RESUMO
Changes of membrane lipid composition contribute to plant adaptation to various abiotic stresses. Here, a comparative study was undertaken to investigate the mechanisms of how lipid alteration affects plant growth and development under nitrogen (N) deficiency. Two wheat cultivars: the N deficiency-tolerant cultivar Xiaoyan 6 (XY) and the N deficiency-sensitive cultivar Aikang 58 (AK) were used to test if the high N-deficiency tolerance was related with lipid metabolism. The results showed that N deficiency inhibited the morpho-physiological parameters in both XY and AK cultivars, which showed a significant decrease in biomass, N content, photosynthetic efficiency, and lipid contents. However, these decreases were more pronounced in AK than XY. In addition, XY showed a notable increase in fatty acid unsaturation, relatively well-maintained chloroplast ultrastructure, and minimized damage of lipid peroxidation and enhanced PSII activity under N-deficient condition, as compared with AK. Transcription levels of many genes involved in lipid biosynthesis and fatty acid desaturation were up-regulated in response to N deficiency in two wheat cultivars, while the expressions were much higher in XY than AK under N deficiency. These results highlight the importance of alterations in lipid metabolism in N deficiency tolerance in wheat. High levels of lipid content and unsaturated fatty acids maintained the membrane structure and function, contributing to high photosynthesis and antioxidant capacities, thereby improved the tolerance to N deficiency.
Assuntos
Metabolismo dos Lipídeos , Nitrogênio , Plântula , Triticum , Metabolismo dos Lipídeos/genética , Nitrogênio/deficiência , Triticum/crescimento & desenvolvimento , Triticum/metabolismo , Plântula/crescimento & desenvolvimento , Plântula/metabolismo , Regulação da Expressão Gênica de Plantas/fisiologia , Ácidos Graxos Insaturados/metabolismo , Fotossíntese/fisiologia , Membrana Celular/fisiologia , Oxidantes/biossíntese , Cloroplastos/fisiologia , Cloroplastos/ultraestruturaRESUMO
Chloroplasts accumulate in regions of plant cells exposed to irradiation to maximize light reception for efficient photosynthesis. This response is mediated by the blue-light receptor phototropin. Upon the perception of blue light, phototropin is photoactivated, an unknown signal is transmitted from the photoactivated phototropin to distant chloroplasts, and the chloroplasts begin their directional movement. How activated phototropin initiates this signal transmission is unknown. Here, using the liverwort Marchantia polymorpha, we analysed whether increased photoactive phototropin levels mediate signal transmission and chloroplast behaviour during the accumulation response. The signal transmission rate was higher in transgenic cells overexpressing phototropin than in wild-type cells. However, the chloroplast directional movement was similar between wild-type and transgenic cells. Consistent with the observation, increasing the amount of photoactivated phototropin through higher blue-light intensity also accelerated signal transmission but did not affect chloroplast behaviour in wild-type cells. Photoactivation of phototropin under weak blue-light led to the greater protein level of phosphorylated phototropin in cells overexpressing phototropin than in wild-type cells, whereas the autophosphorylation level within each phototropin molecule was similar. These results indicate that the abundance of photoactivated phototropin modulates the signal transmission rate to distant chloroplasts but does not affect chloroplast behaviour during the accumulation response.
