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Advances in Plant Cell Imaging
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Advances in Plant Cell Imaging

A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Plant Sciences".

Deadline for manuscript submissions: closed (20 January 2022) | Viewed by 22097

Special Issue Editor

Dr. George Komis

E-Mail Website
Guest Editor
Center of the Region Hana for Biotechnological and Agricultural Research, Palacky University, 78371 Olomouc, Czech Republic
Interests: cell division plane orientation; cell morphogenesis; confocal laser scanning microscopy; microtubule dynamics; mitogen activated protein kinases; mitotic spindle; phragmoplast; plant cytoskeleton; spinning disk microscopy; structured illumination microscopy
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

The visual documentation of the life events of plants requires imaging over a broad spatiotemporal resolution range. Recent advances in microscopy and their successful adaptation in plant biology allow the tracking of individual plasma membrane proteins at a millisecond time scale andthe documentation of thousands of cell fates during primary and lateral root development over periods of hours to days. This Special Issue, entitled “Advances in Plant Cell Imaging”, aims to become a compendium of subcellular imaging of plants and provide a wide range of imaging approaches and applications.

The content of the Special Issue aims to cover significant biological output by applying a certain microscopy method, a novel labeling approach, or a new computational post-acquisition image analysis tool; novel and contemporary approaches on 3D and 4D imaging of fixed or living, cleared, or native plant samples; technical reviews or protocol articles on specific methods of microscopy, such as superresolution or light-sheet and quantitative analytical methods, including but not limited to FRET, FRAP, FLIM, and FCS; single molecule imaging, tracking, and manipulation; development of biosensors for monitoring physiological processes (e.g., Ca2+ signaling, intracellular pH or membrane potential, production of reactive oxygen species, etc.); high throughput screening approaches linking imaging to chemical biology or microfluidics approaches to address plant-microbe interactions and physiological plant processes; and how microscopy-based data can be used to generate computational models or simulations of processes ranging from subcellular organelle dynamics to the dynamics of tissue growth and organ formation.

Contributions spanning the entire diversity of the plant kingdom are expected to include a wide range of model and non-model species, with challenges in sample preparation and image acquisition.

Yours sincerely,
Dr. George Komis
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. International Journal of Molecular Sciences is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. There is an Article Processing Charge (APC) for publication in this open access journal. For details about the APC please see here. Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • Biosensors
  • fluorescence correlation spectroscopy
  • fluorescence lifetime imaging
  • fluorescence recovery after photobleaching
  • fluorescence resonance energy transfer
  • fluorescent protein tags
  • light sheet microscopy
  • microfluidics
  • single molecule localization microscopy
  • single particle tracking
  • structured illumination microscopy
  • subcellular imaging
  • superresolution microscopy
  • tissue clearing
  • two-photon microscopy

Published Papers (7 papers)

