Molecular Species Identification of Newly Hatched Hawaiian Amphidromous Gobioid Larvae |
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Authors: | Daniel P Lindstrom |
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Institution: | (1) Hawaii Institute of Marine Biology/Department of Zoology, University of Hawaii at Manoa, Honolulu, and Department of Biology, Gordon College, Wenham, MA 01984, U.S.A., US |
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Abstract: | This report describes a method for the determination of species identity of newly hatched larvae of five sympatric Hawaiian
amphidromous gobioids (Lentipes concolor, Sicyopterus stimpsoni, Awaous guamensis, Stenogobius hawaiiensis, and Eleotris sandwichensis). Polymerase chain reaction (PCR) was used to amplify a homologous section of the cytochrome b (Cyt b) region of the mitochondrial genome (mtDNA) from adults of all five species. The resulting PCR-amplified DNA was subjected
to restriction fragment length polymorphism (RFLP) analysis producing species-specific restriction patterns. PCR products
from the five species were sequenced to substantiate correct amplification, restriction site locations, and fragment sizes.
The sequence data were also used to construct a phylogenetic tree. Individual, newly hatched, wild-caught larvae of amphidromous
gobioids of unknown species affinity were sorted into six morphotypes based on physical characteristics. These typed larvae
and those from two species that spawned in captivity were subjected to the same molecular analysis as the adults. The RFLP
results from adults and larvae were compared, allowing larval morphotypes to be assigned to the appropriate species. These
comparisons permitted construction of an identification key to the newly hatched larvae of these species based solely on physical
characteristics for use in future field studies.
Received April 29, 1998; accepted September 30, 1998. |
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Keywords: | : fish larvae species identification Hawaii gobioidea PCR RFLP |
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