Identification of flatfish (Pleuronectiforme) species using DNA-based techniques.
Identification of flatfish species using a DNA-based methodology was studied. The polymerase chain reaction was employed to obtain a 464 bp amplicon from mitochondrial cytochrome b gene. The sequences from this fragment belonging to 24 species were analyzed using a genetic distance method, and polymorphic sites were determined. The fragment was found to be highly polymorphic (231 sites), and this permitted the differentiation of most of the species. Phylogenetic tree construction was employed to allow the identification of flatfish species. As a result, each species was grouped in a well-differentiated clade, except for two pairs: Limanda ferruginea and L. limanda, and Solea impar and S. lascaris, which could not be differentiated. On the basis of the sequences obtained, restriction enzymes were selected to provide specific restriction profiles, which allow the differentiation of 21 species of flatfish in a faster and less expensive manner than sequencing. This polymerase chain reaction-restriction fragment length polymorphism methodology (PCR-RFLP) was tested using commercial samples.
Sotelo
C G
CG
Instituto de Investigaciones Marinas, Consejo Superior de Investigaciones Cientificas, Eduardo Cabello 6, 36208 Vigo, Spain. carmen@iim.csic.es
Calo-Mata
P
P
Chapela
M J
MJ
Pérez-Martín
R I
RI
Rehbein
H
H
Hold
G L
GL
Russell
V J
VJ
Pryde
S
S
Quinteiro
J
J
Izquierdo
M
M
Rey-Méndez
M
M
Rosa
C
C
Santos
A T
AT
D000818:Animals; D003573:Cytochrome b Group / Q000235:genetics; D004272:DNA, Mitochondrial / Q000032:analysis / Q000737:chemistry; D015252:Deoxyribonucleases, Type II Site-Specific; D005412:Flatfishes / Q000145:classification* / Q000235:genetics*; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D017422:Sequence Analysis, DNA
D003573:Cytochrome b Group; D004272:DNA, Mitochondrial; C048422:endodeoxyribonuclease NlaIII; D015252:Deoxyribonucleases, Type II Site-Specific; C032229:GANTC-specific type II deoxyribonucleases; C024400:GTYRAC-specific type II deoxyribonucleases