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Measurement of Chloroplastic NAD Kinase Activity and Whole Tissue NAD Kinase Assay

Authors: Yuuma Ishikawa, Maki Kawai-Yamada and Shin-nosuke Hashida, date: 01/05/2020, view: 192, Q&A: 0

[Abstract] Nicotinamide adenine dinucleotide phosphate (NADP) synthesis requires nicotinamide adenine dinucleotide (NAD) kinase activity, substrate NAD and ATP. The NAD kinase responds to various environmental stimuli and its activity is regulated via various regulatory pathways, such as Ca²⁺-dependent and redox-dependent signals. Conventional in ...

Non-aqueous Fractionation (NAF) for Metabolite Analysis in Subcellular Compartments of Arabidopsis Leaf Tissues

Authors: David B. Medeiros, Stéphanie Arrivault, Jessica Alpers, Alisdair R. Fernie and Fayezeh Arabi, date: 10/20/2019, view: 473, Q&A: 0

[Abstract] The accurate determination of metabolite distribution in subcellular compartments is still challenging in plant science. Various methodologies, such as fluorescence resonance energy transfer-based technology, nuclear magnetic resonance spectroscopy and protoplast fractionation allow the study of metabolite compartmentation. However, large changes ...

Simple Method to Determine Protein Redox State in Arabidopsis thaliana

Authors: Keisuke Yoshida and Toru Hisabori, date: 06/05/2019, view: 1735, Q&A: 0

[Abstract] Thiol-based redox regulation is a posttranslational protein modification that plays a key role in many biological aspects. To understand its regulatory functions, we need a method to directly assess protein redox state in vivo. Here we present a simple procedure to determine protein redox state in a model plant Arabidopsis thaliana ...

An Adjustable Protocol to Analyze Chemical Profiles of Non-sterile Rhizosphere Soil

Authors: Alex Williams, Jurriaan Ton and Pierre Pétriacq, date: 05/20/2019, view: 1814, Q&A: 0

[Abstract] The analysis of chemical diversity in non-sterile rhizosphere soil has been a pressing methodological challenge for years. Rhizosphere-enriched chemicals (i.e., rhizochemicals) include root exudation chemicals, (microbial) breakdown products thereof, and de novo produced metabolites by rhizosphere-inhabiting microbes, all of ...

Analysis of Indole-3-acetic Acid (IAA) Production in Klebsiella by LC-MS/MS and the Salkowski Method

Authors: Shraddha Gang, Sheetal Sharma, Meenu Saraf, Martin Buck and Jörg Schumacher, date: 05/05/2019, view: 4246, Q&A: 0

[Abstract] Many rhizobacteria isolated from plant rhizosphere produce various phytohormones in the form of secondary metabolites, the most common of which is Indole-3-acetic acid (IAA). Here, we detail analytical protocols of IAA detection and quantification, in vitro and in situ, as recently applied to Klebsiella SGM 81, a ...

Determination of Root Exudate Concentration in the Rhizosphere Using ¹³C Labeling

Authors: Laurent Simon and Feth el Zahar Haichar, date: 05/05/2019, view: 1726, Q&A: 0

[Abstract] One of the most remarkable metabolic features of plant roots is their ability to secrete a wide range of compounds into the rhizosphere, defined as the volume of soil around living roots. Around 5%-21% of total photosynthetically fixed carbon is transferred into the rhizosphere through root exudates. Until recently, studies on the quantity and ...

Extraction and Purification of Laccases from Rice Stems

Authors: Chenna Swetha and P. V. Shivaprasad, date: 04/05/2019, view: 1612, Q&A: 0

[Abstract] Laccases are found in cell walls of plants in very low amounts. This protocol provides an efficient method to purify laccases from rice stems. The method involves three steps: 1) Isolation of total protein from rice stems using buffers with high salt concentration to extract protein from cell walls; 2) Purification of laccases using concanavalin-A ...

Detection of Disulfides in Protein Extracts of Arabidopsis thaliana Using Monobromobimane (mBB)

Authors: Shin‐nosuke Hashida and Maki Kawai-Yamada, date: 03/05/2019, view: 1520, Q&A: 0

[Abstract] Thiol-disulfide exchange is a key posttranslational modification, determining the folding process of intra- and inter-protein structures. Thiols can be detected by colorimetric reagents, which are stoichiometrically reduced by free thiols, and by fluorescent adducts, showing fluorescence only after thioester formation. We adapted a simple ...

Isolation of Thylakoid Membranes from the Cyanobacterium Synechocystis sp. PCC 6803 and Analysis of Their Photosynthetic Pigment-protein Complexes by Clear Native-PAGE

Authors: Josef Komenda, Vendula Krynická and Tomas Zakar, date: 01/05/2019, view: 2451, Q&A: 0

[Abstract] Cyanobacteria represent a frequently used model organism for the study of oxygenic photosynthesis. They belong to prokaryotic microorganisms but their photosynthetic apparatus is quite similar to that found in algal and plant chloroplasts. The key players in light reactions of photosynthesis are Photosystem I and Photosystem II complexes (PSI and ...

GC-MS-Based Analysis of Methanol: Chloroform-extracted Fatty Acids from Plant Tissues

Authors: Manish Kumar Patel, Shubhashis Das and Jitendra Kumar Thakur, date: 09/20/2018, view: 4841, Q&A: 0

[Abstract] Fatty acids (FAs) are carboxylic acids with long aliphatic chains that may be straight, branched and saturated or unsaturated. Most of the naturally occurring plant FAs contains an even number of carbon (C4-C24). FAs are used in food and pharmacological industries due to their nutritional importance. In addition, FAs are considered as a promising ...

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