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DNA and Chromatin Fiber-Based Plant Cytogenetics

  • Jason G. Walling
  • Jiming Jiang
Chapter
Part of the Plant Genetics and Genomics: Crops and Models book series (PGG, volume 4)

Abstract

Development of the fluorescence in situ hybridization (FISH) technique revolutionized cytogenetic research. FISH on prepared chromosomes has become the most commonly used technique in plant molecular cytogenetics, especially as a physical mapping tool in plant genome research. Despite its popularity, chromosome-based FISH analysis is limited in its capacity to distinguish DNA probes that separated by less than a few megabases. Development of FISH methods based on extended DNA fibers has dramatically increased the resolving power of this technique to the point where one can identify clones separated by only a few kilobases. In addition to the conventional fiber-FISH analysis, specialized techniques have been developed to prepare DNA or chromatin fibers that are suitable for restriction mapping (optical mapping) or immunofluorescence assays. Fiber-FISH and its derivatives are now used extensively in various mapping and genome research projects.

Keywords

DNA fibers · Fiber-FISH protocol · Plant nuclei isolation · DNA probe · Antibody detection · Fluorescence 

Abbreviations

BAC

Bacterial artificial chromosome

FISH

Fluorescence in situ hybridization

mtDNA

Mitochondrial DNA

Notes

Acknowledgments

We are grateful to Drs. David C. Schwartz and Shiguo Zhou for their input on our summary of optical mapping and for providing the image in Fig. 5.2. The most recent cytogenetic mapping research in the authors’ laboratory has been supported by grants DBI-0421671 and DBI-0603927 from the National Science Foundation.

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Department of HorticultureUniversity of WisconsinMadisonUSA

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