Allozymic Polymorphism and Phylogeny of the Family Channichthyidae
301
WS (11 Sp) + Prydz Bay (6 sp); Dacodraco hunteri Waite 1916, WS (1
sp); Neopagetopsis ionah Nybelin 1947, WS (28 sp) + Prydz Bay (7 sp);
Pagetopsis macropterus Boulenger 1907, WS (1 sp) + Prydz Bay (3 sp);
Pagetopsis maculatus Barsukov and Permitin 1958, WS (3 sp);
Pseudochaenichthys georgian us Norman 1937, WS (1 sp).
Allozymic Electrophoresis
The tissues were homogenized with an equal volume of Tris-HCl-EDTANADP pH 6.8 buffer. Some drops of CCl 4 helped the elimination of lipids
from the enzymatic extract after a 30 min centrifugation at 15000 g. The
extract was then frozen at -80°C, before being analyzed.
Enzyme extraction, horizontal 12% starch gel electrophoresis and the
staining solutions were carried out according to the protocols of Pasteur et
al. [8]. Shaklee et al. [9] nomenclature was used for the loci.
Data Analyses
After interpretation of patterns on the gels, a genotype matrix was
established. The Genetix software [10] has been used to calculate the
allelic frequencies, the heterozygosity, the Nei [11] genetic distances and
the estimators of the F statistics of Wright [7].
The Fis can detect the deviations from the Hardy Weinberg genotype
proportions. Actually, it is the estimator f of Weir and Cockerham [11]
which is calculated. The software Genetix permits estimation of the
significance of the deviation of the estimator from zero by permutation
method generating 1000 artificial matrices. This parameter permits
detection ofpanmixia disequilibrium or Walhund effect in each sample.
The Fst detects the differences in heterozygote proportions between
samples (species or populations). In fact, the estimator e of Weir and
Cockerham [12] is calculated. The significance of the value is also
obtained by permutations.
Phylogenetic Reconstruction
With the Nei [11] genetic distances, a phylogenetic tree is built (Fig. 1)
using the neighbor joining method [13].
A second phylogenetic tree is established (Fig. 2) using the Dollo
parsimony method (the method allows, for each allele, only one
appearance but several losses in the tree). To limit the disadvantage of
small sample, a threshold of 5% is applied, considering an allele as
present when its frequency is over 5%. This matrix [12 taxa, 51
characters, 612 character states among which are 30 question marks
301
WS (11 Sp) + Prydz Bay (6 sp); Dacodraco hunteri Waite 1916, WS (1
sp); Neopagetopsis ionah Nybelin 1947, WS (28 sp) + Prydz Bay (7 sp);
Pagetopsis macropterus Boulenger 1907, WS (1 sp) + Prydz Bay (3 sp);
Pagetopsis maculatus Barsukov and Permitin 1958, WS (3 sp);
Pseudochaenichthys georgian us Norman 1937, WS (1 sp).
Allozymic Electrophoresis
The tissues were homogenized with an equal volume of Tris-HCl-EDTANADP pH 6.8 buffer. Some drops of CCl 4 helped the elimination of lipids
from the enzymatic extract after a 30 min centrifugation at 15000 g. The
extract was then frozen at -80°C, before being analyzed.
Enzyme extraction, horizontal 12% starch gel electrophoresis and the
staining solutions were carried out according to the protocols of Pasteur et
al. [8]. Shaklee et al. [9] nomenclature was used for the loci.
Data Analyses
After interpretation of patterns on the gels, a genotype matrix was
established. The Genetix software [10] has been used to calculate the
allelic frequencies, the heterozygosity, the Nei [11] genetic distances and
the estimators of the F statistics of Wright [7].
The Fis can detect the deviations from the Hardy Weinberg genotype
proportions. Actually, it is the estimator f of Weir and Cockerham [11]
which is calculated. The software Genetix permits estimation of the
significance of the deviation of the estimator from zero by permutation
method generating 1000 artificial matrices. This parameter permits
detection ofpanmixia disequilibrium or Walhund effect in each sample.
The Fst detects the differences in heterozygote proportions between
samples (species or populations). In fact, the estimator e of Weir and
Cockerham [12] is calculated. The significance of the value is also
obtained by permutations.
Phylogenetic Reconstruction
With the Nei [11] genetic distances, a phylogenetic tree is built (Fig. 1)
using the neighbor joining method [13].
A second phylogenetic tree is established (Fig. 2) using the Dollo
parsimony method (the method allows, for each allele, only one
appearance but several losses in the tree). To limit the disadvantage of
small sample, a threshold of 5% is applied, considering an allele as
present when its frequency is over 5%. This matrix [12 taxa, 51
characters, 612 character states among which are 30 question marks