RESUMO
In cereal grains, starch is synthesized by the concerted actions of multiple enzymes on the surface of starch granules within the amyloplast. However, little is known about how starch-synthesizing enzymes access starch granules, especially for amylopectin biosynthesis. Here, we show that the rice (Oryza sativa) floury endosperm9 (flo9) mutant is defective in amylopectin biosynthesis, leading to grains exhibiting a floury endosperm with a hollow core. Molecular cloning revealed that FLO9 encodes a plant-specific protein homologous to Arabidopsis (Arabidopsis thaliana) LIKE EARLY STARVATION1 (LESV). Unlike Arabidopsis LESV, which is involved in starch metabolism in leaves, OsLESV is required for starch granule initiation in the endosperm. OsLESV can directly bind to starch by its C-terminal tryptophan (Trp)-rich region. Cellular and biochemical evidence suggests that OsLESV interacts with the starch-binding protein FLO6, and loss-of-function mutations of either gene impair ISOAMYLASE1 (ISA1) targeting to starch granules. Genetically, OsLESV acts synergistically with FLO6 to regulate starch biosynthesis and endosperm development. Together, our results identify OsLESV-FLO6 as a non-enzymatic molecular module responsible for ISA1 localization on starch granules, and present a target gene for use in biotechnology to control starch content and composition in rice endosperm.
Assuntos
Endosperma , Regulação da Expressão Gênica de Plantas , Oryza , Proteínas de Plantas , Amido , Oryza/genética , Oryza/metabolismo , Oryza/crescimento & desenvolvimento , Endosperma/metabolismo , Endosperma/genética , Amido/metabolismo , Amido/biossíntese , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Amilopectina/metabolismo , Mutação , Plantas Geneticamente ModificadasRESUMO
The regulatory mechanisms of anthocyanin biosynthesis have been well documented at the transcriptional and translational levels. By contrast, how anthocyanin biosynthesis is epigenetically regulated remains largely unknown. In this study, we employed genetic, molecular biology, and chromatin immunoprecipitation-quantitative polymerase chain reaction assays to identify a regulatory module essential for repressing the expression of genes involved in anthocyanin biosynthesis through chromatin remodeling. We found that SILENCING DEFECTIVE 2 (SDE2), which was previously identified as a negative regulator for sucrose-induced anthocyanin accumulation in Arabidopsis, is cleaved into N-terminal SDE2-UBL and C-terminal SDE2-C fragments at the first diglycine motif, and the cleaved SDE2-C, which can fully complement the sde2 mutant, is localized in the nucleus and physically interacts with LIKE HETEROCHROMATIN PROTEIN 1 (LHP1) in vitro and in vivo. Genetic analyses showed that both SDE2 and LHP1 act as negative factors for anthocyanin biosynthesis. Consistently, immunoblot analysis revealed that the level of LHP1-bound histone H3 lysine 27 trimethylation (H3K27me3) significantly decreases in sde2 and lhp1 mutants, compared to wild-type (WT). In addition, we found that sugar can induce expression of SDE2 and LHP1, and enhance the level of the nucleus-localized SDE2-C. Taken together, our data suggest that the SDE2-C-LHP1 module is required for repression of gene expression through H3K27me3 modification during sugar-induced anthocyanin biosynthesis in Arabidopsis thaliana.
Assuntos
Antocianinas , Proteínas de Arabidopsis , Arabidopsis , Núcleo Celular , Regulação da Expressão Gênica de Plantas , Histonas , Mutação , Ligação Proteica , Sacarose , Antocianinas/biossíntese , Antocianinas/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Núcleo Celular/metabolismo , Proteínas Cromossômicas não Histona/metabolismo , Proteínas Cromossômicas não Histona/genética , Histonas/metabolismo , Metilação , Mutação/genética , Ligação Proteica/efeitos dos fármacos , Sacarose/metabolismo , Fatores de TranscriçãoRESUMO
Endosperm is the main storage organ in cereal grain and determines grain yield and quality. The molecular mechanisms of heat shock proteins in regulating starch biosynthesis and endosperm development remain obscure. Here, we report a rice floury endosperm mutant flo24 that develops abnormal starch grains in the central starchy endosperm cells. Map-based cloning and complementation test showed that FLO24 encodes a heat shock protein HSP101, which is localized in plastids. The mutated protein FLO24T296I dramatically lost its ability to hydrolyze ATP and to rescue the thermotolerance defects of the yeast hsp104 mutant. The flo24 mutant develops more severe floury endosperm when grown under high-temperature conditions than normal conditions. And the FLO24 protein was dramatically induced at high temperature. FLO24 physically interacts with several key enzymes required for starch biosynthesis, including AGPL1, AGPL3 and PHO1. Combined biochemical and genetic evidence suggests that FLO24 acts cooperatively with HSP70cp-2 to regulate starch biosynthesis and endosperm development in rice. Our results reveal that FLO24 acts as an important regulator of endosperm development, which might function in maintaining the activities of enzymes involved in starch biosynthesis in rice.
