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JPR Advance Access published online on January 19, 2006
Journal of Plankton Research, doi:10.1093/plankt/fbi122
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© The Author 2006. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
Received October 16, 2005
Accepted January 17, 2006

Article

A new strategy for species identification of planktonic larvae: PCR-RFLP analysis of the internal transcribed spacer region of ribosomal DNA detected by agarose gel electrophoresis or DHPLC

Shi Wang 1, Zhenmin Bao 1 *, Lingling Zhang 1, Ning Li 1, Aibin Zhan 1, Wenbo Guo 2, Xiaolong Wang 1, and Jingjie Hu 1

1 Laboratory of Marine Genetics and Breeding (MGB), Division of Life Science and Technology, Ocean University of China, Qingdao 266003, People’s Republic of China
2 Key Laboratory of Mariculture (KLM), Division of Life Science and Technology, Ocean University of China, Qingdao 266003, People’s Republic of China

* To whom correspondence should be addressed.
Zhenmin Bao, E-mail: zmbao{at}ouc.edu.cn


  Abstract

Planktonic survey is important for understanding the dynamics and structure of populations or communities. However, the small size of early planktonic larvae, usually less than 500µm, make it difficult or impossible to discriminate closely related taxa based on morphological characters. We developed a new strategy for species identification of planktonic larvae, namely, polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) analysis of the internal transcribed spacer (ITS) region detected by agarose gel electrophoresis or denaturing high-performance liquid chromatography (DHPLC). A total of 4 restriction enzymes were selected for PCR-RFLP analysis. Based on any one of 4 restriction maps, 12 commercial shellfish species could be differentiated from each other in agarose gel analysis. But more accurate results were obtained in DHPLC analysis. As an example of larval identification, hybrid larvae were successfully identified by their showing diagnostic peaks of both parents in DHPLC analysis. These results suggest that this strategy can meet the demands for plankton survey and studies of hybridization.

Communicating Editor: KJ Flynn
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