Application of target region amplification polymorphism (TRAP) technique to Porphyra (Bangiales, Rhodophyta) fingerprinting

Abstract

L. Qiao, H. Liu, J. Sun, F. Zhao, B. Guo, M. Weng, T. Liu, J. Dai, and B. Wang. 2007. Application of target region amplification polymorphism (TRAP) technique to Porphyra (Bangiales, Rhodophyta) fingerprinting. Phycologia 46: 450–455. DOI: 10.2216/07-08.1

To demonstrate the applicability of the target region amplification polymorphism (TRAP) marker technique to Porphyra, 15 Porphyra lines, representing three species, were fingerprinted with 32 primer-combinations. Ten primer-combinations that gave a stable and reproducible amplification pattern were selected. In total 432 fragments from 50 to 500 base pairs (bp) in length were amplified with these 10 selected primer-combinations, and all the fragments were polymorphic. The 432 fragments were scored respectively and used to construct a dendrogram of the 15 Porphyra lines with unweighted pair-group method arithmetic average (UPGMA). These Porphyra lines were divided into two major groups at the 0.71 similarity level. This result is basically consistent with the conventional Porphyra taxonomy. Twelve of the 432 fragments, which were amplified by two primer-combinations, Tps/1F and Tps/3F, were chosen and used to develop the DNA fingerprints of these Porphyra lines. The developed DNA fingerprints then were converted into binary codes and resulted in one unique binary code for each of the 15 Porphyra lines. This paper is the first report concerning the application of TRAP technique to seaweeds. Our results suggest that TRAP is a simple, stable, polymorphic and reproducible molecular marker technique for the identification, classification and resource protection of Porphyra lines.

Key Words::

• DNA fingerprinting
• Porphyra lines
• Target region amplification polymorphism (TRAP)

ACKNOWLEDGMENTS

This work was partially supported by the Hi-Tech and Development Program of China (2006AA10A412). The authors acknowledge Drs Delin Duan and Pu Xu for providing Porphyra materials.