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Peanut (Arachis hypogaea L.)

Arachis hypogea (peanut, groundnut), an annual oil seed belonging to the Leguminosae family and the Papillionacea subfamily, is a legume native to South America but now grown in diverse environments in six continents between latitudes 40�N and 40�S. Arachis hypogea can grow in a wide range of cli ...

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Pigeonpea (Cajanus cajn (L.) Millsp.)

Pigeonpea , also known as redgram, is one of the major grain legume (pulses) crops grown in the semiarid tropics (SAT) extending between 30�N and 30�S; it is the second most important food legume of India. It is cultivated in about 50 countries of Asia, Africa, and the Americas for a variety of uses (food, fodder, f ...

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Soybean (Glycine max) Transformation Using Mature Cotyledonary Node Explants

Agrobacterium tumefaciens-mediated transformation of soybeans has been steadily improved since its development in 1988. Soybean transformation is now possible in a range of genotypes from different maturity groups using different explants as sources of regenerable cells, v ...

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Red Clover (Trifolium pratense)

Genetic modification of plants by the insertion of transgenes can be a powerful experimental approach to answer basic questions about gene product function. This technology can also be used to make improved crop varieties for use in the field. To apply this powerful tool to red clover, an importa ...

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Soybean (Glycine max) Transformation Using Immature Cotyledon Explants

Agrobacterium tumefaciens-mediated transformation of soybeans can be accomplished using immature zygotic cotyledons as target tissues providing an alternate explant to embryogenic tissue cultures, proliferating meristems, and cotyledonary nodes. The immature cot ...

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Tepary Bean (Phaseolus acutifolius)

Phaseolus beans are among the major legumes for food consumption, especially in Latin America, Africa, and Asia. Tepary bean (Phaseolus acutifolius L. Gray) is one of the five cultivated species of the genus Phaseolus. This chapter describes an Agrobacterium-mediated transformation p ...

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Brassica oleracea

A better understanding of the genetic basis underlying the genotype dependence of Brassica oleracea transformation is enabling researchers to distinguish between recalcitrant and successful candidate genotypes for routine transformation. In this chapter we outline an A. tu ...

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Cucumber (Cucumis sativus L.)

We describe two novel Agrobacterium tumefaciens-based methods of cucumber transformation. The first involves direct regeneration from leaf microexplants selected on kanamycin-containing medium. The second involves regeneration from a long-term established embryo ...

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Eggplant (Solanum melongena L.)

Eggplant is an economically important vegetable crop in Asia and Africa, and although it is grown in Europe and the United States, it does not account for a significant percentage of agricultural production. It is susceptible to a number of pathogens and insects, with bacterial and fungal wilts be ...

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Tomato (Lycopersicum esculentum)

Tomato (Lycopersicum esculentum) is an important fruit crop in the Americas, southern Europe, the Middle East, and India, with increasing production in China, Japan, and Southeast Asia. It is amenable to producing pharmaceuticals, particularly for oral delivery; for many of the same reaso ...

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Prevention and Control of Pests and Diseases

A well-controlled growth environment with plants that are not unduly stressed is essential for Arabidopsis molecular biology research. Even if they do not kill the plants outright, insect pests and microbial pathogens can cause subtle changes in gene expression or plant metabolism that ...

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Growth of Other Species Related to Arabidopsis thaliana

Recent interest in the study of biological phenomena in Brassicaceae members (crucifers) related to the model plant, Arabidopsis thaliana, fuels the need for standard protocols for growth and maintenance of these plant species. Like A. thaliana, many wild crucifers are amenable to growth ...

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Grafting

Grafting provides a simple way to generate chimeric plants with regions of different genotypes, and thus to assess the cell autonomy of gene action. The technique of grafting has been widely used in other species, but in Arabidopsis, its small size makes the process rather more complicated. Howe ...

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Synchronization, Transformation, and Cryopreservation of Suspension-Cultured Cells

We have recently described the selection of rapidly dividing Arabidopsis cell suspension cultures MM1 and MM2d that provide a powerful platform for plant cell-cycle research. Here we provide detailed protocols and procedures to achieve high levels of synchronization, either by sta ...

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Gene Identification and Cloning by Molecular Marker Mapping

In the course of map-based cloning, mutant genes are identified through linkage to a sufficiently small region of the genetic map and subsequent DNA sequencing. This process has become fairly straightforward for Arabidopsis mutations, owing to the completed genome sequence and the disc ...

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High-Throughput TILLING for Arabidopsis

Targeting induced local lesions in genomes (TILLING) is a general strategy for identifying induced point mutations that can be applied to almost any organism. In this chapter, we describe the basic methodology for high-throughput TILLING. Gene segments are amplified using fluoresce ...

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Genotyping and Mapping With High-Density Oligonucleotide Arrays

This chapter discusses the specific details of how oligonucleotide arrays originally designed for expression analysis can be used for high-density genotyping to reveal tens of thousands of single-feature polymorphisms (SFPs). Cluster of SFPs reveal potential natural deletion ...

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Forward Genetic Screening of Insertional Mutants

Insertional mutagenesis has contributed to the success of forward genetics in Arabidopsis thaliana. The availability of large collections of lines mutagenized by either transposon or T-DNA insertions, in combination with the systematic sequencing of insertion/genome junct ...

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PCR-Based Screening for Insertional Mutants

In Arabidopsis, where homologous recombination is highly inefficient, insertional mutagenesis has provided a great alternative means for identifying loss-of-function mutants in genes of interest. Many collections of T-DNA and transposon-tagged plants have been generated ...

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Cytogenetic Analyses of Arabidopsis

For a long time, Arabidopsis thaliana was not a favorite of cytogeneticists because of its small chromosomes. Nevertheless, most cytogenetic approaches have become applicable to this model plant during the past decade. Recently, the very small genome, the low content, and strong cluster ...

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