Assuntos
Endosperma , Oryza , Proteínas de Plantas , Amido , Trifosfato de Adenosina/metabolismo , Trifosfato de Adenosina/biossíntese , Endosperma/genética , Endosperma/crescimento & desenvolvimento , Endosperma/metabolismo , Regulação da Expressão Gênica de Plantas , Teste de Complementação Genética , Mutação/genética , Oryza/genética , Oryza/metabolismo , Oryza/crescimento & desenvolvimento , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Plastídeos/metabolismo , Ligação Proteica , Amido/biossíntese , Amido/genética , Termotolerância , Fatores de TranscriçãoRESUMO
Glyphosate (GLY) is a widely used herbicide that has been revealed to inhibit testosterone synthesis in humans and animals. Melatonin (MET) is an endogenous hormone that has been demonstrated to promote mammalian testosterone synthesis via protecting mitochondrial function. However, it remains unclear whether MET targets mitochondria to alleviate GLY-inhibited testosterone synthesis in avian. In this study, an avian model using 7-day-old rooster upon chronic exposure to GLY with the treatment of MET was designed to clarify this issue. Data first showed that GLY-induced testicular Leydig cell damage, structural damage of the seminiferous tubule, and sperm quality decrease were mitigated by MET. Transcriptomic analyses of the testicular tissues revealed the potentially critical role of mitophagy and steroid hormone biosynthesis in the process of MET counteracting GLY-induced testicular damage. Also, validation data demonstrated that the inhibition of testosterone synthesis due to GLY-induced mitochondrial dynamic imbalance and concomitant Parkin-dependent mitophagy activation is alleviated by MET. Moreover, GLY-induced oxidative stress in serum and testicular tissue were significantly reversed by MET. In summary, these findings demonstrate that MET effectively ameliorates GLY-inhibited testosterone synthesis by inhibiting mitophagy activation, which provides a promising remedy for the application of MET as a potential therapeutic agent to antagonize reproductive toxicity induced by GLY and similar contaminants.
Assuntos
Glifosato , Melatonina , Humanos , Masculino , Animais , Testosterona , Melatonina/farmacologia , Galinhas , Sêmen , Mitocôndrias , MamíferosRESUMO
INTRODUCTION AND OBJECTIVE: As globalization and modernization continue to impact people's lives, a significant shift in lifestyle has taken place, resulting in a worldwide decrease in physical activity and an increase in unhealthy eating patterns. Physical inactivity has become the fourth leading cause of death globally. The aim of this scoping review is to analyze the concept and development of integrating physical activity into healthcare (IPAHc), based on the principles of sports and exercise medicine (SEM) and exercise is medicine (EIM). REVIEW METHODS: A systematic search was conducted of relevant published studies with full text using PubMed, Scopus, Web of Science, Academic Search Ultimate, Medline, and SPORTDiscus, via the EBSCO search platform. BRIEF DESCRIPTION OF THE STATE OF KNOWLEDGE: Twenty-nine studies met the inclusion criteria. The integration pathway centres around physical activity consultation and/or referral, and information technology which has been extensively utilized in IPAHc, including websites, electronic medical records, social media, wearable devices, mobile software, and referral tools. SEM and EIM face numerous implementation challenges, such as time constraints, education/training, resources, and tools. SUMMARY: The concept of IPAHc involves the integration of Physical Activity Vital Signs (PAVS) into electronic medical records to evaluate the physical activity levels of the general population. This can assist individuals in achieving fitness goals, preventing diseases, treating existing illnesses, and undergoing rehabilitation. IPAHc has been in development for many years and is now being explored in practice. Despite the widespread use of information technology in this integration process, a number of challenges still need addressing.
Assuntos
Exercício Físico , Humanos , Atenção à SaúdeRESUMO
Corrugated damage to bearings is a common fault in electrical facilities such as new energy vehicles, wind power, and high-speed railways. The aim of this article is to reveal the microscopic characteristics and formation mechanism of such damages. The corrugation with alternating "light" and "dark" shape was produced on GCr15 bearing races in the experimental conditions. Compared to the light area, the dark area (in the images generated by optical microscope) has more severe electrical erosion, lower hardness, more concave morphology, and lower oxidation. As the voltage increases, the width of the corrugation, the height difference between corrugation, and surface roughness all increase. It is believed that the formation of corrugated damage requires a sufficiently high voltage to induce the periodic destruction and reconstruction of the lubrication film. When the bearing is in a metal-lubrication film-metal contact state, the high voltage causes the lubrication film to break down and induce electrical erosion. Then, the contact area is in metal-metal contact, and the surface is mainly damaged by mechanical rolling. After the reconstruction of lubrication film, the next round of electrical erosion begins. The results are helpful for a deeper understanding of the mechanism of bearing erosion in electrical application.
