Quantitative analysis of two-dimensional gel electrophoresis protein patterns: a method for studying genetic relationships among Globodera pallida populations

A method based in two-dimensional protein gel electrophoresis has been developed in order to improve the analysis of genetic relationships among populations of Globodera. It has been used to estimate genetic divergence among nine Globodera pallida nematode populations. Sixty-one anonymous polypeptide spots were resolved using silver-stained high-resolution 2D gels and they were quantified in each population to establish genetic variation among G. pallida populations. The results of this analysis were compared with those obtained after a study of allelic frequency variation, which was carried out using seven previously described loci. Genetic distances among populations were calculated by means of both studies, the quantitative analysis and the allelic frequency variation, and phylogenetic trees were constructed for each type of analysis. A correlation analysis between the two distance matrices was carried out and a bootstrap analysis was performed to determine the strength of the clusters obtained with each method. The results obtained support the idea that quantitative protein analysis can be successfully applied to phylogenetic analysis of G. pallida populations.

[1]  M. Nei,et al.  Statistical Studies on Protein Polymorphism in Natural Populations. III. Distribution of Allele Frequencies and the Number of Alleles per Locus. , 1980, Genetics.

[2]  D. Mugniéry,et al.  Intra- and interspecific variability in Globodera, parasites of Solanaceous plants, revealed by Random Amplified Polymorphic DNA (RAPD) and correlation with biological features , 1997 .

[3]  D. Buth The Application of Electrophoretic Data in Systematic Studies , 1984 .

[4]  D. Mugniéry,et al.  Specific status of six Globodera parasites of solanaceous plants studied by means of two-dimensional gel electrophoresis with a comparison of gel patterns by a computed system. , 1993 .

[5]  Philippe Picard,et al.  Potential of two‐dimensional electrophoresis in routine identification of closely related durum wheat lines , 1997, Electrophoresis.

[6]  B S Weir,et al.  Estimation of the coancestry coefficient: basis for a short-term genetic distance. , 1983, Genetics.

[7]  R. Singh,et al.  A comprehensive study of genic variation in natural populations of Drosophila melanogaster. VII. Varying rates of genic divergence as revealed by two-dimensional electrophoresis. , 1992, Molecular biology and evolution.

[8]  S. Nadler Molecular approaches to studying helminth population genetics and phylogeny. , 1990, International journal for parasitology.

[9]  S. O’Brien,et al.  Molecular genetic divergence of orang utan (Pongo pygmaeus) subspecies based on isozyme and two-dimensional gel electrophoresis. , 1990, The Journal of heredity.

[10]  F. Rohlf An Empirical Comparison of Three Ordination Techniques in Numerical Taxonomy , 1972 .

[11]  Comparison of British populations of potato cyst nematodes with populations from continental Europe and South America using RAPDs. , 1997, Genome.

[12]  J. Felsenstein CONFIDENCE LIMITS ON PHYLOGENIES: AN APPROACH USING THE BOOTSTRAP , 1985, Evolution; international journal of organic evolution.

[13]  E. Platzer CHAPTER 1 – Potential Use of Protein Patterns and DNA Nucleotide Sequences in Nematode Taxonomy , 1981 .

[14]  T J White,et al.  Biochemical evolution. , 1977, Annual review of biochemistry.

[15]  M. Nei,et al.  Statistical Studies on Protein Polymorphism in Natural Populations II. Gene Differentiation between Populations. , 1978, Genetics.

[16]  C. Zijlstra,et al.  Genetic variation among parthenogenetic Meloidogyne species revealed by AFLPs and 2D protein electrophoresis contrasted to morphology , 1998 .

[17]  A. Görg,et al.  Genetic variability of pepper (Capsicum annuum L.) seed proteins studied by 2‐D electrophoresis with immobilized pH gradients , 1992, Electrophoresis.

[18]  Fred R. McMorris,et al.  Consensusn-trees , 1981 .

[19]  P. O’Farrell High resolution two-dimensional electrophoresis of proteins. , 1975, The Journal of biological chemistry.

[20]  V. Blok,et al.  Intraspecific Variation in Ribosomal DNA in Populations of the Potato Cyst Nematode Globodera pallida. , 1998, Journal of nematology.

[21]  J. Gauthier,et al.  Meloidogyne chitwoodi and M. fallax protein variation assessed by two-dimensional electrophoregram computed analysis , 1999 .

[22]  D. Smith,et al.  Allozyme polymorphisms in Spanish honeybees (Apis mellifera iberica). , 1995, The Journal of heredity.

[23]  A. Schots,et al.  Gene pool similarities of potato cyst nematode populations assessed by AFLP analysis. , 1996, Molecular plant-microbe interactions : MPMI.

[24]  J. Bakker Protein variation in cyst nematodes , 1987 .

[25]  M. Moens,et al.  Meloidogyne chitwoodi and M. fallax in Belgium , 2001 .

[26]  J. Bakker,et al.  Molecular evidence that Meloidogyne hapla, M. Chitwoodi and M. Fallax are distinct biological entities , 1997 .

[27]  L. Rhomberg,et al.  A Comprehensive Study of Genic Variation in Natural Populations of Drosophila melanogaster. I. Estimates of Gene Flow from Rare Alleles. , 1987, Genetics.

[28]  M. Coulthart,et al.  A comprehensive study of genic variation in natural populations of Drosophila melanogaster. VI. Patterns and processes of genic divergence between D. melanogaster and its sibling species, Drosophila simulans. , 1992, Genetics.

[29]  F. Rohlf,et al.  NTSYS-pc Numerical Taxonomy and Multivariate Analysis System, version 2.1: Owner manual , 1992 .