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Research

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16 pages, 5104 KiB  
Article
Revising the Role of Cortical Cytoskeleton during Secretion: Actin and Myosin XI Function in Vesicle Tethering
by Weiwei Zhang and Christopher J. Staiger
Int. J. Mol. Sci. 2022, 23(1), 317; https://doi.org/10.3390/ijms23010317 - 28 Dec 2021
Cited by 4 | Viewed by 1882
Abstract
In plants, secretion of cell wall components and membrane proteins plays a fundamental role in growth and development as well as survival in diverse environments. Exocytosis, as the last step of the secretory trafficking pathway, is a highly ordered and precisely controlled process [...] Read more.
In plants, secretion of cell wall components and membrane proteins plays a fundamental role in growth and development as well as survival in diverse environments. Exocytosis, as the last step of the secretory trafficking pathway, is a highly ordered and precisely controlled process involving tethering, docking, and fusion of vesicles at the plasma membrane (PM) for cargo delivery. Although the exocytic process and machinery are well characterized in yeast and animal models, the molecular players and specific molecular events that underpin late stages of exocytosis in plant cells remain largely unknown. Here, by using the delivery of functional, fluorescent-tagged cellulose synthase (CESA) complexes (CSCs) to the PM as a model system for secretion, as well as single-particle tracking in living cells, we describe a quantitative approach for measuring the frequency of vesicle tethering events. Genetic and pharmacological inhibition of cytoskeletal function, reveal that the initial vesicle tethering step of exocytosis is dependent on actin and myosin XI. In contrast, treatments with the microtubule inhibitor, oryzalin, did not significantly affect vesicle tethering or fusion during CSC exocytosis but caused a minor increase in transient or aborted tethering events. With data from this new quantitative approach and improved spatiotemporal resolution of single particle events during secretion, we generate a revised model for the role of the cortical cytoskeleton in CSC trafficking. Full article
(This article belongs to the Special Issue Advances in Plant Cell Imaging)
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19 pages, 2640 KiB  
Article
Specific Mycoparasite-Fusarium Graminearum Molecular Signatures in Germinating Seeds Disabled Fusarium Head Blight Pathogen’s Infection
by Seon Hwa Kim, Rachid Lahlali, Chithra Karunakaran and Vladimir Vujanovic
Int. J. Mol. Sci. 2021, 22(5), 2461; https://doi.org/10.3390/ijms22052461 - 28 Feb 2021
Cited by 9 | Viewed by 2760
Abstract
Advances in Infrared (IR) spectroscopies have entered a new era of research with applications in phytobiome, plant microbiome and health. Fusarium graminearum 3-ADON is the most aggressive mycotoxigenic chemotype causing Fusarium head blight (FHB) in cereals; while Sphaerodes mycoparasitica is the specific Fusarium [...] Read more.
Advances in Infrared (IR) spectroscopies have entered a new era of research with applications in phytobiome, plant microbiome and health. Fusarium graminearum 3-ADON is the most aggressive mycotoxigenic chemotype causing Fusarium head blight (FHB) in cereals; while Sphaerodes mycoparasitica is the specific Fusarium mycoparasite with biotrophic lifestyle discovered in cereal seeds and roots. Fourier transform infrared (FTIR) spectroscopy analyses depicted shifts in the spectral peaks related to mycoparasitism mainly within the region of proteins, lipids, also indicating a link between carbohydrates and protein regions, involving potential phenolic compounds. Especially, S. mycoparasitica contributes to significant changes in lipid region 3050–2800 cm−1, while in the protein region, an increasing trend was observed for the peaks 1655–1638 cm−1 (amide I) and 1549–1548 cm−1 (amide II) with changes in indicative protein secondary structures. Besides, the peak extending on the region 1520–1500 cm−1 insinuates a presence of aromatic compounds in presence of mycoparasite on the F. graminearum root sample. Monitoring shift in improved seed germination, fungus-fungus interface through scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM), and FTIR molecular signatures combined with principal component analysis (PCA) proved useful tools to detect an early mycoparasitism as a vital asset of the preventive biocontrol strategy against plant pathogens. Full article
(This article belongs to the Special Issue Advances in Plant Cell Imaging)
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19 pages, 3661 KiB  
Article
Comparing Super-Resolution Microscopy Techniques to Analyze Chromosomes
by Ivona Kubalová, Alžběta Němečková, Klaus Weisshart, Eva Hřibová and Veit Schubert
Int. J. Mol. Sci. 2021, 22(4), 1903; https://doi.org/10.3390/ijms22041903 - 14 Feb 2021
Cited by 16 | Viewed by 4541
Abstract
The importance of fluorescence light microscopy for understanding cellular and sub-cellular structures and functions is undeniable. However, the resolution is limited by light diffraction (~200–250 nm laterally, ~500–700 nm axially). Meanwhile, super-resolution microscopy, such as structured illumination microscopy (SIM), is being applied more [...] Read more.
The importance of fluorescence light microscopy for understanding cellular and sub-cellular structures and functions is undeniable. However, the resolution is limited by light diffraction (~200–250 nm laterally, ~500–700 nm axially). Meanwhile, super-resolution microscopy, such as structured illumination microscopy (SIM), is being applied more and more to overcome this restriction. Instead, super-resolution by stimulated emission depletion (STED) microscopy achieving a resolution of ~50 nm laterally and ~130 nm axially has not yet frequently been applied in plant cell research due to the required specific sample preparation and stable dye staining. Single-molecule localization microscopy (SMLM) including photoactivated localization microscopy (PALM) has not yet been widely used, although this nanoscopic technique allows even the detection of single molecules. In this study, we compared protein imaging within metaphase chromosomes of barley via conventional wide-field and confocal microscopy, and the sub-diffraction methods SIM, STED, and SMLM. The chromosomes were labeled by DAPI (4′,6-diamidino-2-phenylindol), a DNA-specific dye, and with antibodies against topoisomerase IIα (Topo II), a protein important for correct chromatin condensation. Compared to the diffraction-limited methods, the combination of the three different super-resolution imaging techniques delivered tremendous additional insights into the plant chromosome architecture through the achieved increased resolution. Full article
(This article belongs to the Special Issue Advances in Plant Cell Imaging)
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12 pages, 12081 KiB  
Article
3D Dissection of Structural Membrane-Wall Contacts in Filamentous Moss Protonemata
by Dominik Harant and Ingeborg Lang
Int. J. Mol. Sci. 2021, 22(1), 158; https://doi.org/10.3390/ijms22010158 - 26 Dec 2020
Cited by 1 | Viewed by 2792
Abstract
In conventional light microscopy, the adjacent cell walls of filamentous moss protonemata are seen from its narrow side thereby obscuring the major area of cell–cell connection. Optical sectioning, segmentation and 3D reconstructions allow the tilting and rotation of intracellular structures thereby greatly improving [...] Read more.
In conventional light microscopy, the adjacent cell walls of filamentous moss protonemata are seen from its narrow side thereby obscuring the major area of cell–cell connection. Optical sectioning, segmentation and 3D reconstructions allow the tilting and rotation of intracellular structures thereby greatly improving our understanding of interaction between organelles, membranes and the cell wall. Often, the findings also allow for conclusions on the respective functions. The moss Physcomitrium (Physcomitrella) patens is a model organism for growth, development and morphogenesis. Its filamentous protonemata are ideal objects for microscopy. Here, we investigated the cell wall between two neighboring cells and the connection of membranes towards this wall after plasmolysis in 0.8 M mannitol. An m-green fluorescent protein (GFP)-HDEL cell line was used to visualize the endoplasmatic reticulum (ER), the plasma membrane (PM) was stained with FM4-64. Our studies clearly show the importance of cell–cell contacts in P. patens protonemata. In 86% of the investigated cell pairs, at least one of the protoplasts remained fully attached to the adjacent cell wall. By tilting of z-stacks, volume renderings and 3D reconstructions, we visualized the amount of attached/detached PM and ER components after plasmolysis and membrane piercings through the wall of cell neighbors. Full article
(This article belongs to the Special Issue Advances in Plant Cell Imaging)
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Review

