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Microscopy Techniques in Plant Studies
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Microscopy Techniques in Plant Studies

A special issue of Plants (ISSN 2223-7747).

Deadline for manuscript submissions: 20 June 2024 | Viewed by 8152

Special Issue Editors

Dr. Emilio de Castro Miguel

E-Mail Website
Guest Editor
Department of Metallurgical and Materials Engineering, Federal University of Ceará, Fortaleza, Brazil
Interests: microscopy; plant ultrastructure; cell biology; nanomaterials
Dr. Maura Da Cunha

E-Mail Website
Guest Editor
Laboratório de Biologia Celular e Tecidual, Universidade Estadual do Norte Fluminense, Campos dos Goitacazes, Brazil
Interests: plant science; microscopy
Dr. Thaiz Batista Azevedo Rangel Miguel

E-Mail Website
Guest Editor
Department Food Engineering, Federal University of Ceará, 60020-181 Fortaleza, Brazil
Interests: microscopy; plant cell biology; non-thermal food processing

Special Issue Information

Dear Colleagues,

Plant biology studies include several approaches that range from gene expression experiments to the evaluation of plant growth in contact with nanostructured materials. Undoubtedly, all studies contribute to the accumulation of knowledge in plant physiology, anatomy and ultrastructure. While many techniques can be used to advance research, microscopy techniques can be applied to solve problems in many fields of plant biology and are extremely versatile. In this sense, microscopy techniques are capable of being applied in all fields of plant biology.

Recently, the investigation of plants using microscopy has surpassed traditional techniques such as light and scanning electron microscopy, and has even focused on techniques that have not been applied before, such as Raman microscopy, SIMS microscopy, focused ion beam, and others.

With the aim of strengthening the scientific basis for plant science, we welcome articles that enhance our understanding of all aspects of plant biology that include microscopic investigation. Review articles are also welcome.

Dr. Emilio de Castro Miguel
Dr. Maura Da Cunha
Dr. Thaiz Batista Azevedo Rangel Miguel
Guest Editors

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. Plants is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). 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

  • plant microscopy
  • plant cell
  • ultrastructure cell wall
  • plant anatomy
  • nanotechnology

Published Papers (7 papers)

