ABSTRACT
Macroautophagy, hereafter autophagy, plays a crucial role in the degradation of harmful or unwanted cellular components through a double-membrane autophagosome. Upon autophagosome fusion with the vacuole, the degraded materials are subsequently recycled to generate macromolecules, contributing to cellular homeostasis, metabolism, and stress tolerance in plants. A hallmark during autophagy is the formation of isolation membrane structure named as phagophore, which undergoes multiple steps to become as a complete double-membrane autophagosome. Methodologies have been developed in recent years to observe and quantify the autophagic process, which greatly advance knowledge of autophagosome biogenesis in plant cells. In this chapter, we will introduce two methods to dissect the autophagosome-related structures in the Arabidopsis plant cells, including the correlative light and electron microscopy, to map the ultrastructural feature of autophagosomal structures, and time-lapse imaging to monitor the temporal recruitment of autophagy machinery during autophagosome formation.
Subject(s)
Arabidopsis , Autophagosomes , Autophagy , Plant Cells , Autophagosomes/metabolism , Autophagosomes/ultrastructure , Arabidopsis/metabolism , Arabidopsis/ultrastructure , Autophagy/physiology , Plant Cells/metabolism , Plant Cells/ultrastructure , Time-Lapse Imaging/methods , Phagosomes/metabolism , Phagosomes/ultrastructure , Microscopy, Electron/methodsABSTRACT
Biomolecular condensates are triggered by multivalent interactions conferred by the intrinsically disordered regions and/or interacting domains within the constituents. While light microscopy has provided powerful tools to study the dynamics of intracellular condensates, electron microscopy (EM) gives more detailed insights into their ultrastructure and spatial connectivity with membrane system. In this chapter, we describe the methods for detecting the membraneless condensates in plant cells by high-pressure freezing -based EM coupled with immuno-gold labeling and correlative light electron microscopy techniques, which may benefit researchers in future studies.
Subject(s)
Plant Cells , Plant Cells/ultrastructure , Plant Cells/metabolism , Microscopy, Electron/methodsABSTRACT
Germline cells are critical for transmitting genetic information to subsequent generations in biological organisms. While their differentiation from somatic cells during embryonic development is well-documented in most animals, the regulatory mechanisms initiating plant germline cells are not well understood. To thoroughly investigate the complex morphological transformations of their ultrastructure over developmental time, nanoscale 3D reconstruction of entire plant tissues is necessary, achievable exclusively through electron microscopy imaging. This paper presents a full-process framework designed for reconstructing large-volume plant tissue from serial electron microscopy images. The framework ensures end-to-end direct output of reconstruction results, including topological networks and morphological analysis. The proposed 3D cell alignment, denoise, and instance segmentation pipeline (3DCADS) leverages deep learning to provide a cell instance segmentation workflow for electron microscopy image series, ensuring accurate and robust 3D cell reconstructions with high computational efficiency. The pipeline involves five stages: the registration of electron microscopy serial images; image enhancement and denoising; semantic segmentation using a Transformer-based neural network; instance segmentation through a supervoxel-based clustering algorithm; and an automated analysis and statistical assessment of the reconstruction results, with the mapping of topological connections. The 3DCADS model's precision was validated on a plant tissue ground-truth dataset, outperforming traditional baseline models and deep learning baselines in overall accuracy. The framework was applied to the reconstruction of early meiosis stages in the anthers of Arabidopsis thaliana, resulting in a topological connectivity network and analysis of morphological parameters and characteristics of cell distribution. The experiment underscores the 3DCADS model's potential for biological tissue identification and its significance in quantitative analysis of plant cell development, crucial for examining samples across different genetic phenotypes and mutations in plant development. Additionally, the paper discusses the regulatory mechanisms of Arabidopsis thaliana's germline cells and the development of stamen cells before meiosis, offering new insights into the transition from somatic to germline cell fate in plants.