RESUMO
Glyphosate (GLY) is the most universally used herbicide worldwide and its application has caused extensive pollution to the ecological environment. Increasing evidence has revealed the multi-organ toxicity of GLY in different species, but its male reproductive toxicity in avian species remains unknown. Thus, in vivo and in vitro studies were conducted to clarify this issue. Data firstly showed that chronic GLY exposure caused testicular pathological damage. Intriguingly, we identified and verified a marked down-regulation gap junction gene Connexin 43 (Cx43) in GLY-exposed rooster testis by transcriptome analysis. Cx43 generated by Sertoli cells acts as a key component of blood-testis barrier (BTB). To further investigate the cause of GLY-induced downregulation of Cx43 to disrupt BTB, we found that autophagy activation is revealed in GLY-exposed rooster testis and primary avian Sertoli cells. Moreover, GLY-induced Cx43 downregulation was significantly alleviated by ATG5 knockdown or CQ administration, respectively, demonstrating that GLY-induced autophagy activation contributed to Cx43 degradation. Mechanistically, GLY-induced autophagy activation and resultant Cx43 degradation was due to its direct interaction with ER-α. In summary, these findings demonstrate that chronic GLY exposure activates autophagy to induce Cx43 degradation, which causes BTB damage and resultant reproductive toxicity in roosters.
RESUMO
Although both protein arginine methylation (PRMT) and jasmonate (JA) signaling are crucial for regulating plant development, the relationship between these processes in the control of spikelet development remains unclear. In this study, we used the CRISPR/Cas9 technology to generate two OsPRMT6a loss-of-function mutants that exhibit various abnormal spikelet structures. Interestingly, we found that OsPRMT6a can methylate arginine residues in JA signal repressors OsJAZ1 and OsJAZ7. We showed that arginine methylation of OsJAZ1 enhances the binding affinity of OsJAZ1 with the JA receptors OsCOI1a and OsCOI1b in the presence of JAs, thereby promoting the ubiquitination of OsJAZ1 by the SCFOsCOI1a/OsCOI1b complex and degradation via the 26S proteasome. This process ultimately releases OsMYC2, a core transcriptional regulator in the JA signaling pathway, to activate or repress JA-responsive genes, thereby maintaining normal plant (spikelet) development. However, in the osprmt6a-1 mutant, reduced arginine methylation of OsJAZ1 impaires the interaction between OsJAZ1 and OsCOI1a/OsCOI1b in the presence of JAs. As a result, OsJAZ1 proteins become more stable, repressing JA responses, thus causing the formation of abnormal spikelet structures. Moreover, we discovered that JA signaling reduces the OsPRMT6a mRNA level in an OsMYC2-dependent manner, thereby establishing a negative feedback loop to balance JA signaling. We further found that OsPRMT6a-mediated arginine methylation of OsJAZ1 likely serves as a switch to tune JA signaling to maintain normal spikelet development under harsh environmental conditions such as high temperatures. Collectively, our study establishes a direct molecular link between arginine methylation and JA signaling in rice.
Assuntos
Arginina , Ciclopentanos , Oryza , Oxilipinas , Proteínas de Plantas , Proteína-Arginina N-Metiltransferases , Transdução de Sinais , Ciclopentanos/metabolismo , Oxilipinas/metabolismo , Oryza/crescimento & desenvolvimento , Oryza/genética , Oryza/metabolismo , Arginina/metabolismo , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Metilação , Proteína-Arginina N-Metiltransferases/metabolismo , Proteína-Arginina N-Metiltransferases/genética , Regulação da Expressão Gênica de PlantasRESUMO
Endosperm, the major storage organ in cereal grains, determines the grain yield and quality. Mitochondria provide the energy for dry matter accumulation, in the endosperm development. Although mitochondrial single-stranded DNA-binding proteins (mtSSBs) play a canonical role in the maintenance of single-stranded mitochondrial DNA, their molecular functions in RNA processing and endosperm development remain obscure. Here, we report a defective rice endosperm mutant, floury endosperm26 (flo26), which develops abnormal starch grains in the endosperm. Map-based cloning and complementation experiments showed that FLO26 allele encodes a mitochondrial single-stranded DNA-binding protein, named as mtSSB1.1. Loss of function of mtSSB1.1 affects the transcriptional level of many mitochondrially-encoded genes and RNA splicing of nad1, a core component of respiratory chain complex I in mitochondria. As a result, dysfunctional mature nad1 led to dramatically decreased complex I activity, thereby reducing ATP production. Our results reveal that mtSSB1.1 plays an important role in the maintenance of mitochondrial function and endosperm development by stabilizing the splicing of mitochondrial RNA in rice.