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Research

16 pages, 4499 KiB  
Article
Unveiling the Anatomical and Functional Attributes of Stipular Colleters in Palicourea tetraphylla Cham. & Schltdl. and Palicourea rudgeoides (Müll. Arg.) Standl. (Rubiaceae)
by Laís de Almeida Bezerra, Emilio Castro Miguel, Camilla Ribeiro Alexandrino, Thaiz Batista de Azevedo Rangel Miguel, Valdirene Moreira Gomes and Maura Da Cunha
Plants 2024, 13(9), 1206; https://doi.org/10.3390/plants13091206 - 26 Apr 2024
Viewed by 155
Abstract
The characterization of colleters in Rubiaceae is crucial for understanding their role in plant function. Analyzing colleters in Palicourea tetraphylla and Palicourea rudgeoides aims to deepen the understanding of these structures morphoanatomical and functional characteristics. The study reveals colleters with palisade epidermis and [...] Read more.
The characterization of colleters in Rubiaceae is crucial for understanding their role in plant function. Analyzing colleters in Palicourea tetraphylla and Palicourea rudgeoides aims to deepen the understanding of these structures morphoanatomical and functional characteristics. The study reveals colleters with palisade epidermis and a parenchymatic central axis, classified as standard type, featuring vascularization and crystals. Colleter secretion, abundant in acidic mucopolysaccharides, proteins, and phenolic compounds, protects against desiccation. The ontogenesis, development, and senescence of the colleters are quite rapid and fulfill their role well in biotic and abiotic protection because these structures are present at different stages of development in the same stipule. Pronounced protrusions on the colleters surface, coupled with the accumulation of secretion in the intercellular and subcuticular spaces, suggest that the secretory process occurs through the wall, driven by pressure resulting from the accumulation of secretion. The microorganisms in the colleters’ secretion, especially in microbiota-rich environments such as the Atlantic Forest, provide valuable information about plant–microorganism interactions, such as resistance to other pathogens and organisms and ecological balance. This enhanced understanding of colleters contributes to the role of these structures in the plant and enriches knowledge about biological interactions within specific ecosystems and the family taxonomy. Full article
(This article belongs to the Special Issue Microscopy Techniques in Plant Studies)
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21 pages, 3410 KiB  
Article
Freeze Substitution Accelerated via Agitation: New Prospects for Ultrastructural Studies of Lichen Symbionts and Their Extracellular Matrix
by Siegfried Reipert, Daniela Gruber, Norbert Cyran, Brigitte Schmidt, Rosa de la Torre Noetzel, Leopoldo G. Sancho, Michal Goga, Martin Bačkor and Katy Schmidt
Plants 2023, 12(23), 4039; https://doi.org/10.3390/plants12234039 - 30 Nov 2023
Viewed by 900
Abstract
(1) Background: Lichens, as an important part of the terrestrial ecosystem, attract the attention of various research disciplines. To elucidate their ultrastructure, transmission electron microscopy of resin-embedded samples is indispensable. Since most observations of lichen samples are generated via chemical fixation and processing [...] Read more.
(1) Background: Lichens, as an important part of the terrestrial ecosystem, attract the attention of various research disciplines. To elucidate their ultrastructure, transmission electron microscopy of resin-embedded samples is indispensable. Since most observations of lichen samples are generated via chemical fixation and processing at room temperature, they lack the rapid immobilization of live processes and are prone to preparation artefacts. To improve their preservation, cryoprocessing was tested in the past, but never widely implemented, not least because of an extremely lengthy protocol. (2) Methods: Here, we introduce an accelerated automated freeze substitution protocol with continuous agitation. Using the example of three lichen species, we demonstrate the preservation of the native state of algal photobionts and mycobionts in association with their extracellular matrix. (3) Results: We bring to attention the extent and the structural variability of the hyphae, the extracellular matrix and numerous crystallized metabolites. Our findings will encourage studies on transformation processes related to the compartmentation of lichen thalli. They include cryopreserved aspects of algal photobionts and observations of putative physiological relevance, such as the arrangement of numerous mitochondria within chloroplast pockets. (4) Conclusions: In summary, we present accelerated freeze substitution as a very useful tool for systematic studies of lichen ultrastructures. Full article
(This article belongs to the Special Issue Microscopy Techniques in Plant Studies)
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18 pages, 5366 KiB  
Article
Vessel Anatomical Features of ‘Picual’ and ‘Frantoio’, Two Olive Cultivars Different in Resistance against Verticillium Wilt of Olive
by Antonio Santos-Rufo, Martín Molina-Molina, Esteban Alcántara-Vara, Carlos Weiland-Ardáiz and Fco. Javier López-Escudero
Plants 2023, 12(16), 2910; https://doi.org/10.3390/plants12162910 - 10 Aug 2023
Cited by 2 | Viewed by 870
Abstract
The olive tree (Olea europaea), a non-tropical woody crop that occupies the largest area in the world, is severely affected by the fungus Verticillium dahliae worldwide. In this regard, there is currently detailed information on the level of resistance to this [...] Read more.