Subject(s)
Imaging, Three-Dimensional , Imaging, Three-Dimensional/methods , Microscopy, Electron/methods , Arabidopsis/ultrastructure , Arabidopsis/growth & development , Arabidopsis/cytology , Algorithms , Plant Cells/ultrastructure , Image Processing, Computer-Assisted/methodsABSTRACT
The influence of cell morphology on the textural characteristic of freeze-dried apple, strawberry, and mango cubes was evaluated. Corresponding restructured cube samples without intact cell morphology were prepared as controls. Results indicated that the presence of cell morphology strengthened the shrinkage and collapse of samples during freeze-drying, especially in mangoes due to the high content of sugar. Intact cell morphology was found in natural fruit cubes after freeze-drying by scanning electron microscopy (SEM) observation, making them exhibit a more regular microporous structure, further resulting in higher hardness than the restructured cubes. However, the intact cell morphology negatively affected the crispness of freeze-dried cubes since it enhanced structural collapse. The freeze-dried samples without cell morphology would destroy the cellulose structure and form a continuous open-pore structure under the concentration effect of ice crystals during freezing, which accelerates the escape of water molecules, increases the drying rate, and avoid collapse. Sensory experiments found that restructured cubes without intact cell morphology exhibited greater comprehensive acceptance, suggesting the potential application of cell morphology disruption in the future freeze-drying industry.
Subject(s)
Fragaria , Freeze Drying , Malus , Mangifera , Plant Cells , Fragaria/chemistry , Fragaria/ultrastructure , Freeze Drying/methods , Fruit/chemistry , Fruit/ultrastructure , Malus/chemistry , Malus/ultrastructure , Mangifera/chemistry , Mangifera/ultrastructure , Plant Cells/chemistry , Plant Cells/ultrastructure , Microscopy, Electrochemical, ScanningABSTRACT
Fruit cells are living irregular three-dimensional (3D) transparent objects which makes them challenging to determine their real 3D size and shape through only two-dimensional (2D) images using the existing biological microscope. This study deals with a newly self-developed biological microscope including a microscope imaging system, a light source system, a stage and a support base for the 3D size characterization of fruit single cells. The main design concept is based on two optical path systems set up at the front (x-axis) and bottom (z-axis) directions of a transparent chamber containing single cells that allow the front view and bottom view of the single cell to be observed. Performance indicators such as mass, size, observation range, objective magnification, total magnification, focal range, focal accuracy, and resolution of the developed biological microscope were estimated. Finally, the 3D geometry size of single tomato cells was measured by the new biological microscope to demonstrate the relative ease at which accurate real 3D geometry information of single fruit cells could be obtained, which echoes its scientific value.
Subject(s)
Fruit , Plant Cells , Fruit/cytology , Plant Cells/ultrastructure , MicroscopyABSTRACT
The documentation of plant growth and development requires integrative and scalable approaches to investigate and spatiotemporally resolve various dynamic processes at different levels of plant body organization. The present update deals with vigorous developments in mesoscopy, microscopy and nanoscopy methods that have been translated to imaging of plant subcellular compartments, cells, tissues and organs over the past 3 years with the aim to report recent applications and reasonable expectations from current light-sheet fluorescence microscopy (LSFM) and super-resolution microscopy (SRM) modalities. Moreover, the shortcomings and limitations of existing LSFM and SRM are discussed, particularly for their ability to accommodate plant samples and regarding their documentation potential considering spherical aberrations or temporal restrictions prohibiting the dynamic recording of fast cellular processes at the three dimensions. For a more comprehensive description, advances in living or fixed sample preparation methods are also included, supported by an overview of developments in labeling strategies successfully applied in plants. These strategies are practically documented by current applications employing model plant Arabidopsis thaliana (L.) Heynh., but also robust crop species such as Medicago sativa L. and Hordeum vulgare L. Over the past few years, the trend towards designing of integrative microscopic modalities has become apparent and it is expected that in the near future LSFM and SRM will be bridged to achieve broader multiscale plant imaging with a single platform.
