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Characterization and Phylogenetic Analysis of Four Root-Knot Nematode Species Using DNA Amplification Fingerprinting and Automated Polyacrylamide Gel Electrophoresis
Molecular Plant Microbe Interactions (1994)
  • Thomas J. Baum, Clemson University
  • Peter M. Gresshoff, University of Tennessee, Knoxville
  • Stephen A. Lewis, Clemson University
  • Ralph A. Dean, Clemson University
Abstract
Species identification and quantification of genotypic diversity in root-knot nematodes were achieved by DNA
amplification fingerprinting (DAF). Purified DNA from isolates of Meloidogyne arenaria, M. incognita, M. javanica,
and M. hap/a was amplified by low-stringency amplification reactions with 20 single-octamer primers of arbitrary
sequence. Amplification products were separated by electrophoresis in precast polyacrylamide minigels and
visualized by silver staining with an automated electrophoresis and staining unit (PhastSystem). Fingerprinting
data were analyzed with PAUP 3.1.1 using the Wagner parsimony method. Fourteen primers successfully amplified nematode DNA. Five primers revealed very conspicuous polymorphisms between species, while the remaining primers showed moderate to small differences. Polymorphisms between populations of the four species from the southeastern United States could also be detected. Phylogenetic analyses for the four species showed closer relatedness between M. arenaria and M.javanica than between M. arenaria and M. incognita. M. hap/a was most distant from M. arenaria, M. javanica, and M. incognita. These results are in agreement with phylogenetic trees derived from different characters. DAF utilizing automated polyacrylamide gel electrophoresis and silver staining is highly reproducible and offers excellent resolution, optimal sensitivity, unequaled speed, and the simplicity of automation.
Publication Date
1994
Publisher Statement
This article is in the public domain and not copyrightable. It may be freely reprinted with customary crediting of the source. The American Phytopathological Society, 1994.
Citation Information
Thomas J. Baum, Peter M. Gresshoff, Stephen A. Lewis and Ralph A. Dean. "Characterization and Phylogenetic Analysis of Four Root-Knot Nematode Species Using DNA Amplification Fingerprinting and Automated Polyacrylamide Gel Electrophoresis" Molecular Plant Microbe Interactions Vol. 7 Iss. 1 (1994) p. 39 - 47
Available at: http://works.bepress.com/thomas-baum/6/