles in rice flowering regulation. Notably, polygene deletion resulted in an additive effect on delaying flowering. Our findings corroborate the previous studies indicating the positive regulatory role of SAPK10 in rice flowering, as evidenced by delayed flowering observed in sapk9/10 double mutants. Moving forward, our future research will focus on analyzing the molecular mechanisms underlying SAPKs involvement in rice flowering regulation, aiming to enhance our understanding of the rice heading date relationship network and lay a theoretical foundation for breeding efforts to alter rice ripening dates.
Assuntos
Flores , Regulação da Expressão Gênica de Plantas , Oryza , Proteínas de Plantas , Oryza/genética , Oryza/crescimento & desenvolvimento , Oryza/fisiologia , Oryza/enzimologia , Flores/genética , Flores/crescimento & desenvolvimento , Flores/fisiologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Mutação , Edição de Genes , Estresse Fisiológico/genética , Proteínas Quinases/genética , Proteínas Quinases/metabolismo , Ácido Abscísico/metabolismo , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/metabolismoRESUMO
In rice (Oryza sativa) and other plants, plant architecture and seed size are closely related to yield. Brassinosteroid (BR) signaling and the mitogen-activated protein kinase (MAPK) pathway (MAPK kinase kinase 10 [MAPKKK10]-MAPK kinase 4 [MAPKK4]-MAPK6) are two major regulatory pathways that control rice architecture and seed size. However, their possible relationship and crosstalk remain elusive. Here, we show that WRKY53 mediated the crosstalk between BR signaling and the MAPK pathway. Biochemical and genetic assays demonstrated that glycogen synthase kinase-2 (GSK2) phosphorylates WRKY53 and lowers its stability, indicating that WRKY53 is a substrate of GSK2 in BR signaling. WRKY53 interacted with BRASSINAZOLE-RESISTANT 1(BZR1); they function synergistically to regulate BR-related developmental processes. We also provide genetic evidence showing that WRKY53 functions in a common pathway with the MAPKKK10-MAPKK4-MAPK6 cascade in leaf angle and seed size control, suggesting that WRKY53 is a direct substrate of this pathway. Moreover, GSK2 phosphorylated MAPKK4 to suppress MAPK6 activity, suggesting that GSK2-mediated BR signaling might also regulated MAPK pathway. Together, our results revealed a critical role for WRKY53 and uncovered sophisticated levels of interplay between BR signaling and the MAPK pathway in regulating rice architecture and seed size.
Assuntos
Brassinosteroides/metabolismo , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Oryza/fisiologia , Proteínas de Plantas/metabolismo , Sementes/fisiologia , Regulação da Expressão Gênica de Plantas , Quinases da Glicogênio Sintase/genética , Quinases da Glicogênio Sintase/metabolismo , Proteínas Quinases Ativadas por Mitógeno/genética , Oryza/genética , Fosforilação , Folhas de Planta/fisiologia , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Estabilidade Proteica , Transdução de Sinais , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
The progression of liver fibrosis is determined by the interaction of damaged hepatocytes, active hepatic stellate cells, and macrophages, contributing to the development of oxidative stress and inflammatory environments within the liver. Unfortunately, the current pharmacological treatment for liver fibrosis is limited by its inability to regulate inflammation and oxidative stress concurrently. In this study, we developed a cell membrane biomaterial for the treatment of liver fibrosis, which we designated as PM. PM is a biomimetic nanomaterial constructed by encapsulating polydopamine (PDA) with a macrophage membrane (MM). It is hypothesized that PM nanoparticles (NPs) can successfully target the site of inflammation, simultaneously inhibit inflammation, and scavenge reactive oxygen species (ROS). In vitro experiments demonstrated that PM NPs exhibited strong antioxidant properties and the ability to neutralize pro-inflammatory cytokines (TNF-α, IL-6, and IL-1ß). Moreover, the capacity of PM NPs to safeguard cells from oxidative stress and their anti-inflammatory efficacy in an inflammatory model were validated in subsequent cellular experiments. Additionally, PM NPs exhibited a high biocompatibility. In a mouse model of hepatic fibrosis, PM NPs were observed to aggregate efficiently in the fibrotic liver, displaying excellent antioxidant and anti-inflammatory properties. Notably, PM NPs exhibited superior targeting, anti-inflammatory, and ROS scavenging abilities in inflamed tissues compared to MM, PDA, or erythrocyte membrane-encapsulated PDA. Under the synergistic effect of anti-inflammation and antioxidant, PM NPs produced significant therapeutic effects on liver fibrosis in mice. In conclusion, the synergistic alleviation of inflammation and ROS scavenging by this specially designed nanomaterial, PM NPs, provides valuable insights for the treatment of liver fibrosis and other inflammatory- or oxidative stress-related diseases.
