Protocols in Current Issue
    Spatiotemporal Quantification of Cytosolic pH in Arabidopsis Pollen Tubes
    Authors:  Maria Teresa Portes, Daniel S.C. Damineli and José A. Feijó, date: 07/20/2021, view: 1526, Q&A: 0

    Ion-specific probes and fluorescent indicators have been key in establishing the role of ion signaling, namely calcium, protons, and anions, in plant development, providing a robust approach for monitoring spatiotemporal changes in intracellular ion dynamics. The integration of protons/pH in signaling mechanisms is especially important as reports

    Measuring Extracellular Proton and Anionic Fluxes in Arabidopsis Pollen Tubes
    Authors:  Maria Teresa Portes and José A. Feijó, date: 02/05/2021, view: 2696, Q&A: 0

    The ion-selective vibrating probe has been used to detect and quantify the magnitude and direction of transmembrane fluxes of several ions in a wide range of biological systems. Inherently non-invasive, vibrating probes have been essential to access relevant electrophysiological parameters related to apical growth and morphogenesis in pollen

    Visualization of Actin Organization and Quantification in Fixed Arabidopsis Pollen Grains and Tubes
    Authors:  Xiaolu Qu, Qiannan Wang, Haiyan Wang and Shanjin Huang, date: 01/05/2020, view: 3305, Q&A: 0
    [Abstract] Although it is widely accepted that actin plays an important role in regulating pollen germination and pollen tube growth, how actin exactly performs functions remains incompletely understood. As the function of actin is dictated by its spatial organization, it is the key to reveal how exactly actin distributes in space in pollen cells. Here we ...
    Pollen Germination and Pollen Tube Growth of Arabidopsis thaliana: In vitro and Semi in vivo Methods
    Authors:  Hugh Dickinson, Josefina Rodriguez-Enriquez and Robert Grant-Downton , date: 08/20/2018, view: 12698, Q&A: 2
    [Abstract] Studies of pollen germination and post-germination development are not only essential for understanding plant reproduction but also are an excellent model system for tip-based growth. Here we describe easy, reproducible methods for germination and growth of pollen from the model plant Arabidopsis thaliana in artificial conditions. Our ...
    Rapid Isolation of Total Protein from Arabidopsis Pollen
    Authors:  Ming Chang and Shanjin Huang, date: 04/20/2017, view: 9078, Q&A: 0
    [Abstract] Arabidopsis pollen is an excellent system for answering important biological questions about the establishment and maintenance of cellular polarity and polar cell growth, because these processes are amenable to the genetic and genomic approaches that are readily available in Arabidopsis. Given that proteins are the direct ...
    Semi-thin Sectioning, Light and Fluorescence Microscopy of Floral Bud to Study Microspore Development in Arabidopsis
    Authors:  Min-Jung Kim and Jungmook Kim, date: 03/05/2016, view: 8599, Q&A: 0
    [Abstract] Pollen grains are male gametophytes produced within the pollen sacs of the anthers of the flower. Recent genetic studies have revealed several components involved in microspore development (Borg et al., 2009; Berger and Twell, 2011), and yet many components controlling microspore development remain to be identified. Semi-thin sectioning ...
    Analysis of Developing Pollen Grains within Intact Arabidopsis thaliana Anthers by Olympus Two-Photon Laser Scanning Microscopy
    Authors:  Teagen D. Quilichini, A. Lacey Samuels and Carl J. Douglas, date: 12/05/2015, view: 8819, Q&A: 0
    [Abstract] The method consists of imaging developing pollen grains as they form within intact, immature Arabidopsis thaliana anthers. Using two-photon excitation in the infrared wavelength range, the intrinsic fluorescence (autofluorescence) of developing pollen grains and surrounding sporophytic tissues of the anther wall, including the tapetum, ...
    Sample Preparation and Fractionation of Arabidopsis thaliana Sperm and Vegetative Cell Nuclei by FACS
    Authors:  Nina Chumak, Magdalena Mosiolek and Vera Karolina Schoft, date: 11/20/2015, view: 11656, Q&A: 1
    [Abstract] One of the major topics in plant and animal biology is sexual reproduction. It is, therefore, of great interest to isolate and study germ cells and accessory cells. The male gametophyte of the flowering plant Arabidopsis thaliana (A. thaliana), pollen, is the product of two post-meiotic mitotic divisions. Each mature pollen grain ...
    Analysis of Sugar Component of a Hot Water Extract from Arabidopsis thaliana Pollen Tubes Using GC-EI-MS
    [Abstract] Extraction with hot water is the oldest and simplest method used to recover pectin from an alcohol insoluble residue extract, although this method has not been widely used for the cell wall analysis of pollen tube, a model used to study cell wall. This protocol described this method applied for pectin extraction from 6 h-old Arabidopsis ...
    A Simple Protocol for the Immunolabelling of Arabidopsis Pollen Tube Membranes and Cell Wall Polymers
    [Abstract] The pollen tube, a fast tip-growing cell, is an excellent model to study membrane and cell wall biosynthesis. Here, we describe a simple protocol using an easy to use device to perform immunofluorescence labelling of pollen tube membrane and cell wall. The use of the NucleoSpin column to perform all the steps of the immunolabelling procedure ...

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