Assuntos
Cloroplastos , Luz , Marchantia , Fototropinas , Plantas Geneticamente Modificadas , Transdução de Sinais , Cloroplastos/metabolismo , Cloroplastos/efeitos da radiação , Cloroplastos/fisiologia , Fototropinas/metabolismo , Fototropinas/genética , Marchantia/fisiologia , Marchantia/efeitos da radiação , Marchantia/genética , Marchantia/metabolismo , Fosforilação , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genéticaRESUMO
Much progress has been made in understanding the molecular mechanisms of plant adaptation to heat stress. However, the great diversity of models and stress conditions, and the fact that analyses are often limited to a small number of approaches, complicate the picture. We took advantage of a liquid culture system in which Arabidopsis seedlings are arrested in their development, thus avoiding interference with development and drought stress responses, to investigate through an integrative approach seedlings' global response to heat stress and acclimation. Seedlings perfectly tolerate a noxious heat shock (43°C) when subjected to a heat priming treatment at a lower temperature (38°C) the day before, displaying a thermotolerance comparable to that previously observed for Arabidopsis. A major effect of the pre-treatment was to partially protect energy metabolism under heat shock and favor its subsequent rapid recovery, which was correlated with the survival of seedlings. Rapid recovery of actin cytoskeleton and mitochondrial dynamics were another landmark of heat shock tolerance. The omics confirmed the role of the ubiquitous heat shock response actors but also revealed specific or overlapping responses to priming, heat shock, and their combination. Since only a few components or functions of chloroplast and mitochondria were highlighted in these analyses, the preservation and rapid recovery of their bioenergetic roles upon acute heat stress do not require extensive remodeling of the organelles. Protection of these organelles is rather integrated into the overall heat shock response, thus allowing them to provide the energy required to elaborate other cellular responses toward acclimation.
Assuntos
Aclimatação , Arabidopsis , Resposta ao Choque Térmico , Plântula , Arabidopsis/fisiologia , Arabidopsis/genética , Plântula/fisiologia , Plântula/genética , Resposta ao Choque Térmico/fisiologia , Metabolismo Energético , Termotolerância/fisiologia , Cloroplastos/metabolismo , Cloroplastos/fisiologia , Mitocôndrias/metabolismo , Regulação da Expressão Gênica de Plantas , Organelas/fisiologia , Organelas/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Temperatura Alta , Dinâmica Mitocondrial/fisiologiaRESUMO
Chloroplast-actin (cp-actin) filaments are crucial for light-induced chloroplast movement, and appear in the front region of moving chloroplasts when visualized using GFP-mouse Talin. They are short and thick, exist between a chloroplast and the plasma membrane, and move actively and rapidly compared to cytoplasmic long actin filaments that run through a cell. The average period during which a cp-actin filament was observed at the same position was less than 0.5 s. The average lengths of the cp-actin filaments calculated from those at the front region of the moving chloroplast and those around the chloroplast periphery after stopping the movement were almost the same, approximately 0.8 µm. Each cp-actin filament is shown as a dotted line consisting of 4-5 dots. The vector sum of cp-actin filaments in a moving chloroplast is parallel to the moving direction of the chloroplast, suggesting that the direction of chloroplast movement is regulated by the vector sum of cp-actin filaments. However, once the chloroplasts stopped moving, the vector sum of the cp-actin filaments around the chloroplast periphery was close to zero, indicating that the direction of movement was undecided. To determine the precise structure of cp-actin filaments under electron microscopy, Arabidopsis leaves and fern Adiantum capillus-veneris gametophytes were frozen using a high-pressure freezer, and observed under electron microscopy. However, no bundled microfilaments were found, suggesting that the cp-actin filaments were unstable even under high-pressure freezing.