Subject(s)
Microscopy, Fluorescence/methods , Plant Cells/ultrastructure , Plant DevelopmentABSTRACT
A fundamental question in biology concerns how molecular and cellular processes become integrated during morphogenesis. In plants, characterization of 3D digital representations of organs at single-cell resolution represents a promising approach to addressing this problem. A major challenge is to provide organ-centric spatial context to cells of an organ. We developed several general rules for the annotation of cell position and embodied them in 3DCoordX, a user-interactive computer toolbox implemented in the open-source software MorphoGraphX. 3DCoordX enables rapid spatial annotation of cells even in highly curved biological shapes. Using 3DCoordX, we analyzed cellular growth patterns in organs of several species. For example, the data indicated the presence of a basal cell proliferation zone in the ovule primordium of Arabidopsis (Arabidopsis thaliana). Proof-of-concept analyses suggested a preferential increase in cell length associated with neck elongation in the archegonium of Marchantia (Marchantia polymorpha) and variations in cell volume linked to central morphogenetic features of a trap of the carnivorous plant Utricularia (Utricularia gibba). Our work demonstrates the broad applicability of the developed strategies as they provide organ-centric spatial context to cellular features in plant organs of diverse shape complexity.
Subject(s)
Imaging, Three-Dimensional , Plant Cells , Arabidopsis/ultrastructure , Lamiales/ultrastructure , Marchantia/ultrastructure , Morphogenesis , Plant Cells/ultrastructure , SoftwareABSTRACT
The acquisition of photosynthesis is a fundamental step in the evolution of eukaryotes. However, few phototrophic organisms are unambiguously recognized in the Precambrian record. The in situ detection of metabolic byproducts in individual microfossils is the key for the direct identification of their metabolisms. Here, we report a new integrative methodology using synchrotron-based X-ray fluorescence and absorption. We evidence bound nickel-geoporphyrins moieties in low-grade metamorphic rocks, preserved in situ within cells of a ~1 Gyr-old multicellular eukaryote, Arctacellularia tetragonala. We identify these moieties as chlorophyll derivatives, indicating that A. tetragonala was a phototrophic eukaryote, one of the first unambiguous algae. This new approach, applicable to overmature rocks, creates a strong new proxy to understand the evolution of phototrophy and diversification of early ecosystems.
Subject(s)
Chlorophyll/chemistry , Chlorophyta/ultrastructure , Coordination Complexes/chemistry , Fossils , Photosynthesis/physiology , Biological Evolution , Chlorophyll/history , Chlorophyta/anatomy & histology , Chlorophyta/classification , Chlorophyta/physiology , Democratic Republic of the Congo , Ecosystem , Eukaryotic Cells , Geologic Sediments/analysis , History, Ancient , Microscopy, Electron, Transmission , Nickel/chemistry , Phylogeny , Plant Cells/physiology , Plant Cells/ultrastructure , Tetrapyrroles/chemistry , X-Ray Absorption SpectroscopyABSTRACT
Capturing complete internal anatomies of plant organs and tissues within their relevant morphological context remains a key challenge in plant science. While plant growth and development are inherently multiscale, conventional light, fluorescence, and electron microscopy platforms are typically limited to imaging of plant microstructure from small flat samples that lack a direct spatial context to, and represent only a small portion of, the relevant plant macrostructures. We demonstrate technical advances with a lab-based X-ray microscope (XRM) that bridge the imaging gap by providing multiscale high-resolution three-dimensional (3D) volumes of intact plant samples from the cell to the whole plant level. Serial imaging of a single sample is shown to provide sub-micron 3D volumes co-registered with lower magnification scans for explicit contextual reference. High-quality 3D volume data from our enhanced methods facilitate sophisticated and effective computational segmentation. Advances in sample preparation make multimodal correlative imaging workflows possible, where a single resin-embedded plant sample is scanned via XRM to generate a 3D cell-level map, and then used to identify and zoom in on sub-cellular regions of interest for high-resolution scanning electron microscopy. In total, we present the methodologies for use of XRM in the multiscale and multimodal analysis of 3D plant features using numerous economically and scientifically important plant systems.
