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    Protocols in Current Issue
    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: 393, 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 ...
    Real-time Analysis of Auxin Response, Cell Wall pH and Elongation in Arabidopsis thaliana Hypocotyls
    Authors:  Lanxin Li, S. F. Gabriel Krens, Matyáš Fendrych and Jiří Friml, date: 01/05/2018, view: 522, Q&A: 0
    [Abstract] The rapid auxin-triggered growth of the Arabidopsis hypocotyls involves the nuclear TIR1/AFB-Aux/IAA signaling and is accompanied by acidification of the apoplast and cell walls (Fendrych et al., 2016). Here, we describe in detail the method for analysis of the elongation and the TIR1/AFB-Aux/IAA-dependent auxin response in ...
    Polyamine and Paraquat Transport Assays in Arabidopsis Seedling and Callus
    Authors:  Haoxi Chai, Yun Shen and Huazhong Shi, date: 08/05/2017, view: 1054, Q&A: 0
    [Abstract] Polyamines (PAs) are polycationic compounds found in all living organisms and play crucial roles in growth and survival. We here show the ‘Polyamine and paraquat (PQ) transport assay’ protocol, which can be used to examine the uptake activity of PA/PQ transporters. We have used this protocol to demonstrate that PUT3 in Arabidopsis is a ...
    Assessing Plant Tolerance to Acute Heat Stress
    Authors:  Minsoo Kim, Fionn McLoughlin, Eman Basha and Elizabeth Vierling, date: 07/20/2017, view: 1387, Q&A: 0
    [Abstract] It is well-established that plants are able to acclimate to temperatures above or below the optimal temperature for their growth. Here, we provide protocols for assays that can be used quantitatively or qualitatively to assess the relative ability of plants to acquire tolerance to high temperature stress. The hypocotyl elongation assay described ...
    Separation of Plant 6-Phosphogluconate Dehydrogenase (6PGDH) Isoforms by Non-denaturing Gel Electrophoresis
    [Abstract] 6-Phosphogluconate dehydrogenase (6PGDH; EC 1.1.1.44) catalyzes the third and irreversible reaction of the pentose phosphate pathway (PPP). It carries out the oxidative decarboxylation of the 6-phosphogluconate to yield ribulose-5-phosphate, carbon dioxide and NADPH. In higher plants, 6PGDH has several subcellular localizations including cytosol, ...
    In Gel Kinase Assay
    Authors:  Gaston A. Pizzio and Pedro L. Rodriguez, date: 03/05/2017, view: 2942, Q&A: 0
    [Abstract] Proper spatiotemporal regulation of protein phosphorylation in cells and tissues is required for normal development and homeostasis. We present the protocol ‘In Gel Kinase Assay’, which is useful for protein kinase activity measurements from crude protein extracts. We have successfully used ‘In Gel Kinase Assay’ protocol to show that the ...
    A Ribosome Footprinting Protocol for Plants
    Authors:  Catharina Merchante, Qiwen Hu, Steffen Heber, Jose Alonso and Anna N. Stepanova, date: 11/05/2016, view: 3200, Q&A: 0
    [Abstract] Ribosome footprinting, or Ribo-seq, has revolutionized the studies of translation. It was originally developed for yeast and mammalian cells in culture (Ingolia et al., 2009). Herein, we describe a plant-optimized hands-on ribosome footprinting protocol derived from previously published procedures of polysome isolation (Ingolia et al ...
    Rapid Determination of Cellulose, Neutral Sugars, and Uronic Acids from Plant Cell Walls by One-step Two-step Hydrolysis and HPAEC-PAD
    Authors:  Trevor Yeats, Tamara Vellosillo, Nadav Sorek, Ana B. Ibáñez and Stefan Bauer, date: 10/20/2016, view: 3581, Q&A: 1
    [Abstract] The plant cell wall is primarily composed of the polysaccharides cellulose, hemicellulose and pectin. The structural and compositional complexity of these components are important for determining cell wall function during plant growth. Moreover, cell wall structure defines a number of functional properties of plant-derived biomass, such as ...
    15N-nitrate Uptake Activity and Root-to-shoot Transport Assay in Rice
    Authors:  Yongqiang Liu, Bin Hu and Chengcai Chu, date: 08/20/2016, view: 2818, Q&A: 0
    [Abstract] 15N is a nonradioactive heavy isotope of nitrogen, widely used for biochemical and physiological research in plants. For instance, 15N-KNO3 was used as the nitrogen source in plants in order to investigate nitrate uptake activity and transport from roots to shoots (Lin et al., 2008). Here, we describe a ...
    Quantification of the Adhesion Strength between Peroxisomes and Chloroplasts by Femtosecond Laser Technology
    [Abstract] This is the detailed protocol to quantify adhesion strength between peroxisomes and chloroplasts in plant leaf palisade mesophyll cells described by Oikawa et al. (2015). The quantification was performed by utilizing local explosion induced by focusing femtosecond laser pulses into a mesophyll cell under a confocal microscope. When an ...