Assuntos
Citoesqueleto de Actina , Arabidopsis , Cloroplastos , Luz , Cloroplastos/fisiologia , Cloroplastos/metabolismo , Cloroplastos/efeitos da radiação , Cloroplastos/ultraestrutura , Arabidopsis/fisiologia , Arabidopsis/efeitos da radiação , Adiantum/fisiologia , Adiantum/efeitos da radiação , Folhas de Planta/fisiologia , Folhas de Planta/efeitos da radiação , Actinas/metabolismo , MovimentoRESUMO
Plants have unique responses to fluctuating light conditions. One such response involves chloroplast photorelocation movement, which optimizes photosynthesis under weak light by the accumulation of chloroplasts along the periclinal side of the cell, which prevents photodamage under strong light by avoiding chloroplast positioning toward the anticlinal side of the cell. This light-responsive chloroplast movement relies on the reorganization of chloroplast actin (cp-actin) filaments. Previous studies have suggested that CHLOROPLAST UNUSUAL POSITIONING 1 (CHUP1) is essential for chloroplast photorelocation movement as a regulator of cp-actin filaments. In this study, we conducted comprehensive analyses to understand CHUP1 function. Functional, fluorescently tagged CHUP1 colocalized with and was coordinately reorganized with cp-actin filaments on the chloroplast outer envelope during chloroplast movement in Arabidopsis thaliana. CHUP1 distribution was reversibly regulated in a blue light- and phototropin-dependent manner. X-ray crystallography revealed that the CHUP1-C-terminal domain shares structural homology with the formin homology 2 (FH2) domain, despite lacking sequence similarity. Furthermore, the CHUP1-C-terminal domain promoted actin polymerization in the presence of profilin in vitro. Taken together, our findings indicate that CHUP1 is a plant-specific actin polymerization factor that has convergently evolved to assemble cp-actin filaments and enables chloroplast photorelocation movement.
Assuntos
Proteínas de Arabidopsis , Arabidopsis , Actinas , Proteínas de Arabidopsis/genética , Polimerização , Proteínas de Cloroplastos/genética , Arabidopsis/genética , Citoesqueleto de Actina , Cloroplastos/fisiologia , Luz , MovimentoRESUMO
CO2 -induced chloroplast movement was reported in the monograph by Gustav Senn in 1908: unilateral CO2 supply to the one cell-layered moss leaves induced the positively CO2 -tactic periclinal arrangement of chloroplasts. Here, using the model moss plant Physcomitrium patens, we examined basic features of chloroplast CO2 -tactic relocation with a modernized experimental system. The CO2 relocation was light-dependent and, especially, CO2 relocation in red light was substantially dependent on photosynthetic activity. In blue light, CO2 relocation was mainly dependent on microfilaments while microtubule-based movement was insensitive to CO2 , whereas in red light, both cytoskeletons contributed redundantly to CO2 relocation. The CO2 relocation was observed not only when the two leaf surfaces were exposed to CO2 -free air versus CO2 -containing air, but also by exposing them physiologically relevant differences in CO2 concentrations. In the leaves placed on the surface of a gel sheet, chloroplasts avoided the gel side and positioned in the air-facing surface, and this phenomenon was also shown to be photosynthesis dependent. Based on these observations, we propose a hypothesis that the threshold light intensity between the light-accumulation and -avoidance responses of the photorelocation would be increased by CO2 , resulting in the CO2 -tactic relocation of chloroplasts.
Assuntos
Briófitas , Dióxido de Carbono , Luz , Cloroplastos/fisiologia , Folhas de Planta/fisiologia , MovimentoRESUMO
Chloroplasts move to the periclinal walls of cells under weak light to harness light energy for photosynthesis and to anticlinal walls to avoid strong light. These responses involve the cytoskeleton components microtubules and/or actin filaments. In the dark, chloroplasts move to the anticlinal cell walls bordering neighbouring cells (dark-positioning response), but this response in various plants normally requires a prolonged dark incubation period, which has hampered analysis. However, we recently demonstrated the dark-positioning response that can be induced after a short period of dark incubation in the liverwort Apopellia endiviifolia. Here, we investigated whether the cytoskeleton components function in the dark-positioning response of A. endiviifolia cells. Microtubules and actin filaments were fluorescently visualised in A. endiviifolia cells and were disrupted following treatment with the microtubule and actin filament polymerisation inhibitors. The dark-positioning response was unaffected in the cells with disrupted microtubules. By contrast, the dark-positioning response was inhibited by the disruption of actin filaments. The disruption of actin filaments also restricted chloroplast mobility during light- and cold-dependent chloroplast movements in A. endiviifolia. Therefore, the dark-positioning response of A. endiviifolia depends solely on an actin filament-associated motility mechanism, as do the light- and cold-dependent chloroplast responses.
