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Journal Molluscan Studies Advance Access originally published online on March 14, 2009
Journal of Molluscan Studies 2009 75(2):139-146; doi:10.1093/mollus/eyp001
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© The Author 2009. Published by Oxford University Press on behalf of The Malacological Society of London, all rights reserved

Identification of seven Crassostrea oysters from the South China Sea using PCR–RFLP analysis

Jianjun Xia, Ziniu Yu and Xiaoyu Kong

Laboratory of Marine Bio-resource Sustainable Utilization, Laboratory of Applied Marine Biology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou 510301, China

Correspondence: Ziniu Yu; e-mail: carlzyu{at}scsio.ac.cn


  Abstract

We have developed a PCR–RFLP assay of two mitochondrial [16S rDNA and cytochrome oxidase I (COI)] and one nuclear (ITS-1) gene fragment for the identification of seven Crassostrea oysters from the South China Sea. These include the morphologically distinct Crassostrea sikamea and three pairs of morphologically similar species: Crassostrea hongkongensis and Crassostrea ariakensis, Crassostrea gigas and Crassostrea angulata, Crassostrea iredalei and an unidentified Crassostrea sp. All seven species can be distinguished with both mitochondrial markers: DdeI/DraI or AluI/MseI digestion of 16S rDNA and NisI/AluI or NisI/DdeI digestion of COI. Five of the seven species can be distinguished using a single endonuclease digestion of the ITS-1 fragment. Based on genetic distance estimates and on morphology, the newly discovered Crassostrea sp. appears most closely related to C. iredalei. Despite the existence of intra-specific variation at low frequency in some restriction sites, the PCR–RFLP assay developed here can be reliably applied to identify all seven oyster species.

(Received 9 May 2008; accepted 29 December 2008)


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