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16 pages, 7963 KiB  
Review
Differential Polarization Imaging of Plant Cells. Mapping the Anisotropy of Cell Walls and Chloroplasts
by Jasna Simonović Radosavljević, Aleksandra Lj. Mitrović, Ksenija Radotić, László Zimányi, Győző Garab and Gábor Steinbach
Int. J. Mol. Sci. 2021, 22(14), 7661; https://doi.org/10.3390/ijms22147661 - 17 Jul 2021
Viewed by 2636
Abstract
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 [...] Read more.
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. Full article
(This article belongs to the Special Issue Advances in Plant Cell Imaging)
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12 pages, 610 KiB  
Review
Single-Molecule Imaging in Living Plant Cells: A Methodological Review
by Ai-Yu Guo, Ya-Mei Zhang, Liu Wang, Di Bai, Ya-Peng Xu and Wen-Qiang Wu
Int. J. Mol. Sci. 2021, 22(10), 5071; https://doi.org/10.3390/ijms22105071 - 11 May 2021
Cited by 6 | Viewed by 3191
Abstract
Single-molecule imaging is emerging as a revolutionary approach to studying fundamental questions in plants. However, compared with its use in animals, the application of single-molecule imaging in plants is still underexplored. Here, we review the applications, advantages, and challenges of single-molecule fluorescence imaging [...] Read more.
Single-molecule imaging is emerging as a revolutionary approach to studying fundamental questions in plants. However, compared with its use in animals, the application of single-molecule imaging in plants is still underexplored. Here, we review the applications, advantages, and challenges of single-molecule fluorescence imaging in plant systems from the perspective of methodology. Firstly, we provide a general overview of single-molecule imaging methods and their principles. Next, we summarize the unprecedented quantitative details that can be obtained using single-molecule techniques compared to bulk assays. Finally, we discuss the main problems encountered at this stage and provide possible solutions. Full article
(This article belongs to the Special Issue Advances in Plant Cell Imaging)
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19 pages, 1475 KiB  
Review
State-of-the-Art Technologies for Understanding Brassinosteroid Signaling Networks
by Haijiao Wang, Song Song, Huaqiang Cheng and Yan-Wen Tan
Int. J. Mol. Sci. 2020, 21(21), 8179; https://doi.org/10.3390/ijms21218179 - 31 Oct 2020
Viewed by 3039
Abstract
Brassinosteroids, the steroid hormones of plants, control physiological and developmental processes through its signaling pathway. The major brassinosteroid signaling network components, from the receptor to transcription factors, have been identified in the past two decades. The development of biotechnologies has driven the identification [...] Read more.
Brassinosteroids, the steroid hormones of plants, control physiological and developmental processes through its signaling pathway. The major brassinosteroid signaling network components, from the receptor to transcription factors, have been identified in the past two decades. The development of biotechnologies has driven the identification of novel brassinosteroid signaling components, even revealing several crosstalks between brassinosteroid and other plant signaling pathways. Herein, we would like to summarize the identification and improvement of several representative brassinosteroid signaling components through the development of new technologies, including brassinosteroid-insensitive 1 (BRI1), BRI1-associated kinase 1 (BAK1), BR-insensitive 2 (BIN2), BRI1 kinase inhibitor 1 (BKI1), BRI1-suppressor 1 (BSU1), BR signaling kinases (BSKs), BRI1 ethyl methanesulfonate suppressor 1 (BES1), and brassinazole resistant 1 (BZR1). Furthermore, improvement of BR signaling knowledge, such as the function of BKI1, BES1 and its homologous through clustered regularly interspaced short palindromic repeats (CRISPR), the regulation of BIN2 through single-molecule methods, and the new in vivo interactors of BIN2 identified by proximity labeling are described. Among these technologies, recent advanced methods proximity labeling and single-molecule methods will be reviewed in detail to provide insights to brassinosteroid and other phytohormone signaling pathway studies. Full article
(This article belongs to the Special Issue Advances in Plant Cell Imaging)
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