Assuntos
Hepatófitas , Luz , Citoesqueleto de Actina/fisiologia , Microtúbulos , Cloroplastos/fisiologia , ActinasRESUMO
Plants have developed intricate mechanisms to adapt to changing light conditions. Besides phototropism and heliotropism (differential growth toward light and diurnal motion with respect to sunlight, respectively), chloroplast motion acts as a fast mechanism to change the intracellular structure of leaf cells. While chloroplasts move toward the sides of the plant cell to avoid strong light, they accumulate and spread out into a layer on the bottom of the cell at low light to increase the light absorption efficiency. Although the motion of chloroplasts has been studied for over a century, the collective organelle motion leading to light-adapting self-organized structures remains elusive. Here, we study the active motion of chloroplasts under dim-light conditions, leading to an accumulation in a densely packed quasi-2D layer. We observe burst-like rearrangements and show that these dynamics resemble systems close to the glass transition by tracking individual chloroplasts. Furthermore, we provide a minimal mathematical model to uncover relevant system parameters controlling the stability of the dense configuration of chloroplasts. Our study suggests that the meta-stable caging close to the glass transition in the chloroplast monolayer serves a physiological relevance: Chloroplasts remain in a spread-out configuration to increase the light uptake but can easily fluidize when the activity is increased to efficiently rearrange the structure toward an avoidance state. Our research opens questions about the role that dynamical phase transitions could play in self-organized intracellular responses of plant cells toward environmental cues.
Assuntos
Cloroplastos , Células Vegetais , Cloroplastos/fisiologia , Luz Solar , Fototropismo , Folhas de Planta/fisiologia , LuzRESUMO
Silica bodies are commonly found in Selaginella, but their function is unclear. Lens-like appearance and location in many species above giant chloroplasts of dorsal epidermal cells suggest optical functions. Silica body morphology in three Selaginella species was studied by microscopy. Optical effects were assessed by wave-optic simulations. Large convex, approximately hemispherical (papillose) and small approximately conical (concave-convex) silica bodies were found in different species. Both types lead to a concentrated spot of light high in the dorsal epidermal cell. Large convex bodies concentrate light 10-25 times in a shape-dependent manner by refraction, and small silica bodies concentrate light in a shape-insensitive, but wavelength-dependent, manner by diffraction (red light: approx. 2.3 times; blue light: approx. 1.5 times). Due to chloroplast movement, this concentrated light is above the chloroplast under high light, but within it under low light. Beyond the spot of concentration, light is dispersed into the chloroplast. Thin Selaginella leaves mean these effects may enhance light capture and minimize photodamage, but other effects such as inhibition of herbivory, mechanical support, and immune responses need to be considered. Silica bodies undoubtedly have optical effects, but their significance to the functioning of the plant requires direct studies of ecophysiological performance.
Assuntos
Selaginellaceae , Cloroplastos/fisiologia , Folhas de Planta/fisiologia , Dióxido de SilícioRESUMO
How the plasma membrane senses external heat-stress signals to communicate with chloroplasts to orchestrate thermotolerance remains elusive. We identified a quantitative trait locus, Thermo-tolerance 3 (TT3), consisting of two genes, TT3.1 and TT3.2, that interact together to enhance rice thermotolerance and reduce grain-yield losses caused by heat stress. Upon heat stress, plasma membrane-localized E3 ligase TT3.1 translocates to the endosomes, on which TT3.1 ubiquitinates chloroplast precursor protein TT3.2 for vacuolar degradation, implying that TT3.1 might serve as a potential thermosensor. Lesser accumulated, mature TT3.2 proteins in chloroplasts are essential for protecting thylakoids from heat stress. Our findings not only reveal a TT3.1-TT3.2 genetic module at one locus that transduces heat signals from plasma membrane to chloroplasts but also provide the strategy for breeding highly thermotolerant crops.