Assuntos
Antioxidantes , Indóis , Inflamação , Cirrose Hepática , Macrófagos , Nanopartículas , Estresse Oxidativo , Polímeros , Espécies Reativas de Oxigênio , Animais , Polímeros/química , Espécies Reativas de Oxigênio/metabolismo , Indóis/química , Indóis/farmacologia , Indóis/administração & dosagem , Camundongos , Cirrose Hepática/tratamento farmacológico , Cirrose Hepática/patologia , Nanopartículas/química , Estresse Oxidativo/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Antioxidantes/farmacologia , Antioxidantes/química , Inflamação/tratamento farmacológico , Inflamação/patologia , Humanos , Masculino , Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/administração & dosagem , Células RAW 264.7 , Camundongos Endogâmicos C57BL , Modelos Animais de Doenças , Citocinas/metabolismoRESUMO
The chloroplast serves as the primary site of photosynthesis, and its development plays a crucial role in regulating plant growth and morphogenesis. The Pentatricopeptide Repeat Sequence (PPR) proteins constitute a vast protein family that function in the post-transcriptional modification of RNA within plant organelles. In this study, we characterized mutant of rice with pale green leaves (pgl3a). The chlorophyll content of pgl3a at the seedling stage was significantly reduced compared to the wild type (WT). Transmission electron microscopy (TEM) and quantitative PCR analysis revealed that pgl3a exhibited aberrant chloroplast development compared to the wild type (WT), accompanied by significant alterations in gene expression levels associated with chloroplast development and photosynthesis. The Mutmap analysis revealed that a single base deletionin the coding region of Os03g0136700 in pgl3a. By employing CRISPR/Cas9 mediated gene editing, two homozygous cr-pgl3a mutants were generated and exhibited a similar phenotype to pgl3a, thereby confirming that Os03g0136700 was responsible for pgl3a. Consequently, it was designated as OsPGL3A. OsPGL3A belongs to the DYW-type PPR protein family and is localized in chloroplasts. Furthermore, we demonstrated that the RNA editing efficiency of rps8-182 and rpoC2-4106, and the splicing efficiency of ycf3-1 were significantly decreased in pgl3a mutants compared to WT. Collectively, these results indicate that OsPGL3A plays a crucial role in chloroplast development by regulating the editing and splicing of chloroplast genes in rice. Supplementary Information: The online version contains supplementary material available at 10.1007/s11032-024-01468-7.
RESUMO
Rice flowering time determines its geographical distribution and yield traits. As a short-day plant, rice can grow in the northern long-day conditions due to the functional mutations of many photosensitive genes. In this study, to identify novel genes or alleles that regulate flowering time in high latitude region, two cultivar, Dongnong 413 (DN413) and Yukimochi (XN) showing extreme early flowering were used for investigation. DN413 is around 4.0 days earlier than XN, and both cultivars can be grown in II (2500 â-2700 â) to III (2300 â-2500 â) accumulated temperature zones. We found that the two cultivars shared the same genotype of heading date genes, including Hd1/2/4/5/6/16/17/18, Ehd2, DTH2, SE5, Hd3a. Importantly, a novel Ehd3 allele characterized by a A1146C substitution was identified, which results in the E382D substitution, hereafter the 382 position E is defined as Hap_E and the 382 position D is defined as Hap_D. Association analysis showed that Hap_E is earlier flowering than Hap_D. Subsequently, we construct DN413 Hap_D line by three times back-crossing DN413 with XN, and found the heading date of DN413 Hap_D was 1.7-3.5 days later than DN413. Moreover, Hap_E and Hap_D of Ehd3 were transformed into ehd3 mutant, respectively, and the Ehd3pro:Ehd3D/ehd3 flowered later than that Ehd3pro:Ehd3E/ehd3 by around 4.3 days. Furthermore, we showed Ehd3 functions as a transcriptional suppressor and the substitution of Asp-382 lost the inhibition activity in protoplasts. Finally, a CAPS marker was developed and used for genotyping and marker assistant breeding. Collectively, we discovered a novel functional allele of Ehd3, which can used as a valuable breeding target. Supplementary Information: The online version contains supplementary material available at 10.1007/s11032-024-01472-x.