Subject(s)
Imaging, Three-Dimensional/statistics & numerical data , Microscopy, Electron, Scanning/instrumentation , Plant Cells/ultrastructure , Plants/ultrastructure , X-RaysABSTRACT
The plant nucleus plays an irreplaceable role in cellular control and regulation by auxin (indole-3-acetic acid, IAA) mainly because canonical auxin signaling takes place here. Auxin can enter the nucleus from either the endoplasmic reticulum or cytosol. Therefore, new information about the auxin metabolome (auxinome) in the nucleus can illuminate our understanding of subcellular auxin homeostasis. Different methods of nuclear isolation from various plant tissues have been described previously, but information about auxin metabolite levels in nuclei is still fragmented and insufficient. Herein, we tested several published nucleus isolation protocols based on differential centrifugation or flow cytometry. The optimized sorting protocol leading to promising yield, intactness, and purity was then combined with an ultra-sensitive mass spectrometry analysis. Using this approach, we can present the first complex report on the auxinome of isolated nuclei from cell cultures of Arabidopsis and tobacco. Moreover, our results show dynamic changes in auxin homeostasis at the intranuclear level after treatment of protoplasts with free IAA, or indole as a precursor of auxin biosynthesis. Finally, we can conclude that the methodological procedure combining flow cytometry and mass spectrometry offers new horizons for the study of auxin homeostasis at the subcellular level.
Subject(s)
Arabidopsis/metabolism , Cell Fractionation/methods , Cell Nucleus/metabolism , Indoleacetic Acids/metabolism , Indoles/metabolism , Nicotiana/metabolism , Plant Cells/metabolism , Arabidopsis/drug effects , Arabidopsis/ultrastructure , Cell Culture Techniques , Cell Fractionation/instrumentation , Cell Nucleus/drug effects , Cell Nucleus/ultrastructure , Centrifugation/methods , Flow Cytometry , Homeostasis/physiology , Indoles/pharmacology , Mass Spectrometry , Plant Cells/drug effects , Plant Cells/ultrastructure , Plant Growth Regulators/metabolism , Protoplasts/chemistry , Nicotiana/drug effects , Nicotiana/ultrastructureABSTRACT
Pterogyne nitens is commonly known in northeastern Brazil as a lesser-known fast-growing species in the Caatinga biome, which is a difficult place for tree development due to the low natural fertility soils and low availability of water. Due to the importance of expanding information about the anatomical wood properties of Caatinga native species, the aim of this work was to characterize the anatomical elements, to macroscopically describe the wood and make inferences about its possible end-uses. Maceration was performed which enabled measuring fiber dimensions, pore frequency and the following technological indexes: cell wall fraction, slenderness ratio, Runkel index and flexibility coefficient. Histological sections enabled describing the arrangements of the cellular elements in different observation sections and to determine the pore diameter. P. nitens wood has anatomical arrangements characterized by confluent axial parenchyma, being diffuse-porous with the presence of tylosis and heterogeneous/stratified rays (biseriate). The fibers were classified as very short (length 0.81 mm), not flexible and Runkel index 0.82. The pores were few in number with a frequency of 32.9 pores/mm2, distributed in a diffuse format and many were obstructed by tylosis. Based on the anatomical results and considering other technological studies, P. nitens wood is most suitable for charcoal production.
