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Preparation and Characterization of Internally Modified DNA Templates for Chemical Transcription Roadblocking

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A Novel Method to Map Small RNAs with High Resolution

Authors: Kun Huang, Feray Demirci, Blake C. Meyers and Jeffrey L. Caplan, date: 08/20/2021, view: 536, Q&A: 0

[Abstract]

Analyzing cellular structures and the relative location of molecules is essential for addressing biological questions. Super-resolution microscopy techniques that bypass the light diffraction limit have become increasingly popular to study cellular molecule dynamics in situ. However, the application of super-resolution imaging techniques to detect

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A Pulse–chase EdU Method for Detection of Cell Division Orientation in Arabidopsis and Juncus prismatocarpus Leaf Primordia

Authors: Xiaofeng Yin and Hirokazu Tsukaya, date: 01/05/2021, view: 1264, Q&A: 0

[Abstract] In plants, the morphological diversity of leaves is largely determined by cell division, especially cell division orientation. Whereas cell division itself is easily monitored, the detection and quantification of cell division orientation are difficult. The few existing methods for detection and quantification of cell division orientation are ...

Multitarget Immunohistochemistry for Confocal and Super-resolution Imaging of Plant Cell Wall Polysaccharides

Authors: Kalina T. Haas, Methieu Rivière, Raymond Wightman and Alexis Peaucelle, date: 10/05/2020, view: 2752, Q&A: 0

[Abstract] The plant cell wall (PCW) is a pecto-cellulosic extracellular matrix that envelopes the plant cell. By integrating extra-and intra-cellular cues, PCW mediates a plethora of essential physiological functions. Notably, it permits controlled and oriented tissue growth by tuning its local mechano-chemical properties. To refine our knowledge of these ...

Live Cell Imaging of Male Meiosis in Arabidopsis by a Landmark-based System

Authors: Maria Ada Prusicki, Emma Mathilde Keizer, Rik Peter van Rosmalen, Christian Fleck and Arp Schnittger, date: 05/05/2020, view: 3284, Q&A: 0

[Abstract] Live cell imaging has tremendously promoted our understanding of cellular and subcellular processes such as cell division. Here, we present a step-by-step protocol for a robust and easy-to-use live cell imaging approach to study male meiosis in the plant Arabidopsis thaliana as recently established. Our method relies on the concomitant ...

Tandem Tag Assay Optimized for Semi-automated in vivo Autophagic Activity Measurement in Arabidopsis thaliana roots

Authors: Adrian N. Dauphinee, Jonas A. Ohlsson and Elena A. Minina, date: 03/05/2020, view: 2752, Q&A: 0

[Abstract] Autophagy is the main catabolic process in eukaryotes and plays a key role in cell homeostasis. In vivo measurement of autophagic activity (flux) is a powerful tool for investigating the role of the pathway in organism development and stress responses. Here we describe a significant optimization of the tandem tag assay for detection of ...

Increasing the Membrane Permeability of a Fern with DMSO

Author: Marcelo Garcés, date: 06/20/2018, view: 3339, Q&A: 0

[Abstract] Cell membrane prevents the entrance of extra molecules (e.g., transcription and translation inhibitors) into the cell. For studying the physiological effects of transcription and translation inhibitors on Hymenophyllum caudiculatum fronds, we incubate fronds with 0.1% DMSO to test if this increases cell membrane permeability ...

Slow and Fast Desiccation of Single-cell Thick Fronds of Filmy Ferns

Author: Marcelo Garcés, date: 06/05/2018, view: 3391, Q&A: 0

[Abstract] Filmy ferns can desiccate and recover after rehydration to resume photosynthesis. Slow and fast desiccation rates were compared in filmy fern fronds to determine whether structural or physiological differences may occur between desiccation rates. Slow desiccation is considered to be more similar to natural conditions experienced by plants that ...

Fluorescein Transport Assay to Assess Bulk Flow of Molecules Through the Hypocotyl in Arabidopsis thaliana

Authors: Salva Duran-Nebreda and George W. Bassel, date: 04/05/2018, view: 4897, Q&A: 0

[Abstract] The bulk transport of molecules through plant tissues underpins growth and development. The stem acts as a conduit between the upper and low domains of the plant, facilitating transport of solutes and water from the roots to the shoot system, and sugar plus other elaborated metabolites towards the non-photosynthetic organs. In order to perform ...

Electron Tomography to Study the Three-dimensional Structure of Plasmodesmata in Plant Tissues–from High Pressure Freezing Preparation to Ultrathin Section Collection

Authors: William J. Nicolas, Emmanuelle Bayer and Lysiane Brocard, date: 01/05/2018, view: 6976, Q&A: 0

[Abstract] Plasmodesmata (PD) are nanometric (~20 nm wide) membrane lined pores encased in the cell walls of the adjacent plant cells. They allow the cells to exchange all types of molecules ranging from nutrients like sugar, hormones, to RNAs and various proteins. Unfortunately, they are also hijacked by phyto-viruses, enabling them to spread from ...

Preparation of Onion Epidermal Cell Walls for Imaging by Atomic Force Microscopy (AFM)

Authors: Tian Zhang and Daniel J. Cosgrove, date: 12/20/2017, view: 11346, Q&A: 0

[Abstract] The growing plant cell wall is comprised of long, thin cellulose microfibrils embedded in a hydrated matrix of polysaccharides and glycoproteins. These components are typically constructed in layers (lamellae) on the inner surface of the cell wall, i.e., between the existing wall and the plasma membrane. The organization of these ...

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