RESUMO
Panicle exsertion is one of the crucial agronomic traits in rice (Oryza sativa). Shortening of panicle exsertion often leads to panicle enclosure and severely reduces seed production. Gibberellin (GA) plays important roles in regulating panicle exsertion. However, the underlying mechanism and the relative regulatory network remain elusive. Here, we characterized the oswrky78 mutant showing severe panicle enclosure, and found that the defect of oswrky78 is caused by decreased bioactive GA contents. Biochemical analysis demonstrates that OsWRKY78 can directly activate GA biosynthesis and indirectly suppress GA metabolism. Moreover, we found OsWRKY78 can interact with and be phosphorylated by mitogen-activated protein kinase (MAPK) kinase OsMAPK6, and this phosphorylation can enhance OsWRKY78 stability and is necessary for its biological function. Taken together, these results not only reveal the critical function of OsWRKY78, but also reveal its mechanism via mediating crosstalk between MAPK and the GA signaling pathway in regulating panicle exsertion.
Assuntos
Giberelinas , Oryza , Giberelinas/metabolismo , Oryza/genética , Oryza/metabolismo , Transdução de Sinais/genética , Sementes/metabolismo , FenótipoRESUMO
Rice is a chilling-sensitive plant, and extremely low temperatures seriously decrease rice production. Several genes involved in chilling stress have been reported in rice; however, the chilling signaling in rice remains largely unknown. Here, we investigated the chilling tolerance phenotype of overexpression of constitutive active OsMAPK6 (CAMAPK6-OE) and OsMAPK6 mutant dsg1, and demonstrated that OsMAPK6 positively regulated rice chilling tolerance. It was shown that, under cold stress, the survival rate of dsg1 was significantly lower than that of WT, whereas CAMAPK6-OE display higher survival rate than WT. Physiological assays indicate that ion leakage and dead cell in dsg1 was much more severe than those in WT and CAMAPK6-OE. Consistently, expression of chilling responsive genes in dsg1, including OsCBFs and OsTPP1, was significantly lower than that of in WT and CAMAPK6-OE. Biochemical analyses revealed that chilling stress promotes phosphorylation of OsMAPK6. Besides, we found that OsMAPK6 interacts with and phosphorylates two key regulators in rice cold signaling, OsIPA1 and OsICE1, and then enhance their protein stability. Overall, our results revealed a cold-induced OsMAPK6-OsICE1/OsIPA1 signaling cascade by which OsMAPK6 was involved in rice chilling tolerance, which provides novel insights to understand rice cold response at seedling stage.
Assuntos
Oryza , Plântula , Plântula/genética , Plântula/metabolismo , Oryza/metabolismo , Resposta ao Choque Frio/genética , Temperatura Baixa , Fosforilação , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMO
MAIN CONCLUSION: OsWRKY24 functions redundantly with OsWRKY53, while OsWRKY70 functions differently from OsWRKY53 in regulating grain size. Grain size is a key agronomic trait that affects grain yield and quality in rice (Oryza sativa L.). The transcription factor OsWRKY53 positively regulates grain size through brassinosteroid (BR) signaling and Mitogen-Activated Protein Kinase (MAPK) cascades. However, whether the OsWRKY53 homologs OsWRKY24 and OsWRKY70 also contribute to grain size which remains unknown. Here, we report that grain size in OsWRKY24 overexpression lines and oswrky24 mutants is similar to that of the wild type. However, the oswrky24 oswrky53 double mutant produced smaller grains than the oswrky53 single mutant, indicating functional redundancy between OsWRKY24 and OsWRKY53. In addition, OsWRKY70 overexpression lines displayed an enlarged leaf angle, reduced plant height, longer grains, and higher BR sensitivity, phenotypes similar to those of OsWRKY53 overexpression lines. Importantly, a systematic characterization of seed length in the oswrky70 single, the oswrky53 oswrky70 double and the oswrky24 oswrky53 oswrky70 triple mutant indicated that loss of OsWRKY70 also leads to increased seed length, suggesting that OsWRKY70 might play a role distinct from that of OsWRKY53 in regulating grain size. Taken together, these findings suggest that OsWRKY24 and OsWRKY70 regulate rice grain size redundantly and independently from OsWRKY53.
