Development of Species-Specific Markers of the Tropical Oyster (Crassostrea belcheri) in Thailand |
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Authors: | S Klinbunga P Ampayup A Tassanakajon P Jarayabhand W Yoosukh |
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Institution: | (1) Marine Biotechnology Research Unit, National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Bangkok 10400, Thailand, TH;(2) Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand, TH;(3) Angsila Marine Biological Research Station, Department of Marine Science, Faculty of Science, Chulalongkorn University, Chonburi 20210, Thailand; and Aquatic Resources Research Institute, Chulalongkorn University, Bangkok 10330, Thailand, TH;(4) Department of Marine Science, Faculty of Fisheries, Kasetsart University, Bangkok 10900, Thailand, TH |
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Abstract: | Randomly amplified polymorphic DNA (RAPD) analysis was used to identify species-specific markers of 5 oyster species in Thailand:
Crassostrea belcheri, Crassostrea iredalei, Saccostrea cucullata, Saccostrea forskali, and Striostrea (Parastriostrea) mytiloides. Species-specific markers were found in C. belcheri, C. iredalei, and S. cucullata but not in S. forskali and S. mytiloides. Three C. belcheri–specific RAPD fragments were cloned and sequenced. A primer set was designed from each of the recombinant clones (pPACB1,
pPACB2, and pPACB3). The polymerase chain reaction products showed expected sizes of 536, 600, and 500 bp, respectively, with
the sensitivity of detection approximately 30 pg of C. belcheri total DNA template. The specificity of pPACB1 was examined against 135 individuals of indigenous oyster species in Thailand
and against outgroup references S. commercialis (N= 12) and Perna viridis (N= 12). Results indicated the species-specific nature of primers developed from pPACB1. This primer set can be used for broodstock
selection and determination of C. belcheri larvae to assist the selective breeding program for this commercially important species.
Received December 8, 1999; accepted March 16, 2000. |
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Keywords: | : oyster species-specific markers RAPD PCR Crassostrea belcheri |
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