Assuntos
Cloroplastos , Oryza , Proteínas de Plantas , Locos de Características Quantitativas , Termotolerância , Cloroplastos/genética , Cloroplastos/fisiologia , Genes de Plantas , Oryza/genética , Oryza/fisiologia , Melhoramento Vegetal/métodos , Proteínas de Plantas/genética , Termotolerância/genéticaRESUMO
The tocopherol biosynthetic pathway, encoded by VTE genes 1 through 6, is highly conserved in plants but most large effect quantitative trait loci for seed total tocopherols (totalT) lack VTE genes, indicating other activities are involved. A genome-wide association study of Arabidopsis seed tocopherols showed five of seven significant intervals lacked VTE genes, including the most significant, which mapped to an uncharacterized, seed-specific, envelope-localized, alpha/beta hydrolase with esterase activity, designated AtVTE7. Atvte7 null mutants decreased seed totalT 55% while a leaky allele of the maize ortholog, ZmVTE7, decreased kernel and leaf totalT 38% and 49%, respectively. Overexpressing AtVTE7 or ZmVTE7 partially or fully complemented the Atvte7 seed phenotype and increased leaf totalT by 3.6- and 6.9-fold, respectively. VTE7 has the characteristics of an esterase postulated to provide phytol from chlorophyll degradation for tocopherol synthesis, but bulk chlorophyll levels were unaffected in vte7 mutants and overexpressing lines. Instead, levels of specific chlorophyll biosynthetic intermediates containing partially reduced side chains were impacted and strongly correlated with totalT. These intermediates are generated by a membrane-associated biosynthetic complex containing protochlorophyllide reductase, chlorophyll synthase, geranylgeranyl reductase (GGR) and light harvesting-like 3 protein, all of which are required for both chlorophyll and tocopherol biosynthesis. We propose a model where VTE7 releases prenyl alcohols from chlorophyll biosynthetic intermediates, which are then converted to the corresponding diphosphates for tocopherol biosynthesis.
Assuntos
Arabidopsis , Hidrolases , Arabidopsis/genética , Arabidopsis/metabolismo , Cloroplastos/fisiologia , Estudo de Associação Genômica Ampla , Hidrolases/metabolismo , Fitol/metabolismo , Melhoramento Vegetal , Plantas/genética , Plantas/metabolismo , Tocoferóis/metabolismo , Vitamina E/metabolismoRESUMO
Guanosine pentaphosphate and tetraphosphate (together referred to as ppGpp) are hyperphosphorylated nucleotides found in bacteria and the chloroplasts of plants and algae. In plants and algae artificial ppGpp accumulation can inhibit chloroplast gene expression, and influence photosynthesis, nutrient remobilization, growth, and immunity. However, it is so far unknown whether ppGpp is required for abiotic stress acclimation in plants. Here, we demonstrate that ppGpp biosynthesis is necessary for acclimation to nitrogen starvation in Arabidopsis. We show that ppGpp is required for remodeling the photosynthetic electron transport chain to downregulate photosynthetic activity and for protection against oxidative stress. Furthermore, we demonstrate that ppGpp is required for coupling chloroplastic and nuclear gene expression during nitrogen starvation. Altogether, our work indicates that ppGpp is a pivotal regulator of chloroplast activity for stress acclimation in plants.