Assuntos
Oryza , Brassinosteroides/metabolismo , Grão Comestível/genética , Grão Comestível/metabolismo , Regulação da Expressão Gênica de Plantas , Oryza/genética , Oryza/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMO
Flowering time is one of the most fundamental factors that determine the distribution and final yield of rice. Ehd1 (Early heading date 1) is a B-type response regulator which functions as a flowering time activator. Although diverse flowering time genes have been reported as regulatory factors of Ehd1 expression, the potential regulators of Ehd1 largely remain to be identified. Here, we identified a basic leucine zipper transcription factor bZIP65, a homolog of bZIP71, as a new negative regulator of Ehd1. The overexpression of bZIP65 delays flowering, while bzip65 mutants have similar flowering time to SJ2 (Songjing2) in both long-day and short-day conditions. Biochemically, bZIP65 associates with Ehd1 promoter and transcriptionally represses the expression of Ehd1. Moreover, we found that bZIP65 enhances H3K27me3 level of Ehd1. Taken together, we cloned a new gene, bZIP65, regulating rice heading date, and uncovered the mechanism of bZIP65 delaying flowering time, where bZIP65 increases the H3K27me3 level of Ehd1 and transcriptionally represses the expression of Ehd1, similar to its homolog bZIP71. Supplementary Information: The online version contains supplementary material available at 10.1007/s11032-022-01334-4.
RESUMO
OBJECTIVES: Urban-rural comparisons between those who maintain traditional lifestyles such as pastoralist Maasai children, and those who live in cities such as Nairobi, provide implications on how urbanization is associated with children's physical activity (PA) and sedentary behavior (SB) patterns. This study compares PA and SB volumes and patterns across different segments of the week among children in Maasai village and Nairobi city in Kenya. METHOD: A total of 261 children (11.4 ± 1.3 years) from Maasai (n = 118) and Nairobi (n = 143) participated in this cross-sectional study. Moderate- to vigorous-intensity PA (MVPA) and SB on weekdays (before, during, and after school) and weekends (morning, afternoon, and evening) were calculated using accelerometers (ActiGraph). Screen time and sleep duration were assessed using questionnaires. RESULTS: Maasai children were more physically active than Nairobi children with MVPA (min/day) of 166.6 and 81.4 for Maasai and Nairobi boys and 116.4 and 77.4 for Maasai and Nairobi girls, respectively. Our week segments analyses suggested that Maasai children were more active both in and out of school than Nairobi children. Additionally, Nairobi children spent more time watching television and playing computer games than Maasai children. There was no significant difference in sleep duration between Maasai and Nairobi children. CONCLUSION: Our findings suggest that urbanization is negatively associated with activity patterns both in and out of school in Kenyan children. This is concerning given that Kenya is currently undergoing rapid urbanization, which may lead to further reductions in PA among Kenyan children.
Assuntos
Exercício Físico , Comportamento Sedentário , Acelerometria , Criança , Cidades/estatística & dados numéricos , Estudos Transversais , Exercício Físico/estatística & dados numéricos , Feminino , Humanos , Quênia , Masculino , População Rural/estatística & dados numéricosRESUMO
Place recognition is an essential part of simultaneous localization and mapping (SLAM). LiDAR-based place recognition relies almost exclusively on geometric information. However, geometric information may become unreliable when faced with environments dominated by unstructured objects. In this paper, we explore the role of segmentation for extracting key structured information. We propose STV-SC, a novel segmentation and temporal verification enhanced place recognition method for unstructured environments. It contains a range image-based 3D point segmentation algorithm and a three-stage process to detect a loop. The three-stage method consists of a two-stage candidate loop search process and a one-stage segmentation and temporal verification (STV) process. Our STV process utilizes the time-continuous feature of SLAM to determine whether there is an occasional mismatch. We quantitatively demonstrate that the STV process can trigger false detections caused by unstructured objects and effectively extract structured objects to avoid outliers. Comparison with state-of-art algorithms on public datasets shows that STV-SC can run online and achieve improved performance in unstructured environments (Under the same precision, the recall rate is 1.4â¼16% higher than Scan context). Therefore, our algorithm can effectively avoid the mismatching caused by the original algorithm in unstructured environment and improve the environmental adaptability of mobile agents.
