Protocols in Current Issue
    I Plate-based Assay for Studying How Fungal Volatile Compounds (VCs) Affect Plant Growth and Development and the Identification of VCs via SPME-GC-MS
    Authors:  Wenzhao Wang, Ningxiao Li, Xingzhong Liu and Seogchan Kang, date: 02/20/2019, view: 890, Q&A: 0
    [Abstract] Biogenic volatile compounds (VCs) mediate various types of crucial intra- and inter-species interactions in plants, animals, and microorganisms owing to their ability to travel through air, liquid, and porous soils. To study how VCs produced by Verticillium dahliae, a soilborne fungal pathogen, affect plant growth and development, we ...
    An Image Analysis Pipeline to Quantify Emerging Cracks in Materials or Adhesion Defects in Living Tissues
    Authors:  Stéphane Verger, Guillaume Cerutti and Olivier Hamant, date: 10/05/2018, view: 1085, Q&A: 0
    [Abstract] Microcracks in materials reflect their mechanical properties. The quantification of the number or orientation of such cracks is thus essential in many fields, including engineering and geology. In biology, cracks in soft tissues can reflect adhesion defects, and the analysis of their pattern can help to deduce the magnitude and orientation of ...
    Analysis of Autophagic Activity Using ATG8 Lipidation Assay in Arabidopsis thaliana
    Authors:  Mengqian Luo and Xiaohong Zhuang, date: 06/20/2018, view: 2063, Q&A: 0
    [Abstract] As a fundamental metabolic pathway to degrade and recycle cellular cargos, autophagy is highly induced upon stress, starvation and senescence conditions in plants. A double-membrane structure named autophagosome will form during this process for cargo sequestration and delivery into the vacuole.

    A number of regulators have been ...
    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: 3020, 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: 2962, 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: 3050, 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: 4702, Q&A: 1
    [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 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: 7123, 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: 6925, 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 ...

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