RESUMO
BACKGROUND: The endosperm serves as the primary source of nutrients for maize (Zea mays L.) kernel embryo development and germination. Positioned at the base of the endosperm, the transfer cells (TCs) of the basal endosperm transfer layer (BETL) generate cell wall ingrowths, which enhance the connectivity between the maternal plant and the developing kernels. These TCs play a crucial role in nutrient transport and defense against pathogens. The molecular mechanism underlying BETL development in maize remains unraveled. RESULTS: This study demonstrated that the MYB-related transcription factor ZmMYBR29, exhibited specific expression in the basal cellularized endosperm, as evidenced by in situ hybridization analysis. Utilizing the CRISPR/Cas9 system, we successfully generated a loss-of-function homozygous zmmybr29 mutant, which presented with smaller kernel size. Observation of histological sections revealed abnormal development and disrupted morphology of the cell wall ingrowths in the BETL. The average grain filling rate decreased significantly by 26.7% in zmmybr29 mutant in comparison to the wild type, which impacted the dry matter accumulation within the kernels and ultimately led to a decrease in grain weight. Analysis of RNA-seq data revealed downregulated expression of genes associated with starch synthesis and carbohydrate metabolism in the mutant. Furthermore, transcriptomic profiling identified 23 genes that expressed specifically in BETL, and the majority of these genes exhibited altered expression patterns in zmmybr29 mutant. CONCLUSIONS: In summary, ZmMYBR29 encodes a MYB-related transcription factor that is expressed specifically in BETL, resulting in the downregulation of genes associated with kernel development. Furthermore, ZmMYBR29 influences kernels weight by affecting the grain filling rate, providing a new perspective for the complementation of the molecular regulatory network in maize endosperm development.
Assuntos
Grão Comestível , Endosperma , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas , Fatores de Transcrição , Zea mays , Zea mays/genética , Zea mays/crescimento & desenvolvimento , Zea mays/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Grão Comestível/genética , Grão Comestível/crescimento & desenvolvimento , Grão Comestível/metabolismo , Endosperma/genética , Endosperma/crescimento & desenvolvimento , Endosperma/metabolismo , Parede Celular/metabolismo , Parede Celular/genética , Sementes/genética , Sementes/crescimento & desenvolvimento , Sementes/metabolismo , Sistemas CRISPR-CasRESUMO
The mitochondrion is a semi-autonomous organelle that provides energy for cell activities through oxidative phosphorylation. In this study, we identified a defective kernel 66 (dek66)-mutant maize with defective kernels. We characterized a candidate gene, DEK66, encoding a ribosomal assembly factor located in mitochondria and possessing GTPase activity (which belongs to the ribosome biogenesis GTPase A family). In the dek66 mutant, impairment of mitochondrial structure and function led to the accumulation of reactive oxygen species and promoted programmed cell death in endosperm cells. Furthermore, the transcript levels of most of the key genes associated with nutrient storage, mitochondrial respiratory chain complex, and mitochondrial ribosomes in the dek66 mutant were significantly altered. Collectively, the results suggest that DEK66 is essential for the development of maize kernels by affecting mitochondrial function. This study provides a reference for understanding the impact of a mitochondrial ribosomal assembly factor in maize kernel development.
Assuntos
Proteínas de Plantas , Zea mays , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Zea mays/metabolismo , GTP Fosfo-Hidrolases/genética , GTP Fosfo-Hidrolases/metabolismo , Endosperma/metabolismo , Mitocôndrias/genética , Mitocôndrias/metabolismo , Regulação da Expressão Gênica de PlantasRESUMO
KEY MESSAGE: We isolated an MYB-like gene from Korla fragrant pear using differential display RT-PCR. Expression of this gene in flowers appears to be correlated with calyx persistence. Korla fragrant pear (Pyrus brestschneideri Rehd) is an economically important pear cultivar in China. A persistent calyx results in the deformation of the fruit. We used differential display RT-PCR to obtain 42 cDNA fragments from Korla fragrant pear flowers. Alignments of nucleotide and amino acid sequences suggested that two fragments (kfp1and kfp4) were related to calyx persistence. The fragments were 78% homologous with Malus × domestica SPL transcription factor (SPL3) and 83% homologous with Malus × domestica MYB transcription factor (MYB12). The complete cDNA sequence of kfpMYB was determined to clarify the role of MYB in calyx persistence. kfpMYB contained a 116 bp 5'-UTR, a 1122 bp open reading frame encoding 374 amino acids, and a 319 bp 3'-UTR. The nucleotide and amino acid sequences of the cDNA in Korla fragrant pear were highly homologous with those of MYB transcription factors in other plant species, suggesting that the sequence is a MYB transcription factor gene. The abundance of kfpMYB mRNA varied significantly between the second and fourth flowers on the branch. Furthermore, kfpMYB expression changed significantly during anthesis and was significantly higher in Jinfeng pear (persistent calyx) and Korla fragrant pear than in Yali pear (deciduous calyx). Expression of kfpMYB was significantly reduced by naphthalene (NAA), abscisic acid (ABA), PBO, and paclobutrazol (PP333). Uniconazole, ethylene (ETH), and gibberellic acid (GA3) had no signicant effect on kfpMYB expression. In conclusion, the expression of kfpMYB appears to be correlated with calyx persistence in Korla fragrant pear.