Assuntos
Algoritmos , Reconhecimento Psicológico , Cintilografia , CabeçaRESUMO
Cold stress at the booting stage leads to a lower seed setting rate and seriously threatens the production of rice (Oryza sativa L.), which has become a major yield-limiting factor in higher-altitude and -latitude regions. Because cold tolerance at the booting stage (CTB) is a complex trait and is controlled by multiple loci, only a few genes have been reported so far. In this study, a function of OsMKKK70 (Mitogen Activated Protein Kinase Kinase Kinase 70) in response to CTB was characterized. OsMKKK70 expression was rapidly induced by cold stress at the booting stage. OsMKKK70 overexpression (OsMKKK70-OE) plants were more sensitive to cold stress at the booting stage with a lower seed setting and pollen fertility, but there was no significant difference between the osmkkk70 mutant and WT. Considering the effect of functional redundancy, we further tested the CTB response of osmkkk62/70 and osmkkk55/62/70, the double and triple mutants of OsMKKK70 with its closest homologs OsMKKK62 and OsMKKK55, and found that osmkkk62/70 and osmkkk55/62/70 displayed significantly increased CTB with a higher seed setting and pollen fertility, indicating that OsMKKK70 negatively regulates rice CTB. Moreover, under the low-temperature (LT) condition, the osmkkk62/70 mutant had slightly higher Gibberellin (GA) contents, increased expression of GA biosynthesis genes, and lower protein level of OsSLR1 in anthers than those in WT. By contrast, OsMKKK70-OE anther had a lower GA biosynthesis than that of WT. Together, these findings suggest that OsMKKK70 negatively regulates rice CTB by fine-tuning GA levels in anthers.
Assuntos
Oryza , Oryza/genética , Temperatura Baixa , Resposta ao Choque Frio , Tolerância a MedicamentosRESUMO
Flowering time is a fundamental factor determining the global distribution and final yield of rice (Oryza sativa). Although diverse flowering time genes have been reported in this crop, the transcriptional regulation of its key flowering genes are poorly understood. Here, we report that a basic leucine zipper transcription factor, bZIP71, functions as a flowering repressor. The overexpression of bZIP71 delays flowering, while the bzip71 mutant flowers early in both long-day and short-day conditions. A genetic analysis showed that the regulation of flowering by bZIP71 might be independent of Heading date 2 (Hd2), Hd4, and Hd5. Importantly, bZIP71 directly associates with the Early heading date 1 (Ehd1) promoter and represses its transcription, and genetically the function of bZIP71 is impaired in the ehd1 mutant. Moreover, bZIP71 interacts with major components of polycomb repressive complex 2 (PRC2), SET domain group protein 711 (SDG711), and Fertilization independent endosperm 2 (FIE2), through which bZIP71 regulates the H3K27me3 level of Ehd1. Taken together, we present a transcriptional regulatory mechanism in which bZIP71 enhances the H3K27me3 level of Ehd1 and transcriptionally represses its expression, which not only offers a novel insight into a flowering pathway, but also provides a valuable putative target for the genetic engineering and breeding of elite rice cultivars.
