Plant Science


    Protocols in Current Issue
    Fractionation and Extraction of Crude Nuclear Proteins From Arabidopsis Seedlings
    Authors:  Jiajia Zhao, Jinsong Bao and Feifei Xu, date: 01/20/2022, view: 1094, Q&A: 0

    The plant nucleus is an important subcellular organelle that contains the genome, ribosomal RNA, and regulatory proteins, and performs a central role in the functioning and metabolism of the cell. Fractionation of intact nuclei is a crucial process to elucidate the function of nuclear proteins. Here, we present a simple method for the

    Preparation and Transfection of Populus tomentosa Mesophyll Protoplasts
    Authors:  Hou-Ling Wang, Ting Wang, Qi Yang, Weilun Yin, Xinli Xia, Hongwei Guo and Zhonghai Li, date: 11/20/2021, view: 1645, Q&A: 0

    Mesophyll protoplasts freshly isolated from leaves are a useful research system in plants. However, cell walls in woody plants contain more pectin, making mesophyll protoplasts isolation difficult in Populus. This has limited their application in biochemical, molecular, cellular, genetic, genomic, transcriptomic, and proteomic assays. In this

    Enrichment of Cytoplasmic RNA Granules from Arabidopsis Seedlings
    Authors:  Zhen Lei, Eun Yu Kim and Jungnam Cho, date: 11/05/2021, view: 1235, Q&A: 0

    RNA granules (RGs) are membraneless intracellular compartments that play important roles in the post-transcriptional control of gene expression. Stress granules (SGs) are a type of RGs that form under environmental challenges and/or internal cellular stresses. Stress treatments lead to strong mRNAs translational inhibition and storage in SGs until

    Lipid Droplet Isolation from Arabidopsis thaliana Leaves
    Authors:  Yovanny Izquierdo, Rubén Fernández-Santos, Tomás Cascón and Carmen Castresana, date: 12/20/2020, view: 2208, Q&A: 0

    Lipid droplets (LDs) are neutral lipid aggregates surrounded by a phospholipid monolayer and specific proteins. In plants, they play a key role as energy source after seed germination, but are also formed in vegetative tissues in response to developmental or environmental conditions, where their functions are poorly understood. To elucidate these,

    Investigating Localization of Chimeric Transporter Proteins within Chloroplasts of Arabidopsis thaliana
    Authors:  Susumu Uehara, Yasuko Ito-Inaba and Takehito Inaba, date: 02/05/2018, view: 7867, Q&A: 0
    [Abstract] In this protocol, we describe a method to design chimeric proteins for specific targeting to the inner envelope membrane (IEM) of Arabidopsis chloroplasts and the confirmation of their localization by biochemical analysis. Specific targeting to the chloroplast IEM can be achieved by fusing the protein of interest with a transit peptide ...
    Isolation and Quantification of Plant Extracellular Vesicles
    Authors:  Brian D. Rutter, Katie L. Rutter and Roger W. Innes, date: 09/05/2017, view: 17980, Q&A: 4
    [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 ...
    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: 9918, 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 ...
    Plasma Membrane Preparation from Lilium davidii and Oryza sativa Mature and Germinated Pollen
    Authors:  Ning Yang, Bing Han, Lingtong Liu, Hao Yang and Tai Wang, date: 05/20/2017, view: 7290, Q&A: 0
    [Abstract] Pollen germination is an excellent process to study cell polarity establishment. During this process, the tip-growing pollen tube will start elongating. The plasma membrane as the selectively permeable barrier that separates the inner and outer cell environment plays crucial roles in this process. This protocol described an efficient aqueous ...
    Cation (Ca2+ and Mn2+) Partitioning Assays with Intact Arabidopsis Chloroplasts
    Authors:  Anna Harms, Iris Steinberger and Anja Schneider, date: 01/05/2017, view: 7416, Q&A: 0
    [Abstract] Determination of the relative distribution of Ca2+ and Mn2+ is an important tool for analyzing mutants showing altered levels of calcium and/or manganese transporters in the chloroplast envelope or thylakoid membrane. The method described in this protocol allows quantitative analyses of the relative distribution of calcium ...
    A Ribosome Footprinting Protocol for Plants
    Authors:  Catharina Merchante, Qiwen Hu, Steffen Heber, Jose Alonso and Anna N. Stepanova, date: 11/05/2016, view: 12373, 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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