Subject(s)
Fabaceae/anatomy & histology , Trees/anatomy & histology , Wood/analysis , Brazil , Charcoal/chemistry , Ecosystem , Fabaceae/chemistry , Fabaceae/cytology , Humans , Plant Cells/ultrastructure , Trees/chemistry , Trees/cytology , Wood/cytologyABSTRACT
Modern light microscopy imaging techniques have substantially advanced our knowledge about the ultrastructure of plant cells and their organelles. Laser-scanning microscopy and digital light microscopy imaging techniques, in general-in addition to their high sensitivity, fast data acquisition, and great versatility of 2D-4D image analyses-also opened the technical possibilities to combine microscopy imaging with spectroscopic measurements. In this review, we focus our attention on differential polarization (DP) imaging techniques and on their applications on plant cell walls and chloroplasts, and show how these techniques provided unique and quantitative information on the anisotropic molecular organization of plant cell constituents: (i) We briefly describe how laser-scanning microscopes (LSMs) and the enhanced-resolution Re-scan Confocal Microscope (RCM of Confocal.nl Ltd. Amsterdam, Netherlands) can be equipped with DP attachments-making them capable of measuring different polarization spectroscopy parameters, parallel with the 'conventional' intensity imaging. (ii) We show examples of different faces of the strong anisotropic molecular organization of chloroplast thylakoid membranes. (iii) We illustrate the use of DP imaging of cell walls from a variety of wood samples and demonstrate the use of quantitative analysis. (iv) Finally, we outline the perspectives of further technical developments of micro-spectropolarimetry imaging and its use in plant cell studies.
Subject(s)
Plant Cells/ultrastructure , Anisotropy , Cell Wall/ultrastructure , Chloroplasts/ultrastructure , Microscopy, Confocal/methods , Microscopy, Polarization/methods , Thylakoids/ultrastructureABSTRACT
Liquid-liquid phase separation (LLPS) has emerged as a central paradigm for understanding how membraneless organelles compartmentalize diverse cellular activities in eukaryotes1-3. Here we identify a superfamily of plant guanylate-binding protein (GBP)-like GTPases (GBPLs) that assemble LLPS-driven condensates within the nucleus to protect against infection and autoimmunity. In Arabidopsis thaliana, two members of this family-GBPL1 and GBPL3-undergo phase-transition behaviour to control transcriptional responses as part of an allosteric switch that is triggered by exposure to biotic stress. GBPL1, a pseudo-GTPase, sequesters catalytically active GBPL3 under basal conditions but is displaced by GBPL3 LLPS when it enters the nucleus following immune cues to drive the formation of unique membraneless organelles termed GBPL defence-activated condensates (GDACs) that we visualized by in situ cryo-electron tomography. Within these mesoscale GDAC structures, native GBPL3 directly bound defence-gene promoters and recruited specific transcriptional coactivators of the Mediator complex and RNA polymerase II machinery to massively reprogram host gene expression for disease resistance. Together, our study identifies a GBPL circuit that reinforces the biological importance of phase-separated condensates, in this case, as indispensable players in plant defence.
Subject(s)
Arabidopsis/immunology , Cell Nucleus/chemistry , Cell Nucleus/metabolism , GTP-Binding Proteins/metabolism , Intrinsically Disordered Proteins/metabolism , Phase Transition , Plant Immunity , Arabidopsis/genetics , Arabidopsis/metabolism , Arabidopsis/ultrastructure , Cell Nucleus/genetics , Cell Nucleus/ultrastructure , Chromatin/genetics , Cryoelectron Microscopy , GTP-Binding Proteins/chemistry , GTP-Binding Proteins/ultrastructure , Gene Expression Regulation, Plant/genetics , Intrinsically Disordered Proteins/chemistry , Intrinsically Disordered Proteins/ultrastructure , Mediator Complex , Multigene Family/genetics , Organelles/chemistry , Organelles/immunology , Organelles/metabolism , Organelles/ultrastructure , Plant Cells/chemistry , Plant Cells/immunology , Plant Cells/metabolism , Plant Cells/ultrastructure , Plant Diseases/immunology , Plant Immunity/genetics , Promoter Regions, Genetic/genetics , RNA Polymerase II/metabolism , Transcription, GeneticABSTRACT
Plants have evolved complex nanofibril-based cell walls to meet diverse biological and physical constraints. How strength and extensibility emerge from the nanoscale-to-mesoscale organization of growing cell walls has long been unresolved. We sought to clarify the mechanical roles of cellulose and matrix polysaccharides by developing a coarse-grained model based on polymer physics that recapitulates aspects of assembly and tensile mechanics of epidermal cell walls. Simple noncovalent binding interactions in the model generate bundled cellulose networks resembling that of primary cell walls and possessing stress-dependent elasticity, stiffening, and plasticity beyond a yield threshold. Plasticity originates from fibril-fibril sliding in aligned cellulose networks. This physical model provides quantitative insight into fundamental questions of plant mechanobiology and reveals design principles of biomaterials that combine stiffness with yielding and extensibility.
