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Electrogenic Bromosulfalein Transport in Isolated Membrane Vesicles: Implementation in Both Animal and Plant Preparations for the Study of Flavonoid Transporters
Time of Update: 2021-01-16
The spectrophotometric assay of electrogenic bromosulfalein transport in membrane vesicles, isolated from various mammalian organs or tissues, enables to specifically measure the transport activity of bilitranslocase (TCDB 2.A.65.1.1).
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Visual Selection in Rice: A Strategy for the Efficient Identification of Transgenic Calli Accumulating Transgene Products
Time of Update: 2021-01-16
Furthermore, visual selection using GFP is a powerful tool that can be used to isolate transformed cells without antibiotic or herbicide pressure and can be applied to transformation systems in plants hypersensitive to these agents.
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Analyzing Shoot Apical Meristem Development
Time of Update: 2021-01-16
The shoot apical meristem ofArabidopsis thaliana contains a reservoir of pluripotent stem cells that functions as a continuous source of new cells for organ formation during development.
The SAM forms during embryogenesis, when it becomes stratified into specific cell layers and zones that can be delineated based on morphological and molecular criteria.
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Callus Initiation, Maintenance, and Shoot Induction in Rice
Time of Update: 2021-01-16
Embryogenic suspension cultures provide the most widely employed source of totipotent cells for protoplast isolation in rice ( Oryza sativa L.), since mesophyll-derived protoplasts of this important cereal rarely undergo sustained mitotic division leading to the production of tissues capable of plant regeneration.
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Chloramphenicol Acetyl Transferase Assay
Time of Update: 2021-01-16
The chloramphenicol acetyltransferase(cat) gene, isolated from transposon Tn9 ofEscherichia coli ( 1 ), is a convenient genetic marker for studies of transformation.
(For a detailed description of the CAT gene,see Chapter 1 .) In plants, the CAT system is generally used in transient, rather than in stable, gene expression studies.
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Analyzing Floral Meristem Development
Time of Update: 2021-01-16
Flowers contain the male and female sexual organs that are critical for plant reproduction and survival.
Each individual flower is produced from a floral meristem that arises on the flank of the shoot apical meristem and consists of four organ types: sepals, petals, stamens, and carpels.
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Movement Profiles: A Tool for Quantitative Analysis of Cell-to-Cell Movement of Plant Viral Movement Proteins
Time of Update: 2021-01-16
Expressing cells can be directly monitored for subcellular localization and cell-to-cell movement of the MP:GFP fusion protein into neighboring cells by confocal scanning microscopy.
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Agroinoculation and Agroinfiltration: Simple Tools for Complex Gene Function Analyses
Time of Update: 2021-01-16
With the explosive growth of genomic information and the development of advanced vectors to dissect plant gene function, this reliable method of viral gene delivery in plants, has been recruited and morphed into a technique popularly known as agroinfiltration.
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Separation of Plant Proteins by Electrophoresis
Time of Update: 2021-01-16
The present chapter describes methods for the extraction, electrophoresis, and detection of proteins, and for their transfer to membranes for microsequencing.
These can be used for the direct analysis of expressed proteins, or combined with Western blotting as described in the following chapter (Chapter 35 ).
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Isolation of Plant Polysomal mRNA by Differential Centrifugation and Ribosome Immunopurification Methods
Time of Update: 2021-01-15
To evaluate the translation state of an mRNA, ribosomal subunits, ribosomes, and polysomes can be isolated from detergent-treated cell extracts by high-speed differential centrifugation.
Here, we describe methods for the isolation and quantification of polysome complexes from plant tissues.
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Petunia (Petunia hybrida)
Time of Update: 2021-01-15
In this chapter, we describe a simple and reproducible genetic transformation protocol for generating transgenic petunia plants harboring a gene of interest and selectable marker.
Transformation efficiencies of around 10% are achievable with shoots being recovered about 8 wk after transgene insertion and rooted plants transferred to the greenhouse about twelve weeks after inoculation.
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Micropropagation of Citrus spp. by Organogenesis and Somatic Embryogenesis
Time of Update: 2021-01-15
Moreover, in vitro propagation provides a rapid system to multiply the progeny obtained by breeding programs, allows the use of monoembryonic and seedless genotypes as rootstocks, and it is very useful also for breeding and germplasm preservation.
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Establishment and Maintenance of Cell Suspension Cultures
Time of Update: 2021-01-15
The regeneration capability and genetic stability of suspension cells, however, decrease by the length of culture period Therefore, it is recommended to use newly initiated cell suspensions when the applications require the regeneration of dtplotd fertile plants The method described is used to establish morphogenic cell suspensions from the Arabidopszs ecotypes Columbia, C24, RLD, and Wassilewskila.
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Using Reverse Genetics to Develop Small Knockout Collections for Specific Biological Questions
Time of Update: 2021-01-15
With the advent of indexed mutagenized insertion lines in Arabidopsis, it is now possible to order small knockout collections of particular genes to probe a question of biological interest.
This first requires querying Arabidopsis databases to identify lines of interest, ordering them and then verifying homozygous lines to make your collection.
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Transient Gene Expression Analysis in Electroporated Maize Protoplasts
Time of Update: 2021-01-15
This soil bacterium, which naturally acts by transferring its own genes into a host plant, is commonly engineered for transferring foreign genes into plants, but its host range is limited mainly to dicots and noncereal monocots, and generally cannot be used to transfer foreign genes into maize ( 1 , 2 ).
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Clonal Propagation of Woody Species
Time of Update: 2021-01-15
Tissue-culture technology is widely used for the vegetative propagation of selected plants in agriculture and horticulture and, to a lesser extent, in forestry.
However, a major problem in the propagation of woody plants is that most success is achieved with juvenile tissue and not from proven mature trees ( 3 , 4 ).
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Regulation of Agrobacterium Gene Manipulation
Time of Update: 2021-01-15
The regulation of biotechnology within the member states of the European Community has developed very significantly over the last 3 yr, and a comprehensive body of legislation, guidance notes, and co
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Micropropagation of Lavandula spp.
Time of Update: 2021-01-15
Lavandula species are some of the most popular ornamental and medicinal plants with great economic values.
In vitro culture methods are suitable to overcome these limitations.
This chapter describes protocols for in vitro propagation ofLavandula viridis L’H�r andLavandula vera DC.
viridis and plant regeneration from leaf-derived callus ofL.
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Xenopus Oocytes as a Heterologous Expression System
Time of Update: 2021-01-15
Xenopus oocytes were first used to express plant genetic material in 1979 ( 2 ), and in the past decade it has become increasingly evident that they are an appropriate and useful system to study plant proteins.
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Trichovirus Isolation and RNA Extraction
Time of Update: 2021-01-15
The genus Trichovirus, a newly established plant virus genus ( 1 ), contains five viral species (including three tentative members), with similar biological, morphological, physicochemical, and ultra
