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Oxalic Acid

Oxalate is a common constituent of many plant species. Some crop plants accumulate high levels of this dicarboxylic acid anion (oxalate). Depending on the species, oxalate accumulates primarily as soluble oxalate, insoluble calcium oxalate, or a combination of these two forms. Oxalate, b ...

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Nitrate and Nitrite

Occasionally forages accumulate nitrates in quantities that are toxic to some farm animals. Under certain environmental conditions, some plants may accumulate high concentrations of nitrates. The factors that can influence the accumulation of nitrates in the plants are drought, s ...

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L-Mimosine (-(3-Hydroxy-4-Pyridone-1-yl)-L-Alanine)

L-mimosine is a nonprotein amino acid. Despite the consideration of Leucaena leucocephala as a promising alternate source of protein for fodder, the presence of mimosine to the extent of 2% to 10% dry matter in the leaf and 2% to 5% dry matter in the seed has limited its use as a livestock feed since mimosine and its ...

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Canavanine

Canavanine occurs in over 350 species of the papilionoideae, a subfamily of the Leguminosae. It is an analogue of arginine (Fig. 1). The highest concentration of canavanine (13% dry weight) has been reported in Dioclea megacarpa and it occurs up to 5% in seeds of Canavalia ensiformis. L-canavanine ex ...

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Determination of in vivo Protein Phosphorylation in Photosynthetic Membranes

Light- and redox-controlled reversible phosphorylation of thylakoid proteins regulates short- and long-term acclimation of plants to environmental cues. The major phosphoproteins in thylakoids belong to photosystem II and its light-harvesting antenna but phosphorylat ...

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Identification of Thioredoxin Targeted Proteins Using Thioredoxin Single Cysteine Mutant-immobilized Resin

Thioredoxins (Trx) are a ubiquitous family of proteins that modulate the enzymatic activity of their substrate proteins by redox regulation. This is achieved by reduction of a disulfide bond within their target proteins. A conserved pair of cysteine residues in Trx is required for catalys ...

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Imaging of Reactive Oxygen Species in vivo

Reactive oxygen species (ROS) are involved in many signalling pathways and numerous stress responses in plants. Consequently, it is important to be able to identify and localize ROS in vivo to evaluate their roles in signalling. A number of probes that have a high affinity for specific ROS and that are ...

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Dynamic Redox Measurements with Redox-Sensitive GFP in Plants by Confocal Laser Scanning Microscopy

Continuous control of metabolism and development is a key feature of life and is of particular importance under stress conditions. While under normal conditions most cellular compartments maintain a reducing environment, the cellular redox state can be influenced by external facto ...

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Probing Spatio-temporal Intracellular Calcium Variations in Plants

Calcium (Ca2+) ions act as intracellular second messengers in many different signalling processes in plant cells and thus contribute to the amplification step of the signalling pathway and the specificity of the adaptative response. Dynamics of calcium described as spatial and tempo ...

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Accurate Real-time Reverse Transcription Quantitative PCR

Within the last few years real-time quantitative PCR has become the method of choice for the accurate quantification of mRNA levels. Compared to previous methods the sensitivity of real-time quantitative PCR improved to the detection limit of up to one single molecule per reaction tube. Howe ...

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Phenotyping of Abiotic Responses and Hormone Treatments in Arabidopsis

The disruption or modulation of signal transduction pathways does not always lead to drastic changes in plant growth and development. Therefore, many loss- or gain-of-function lines do not exhibit an obvious phenotype under normal greenhouse conditions. To be able to assign biological f ...

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RNA-protein Interaction Mediating Post-transcriptional Regulation in the Circadian System

Post-transcriptional control makes an important contribution to shaping transcript profiles of circadianly regulated genes. In Arabidopsis thaliana, the clock-regulated glycine-rich RNA-binding protein ATGRP7 oscillates with a 24-h rhythm and transmits the rhythmic ...

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Analysis of Plant Regulatory DNA Sequences by Transient Protoplast Assays and Computer Aided Sequence Evaluation

The orchestrated regulation of hundreds of genes responding in a temporal, spatial, and conditional expression is in part mediated by the transient binding of transcription factors to their specific DNA motifs. The analysis of these cis-regulatory DNA sequences is still a challenging t ...

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Analysis of Plant Regulatory DNA sequences by the Yeast-One-Hybrid Assay

Regulatory DNA sequences harbor the essential information to control specific gene expression changes and integrate information derived from upstream signaling cascades. This regulatory potential is mediated by direct binding of proteins, e.g., transcription factors, to de ...

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Fluorescence-based Electrophoretic Mobility Shift Assay in the Analysis of DNA-binding Proteins

Changes in gene expression mediated by DNA-binding protein factors are a crucial part of many signal transduction pathways. Generally, these regulatory proteins are low abundant and thus their purification and characterisation is labour- and time-intensive. Here we describe a work ...

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Chromatin Immunoprecipitation Experiments to Investigate In Vivo Binding of Arabidopsis Transcription Factors to Target Sequences

For understanding the mechanism of transcriptional regulation, it is essential to know which transcription factor is bound in vivo to the promoter to be analysed. If transcription from a given promoter is regulated by developmental or environmental stimuli, the question of inducible v ...

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Functional Complementation of Yeast Mutants to Study Plant Signalling Pathways

The rapidly increasing amount of entirely sequenced genomes generates a need for fast and efficient methods to elucidate gene functions. Functional complementation of yeast mutants, displaying selectable phenotypes, has been used very successfully in the past years to isolate ma ...

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Phosphatase Activities Analyzed by in vivo Expressions

Protein phosphatases act to reverse phosphorylation-related modifications induced by protein kinases. Type 2C protein phosphatases (PP2C) are monomeric Ser/Thr phosphatases that require a metal for their activity and are abundant in prokaryotes and eukaryotes. In plants, such ...

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The Determination of Protein-protein Interactions by the Mating-based Split-ubiquitin system (mbSUS)

Dynamic and reversible protein–protein interactions have a pivotal function in all living cells. For instance, protein–protein interactions are involved in the assembly and regulation of multimeric enzymes and transcription factors, various signal response pathways, intr ...

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Fluorescence Cross-Correlation Spectroscopy of Plant Proteins

Fluorescence cross-correlation spectroscopy (FCCS) is a technique that physically evaluates the molecular interaction between two fluorophore-tagged molecules such as proteins and oligonucleotides in a quantitative manner. Because it simply makes use of the coincident ...

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