Protocols in Current Issue
    Polyamine and Paraquat Transport Assays in Arabidopsis Seedling and Callus
    Authors:  Haoxi Chai, Yun Shen and Huazhong Shi, date: 08/05/2017, view: 4143, 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 ...
    A Ribosome Footprinting Protocol for Plants
    Authors:  Catharina Merchante, Qiwen Hu, Steffen Heber, Jose Alonso and Anna N. Stepanova, date: 11/05/2016, view: 9116, 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: 10806, 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: 7331, 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 ...
    Preparation of Chloroplast Lipid Membrane and Lipid-protein Interaction Assay
    Authors:  Kumiko Okazaki, Shin-ya Miyagishima and Hajime Wada, date: 02/05/2016, view: 6256, Q&A: 0
    [Abstract] Lipid-Protein interaction assay is a method to search lipids, which are bound with proteins in vitro. Since membranes that are spotted with chloroplast lipids such as monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG), and sulfoquinovosyldiacylglycerol (SQDG) are not commercially available, we extracted these lipids ...
    Measurement of PI4P Levels in Intact Chloroplasts Isolated from Arabidopsis thaliana
    Authors:  Kumiko Okazaki, Shin-ya Miyagishima and Hajime Wada, date: 02/05/2016, view: 6208, Q&A: 1
    [Abstract] Phosphatidylinositol 4-phosphate (PI4P), a major species of phosphoinositides, modulates many fundamental cellular processes. We have recently revealed that PI4P plays an important role in chloroplast division as a negative regulator. Despite its importance in chloroplasts, the content of PI4P in chloroplasts is very low and it is difficult to ...
    Quantification of Sodium Accumulation in Arabidopsis thaliana Using Inductively Coupled Plasma Optical Emission Spectrometery (ICP-OES)
    Authors:  Won-Gyu Choi and Simon Gilroy, date: 08/20/2015, view: 6004, Q&A: 0
    [Abstract] Salt stress is a major issue for plants growing in both natural and agricultural settings (Deinlein et al., 2014). For example, irrigation can lead to the build up of salts in the soil as the irrigation water evaporates, leading to salinization, inhibition of plant growth, reduced productivity and eventually to loss of agriculturally ...
    A Chemical Genetic Screening Procedure for Arabidopsis thaliana Seedlings
    [Abstract] Unbiased screening approaches are powerful tools enabling identification of novel players in biological processes. Chemical genetic screening refers to the technique of using a reporter response, such as expression of luciferase driven by a promoter of interest, to discover small molecules that affect a given process when applied to plants. These ...
    Quantification of Anthocyanin Content
    Authors:  Masaru Nakata and Masaru Ohme-Takagi, date: 04/05/2014, view: 19956, Q&A: 2
    [Abstract] Anthocyanins are a class of flavonoids and important plant pigments. They attract insects to pollinate flowers, protect plants from UV irradiation, and act as antimicrobial agents against herbivores and pathogens. Biosynthesis of anthocyanin is stimulated by diverse developmental signals and environmental stresses including drought, wounding, ...

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