Assuntos
Oryza , Flores/fisiologia , Regulação da Expressão Gênica de Plantas/genética , Histonas/metabolismo , Oryza/metabolismo , Fotoperíodo , Melhoramento Vegetal , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMO
Cuticular wax plays crucial roles in protecting plants from environmental stresses, particularly drought stress. Many enzyme-encoding genes and transcription factors involved in wax biosynthesis have been identified, but the underlying posttranslational regulatory mechanisms are poorly understood. Here, we demonstrate that DROUGHT HYPERSENSITIVE (DHS), encoding a Really Interesting New Gene (RING)-type protein, is a critical regulator of wax biosynthesis in rice (Oryza sativa). The cuticular wax contents were significantly reduced in DHS overexpression plants but increased in dhs mutants compared with the wild type, which resulted in a response opposite that of drought stress. DHS exhibited E3 ubiquitin ligase activity and interacted with the homeodomain-leucine zipper IV protein ROC4. Analysis of ROC4 overexpression plants and roc4 mutants indicated that ROC4 positively regulates cuticular wax biosynthesis and the drought stress response. ROC4 is ubiquitinated in vivo and subjected to ubiquitin/26S proteasome-mediated degradation. ROC4 degradation was promoted by DHS but delayed in dhs mutants. ROC4 acts downstream of DHS, and Os-BDG is a direct downstream target of the DHS-ROC4 cascade. These results suggest a mechanism whereby DHS negatively regulates wax biosynthesis by promoting the degradation of ROC4, and they suggest that DHS and ROC4 are valuable targets for the engineering of drought-tolerant rice cultivars.
Assuntos
Vias Biossintéticas , Oryza/metabolismo , Proteínas de Plantas/metabolismo , Proteólise , Fatores de Transcrição/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Ceras/metabolismo , Adaptação Fisiológica , Secas , Mutação/genética , Oryza/genética , Fenótipo , Epiderme Vegetal/metabolismo , Epiderme Vegetal/ultraestrutura , Plantas Geneticamente Modificadas , Complexo de Endopeptidases do Proteassoma/metabolismo , Ligação Proteica , Estabilidade Proteica , Estresse Fisiológico , Ubiquitina/metabolismoRESUMO
Grain size and leaf angle are key agronomic traits that determine final yields in rice. However, the underlying molecular mechanisms are not well understood. Here we demonstrate that the Oryza sativa Mitogen Activated Protein Kinase Kinase Kinase OsMKKK70 regulates grain size and leaf angle in rice. Overexpressing OsMKKK70 caused plants to produce longer seeds. The osmkkk62/70 double mutant and the osmkkk55/62/70 triple mutant displayed significantly smaller seeds and a more erect leaf angle compared to the wild type, indicating that OsMKKK70 functions redundantly with its homologs OsMKKK62 and OsMKKK55. Biochemical analysis demonstrated that OsMKKK70 is an active kinase and that OsMKKK70 interacts with OsMKK4 and promotes OsMAPK6 phosphorylation. In addition, the osmkkk62/70 double mutant showed reduced sensitivity to Brassinosteroids (BRs). Finally, overexpressing constitutively active OsMKK4, OsMAPK6, and OsWRKY53 can partially complement the smaller seed size, erect leaf, and BR hyposensitivity of the osmkkk62/70 double mutant. Taken together, these findings suggest that OsMKKK70 might regulate grain size and leaf angle in rice by activating OsMAPK6 and that OsMKKK70, OsMKK4, OsMAPK6, and OsWRKY53 function in a common signaling pathway that controls grain shape and leaf angle.
Assuntos
Oryza , Brassinosteroides/metabolismo , Grão Comestível/metabolismo , Regulação da Expressão Gênica de Plantas , Oryza/genética , Oryza/metabolismo , Folhas de Planta/genética , Folhas de Planta/metabolismo , Proteínas de Plantas/metabolismo , Transdução de Sinais/genéticaRESUMO
Brassinosteroids (BRs) are plant-specific steroid hormones which regulate plant growth, development, and adaptation. Transcriptional regulation plays key roles in plant hormone signaling. A mediator can serve as a bridge between gene-specific transcription factors and the RNA polymerase machinery, functioning as an essential component in regulating the transcriptional process. However, whether a mediator is involved in BR signaling is unknown. Here, we discovered that Oryza sativa mediator subunit 25 (OsMED25) is an important regulator of rice BR signaling. Phenotypic analyses showed that the OsMED25-RNAi and osmed25 mutant presented erect leaves, as observed in BR-deficient mutants. In addition, the OsMED25-RNAi and osmed25 mutant exhibited decreased BR sensitivity. Genetic analysis indicated that OsMED25-RNAi could suppress the enhanced BR signaling phenotype of Osbzr1-D. Further biochemical analysis showed that OsMED25 interacts with OsBZR1 in vivo, and OsMED25 is enriched on the promoter of OsBZR1 target genes. RNA sequencing analysis indicated that OsMED25 affects the expression of approximately 45% of OsBZR1-regulated genes and mainly functions as a corepressor of OsBZR1. Together, these findings revealed that OsMED25 regulates rice BR signaling by interacting with OsBZR1 and modulating the expression of OsBZR1 target genes, thus expanding our understanding of the roles of mediators in plant hormone signaling.
