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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: 186, Q&A: 0
    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 ...
    In vitro RNA-dependent RNA Polymerase Assay Using Arabidopsis RDR6
    Authors:  Kyungmin Baeg, Yukihide Tomari and Hiro-oki Iwakawa, date: 01/05/2018, view: 117, Q&A: 0
    RNA-dependent RNA polymerases (RdRPs) in eukaryotes convert single-stranded RNAs into double-stranded RNAs, thereby amplifying small interfering RNAs that play crucial roles in the regulation of development, maintenance of genome integrity and ...
    Micro-computed Tomography to Visualize Vascular Networks in Maize Stems
    Authors:  Akiteru Maeno and Katsutoshi Tsuda, date: 01/05/2018, view: 175, Q&A: 0
    Plant vascular systems in the stem connect roots with aerial organs to move solutes containing minerals, nutrients as well as signaling molecules, and therefore, they play pivotal roles in plant growth and development. However, stem vascular ...
    Chromatin Affinity Purification (ChAP) from Arabidopsis thaliana Rosette Leaves Using in vivo Biotinylation System
    Authors:  Weronika Sura and Piotr A. Ziolkowski, date: 01/05/2018, view: 117, Q&A: 0
    Chromatin Affinity Purification (ChAP) is widely used to study chromatin architecture and protein complexes interacting with DNA. Here we present an efficient method for ChAP from Arabidopsis thaliana rosette leaves, in which in vivo ...
    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: 268, Q&A: 0
    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 ...
    Fatty Acid Content and Composition of Triacylglycerols of Chlorella kessleri
    Authors:  Motohide Aoki and Norihiro Sato, date: 01/05/2018, view: 110, Q&A: 0
    Triacylglycerols (TAGs) are esters formed from one glycerol and three fatty acids. TAGs are induced to accumulate in algal cells under environmental stress conditions including nutrient-limitation, hyperosmosis, and low temperature, for the storage ...
    Rolling Circle Amplification to Screen Yam Germplasm for Badnavirus Infections and to Amplify and Characterise Novel Badnavirus Genomes
    Since the first discovery of badnaviruses (family Caulimoviridae, genus Badnavirus) in yam (Dioscorea spp.) germplasm in the 1970s (Harrison and Roberts, 1973), several hundred partial badnavirus reverse transcriptase ...
    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: 186, 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 ...
    In vitro RNA-dependent RNA Polymerase Assay Using Arabidopsis RDR6
    Authors:  Kyungmin Baeg, Yukihide Tomari and Hiro-oki Iwakawa, date: 01/05/2018, view: 117, Q&A: 0
    [Abstract] RNA-dependent RNA polymerases (RdRPs) in eukaryotes convert single-stranded RNAs into double-stranded RNAs, thereby amplifying small interfering RNAs that play crucial roles in the regulation of development, maintenance of genome integrity and antiviral immunity. Here, we describe a method of in vitro RdRP assay using recombinant ...
    Micro-computed Tomography to Visualize Vascular Networks in Maize Stems
    Authors:  Akiteru Maeno and Katsutoshi Tsuda, date: 01/05/2018, view: 175, Q&A: 0
    [Abstract] Plant vascular systems in the stem connect roots with aerial organs to move solutes containing minerals, nutrients as well as signaling molecules, and therefore, they play pivotal roles in plant growth and development. However, stem vascular systems, especially in crop species, have been poorly described since they are deeply embedded in the ...
    Chromatin Affinity Purification (ChAP) from Arabidopsis thaliana Rosette Leaves Using in vivo Biotinylation System
    Authors:  Weronika Sura and Piotr A. Ziolkowski, date: 01/05/2018, view: 117, Q&A: 0
    [Abstract] Chromatin Affinity Purification (ChAP) is widely used to study chromatin architecture and protein complexes interacting with DNA. Here we present an efficient method for ChAP from Arabidopsis thaliana rosette leaves, in which in vivo biotinylation system is used. The chromatin is digested by Micrococcal Nuclease (MNase), hence ...
    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: 268, 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 ...
    Fatty Acid Content and Composition of Triacylglycerols of Chlorella kessleri
    Authors:  Motohide Aoki and Norihiro Sato, date: 01/05/2018, view: 110, Q&A: 0
    [Abstract] Triacylglycerols (TAGs) are esters formed from one glycerol and three fatty acids. TAGs are induced to accumulate in algal cells under environmental stress conditions including nutrient-limitation, hyperosmosis, and low temperature, for the storage of metabolic energy and carbon, and also for the consumption of excess energy (e.g., Hirai ...
    Rolling Circle Amplification to Screen Yam Germplasm for Badnavirus Infections and to Amplify and Characterise Novel Badnavirus Genomes
    [Abstract] Since the first discovery of badnaviruses (family Caulimoviridae, genus Badnavirus) in yam (Dioscorea spp.) germplasm in the 1970s (Harrison and Roberts, 1973), several hundred partial badnavirus reverse transcriptase (RT)-ribonuclease H (RNaseH) sequences have been characterised (Kenyon et al., 2008; Bousalem ...
    Infection of Soybean Plants with the Insect Bacterial Symbiont Burkholderia gladioli and Evaluation of Plant Fitness
    Authors:  Paul Gaube, Martin Kaltenpoth and Laura V. Flórez, date: 12/20/2017, view: 400, Q&A: 0
    [Abstract] To investigate the establishment and consequences of host-microbe interactions, it is important to develop controlled infection assays suitable for each system, as well as appropriate methods to evaluate successful infection and its associated effects. Here, we describe a procedure for bacterial inoculation of soybean plants, followed by the ...
    Biomechanical Characterization of Onion Epidermal Cell Walls
    Authors:  Daniel M. Durachko, Yong Bum Park, Tian Zhang and Daniel J. Cosgrove, date: 12/20/2017, view: 278, Q&A: 0
    [Abstract] Here we describe two experimental protocols to measure the biomechanical properties of primary (growing) plant cell walls, with a focus on analyzing cell wall epidermal strips of onion scales. The first protocol measures cell wall creep (time-dependent irreversible extension) under constant force. Such creep is often mediated by the wall-loosening ...
    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: 422, 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 ...