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Vol. 53, 2014
Genetic identification method for two subspecies of the
Indonesian short-finned eel, Anguilla
bicolor, using an
allelic discrimination technique
Chikaya
Tanaka1, Fumiaki Shirotori1, Masaki Sato1, Mina Ishikawa1, Akira
Shinoda2, Jun Aoyama3 and Tatsuki Yoshinaga1*
1School
of Marine Biosciences, Kitasato University, Kanagawa 252-0373, Japan
2Department of Biology, Tokyo Medical University,
Tokyo 160-8402, Japan
3Atmosphere and Ocean Research Institute, The
University of Tokyo, Chiba 277-8564, Japan
Abstract
Background: There is an increasing
pressure on tropical species of the freshwater eel genus Anguilla for use in aquaculture, so
species identification methods are needed to monitor these activities
and to facilitate conservation efforts. A method was developed to
genetically distinguish two subspecies of the Indonesian short-finned
eel, Anguilla bicolor, based on an allelic
discrimination technique.
Results: A single DNA nucleotide substitution in
the mitochondrial DNA 16S rRNA gene was employed to identify the two
subspecies where Anguilla bicolor bicolor and Anguilla bicolor pacifica possessed adenine and
guanine, respectively. This substitution was highly conserved at 100%
in A. bicolor bicolor
(108/108) and 99.9% in A. bicolor
pacifica (181/182), and the misidentification rate was estimated
to be 0.34%. Subsequently, fluorescent-labeled oligo probes and PCR
primers were designed and succeeded to clearly distinguish the two
subspecies. Further, the other ten anguillid species that may be
sympatrically distributed with A.
bicolor showed negative results.
Conclusions: The method developed
in this study is useful to accurately identify the two subspecies of A. bicolor and can contribute to
ecological studies, stock management, and conservation.
Key words: Allelic discrimination; Anguilla bicolor; Indonesian short-finned
eel; Genetic identification; Red list.
*Correspondence: E-mail: yosinaga@kitasato-u.ac.jp
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