The olive tree (Olea europaea), a non-tropical woody crop that occupies the largest area in the world, is severely affected by the fungus Verticillium dahliae worldwide. In this regard, there is currently detailed information on the level of resistance to this pathogen in the main olive varieties. However, there is little information on quantitative aspects of its anatomy and on the existence of anatomical differences between varieties that could be related to the differential resistance response observed. In the present work, a quantitative study of the xylem of ‘Picual’, susceptible, and ‘Frantoio’, resistant, to V. dahliae is carried out. This study also provides quantitative data on the xylem in different areas of the plant, an aspect on which there is not much information for the olive tree. Among the parameters evaluated, it is probably the greater conductive capacity in the xylem tissue that ‘Frantoio’ presents, mainly due to the greater density of its vessels, which has a more relevant role in the resistance and natural recovery that this cultivar manifests to the disease. In any case, these constitutive anatomical differences, and those others that can be induced in plants during infections, should be investigated in future work that includes inoculation with the pathogen. Full article
(This article belongs to the Special Issue Microscopy Techniques in Plant Studies)
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14 pages, 9667 KiB  
Article
Macroscopical, Microscopical and Histochemical Analysis of Eryngium karatavicum Iljin Growing on the Territory of South Kazakhstan
by Meruyert Amantayeva, Kaldanay Kozhanova, Gulnara Kadyrbayeva, Aigul Medeshova, Yerbolat Tulebayev, Moldir Zhandabayeva, Gulnur Yeleken, Zoya Allambergenova and Szilvia Czigle
Plants 2023, 12(14), 2714; https://doi.org/10.3390/plants12142714 - 21 Jul 2023
Cited by 2 | Viewed by 1322
Abstract
Carrying out macroscopical and microscopical analyses of plants allows determining the species and identifying diagnostic signs of the plant that distinguish the studied object from other related species. Endemic plant species are a specific component of the flora, whose representatives grow in a [...] Read more.
Carrying out macroscopical and microscopical analyses of plants allows determining the species and identifying diagnostic signs of the plant that distinguish the studied object from other related species. Endemic plant species are a specific component of the flora, whose representatives grow in a relatively limited area, represented by a small geographical area. Their diagnostic morphological and anatomical data are insufficiently studied. Such endemic unexplored plant species include Eryngium karatavicum Iljin, which grows in the territory of South Kazakhstan. This article presents the results of macroscopical, microscopical and histochemical analyses of leaves, flowers and stems of Eryngium karatavicum. The results of morphological analysis of Eryngium karatavicum showed that the plant has distinctive features of macroscopical, microscopical and histochemical signs on the upper and lower sides of the leaf, stem, inflorescence, leaves of the wrapper and flower. These results can be used to confirm the authenticity, identification, and standardization of aerial parts of the endemic plant Eryngium karatavicum Iljin. Full article
(This article belongs to the Special Issue Microscopy Techniques in Plant Studies)
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21 pages, 11145 KiB  
Article
Reproductive Capacity and Scanning Electron Microscopy (SEM) Analyses of the Micromorphological Surfaces of Three Endemic Satureja Species from Bulgaria
by Ivanka Semerdjieva, Elina Yankova-Tsvetkova, Valtcho D. Zheljazkov, Lyubka H. Koleva-Valkova and Rozalia Nikolova
Plants 2023, 12(13), 2436; https://doi.org/10.3390/plants12132436 - 24 Jun 2023
Cited by 1 | Viewed by 937
Abstract
Satureja pilosa Velen., S. coerulea Janka and S. kitaibelii Wierzb. ex Heuff. are Balkan endemic species, and they are distributed in restricted territories, primarily found in dry grasslands, rocky slopes, and stony habitats. This study presents the results of the first embryological and [...] Read more.
Satureja pilosa Velen., S. coerulea Janka and S. kitaibelii Wierzb. ex Heuff. are Balkan endemic species, and they are distributed in restricted territories, primarily found in dry grasslands, rocky slopes, and stony habitats. This study presents the results of the first embryological and micromorphological analyses of three Satureja species (S. pilosa, S. kitaibelii, and S. coerulea) from the Bulgarian flora. The aim of this study was to establish the features of the male and female reproductive sphere, as well as surface characteristics of leaves, stem, and calyx in order to understand the mode of reproduction, character, size and state of species populations and delimitation. For the embryological study, flowers and flower buds in different developmental stages were collected from plants of natural populations and treated with the classic paraffin method. Reproductive capacity was assessed using the following approaches: (1) acetocarmine test for pollen viability, (2) tetrazolium test (TTZ) for seed viability, and (3) germination test. The surfaces characteristics of leaves, stem, and calyx were analyzed by scanning electron microscopy (SEM). As a result, the study revealed the structures of the male (M) and female (F) generative spheres as well as the processes of gamete development, pollination, and endosperm and embryo formation. It was found that the three Satureja species exhibited a high pollen viability but low seed viability and germination. The SEM analysis showed both common and distinct micromorphology features regarding epidermis, calyx and stem surfaces among the three Satureja species. Notably, the S. coerulea surfaces (leaves, calyx, stem) were clearly distinguishable from the other two Satureja species. Regarding the nutlet surfaces, it was observed that the exocarp surfaces of S. coerulea and S. kitaibelii have a reticulate convex type surface and comprise two types of cells: (1) highly bulging, bubble-like cells; and (2) flat cells with numerous striations. On the other hand, the exocarp of S. pilosa displayed rectangular or polygonal shapes without bubble-like cells, and it had a tabular to slightly convex type surface. Additionally, nutlets (seeds) of both S. coerulea and S. pilosa exhibited distinct papilla formations resembling non-glandular trichomes seen on the ends of the nutlets for S. coerulea and over the entire surface for S. pilosa. The characteristics of the embryological structures and processes, along with the absence of apomixis, characterized the three studied Satureja species as sexually reproducing. The established balanced processes and stable structures contribute to their high reproductive potential and population stability. However, these traits may also decrease their adaptability to environmental changes. Full article
