Live-Imaging of the Arabidopsis Inflorescence Meristem

Heisler, Marcus G.; Ohno, Carolyn

doi:10.1007/978-1-4614-9408-9_25

Cited by 22 publications

(20 citation statements)

References 4 publications

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“…Imaging nuclei in small, young seedlings (roots or cotyledons, leaf fragments) that can fit on a microscopic-slide coated with vital medium offers an accessible set-up for experienced microscopists. For larger organs, semi-in vitro solutions have been established: embryos, shoot meristems, floral buds, anthers and ovules can be detached from the plant and cultured in microscopyoptimized chambers [146][147][148]149] .…”

Section: Dynamic Live Imagingmentioning

confidence: 99%

Probing the 3D architecture of the plant nucleus with microscopy approaches: challenges and solutions

Dumur

Duncan

Graumann

et al. 2019

Nucleus

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The eukaryotic cell nucleus is a central organelle whose architecture determines genome function at multiple levels. Deciphering nuclear organizing principles influencing cellular responses and identity is a timely challenge. Despite many similarities between plant and animal nuclei, plant nuclei present intriguing specificities. Complementary to molecular and biochemical approaches, 3D microscopy is indispensable for resolving nuclear architecture. However, novel solutions are required for capturing cell-specific, sub-nuclear and dynamic processes. We provide a pointer for utilising high-to-super-resolution microscopy and image processing to probe plant nuclear architecture in 3D at the best possible spatial and temporal resolution and at quantitative and cell-specific levels. High-end imaging and image-processing solutions allow the community now to transcend conventional practices and benefit from continuously improving approaches. These promise to deliver a comprehensive, 3D view of plant nuclear architecture and to capture spatial dynamics of the nuclear compartment in relation to cellular states and responses. Abbreviations: 3D and 4D: Three and Four dimensional; AI: Artificial Intelligence; ant: antipodal nuclei (ant); CLSM: Confocal Laser Scanning Microscopy; CTs: Chromosome Territories; DL: Deep Learning; DLIm: Dynamic Live Imaging; ecn: egg nucleus; FACS: Fluorescence-Activated Cell Sorting; FISH: Fluorescent In Situ Hybridization; FP: Fluorescent Proteins (GFP, RFP, CFP, YFP, mCherry); FRAP: Fluorescence Recovery After Photobleaching; GPU: Graphics Processing Unit; KEEs: KNOT Engaged Elements; INTACT: Isolation of Nuclei TAgged in specific Cell Types; LADs: Lamin-Associated Domains; ML: Machine Learning; NA: Numerical Aperture; NADs: Nucleolar Associated Domains; PALM: Photo-Activated Localization Microscopy; Pixel: Picture element; pn: polar nuclei; PSF: Point Spread Function; RHF: Relative Heterochromatin Fraction; SIM: Structured Illumination Microscopy; SLIm: Static Live Imaging; SMC: Spore Mother Cell; SNR: Signal to Noise Ratio; SRM: Super-Resolution Microscopy; STED: STimulated Emission Depletion; STORM: STochastic Optical Reconstruction Microscopy; syn: synergid nuclei; TADs: Topologically Associating Domains; Voxel: Volumetric pixel

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Section: Dynamic Live Imagingmentioning

confidence: 99%

Probing the 3D architecture of the plant nucleus with microscopy approaches: challenges and solutions

Dumur

Duncan

Graumann

et al. 2019

Nucleus

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“…Treatments were carried out on the inflorescence meristems of whole plants transplanted 528 from soil to boxes containing GM medium supplemented with vitamins. The older 529 flowers were removed as described (Heisler and Ohno, 2014). The plants were chosen 530 such that the stem of the meristem was a few millimeters above the rosette to prevent the 531 drug solution from dispersing into the surrounding medium.…”

Section: Chemical Treatments For Auxin Depletion In the Inflorescencementioning

confidence: 99%

Cell type boundaries organize plant development

Caggiano

Bhatia

et al. 2017

Preprint

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Abstract:In plants the dorsoventral boundary of leaves defines an axis of symmetry 22 through the centre of the organ separating the top (dorsal) and bottom (ventral) tissues. 23Although the positioning of this boundary is critical for leaf morphogenesis, how the 24 boundary is established and how it influences development remains unclear. Using live-25 imaging and perturbation experiments we show that leaf orientation, morphology and 26 position are pre-patterned by HD-ZIPIII and KAN gene expression in the shoot, leading 27 to a model in which dorsoventral genes coordinate to regulate plant development by 28 localizing auxin response between their expression domains. However we also find that 29 auxin levels feedback on dorsoventral patterning by spatially organizing HD-ZIPIII and 30

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“…Confocal microscopy was performed on a Nikon A1 Confocal with a CFI Apo LWD 25× water immersion objective (Nikon Instruments) as described in von Wangenheim et al (52) and Heisler et al (53).…”

Section: Methodsmentioning

confidence: 99%

A mechanistic framework for noncell autonomous stem cell induction in Arabidopsis

Daum

Medzihradszky

Suzaki

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

282

260

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Cell-cell communication is essential for multicellular development and, consequently, evolution has brought about an array of distinct mechanisms serving this purpose. Consistently, induction and maintenance of stem cell fate by noncell autonomous signals is a feature shared by many organisms and may depend on secreted factors, direct cell-cell contact, matrix interactions, or a combination of these mechanisms. Although many basic cellular processes are well conserved between animals and plants, cell-to-cell signaling is one function where substantial diversity has arisen between the two kingdoms of life. One of the most striking differences is the presence of cytoplasmic bridges, called plasmodesmata, which facilitate the exchange of molecules between neighboring plant cells and provide a unique route for cell-cell communication in the plant lineage. Here, we provide evidence that the stem cell inducing transcription factor WUSCHEL (WUS), expressed in the niche, moves to the stem cells via plasmodesmata in a highly regulated fashion and that this movement is required for WUS function and, thus, stem cell activity in Arabidopsis thaliana. We show that cell context-independent mobility is encoded in the WUS protein sequence and mediated by multiple domains. Finally, we demonstrate that parts of the protein that restrict movement are required for WUS homodimerization, suggesting that formation of WUS dimers might contribute to the regulation of apical stem cell activity.transcription factor mobility | cell-to-cell trafficking | shoot apical meristem | SAM

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Live-Imaging of the Arabidopsis Inflorescence Meristem

Cited by 22 publications

References 4 publications

Probing the 3D architecture of the plant nucleus with microscopy approaches: challenges and solutions

Probing the 3D architecture of the plant nucleus with microscopy approaches: challenges and solutions

Cell type boundaries organize plant development

A mechanistic framework for noncell autonomous stem cell induction in Arabidopsis

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