Subject(s)
Cell Wall/physiology , Cell Wall/ultrastructure , Cellulose , Plant Cells/ultrastructure , Plant Epidermis/ultrastructure , Polysaccharides , Biomechanical Phenomena , Carbohydrate Conformation , Cellulose/chemistry , Elasticity , Models, Biological , Molecular Dynamics Simulation , Onions/ultrastructure , Stress, MechanicalABSTRACT
The endoplasmic reticulum (ER) is an organelle with remarkable plasticity, capable of rapidly changing its structure to accommodate different functions based on intra- and extracellular cues. One of the ER structures observed in plants is known as "organized smooth endoplasmic reticulum" (OSER), consisting of symmetrically stacked ER membrane arrays. In plants, these structures were first described in certain specialized tissues, e.g. the sieve elements of the phloem, and more recently in transgenic plants overexpressing ER membrane resident proteins. To date, much of the investigation of OSER focused on yeast and animal cells but research into plant OSER has started to grow. In this update, we give a succinct overview of research into the OSER phenomenon in plant cells with case studies highlighting both native and synthetic occurrences of OSER. We also assess the primary driving forces that trigger the formation of OSER, collating evidence from the literature to compare two competing theories for the origin of OSER: that OSER formation is initiated by oligomerizing protein accumulation in the ER membrane or that OSER is the result of ER membrane proliferation. This has long been a source of controversy in the field and here we suggest a way to integrate arguments from both sides into a single unifying theory. Finally, we discuss the potential biotechnological uses of OSER as a tool for the nascent plant synthetic biology field with possible applications as a synthetic microdomain for metabolic engineering and as an extensive membrane surface for synthetic chemistry or protein accumulation.
Subject(s)
Biosynthetic Pathways , Endoplasmic Reticulum, Smooth/physiology , Endoplasmic Reticulum, Smooth/ultrastructure , Intracellular Membranes/physiology , Intracellular Membranes/ultrastructure , Plant Cells/physiology , Plant Cells/ultrastructureABSTRACT
Organelles of the plant cell cooperate to synthesize and secrete a strong yet flexible polysaccharide-based extracellular matrix: the cell wall. Cell wall composition varies among plant species, across cell types within a plant, within different regions of a single cell wall, and in response to intrinsic or extrinsic signals. This diversity in cell wall makeup is underpinned by common cellular mechanisms for cell wall production. Cellulose synthase complexes function at the plasma membrane and deposit their product into the cell wall. Matrix polysaccharides are synthesized by a multitude of glycosyltransferases in hundreds of mobile Golgi stacks, and an extensive set of vesicle trafficking proteins govern secretion to the cell wall. In this review, we discuss the different subcellular locations at which cell wall synthesis occurs, review the molecular mechanisms that control cell wall biosynthesis, and examine how these are regulated in response to different perturbations to maintain cell wall homeostasis.
Subject(s)
Cell Wall/metabolism , Plant Cells/metabolism , Cell Membrane/enzymology , Cell Membrane/metabolism , Cell Wall/chemistry , Cell Wall/ultrastructure , Endocytosis , Endoplasmic Reticulum/metabolism , Glucosyltransferases/metabolism , Golgi Apparatus/metabolism , Homeostasis , Plant Cells/enzymology , Plant Cells/ultrastructure , Polysaccharides/biosynthesisABSTRACT
Soil salinization is a serious and growing problem around the world. Some plants, recognized as the recretohalophytes, can normally grow on saline-alkali soil without adverse effects by secreting excessive salt out of the body. The elucidation of the salt secretion process is of great significance for understanding the salt tolerance mechanism adopted by the recretohalophytes. Between the 1950s and the 1970s, three hypotheses, including the osmotic potential hypothesis, the transfer system similar to liquid flow in animals, and vesicle-mediated exocytosis, were proposed to explain the salt secretion process of plant salt glands. More recently, increasing evidence has indicated that vesicular transport plays vital roles in salt secretion of recretohalophytes. Here, we summarize recent findings, especially regarding the molecular evidence on the functional roles of vesicular trafficking in the salt secretion process of plant salt glands. A model of salt secretion in salt gland is also proposed.