Assuntos
Arabidopsis/metabolismo , Guanosina Pentafosfato/metabolismo , Guanosina Tetrafosfato/metabolismo , Nitrogênio/metabolismo , Fotossíntese , Aclimatação , Arabidopsis/genética , Cloroplastos/fisiologia , Cianobactérias/citologia , Regulação da Expressão Gênica de Plantas , Células Vegetais , Estresse FisiológicoRESUMO
BACKGROUND: Mitogen-activated protein kinase (MAPK) cascades are conserved signaling modules in eukaryotic organisms and play essential roles in immunity and stress responses. However, the role of MAPKs in chloroplast development remains to be evidently established. RESULTS: In this study, a rice chlorosis seedling lethality 1 (csl1) mutant with a Zhonghua11 (ZH11, japonica) background was isolated. Seedlings of the mutant were characterized by chlorotic leaves and death after the trefoil stage, and chloroplasts were observed to contain accumulated starch granules. Molecular cloning revealed that OsCSL1 encoded a MAPK kinase kinase22 (MKKK22) targeted to the endoplasmic reticulum (ER), and functional complementation of OsCSL1 was found to restore the normal phenotype in csl1 plants. The CRISPR/Cas9 technology was used for targeted disruption of OsCSL1, and the OsCSL1-Cas9 lines obtained therein exhibited yellow seedlings which phenocopied the csl1 mutant. CSL1/MKKK22 was observed to establish direct interaction with MKK4, and altered expression of MKK1 and MKK4 was detected in the csl1 mutant. Additionally, disruption of OsCSL1 led to reduced expression of chloroplast-associated genes, including chlorophyll biosynthetic genes, plastid-encoded RNA polymerases, nuclear-encoded RNA polymerase, and nuclear-encoded chloroplast genes. CONCLUSIONS: The findings of this study revealed that OsCSL1 played roles in regulating the expression of multiple chloroplast synthesis-related genes, thereby affecting their functions, and leading to wide-ranging defects, including chlorotic seedlings and severely disrupted chloroplasts containing accumulated starch granules.
Assuntos
Cloroplastos/fisiologia , Proteínas Quinases Ativadas por Mitógeno/fisiologia , Biogênese de Organelas , Oryza/crescimento & desenvolvimento , Proteínas de Plantas/fisiologia , Clorofila/genética , Retículo Endoplasmático/metabolismo , Genes de Cloroplastos , Genes Letais , Proteínas Quinases Ativadas por Mitógeno/genética , Mutação , Oryza/genética , Oryza/ultraestrutura , Proteínas de Plantas/genéticaRESUMO
The recovery and maintenance of plant homeostasis under stressful environments are complex processes involving organelle crosstalk for a coordinated cellular response. Here, we revealed through nuclear and chloroplast subcellular proteomics, biochemical cell profiles and targeted transcriptomics how chloroplasts and nuclei developed their responses under increased temperatures in a long-lived species (Pinus radiata). Parallel to photosynthetic impairment and reactive oxygen species production in the chloroplast, a DNA damage response was triggered in the nucleus followed by an altered chromatin conformation. In addition, in the nuclei, we found several proteins, such as HEMERA or WHIRLY, which change their locations from the chloroplasts to the nuclei carrying the stress message. Additionally, our data showed a deep rearrangement of RNA metabolism in both organelles, revealing microRNAs and AGO1 as potential regulators of the acclimation mechanisms. Altogether, our study highlights the synchronisation among the different stages required for thermotolerance acquisition in P. radiata, pointing out the role of chromatin conformation and posttranscriptional gene regulation in overcoming heat stress and assuring plant survival for the following years.
Assuntos
Núcleo Celular/fisiologia , Cloroplastos/fisiologia , Resposta ao Choque Térmico , Pinus/fisiologia , Proteínas de Plantas/fisiologia , Proteoma/fisiologia , MicroRNAs/metabolismo , RNA de Plantas/metabolismo , Transdução de SinaisRESUMO
In bacteria and chloroplasts, the GTPase filamentous temperature-sensitive Z (FtsZ) is essential for division and polymerizes to form rings that mark the division site. Plants contain two FtsZ subfamilies (FtsZ1 and FtsZ2) with different assembly dynamics. FtsZ1 lacks the C-terminal domain of a typical FtsZ protein. Here, we show that the conserved short motif FtsZ1Carboxyl-terminus (Z1C) (consisting of the amino acids RRLFF) with weak membrane-binding activity is present at the C-terminus of FtsZ1 in angiosperms. For a polymer-forming protein such as FtsZ, this activity is strong enough for membrane tethering. Arabidopsis thaliana plants with mutated Z1C motifs contained heterogeneously sized chloroplasts and parallel FtsZ rings or long FtsZ filaments, suggesting that the Z1C motif plays an important role in regulating FtsZ ring dynamics. Our findings uncover a type of amphiphilic beta-strand motif with weak membrane-binding activity and point to the importance of this motif for the dynamic regulation of protein complex formation.
Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/fisiologia , Cloroplastos/fisiologia , Sequência de Aminoácidos , Arabidopsis/genética , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/metabolismoRESUMO
The chloroplast has a central position in oxygenic photosynthesis and primary metabolism. In addition to these functions, the chloroplast has recently emerged as a pivotal regulator of plant responses to abiotic and biotic stress conditions. Chloroplasts have their own independent genomes and gene-expression machinery and synthesize phytohormones and a diverse range of secondary metabolites, a significant portion of which contribute the plant response to adverse conditions. Furthermore, chloroplasts communicate with the nucleus through retrograde signaling, for instance, reactive oxygen signaling. All of the above facilitate the chloroplast's exquisite flexibility in responding to environmental stresses. In this review, we summarize recent findings on the involvement of chloroplasts in plant regulatory responses to various abiotic and biotic stresses including heat, chilling, salinity, drought, high light environmental stress conditions, and pathogen invasions. This review will enrich the better understanding of interactions between chloroplast and environmental stresses, and will lay the foundation for genetically enhancing plant-stress acclimatization.
Assuntos
Cloroplastos/fisiologia , Estresse Fisiológico/fisiologia , Aclimatação , Cloroplastos/metabolismo , Resposta ao Choque Frio/fisiologia , Secas , Expressão Gênica/genética , Regulação da Expressão Gênica de Plantas/genética , Resposta ao Choque Térmico/fisiologia , Fotossíntese , Reguladores de Crescimento de Plantas/metabolismo , Fenômenos Fisiológicos Vegetais/genética , Proteínas de Plantas/genética , Plantas/genética , Plantas/metabolismo , Salinidade , Transdução de SinaisRESUMO
The riboflavin derivatives flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD) are essential cofactors for enzymes in multiple cellular processes. Characterizing mutants with impaired riboflavin metabolism can help clarify the role of riboflavin in plant development. Here, we characterized a rice (Oryza sativa) white and lesion-mimic (wll1) mutant, which displays a lesion-mimic phenotype with white leaves, chlorophyll loss, chloroplast defects, excess reactive oxygen species (ROS) accumulation, decreased photosystem protein levels, changes in expression of chloroplast development and photosynthesis genes, and cell death. Map-based cloning and complementation test revealed that WLL1 encodes lumazine synthase, which participates in riboflavin biosynthesis. Indeed, the wll1 mutant showed riboflavin deficiency, and application of FAD rescued the wll1 phenotype. In addition, transcriptome analysis showed that cytokinin metabolism was significantly affected in wll1 mutant, which had increased cytokinin and δ-aminolevulinic acid contents. Furthermore, WLL1 and riboflavin synthase (RS) formed a complex, and the rs mutant had a similar phenotype to the wll1 mutant. Taken together, our findings revealed that WLL1 and RS play pivotal roles in riboflavin biosynthesis, which is necessary for ROS balance and chloroplast development in rice.
