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Subcellular Fractionation of Hela Cells for Lysosome Enrichment Using a Continuous Percoll-density Gradient

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Quantification of Starch in Guard Cells of Arabidopsis thaliana

Authors: Sabrina Flütsch, Luca Distefano and Diana Santelia, date: 07/05/2018, view: 3132, Q&A: 0

[Abstract] In this protocol, we describe how to quantify starch in guard cells of Arabidopsis thaliana using the fluorophore propidium iodide and confocal laser scanning microscopy. This simple method enables monitoring, with unprecedented resolution, the dynamics of starch in guard cells.

Isolation of Intact Vacuoles from Petunia Petals and Extraction of Sequestered Glycosylated Phenylpropanoid Compounds

Authors: Oded Skaliter , Jasmin Ravid, Alon Cna'ani, Gony Dvir, Rafael Knafo and Alexander Vainstein, date: 07/05/2018, view: 2229, Q&A: 0

[Abstract] Plant vacuoles are the largest compartment in plant cells, occupying more than 80% of the cell volume. A variety of proteins, sugars, pigments and other metabolites are stored in these organelles (Paris et al., 1996; Olbrich et al., 2007). Flowers produce a variety of specialized metabolites, some of which are unique to this ...

Radioactive Tracer Feeding Experiments and Product Analysis to Determine the Biosynthetic Capability of Comfrey (Symphytum officinale) Leaves for Pyrrolizidine Alkaloids

Authors: Thomas Stegemann, Lars H. Kruse and Dietrich Ober, date: 02/05/2018, view: 2301, Q&A: 0

[Abstract] This protocol delivers a method to determine the biosynthetic capability of comfrey leaves for pyrrolizidine alkaloids independently from other organs like roots or flowers.

The protocol applies and combines radioactive tracer experiments with standard and modern techniques like thin layer chromatography (TLC), solid-phase extraction ...

1-MCP (1-methylcyclopropene) Treatment Protocol for Fruit or Vegetables

Authors: Gamrasni Dan, Goldway Martin, Stern Yosi, Breitel Dario and Aharoni Asaph, date: 05/20/2017, view: 7372, Q&A: 4

[Abstract] 1-MCP (1-methylcyclopropene) is a simple synthetic hydrocarbon molecule that interacts with the ethylene receptor and inhibits the response of fruit or plant to ethylene. 1-MCP has opened new opportunities in handling harvested crops and serves as a powerful tool to learn about plant response to ethylene (Watkins and Miller, 2006). 1-MCP is ...

Simple Spectroscopic Determination of Nitrate, Nitrite, and Ammonium in Arabidopsis thaliana

Authors: Takushi Hachiya and Yuki Okamoto, date: 05/20/2017, view: 8895, Q&A: 1

[Abstract] Plants use nitrate, nitrite, and ammonium as inorganic nitrogen (N) sources. These N compounds are included in plant tissues at various concentrations depending on the balance between their uptake and assimilation. Thus, the contents of nitrate, nitrite, and ammonium are physiological indicators of plant N economy. Here, we describe a protocol for ...

Extraction and Analysis of Carotenoids from Escherichia coli in Color Complementation Assays

Authors: Andreas Blatt and Martin Lohr, date: 03/20/2017, view: 6436, Q&A: 0

[Abstract] A common method to investigate the function of genes putatively involved in carotenoid biosynthesis is the so called color complementation assay in Escherichia coli (see, e.g., Cunningham and Gantt, 2007). In this assay, the gene under investigation is expressed in E. coli strains genetically engineered to synthesize ...

Acetyl Bromide Soluble Lignin (ABSL) Assay for Total Lignin Quantification from Plant Biomass

Authors: William J. Barnes and Charles T. Anderson, date: 03/05/2017, view: 8191, Q&A: 0

[Abstract] Lignin is the second most abundant biopolymer on Earth, providing plants with mechanical support in secondary cell walls and defense against abiotic and biotic stresses. However, lignin also acts as a barrier to biomass saccharification for biofuel generation (Carroll and Somerville, 2009; Zhao and Dixon, 2011; Wang et al., 2013). For ...

Rice Root Organic Acid Efflux Measurement by Using Ion Chromatography

Authors: Chun-quan Zhu, Xiao-fang Zhu and Ren-fang Shen, date: 02/20/2017, view: 4617, Q&A: 0

[Abstract] Organic acids secreted from plant roots play important roles in various biological processes including nutrient acquisition, metal detoxification, and pathogen attraction. The secretion of organic acids may be affected by various conditions such as plant growth stage, nutrient deficiency, and abiotic stress. For example, when white lupin (Lupinus ...

Metabolite Profiling of Mature Arabidopsis thaliana Seeds Using Gas Chromatography-Mass Spectrometry (GC-MS)

Authors: Hagai Cohen, Ifat Matityahu and Rachel Amir, date: 11/05/2016, view: 6251, Q&A: 0

[Abstract] Metabolite profiling using gas chromatography-mass spectrometry (GC-MS) permits the annotation and quantification of a relatively wide variety of metabolites, covering a wide range of biochemical groups of metabolites. Lisec et al. (2006) established a method for GC-MS profiling in plants. Based on this protocol, we provide here a ...

Hydrogen Peroxide Measurement in Arabidopsis Root Tissue Using Amplex Red

Authors: Tzvetina Brumbarova, Cham Thi Tuyet Le and Petra Bauer, date: 11/05/2016, view: 8298, Q&A: 0

[Abstract] This protocol describes the measurement of hydrogen peroxide (H₂O₂) content in Arabidopsis root tissue by using the Amplex^® Red Hydrogen Peroxide/Peroxidase Assay Kit. When root tissue is disrupted and resuspended in phosphate buffer, H₂O₂ is released from the cells. The obtained root ...

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