Combining Clearing and Fluorescence Microscopy for Visualising Changes in Gene Expression and Physiological Responses to &lt;em&gt;Plasmodiophora brassicae&lt;/em&gt;

Singh, Deeksha; Blicharz, Sara; Stefanowicz, Karolina; Ragni, Laura; Michalak, K.; Bagniewska‐Zadworna, Agnieszka; Malinowski, Robert

doi:10.3791/64297-v

Cited by 2 publications

(1 citation statement)

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“…It is noteworthy that Nile Red is compatible with ClearSee clearing technique and, due to its convenient excitation and emission spectra, can be combined with other stains, such as CW and fluorescent proteins in the blue and green emission spectra (Table 1). 14 For instance, a combination of Nile Red and CW was used to visualize the Plasmodiophora brassicae –caused gall formation in A. thaliana roots 136 …”

Section: Suberin and Cutinmentioning

confidence: 99%

Imaging plant cell walls using fluorescent stains: The beauty is in the details

Piccinini,

Nirina Ramamonjy,

Ursache

2024

Journal of Microscopy

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Plants continuously face various environmental stressors throughout their lifetime. To be able to grow and adapt in different environments, they developed specialized tissues that allowed them to maintain a protected yet interconnected body. These tissues undergo specific primary and secondary cell wall modifications that are essential to ensure normal plant growth, adaptation and successful land colonization. The composition of cell walls can vary among different plant species, organs and tissues. The ability to remodel their cell walls is fundamental for plants to be able to cope with multiple biotic and abiotic stressors. A better understanding of the changes taking place in plant cell walls may help identify and develop new strategies as well as tools to enhance plants’ survival under environmental stresses or prevent pathogen attack. Since the invention of microscopy, numerous imaging techniques have been developed to determine the composition and dynamics of plant cell walls during normal growth and in response to environmental stimuli. In this review, we discuss the main advances in imaging plant cell walls, with a particular focus on fluorescent stains for different cell wall components and their compatibility with tissue clearing techniques.Lay Description: Plants are continuously subjected to various environmental stresses during their lifespan. They evolved specialized tissues that thrive in different environments, enabling them to maintain a protected yet interconnected body. Such tissues undergo distinct primary and secondary cell wall alterations essential to normal plant growth, their adaptability and successful land colonization. Cell wall composition may differ among various plant species, organs and even tissues. To deal with various biotic and abiotic stresses, plants must have the capacity to remodel their cell walls. Gaining insight into changes that take place in plant cell walls will help identify and create novel tools and strategies to improve plants’ ability to withstand environmental challenges. Multiple imaging techniques have been developed since the introduction of microscopy to analyse the composition and dynamics of plant cell walls during growth and in response to environmental changes. Advancements in plant tissue cleaning procedures and their compatibility with cell wall stains have significantly enhanced our ability to perform high‐resolution cell wall imaging. At the same time, several factors influence the effectiveness of cleaning and staining plant specimens, as well as the time necessary for the process, including the specimen's size, thickness, tissue complexity and the presence of autofluorescence. In this review, we will discuss the major advances in imaging plant cell walls, with a particular emphasis on fluorescent stains for diverse cell wall components and their compatibility with tissue clearing techniques. We hope that this review will assist readers in selecting the most appropriate stain or combination of stains to highlight specific cell wall components of interest.

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Section: Suberin and Cutinmentioning

confidence: 99%

Imaging plant cell walls using fluorescent stains: The beauty is in the details

Piccinini,

Nirina Ramamonjy,

Ursache

2024

Journal of Microscopy

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MYB68 orchestrates cork differentiation by regulating stem cell proliferation and suberin deposition

Molina,

Horvath,

Zhang

et al. 2024

Preprint

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Plants have developed specialized barriers to protect and isolate the inner tissues from the environment while maintaining homeostasis. Different barriers are present in various organs and at different growth stages. During secondary growth, the periderm acts as the protective tissue, covering roots, stems, and branches as they become thick. The periderm is a dynamic barrier comprising a stem cell niche known as the cork cambium, which bifacially divides to generate the phelloderm inward and the cork outward. Cork cells have a unique cell wall impregnated with suberin and lignin polymers, essential for the barrier function. Despite its importance, the differentiation process that forms new cork cells from the stem cell is largely unknown. In this work, we identify members of the MYB36-subclade transcription factors as key regulators of cork differentiation. On the one hand, this set of transcription factors promotes suberin deposition by inducing the expression of enzymes involved in all steps of suberin biosynthesis, including the recently discovered suberin-polymerizing enzymes GDS Lipases; on the other hand, it represses cork cambium proliferation. Furthermore, we demonstrated that suberin deposition in the cork is a robust process regulated by a complex network of transcription factors, including other MYB transcription factors that activate suberin deposition in the endodermis. However, only members of the MYB36 subclade can repress cell proliferation in different developmental contexts, highlighting general and specific functions for MYB transcription factors. These findings have broad applicability, as tissue-specific manipulation of MYB activity has the potential for improving traits of biotechnological interest, such as thicker periderms and more suberized cork layers, and for assessing how these traits affect plant performance in response to stresses.

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Combining Clearing and Fluorescence Microscopy for Visualising Changes in Gene Expression and Physiological Responses to <em>Plasmodiophora brassicae</em>

Cited by 2 publications

References 0 publications

Imaging plant cell walls using fluorescent stains: The beauty is in the details

Imaging plant cell walls using fluorescent stains: The beauty is in the details

MYB68 orchestrates cork differentiation by regulating stem cell proliferation and suberin deposition

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