Assuntos
Cloroplastos/fisiologia , Complexos Multienzimáticos/metabolismo , Oryza/fisiologia , Proteínas de Plantas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Clorofila/genética , Clorofila/metabolismo , Citocininas/genética , Citocininas/metabolismo , Dano ao DNA , Evolução Molecular , Flavina-Adenina Dinucleotídeo/genética , Flavina-Adenina Dinucleotídeo/metabolismo , Regulação da Expressão Gênica de Plantas , Complexos Multienzimáticos/genética , Mutação , Fenótipo , Filogenia , Folhas de Planta/citologia , Folhas de Planta/genética , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Riboflavina/genética , Riboflavina/metabolismo , Técnicas do Sistema de Duplo-HíbridoRESUMO
Chloroplasts play an essential role in plant growth and development. Any factors affecting chloroplast development will lead to abnormal plant growth. Here, we characterized a new maize mutant, albino seedling mutant 81647 (as-81647), which exhibits an entirely albino phenotype in leaves and eventually died before the three-leaf stage. Transmission electron microscopy (TEM) demonstrated that the chloroplast thylakoid membrane was impaired and the granum lamellae significantly decreased in as-81647. Map-based cloning and transgenic analysis confirmed that PPR647 encodes a new chloroplast protein consisting of 11 pentratricopeptide repeat domains. Quantitative real-time PCR (qRT-PCR) assays and transcriptome analysis (RNA-seq) showed that the PPR647 mutation significantly disrupted the expression of PEP-dependent plastid genes. In addition, RNA splicing and RNA editing of multiple chloroplast genes showed severe defects in as-81647. These results indicated that PPR647 is crucial for RNA editing, RNA splicing of chloroplast genes, and plays an essential role in chloroplast development.
Assuntos
Cloroplastos/fisiologia , Proteínas de Plantas/genética , Edição de RNA , Splicing de RNA , RNA de Cloroplastos/metabolismo , Zea mays/genética , Zea mays/metabolismo , Cloroplastos/ultraestrutura , Regulação da Expressão Gênica de Plantas , Genes de Cloroplastos , Mutação , Fenótipo , Filogenia , Folhas de Planta/citologia , Proteínas de Plantas/metabolismo , Domínios Proteicos , Plântula/genética , Plântula/metabolismo , Tilacoides/fisiologia , Tilacoides/ultraestruturaRESUMO
BACKGROUND: Plant mitochondrial transcription termination factor (mTERF) family members play important roles in development and stress tolerance through regulation of organellar gene expression. However, their molecular functions have yet to be clearly defined. RESULTS: Here an mTERF gene V14 was identified by fine mapping using a conditional albino mutant v14 that displayed albinism only in the first two true leaves, which was confirmed by transgenic complementation tests. Subcellular localization and real-time PCR analyses indicated that V14 encodes a chloroplastic protein ubiquitously expressed in leaves while spiking in the second true leaf. Chloroplastic gene expression profiling in the pale leaves of v14 through real-time PCR and Northern blotting analyses showed abnormal accumulation of the unprocessed transcripts covering the rpoB-rpoC1 and/or rpoC1-rpoC2 intercistronic regions accompanied by reduced abundance of the mature rpoC1 and rpoC2 transcripts, which encode two core subunits of the plastid-encoded plastid RNA polymerase (PEP). Subsequent immunoblotting analyses confirmed the reduced accumulation of RpoC1 and RpoC2. A light-inducible photosynthetic gene psbD was also found down-regulated at both the mRNA and protein levels. Interestingly, such stage-specific aberrant posttranscriptional regulation and psbD expression can be reversed by high temperatures (30 ~ 35 °C), although V14 expression lacks thermo-sensitivity. Meanwhile, three V14 homologous genes were found heat-inducible with similar temporal expression patterns, implicating their possible functional redundancy to V14. CONCLUSIONS: These data revealed a critical role of V14 in chloroplast development, which impacts, in a stage-specific and thermo-sensitive way, the appropriate processing of rpoB-rpoC1-rpoC2 precursors and the expression of certain photosynthetic proteins. Our findings thus expand the knowledge of the molecular functions of rice mTERFs and suggest the contributions of plant mTERFs to photosynthesis establishment and temperature acclimation.