Assuntos
Brassinosteroides/metabolismo , Oryza/anatomia & histologia , Oryza/metabolismo , Subunidades Proteicas/metabolismo , Transdução de Sinais , Brassinosteroides/farmacologia , Proteínas Correpressoras/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Modelos Biológicos , Mutação/genética , Oryza/efeitos dos fármacos , Oryza/genética , Fenótipo , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/metabolismo , Plantas Geneticamente Modificadas , Ligação Proteica/efeitos dos fármacos , Interferência de RNA , Transdução de Sinais/efeitos dos fármacosRESUMO
The grain boundary and dislocation motion characteristics on the atomic scale are significant for the study of material failure mechanisms. In the present work, by theoretical analysis and numerical simulation, the most stable phase of Fe crystal under given conditions is confirmed. Distribution of dislocation potential under different torsion angles is studied for BCC-Fe (001) twist grain boundary. The dislocation motion in Fe (001), Fe (110) and Fe (111) twist grain boundary under tension, compression and shear loading are also investigated.
RESUMO
Brassinosteroids (BRs) are a class of steroid hormones regulating multiple aspects of plant growth, development, and adaptation. Compared with extensive studies in Arabidopsis (Arabidopsis thaliana), the mechanism of BR signaling in rice (Oryza sativa) is less understood. Here, we identified OsWRKY53, a transcription factor involved in defense responses, as an important regulator of rice BR signaling. Phenotypic analyses showed that OsWRKY53 overexpression led to enlarged leaf angles and increased grain size, in contrast to the erect leaves and smaller seeds in oswrky53 mutant. In addition, the oswrky53 exhibited decreased BR sensitivity, whereas OsWRKY53 overexpression plants were hypersensitive to BR, suggesting that OsWRKY53 positively regulates rice BR signaling. Moreover, we show that OsWRKY53 can interact with and be phosphorylated by the OsMAPKK4-OsMAPK6 cascade, and the phosphorylation is required for the biological function of OsWRKY53 in regulating BR responses. Furthermore, we found that BR promotes OsWRKY53 protein accumulation but represses OsWRKY53 transcript level. Taken together, this study revealed the novel role of OsWRKY53 as a regulator of rice BR signaling and also suggested a potential role of OsWRKY53 in mediating the cross talk between the hormone and other signaling pathways.
Assuntos
Brassinosteroides/metabolismo , Oryza/anatomia & histologia , Oryza/metabolismo , Proteínas de Plantas/metabolismo , Transdução de Sinais , Fatores de Transcrição/metabolismo , Retroalimentação Fisiológica , Regulação da Expressão Gênica de Plantas , Mutação/genética , Oryza/genética , Fosforilação , Folhas de Planta/anatomia & histologia , Folhas de Planta/metabolismo , Plantas Geneticamente Modificadas , Ligação Proteica , Transcrição GênicaRESUMO
BACKGROUND: Hereditary spastic paraplegia (HSP) is a neurodegenerative disease that is characterized by progressive weakness and spasticity of the lower extremities; HSP can present as complicated forms with additional neurological signs. More than 70 disease loci have been described with different modes of inheritance. METHODS: In this study, nine subjects from a Chinese family that included two individuals affected by HSP were examined through detailed clinical evaluations, physical examinations, and genetic tests. Targeted exome capture technology was used to identify gene mutations. RESULTS: Two novel compound heterozygous mutations in the SPG 11 gene were identified, c.4001_4002insATAAC and c.4057C>G. The c.4001_4002insATAAC mutation leads to a reading frame shift during transcription, resulting in premature termination of the protein product. The missense mutation c.4057C>G (p.H1353D) is located in a highly conserved domain and is predicted to be a damaging substitution. CONCLUSIONS: Based on the results described here, we propose that these novel compound heterozygous mutations in SPG 11 are the genetic cause of autosomal recessive HSP in this Chinese family.