Subject(s)
Salt-Tolerant Plants/anatomy & histology , Salt-Tolerant Plants/physiology , Salts/metabolism , Biological Transport , Gene Expression Regulation, Plant , Plant Cells/ultrastructure , Plant Proteins/genetics , Plant Proteins/metabolism , Salt-Tolerant Plants/cytologyABSTRACT
KEY MESSAGE: Macroscopic, ultrastructural, and molecular features-like a ball shape, the presence of starch granules, and the up-regulation of genes involved in carbohydrate metabolism and secondary metabolite biosynthesis-distinguish PT regions within a callus. The modification of the mass of pluripotent cells into de novo shoot bud regeneration is highly relevant to developmental biology and for agriculture and biotechnology. This study deals with protuberances (PT), structures that appear during the organogenic long-term culturing of callus (OC) in kiwifruit. These ball-shaped regions of callus might be considered the first morphological sign of the subsequent shoot bud development. Sections of PT show the regular arrangement of some cells, especially on the surface, in contrast to the regions of OC beyond the PT. The cells of OC possess chloroplasts; however, starch granules were observed only in PTs' plastids. Transcriptomic data revealed unique gene expression for each kind of sample: OC, PT, and PT with visible shoot buds (PT-SH). Higher expression of the gene involved in lipid (glycerol-3-phosphate acyltransferase 5 [GPAT5]), carbohydrate (granule-bound starch synthase 1 [GBSS1]), and secondary metabolite (beta-glucosidase 45 [BGL45]) pathways were detected in PT and could be proposed as the markers of these structures. The up-regulation of the regulatory associated protein of TOR (RAPTOR1) was found in PT-SH. The highest expression of the actinidain gene in leaves from two-year-old regenerated plants suggests that the synthesis of this protein takes place in fully developed organs. The findings indicate that PT and PT-SH are specific structures within OC but have more features in common with callus tissue than with organs.
Subject(s)
Actinidia/cytology , Actinidia/genetics , Gene Expression Regulation, Plant , Plant Proteins/genetics , Carbohydrate Metabolism/genetics , Gene Expression Profiling , Lipid Metabolism/genetics , Microscopy, Electron, Scanning , Plant Cells/ultrastructure , Plant Proteins/metabolism , Secondary Metabolism/genetics , Tissue Culture Techniques/methodsABSTRACT
This study aimed to clarify whether the light condition-dependent changes in the redox state and subcellular distribution of glutathione were similar in the dicotyledonous model plant Arabidopsis (wild-type, ascorbate- and glutathione-deficient mutants) and the monocotyledonous crop species wheat (Chinese Spring variety). With increasing light intensity, the amount of its reduced (GSH) and oxidized (GSSG) form and the GSSG/GSH ratio increased in the leaf extracts of both species including all genotypes, while far-red light increased these parameters only in wheat except for GSH in the GSH-deficient Arabidopsis mutant. Based on the expression changes of the glutathione metabolism-related genes, light intensity influences the size and redox state of the glutathione pool at the transcriptional level in wheat but not in Arabidopsis. In line with the results in leaf extracts, a similar inducing effect of both light intensity and far-red light was found on the total glutathione content at the subcellular level in wheat. In contrast to the leaf extracts, the inducing influence of light intensity on glutathione level was only found in the cell compartments of the GSH-deficient Arabidopsis mutant, and far-red light increased it in both mutants. The observed general and genotype-specific, light-dependent changes in the accumulation and subcellular distribution of glutathione participate in adjusting the redox-dependent metabolism to the actual environmental conditions.