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Deducing Intracellular Distributions of Metabolic Pathways from Genomic Data
Time of Update: 2021-01-14
Additionally, we describe strategies suitable for those groups of algae with secondary plastids (for instance diatoms), which are characterized by a different cellular topology and a larger number of intracellular compartments compared to plants.
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Reporter Genes and Transient Assays for Plants
Time of Update: 2021-01-14
A series of DNA plasmids containing reporter genes encoding readily assayed enzymes are available for this purpose.
In general, these reporter genes encode enzymes whose activities can be detected through assays and stains, thus facilitating the identification of transformed cells and quantification of the transformation process.
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Noncryogenic, Long-Term Germplasm Storage
Time of Update: 2021-01-14
Germplasm conservation is an important goal in the preservation of genetic diversity. Conventional methods, such as field and seed conservation, are used for most crops, although nonconventional meth
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Purification of Rubisco Activase from Leaves or after Expression in Escherichia coli
Time of Update: 2021-01-14
Rubisco activase is a molecular chaperone that modulates the activation state of Rubisco by catalyzing the ATP-dependent removal of tightly-bound inhibitory sugar-phosphates from Rubisco’s catalytic
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Pollen Tube Development
Time of Update: 2021-01-14
Pollen tubes grow rapidly in a strictly polarized manner as they transport male reproductive cells through female flower tissues to bring about fertilization.
Vegetative pollen tube cells are an excellent model system to investigate processes underlying directional cell expansion.
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Selected Simple Methods of Plant Cell Wall Histochemistry and Staining for Light Microscopy
Time of Update: 2021-01-14
Histochemical methods allow for identification and localization of various components within the tissue.
Such information on the spatial heterogeneity is not available with biochemical methods.
Hereby we present set of selected simple staining and histochemical methods with comments based on our laboratory experience.
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Statistical Issues in Microarray Data Analysis
Time of Update: 2021-01-14
Any microarray experiment consists of several components: (1) carrying out an appropriately designed (replicated) plant experiment; (2) array processing, which includes several steps of data acquisition and normalization; and (3) analysis of expression data to identify differentially expressed genes and overall patterns of expression.
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Maintaining Collections of Mutants for Plant Functional Genomics
Time of Update: 2021-01-14
Information on most plant genomic resources can be obtained through simple yet powerful Web searches, and ordering mechanisms are linked to the information.
Currently, available genomic resources are described, and references, links for Web data, and ordering information are also included.
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The Use of Multidrug Approach to Uncover New Players of the Endomembrane System Trafficking Machinery
Time of Update: 2021-01-14
Today, several of these chemical tools are widely used to perform mutant screens and establish the trafficking pathway of a given cellular component.
On the other hand, a multidrug approach can address the characterization of a new bioactive molecule and find its cellular pathway target.
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Brassica oleracea
Time of Update: 2021-01-13
Transformation efficiencies in the region of 10 to 25% (based on 50-125PCR -positive independent shoots from 500 infected explants) are typically achieved.
For researchers wishing to use their own plant genotype, we highlight the tissue culture phenotypes that are conducive to efficient transformation.
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Whole-Genome Microarrays: Applications and Technical Issues
Time of Update: 2021-01-13
This chapter attempts to summarize such applications as well as discusses some technical and strategic issues that are particular to the use of tiling microarrays.
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Plant Proteomics Methods to Reach Low-Abundance Proteins
Time of Update: 2021-01-13
The question of low-abundance proteins from biological tissues is still a major issue.
Technologies have been devised to improve the situation and in the last few years a method based on solid-phase combinatorial peptide ligand libraries has been extensively applied to animal extracts.
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Production of Monoclonal Antibodies to Plant Pathogens
Time of Update: 2021-01-13
The use of monoclonal antibodies in plant pathology has improved the quality and specificity of detection methods for diseases. Hybridoma technology allows the limitless production of highly specific
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Genome-Scale Models of Plant Metabolism
Time of Update: 2021-01-13
A complete, accurate GSM, in conjunction with a simulation technique such as flux balance analysis (FBA), can be used to comprehensively predict cellular metabolic flux distributions for a given genotype and given environmental conditions.
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Analysis of Viroid Replication
Time of Update: 2021-01-13
Replication of these minimal genomes, composed exclusively by a circular RNA of 246–401 nt, occurs in the nucleus (family Pospiviroidae) or in the chloroplast (family Avsunviroidae) by an RNA-based rolling-circle mechanism with three steps: ( 1 ) synthesis of longer-than-unit strands catalyzed by hostDNA -dependent RNA polymerases recruited and redirected to transcribe RNA templates, ( 2 ) cleavage to unit-length, which in family Avsunviroidae is mediated by hammerhead ribozymes, and ( 3 ) circularization through an RNA ligase or autocatalytically.
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Manipulating Photosynthesis in Transgenic Plants
Time of Update: 2021-01-13
Agrobacterium tumefaciens -mediated transformation of plants provides a relatively straightforward means of altering the amounts of individual proteins involved in photosynthetic processes.
These plants can then be analyzed to examine the effects of the altered protein levels on a wide range of physiological or biochemical processes.
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Quantifying Phytohormones in Transformed Plants
Time of Update: 2021-01-13
Liquid chromatography mass spectrometry (LC-MS) or gas chromatography-coupled mass spectrometry (GC-MS) in particular, recently have become more important for both qualitative and quantitative analyzes of all phytohormones, except ethylene (for a recent reviewsee ref.
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Yeast (Saccharomyces cerevisiae)
Time of Update: 2021-01-13
This species has enabled a detailed study of the (genetic) requirements forAgrobacterium -mediatedDNA transformation.
cerevisiae methodology has been developed for the transformation of many other yeast and fungal species.
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Heterologous Expression in Budding Yeast as a Tool for Studying the Plant Cell Morphogenesis Machinery
Time of Update: 2021-01-13
The budding yeast ( Saccharomyces cerevisiae ) can serve as a unique experimental system for functional studies of heterologous genes, allowing not only complementation of readily available yeast mu
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Constructing Gene-Enriched Plant Genomic Libraries Using Methylation Filtration Technology
Time of Update: 2021-01-13
For this reason, gene targeted partial genomic sequencing becomes a realistic option.
The method reported here is a simple approach to generate gene-enriched plant genomic libraries called methylation filtration.