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    Protocols in Current Issue
    Visualization and Quantification of Cell-to-cell Movement of Proteins in Nicotiana benthamiana
    Authors:  Mila C Blekemolen, Mariliis Tark-Dame and Frank L.W Takken, date: 12/20/2018, view: 1202, Q&A: 0
    [Abstract] Cell-to-cell movement of proteins through plasmodesmata is a widely-established mechanism for intercellular signaling in plants. Current techniques to study intercellular protein translocation rely on single-cell transformation using particle bombardment or transgenic lines expressing photo-inducible fluorophores. The method presented here allows ...
    Examining Autophagy in Plant by Transmission Electron Microscopy (TEM)
    Authors:  Xiyin Zheng, Chenguang Zhao and Yule Liu, date: 10/20/2018, view: 1144, Q&A: 0
    [Abstract] In plants, macroautophagy, here referred as autophagy, is a degradation pathway during which the double-membrane structure named autophagosome engulfs the cargo and then fuses with vacuole for material recycling.

    To investigate the process of autophagy, transmission electron microscopy (TEM) was used to monitor the ultrastructure of ...
    Real-time PCR Analysis of PAMP-induced Marker Gene Expression in Nicotiana benthamiana
    Authors:  Fan Liu, Yuanpeng Xu, Yan Wang and Yuanchao Wang, date: 10/05/2018, view: 1464, Q&A: 0
    [Abstract] Perception of pathogen-associated molecular patterns (PAMPs) often triggers various innate immune responses in plants. The transcriptional changes of defense-related genes are often used as a marker to assay PAMP-triggered plant immune response. Here we described a protocol to monitor the relative expression level of marker genes in Nicotiana ...
    Measuring Secretion of Capsidiol in Leaf Tissues of Nicotiana benthamiana
    Authors:  Teruhiko Kuroyanagi, Maurizio Camagna and Daigo Takemoto, date: 08/05/2018, view: 1263, Q&A: 0
    [Abstract] Plant species produce a wide variety of antimicrobial metabolites to protect themselves against potential pathogens in natural environments. Phytoalexins are low molecular weight compounds produced by plants in response to attempted attacks of pathogens. Accumulation of phytoalexins in attacked plant tissues can inhibit the growth of penetrating ...
    Histone Deubiquitination Assay in Nicotiana benthamiana
    Authors:  Shujing Liu and Lars Hennig, date: 03/05/2018, view: 1487, Q&A: 0
    [Abstract] Histone modifications are a group of post-translational modifications on histones which can alter chromatin structure and affect gene expression. Histone ubiquitination is a histone modification found in particular on histone H2A and H2B. Histone ubiquitination can be reversed by ubiquitin-specific proteases (UBP). Here, we describe an in vivo ...
    Transient Gene Expression for the Characteristic Signal Sequences and the Estimation of the Localization of Target Protein in Plant Cell
    [Abstract] We have proposed and tested a method for characterization of the signal sequences and determinations of target protein localization in a plant cell. This method, called the AgI-PrI, implies extraction of protoplasts from plant tissues after agroinfiltration. The suggested approach combines the advantages of two widely used methods for transient ...
    Trimolecular Fluorescence Complementation (TriFC) Assay for Direct Visualization of RNA-Protein Interaction in planta
    Authors:  Jun Sung Seo and Nam-Hai Chua, date: 10/20/2017, view: 4574, Q&A: 0
    [Abstract] RNA-Protein interactions play important roles in various eukaryotic biological processes. Molecular imaging of subcellular localization of RNA/protein complexes in plants is critical for understanding these interactions. However, methods to image RNA-Protein interactions in living plants have not yet been developed until now. Recently, we have ...
    Ubiquitin Proteasome Activity Measurement in Total Plant Extracts
    Authors:  Suayib Üstün and Frederik Börnke, date: 09/05/2017, view: 3763, Q&A: 0
    [Abstract] The fine-tuned balance of protein level, conformation and location within the cell is vital for the dynamic changes required for a cell to respond to a given stimulus. This requires the regulated turnover of damaged or short-lived proteins through the ubiquitin proteasome system (UPS). Thus, the protease activity of the proteasome is adjusted to ...
    Use of Geminivirus for Delivery of CRISPR/Cas9 Components to Tobacco by Agro-infiltration
    Authors:  Kangquan Yin, Ting Han and Yule Liu, date: 04/05/2017, view: 6531, Q&A: 0
    [Abstract] CRISPR/Cas9 system is a recently developed genome editing tool, and its power has been demonstrated in many organisms, including some plant species (Wang et al., 2016). In eukaryotes, the Cas9/gRNA complexes target genome sites specifically and cleave them to produce double-strand breaks (DSBs), which can be repaired by non-homologous end ...
    Infection of Nicotiana benthamiana Plants with Potato Virus X (PVX)
    [Abstract] Potato Virus X (PVX) is the type member of Potexvirus genus, a group of plant viruses with a positive-strand RNA genome (~6.4 kb). PVX is able to establish compatible infections in Nicotiana benthamiana, a commonly used host in plant virology, leading to mild symptoms, such as chlorotic mosaic and mottling. PVX has been ...



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