Assuntos
Regulação da Expressão Gênica de Plantas , Pyrus/genética , Fatores de Transcrição/genética , Ácido Abscísico/farmacologia , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , DNA Complementar/genética , Flores/genética , Frutas/genética , Frutas/crescimento & desenvolvimento , Perfilação da Expressão Gênica , Dados de Sequência Molecular , Naftalenos/farmacologia , Filogenia , Reguladores de Crescimento de Plantas/farmacologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Estrutura Secundária de Proteína , Pyrus/crescimento & desenvolvimento , RNA de Plantas/genética , Alinhamento de Sequência , Análise de Sequência de DNA , Fatores de Transcrição/metabolismo , Triazóis/farmacologiaRESUMO
Nitrogen is an essential macronutrient for plants and regulates many aspects of plant growth and development. Nitrate is one of the major forms of nitrogen in plants. However, the role of nitrate uptake and allocation in seed development is not fully understood. Here, we identified the maize (Zea mays) small-kernel mutant zmnpf7.9 and characterized the candidate gene, ZmNPF7.9, which was the same gene as nitrate transport 1.5 (NRT1.5) in maize. This gene is specifically expressed in the basal endosperm transfer layer cells of maize endosperm. Dysfunction of ZmNPF7.9 resulted in delayed endosperm development, abnormal starch deposition and decreased hundred-grain weight. Functional analysis of cRNA-injected Xenopus oocytes showed that ZmNPF7.9 is a low-affinity, pH-dependent bidirectional nitrate transporter. Moreover, the amount of nitrate in mature seeds of the zmnpf7.9 mutant was reduced. These suggest that ZmNPF7.9 is involved in delivering nitrate from maternal tissues to the developing endosperm. Moreover, most of the key genes associated with glycolysis/gluconeogenesis, carbon fixation, carbon metabolism and biosynthesis of amino acids pathways in the zmnpf7.9 mutant were significantly down-regulated. Thus, our results demonstrate that ZmNPF7.9 plays a specific role in seed development and grain weight by regulating nutrition transport and metabolism, which might provide useful information for maize genetic improvement.
Assuntos
Proteínas de Transporte de Ânions/genética , Proteínas de Plantas/genética , Sementes/crescimento & desenvolvimento , Zea mays/crescimento & desenvolvimento , Zea mays/genética , Proteínas de Transporte de Ânions/metabolismo , Endosperma/crescimento & desenvolvimento , Transportadores de Nitrato , Proteínas de Plantas/metabolismo , Amido/metabolismo , Zea mays/metabolismoRESUMO
Facilitating navigation in pedestrian environments is critical for enabling people who are blind and visually impaired (BVI) to achieve independent mobility. A deep reinforcement learning (DRL)-based assistive guiding robot with ultrawide-bandwidth (UWB) beacons that can navigate through routes with designated waypoints was designed in this study. Typically, a simultaneous localization and mapping (SLAM) framework is used to estimate the robot pose and navigational goal; however, SLAM frameworks are vulnerable in certain dynamic environments. The proposed navigation method is a learning approach based on state-of-the-art DRL and can effectively avoid obstacles. When used with UWB beacons, the proposed strategy is suitable for environments with dynamic pedestrians. We also designed a handle device with an audio interface that enables BVI users to interact with the guiding robot through intuitive feedback. The UWB beacons were installed with an audio interface to obtain environmental information. The on-handle and on-beacon verbal feedback provides points of interests and turn-by-turn information to BVI users. BVI users were recruited in this study to conduct navigation tasks in different scenarios. A route was designed in a simulated ward to represent daily activities. In real-world situations, SLAM-based state estimation might be affected by dynamic obstacles, and the visual-based trail may suffer from occlusions from pedestrians or other obstacles. The proposed system successfully navigated through environments with dynamic pedestrians, in which systems based on existing SLAM algorithms have failed.
RESUMO
Transformation of perfluorooctane sulfonate (PFOS) precursors (PreFOSs) is considered an additional source of PFOS in the environment and biota. A PreFOSs-degrading bacterium PF1, which was able to utilize PreFOSs as the sole carbon and energy source for growth, was isolated from contaminated soil collected from the surroundings of a fluoride factory. According to its morphology and 16S rDNA gene sequence analysis, strain PF1 was identified as Hyphomicrobium sp. The degradation rates of perfluorooctane sulfonamide (PFOSA) and N-ethyl perfluorooctane sulfonamide (N-EtFOSA) by PF1 were 14.6% and 8.2% (30â; pH=7.0-7.2), respectively, whereas PF1 was unable to degrade PFOS. PFOSA could be biodegraded to PFOS. N-EtFOSA could be biodegraded to perfluorooctane sulfonamide acetic acid (FOSAA), PFOSA, and PFOS; PFOS was the predominant metabolite. Based on the above analysis, the proposed metabolic pathway of PFOSA by strain PF1 is deamination to form PFOS. Two possible degradation pathways are proposed for N-EtFOSA: â deethylation of N-EtFOSA to produce PFOSA, followed by deamination to form PFOS, and â¡oxidation of N-EtFOSA to FOSAA followed by sequential dealkylation to produce PFOSA, and then transformation to PFOS by deamination.