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    Protocols in Current Issue
    Isolation and Quantification of Plant Extracellular Vesicles
    Authors:  Brian D. Rutter, Katie L. Rutter and Roger W. Innes, date: 09/05/2017, view: 325, Q&A: 0
    [Abstract] Extracellular vesicles (EVs) play an important role in intercellular communication by transporting proteins and RNA. While plant cells secrete EVs, they have only recently been isolated and questions regarding their biogenesis, release, uptake and function remain unanswered. Here, we present a detailed protocol for isolating EVs from the ...
    [Bio101] Leaf Clearing Protocol to Observe Stomata and Other Cells on Leaf Surface
    Author:  Nidhi Sharma, date: 09/05/2017, view: 157, Q&A: 0
    [Abstract] In this protocol, leaves are cleared and fixed in an ethanol and acetic acid solution, and mounted in Hoyer’s solution. The cleared leaves are imaged under differential interference contrast (DIC) microscope. This protocol is beneficial for studying stomata, hair cells, and other epidermal cells in plants.
    Using Silicon Polymer Impression Technique and Scanning Electron Microscopy to Measure Stomatal Aperture, Morphology, and Density
    Authors:  Hui-Chen Wu, Ya-Chen Huang, Chia-Hung Liu and Tsung-Luo Jinn, date: 08/20/2017, view: 409, Q&A: 0
    [Abstract] The number of stomata on leaves can be affected by intrinsic development programming and various environmental factors, in addition the control of stomatal apertures is extremely important for the plant stress response. In response to elevated temperatures, transpiration occurs through the stomatal apertures, allowing the leaf to cool through ...
    Quantification of Membrane Damage/Cell Death Using Evan’s Blue Staining Technique
    [Abstract] Membrane damage is a hallmark of both biotic and abiotic stress responses. The membrane determines the ability of a cell to sustain altered environmental conditions and hence can be used as a biomarker to assess stress-induced cell damage or death. We present an easy, quick, cost-effective, staining and spectrophotometric method to assess membrane ...
    Isolation of Cytosol, Microsome, Free Polysomes (FPs) and Membrane-bound Polysomes (MBPs) from Arabidopsis Seedlings
    Authors:  Yonghui Zhao and Shengben Li, date: 08/05/2017, view: 480, Q&A: 0
    [Abstract] The plant endomembrane system plays vital roles for synthesis, modification and secretion of proteins and lipids. From the classic view, only mRNAs encoding secreted proteins could be targeted to the endoplasmic reticulum (ER) for translation via a co-translational translocation manner, however, recently this model has been challenged by ...
    Isolation of Guard-cell Enriched Tissue for RNA Extraction
    Authors:  Pirko Jalakas, Dmitry Yarmolinsky , Hannes Kollist and Mikael Brosche, date: 08/05/2017, view: 483, Q&A: 0
    [Abstract] This is a protocol for isolation of guard cell enriched samples from Arabidopsis thaliana plants for RNA extraction. Leaves are blended in ice-water and filtered through nylon mesh to obtain guard cell enriched fragments. With guard cell enriched samples, gene expression analysis can be done, e.g., comparing different gene ...
    Isolation of Fucus serratus Gametes and Cultivation of the Zygotes
    Authors:  Amandine Siméon and Cécile Hervé, date: 07/20/2017, view: 320, Q&A: 0
    [Abstract] Zygotes of the Fucale species are a powerful model system to study cell polarization and asymmetrical cell division (Bisgrove and Kropf, 2008). The Fucale species of brown algae grow in the intertidal zone where they reproduce by releasing large female eggs and mobile sperm in the surrounding seawater. The gamete release can be induced from ...
    Phototaxis Assay for Chlamydomonas reinhardtii
    Authors:  Noriko Ueki and Ken-ichi Wakabayashi, date: 06/20/2017, view: 789, Q&A: 0
    [Abstract] Phototaxis is a behavior in which organisms move toward or away from the light source (positive or negative phototaxis, respectively). It is crucial for phototrophic microorganisms to inhabit under proper light conditions for phototaxis. The unicellular green alga Chlamydomonas reinhardtii rapidly changes its swimming direction upon light ...
    Analysis of 3D Cellular Organization of Fixed Plant Tissues Using a User-guided Platform for Image Segmentation
    Authors:  Ethel Mendocilla Sato and Célia Baroux, date: 06/20/2017, view: 727, Q&A: 0
    [Abstract] The advent of non-invasive, high-resolution microscopy imaging techniques and computational pipelines for high-throughput image processing has contributed to gain insights in plant organ morphogenesis at the cellular level. Confocal scanning laser microscopy (CSLM) allows the generation of three dimensional images constituted of serial optical ...
    Root Aliphatic Suberin Analysis Using Non-extraction or Solvent-extraction Methods
    Authors:  Camille Delude, Sollapura J. Vishwanath, Owen Rowland and Frédéric Domergue, date: 06/20/2017, view: 623, Q&A: 0
    [Abstract] Here we describe both non-extraction and solvent-extraction methods for root aliphatic suberin analysis. The non-extraction method is fast as roots are directly depolymerized using acidic transmethylation. However, suberin aliphatic components are isolated together with all the other acyl chains making up the lipids (e.g., membranes) ...