(This article belongs to the Special Issue Microscopy Techniques in Plant Studies)
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12 pages, 2277 KiB  
Article
Confocal Microscopy Investigations of Biopolymeric PLGA Nanoparticle Uptake in Arabidopsis thaliana L. Cultured Cells and Plantlet Roots
by Giulia De Angelis, Camilla Badiali, Laura Chronopoulou, Cleofe Palocci and Gabriella Pasqua
Plants 2023, 12(13), 2397; https://doi.org/10.3390/plants12132397 - 21 Jun 2023
Cited by 2 | Viewed by 1320
Abstract
To date, most endocytosis studies in plant cells have focused on clathrin-dependent endocytosis, while limited evidence is available on clathrin-independent pathways. Since dynamin a is a key protein both in clathrin-mediated endocytosis and in clathrin-independent endocytic processes, this study investigated its role in [...] Read more.
To date, most endocytosis studies in plant cells have focused on clathrin-dependent endocytosis, while limited evidence is available on clathrin-independent pathways. Since dynamin a is a key protein both in clathrin-mediated endocytosis and in clathrin-independent endocytic processes, this study investigated its role in the uptake of poly-(lactic-co-glycolic) acid (PLGA) nanoparticles (NPs). The experiments were performed on cultured cells and roots of Arabidopsis thaliana. Dynasore was used to inhibit the activity of dynamin-like proteins to investigate whether PLGA NPs enter plant cells through a dynamin-like-dependent or dynamin-like-independent endocytic pathway. Observations were performed by confocal microscopy using a fluorescent probe, coumarin 6, loaded in PLGA NPs. The results showed that both cells and roots of A. thaliana rapidly take up PLGA NPs. Dynasore was administered at different concentrations and exposure times in order to identify the effective ones for inhibitory activity. Treatments with dynasore did not prevent the NPs uptake, as revealed by the presence of fluorescence emission detected in the cytoplasm. At the highest concentration and the longest exposure time to dynasore, the fluorescence of NPs was not visible due to cell death. Thus, the results suggest that, because the NPs’ uptake is unaffected by dynasore exposure, NPs can enter cells and roots by following a dynamin-like-independent endocytic pathway. Full article
(This article belongs to the Special Issue Microscopy Techniques in Plant Studies)
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16 pages, 12285 KiB  
Article
Microscopical Analysis of Autofluorescence as a Complementary and Useful Method to Assess Differences in Anatomy and Structural Distribution Underlying Evolutive Variation in Loss of Seed Dispersal in Common Bean
by Ana M. Santos, Ana M. González, Juan De Dios Alche and Marta Santalla
Plants 2023, 12(11), 2212; https://doi.org/10.3390/plants12112212 - 03 Jun 2023
Cited by 1 | Viewed by 1487
Abstract
The common bean has received attention as a model plant for legume studies, but little information is available about the morphology of its pods and the relation of this morphology to the loss of seed dispersal and/or the pod string, which are key [...] Read more.
The common bean has received attention as a model plant for legume studies, but little information is available about the morphology of its pods and the relation of this morphology to the loss of seed dispersal and/or the pod string, which are key agronomic traits of legume domestication. Dehiscence is related to the pod morphology and anatomy of pod tissues because of the weakening of the dorsal and ventral dehiscence zones and the tensions of the pod walls. These tensions are produced by the differential mechanical properties of lignified and non-lignified tissues and changes in turgor associated with fruit maturation. In this research, we histologically studied the dehiscence zone of the ventral and dorsal sutures of the pod in two contrasting genotypes for the dehiscence and string, by comparing different histochemical methods with autofluorescence. We found that the secondary cell wall modifications of the ventral suture of the pod were clearly different between the dehiscence-susceptible and stringy PHA1037 and the dehiscence-resistant and stringless PHA0595 genotypes. The susceptible genotype had cells of bundle caps arranged in a more easily breakable bowtie knot shape. The resistant genotype had a larger vascular bundle area and larger fibre cap cells (FCCs), and due to their thickness, the external valve margin cells were significantly stronger than those from PHA1037. Our findings suggest that the FCC area, and the cell arrangement in the bundle cap, might be partial structures involved in the pod dehiscence of the common bean. The autofluorescence pattern at the ventral suture allowed us to quickly identify the dehiscent phenotype and gain a better understanding of cell wall tissue modifications that took place along the bean’s evolution, which had an impact on crop improvement. We report a simple autofluorescence protocol to reliably identify secondary cell wall organization and its relationship to the dehiscence and string in the common bean. Full article
(This article belongs to the Special Issue Microscopy Techniques in Plant Studies)
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