Journal Article

<i>trnL-F</i> is a powerful marker for DNA identification of field vittarioid gametophytes (Pteridaceae)

Cheng Wei Chen, Yao Moan Huang, Li Yaung Kuo, Quoc Dat Nguyen, Hong Truong Luu, John Rey Callado, Donald R. Farrar and Wen Liang Chiou

in Annals of Botany

Published on behalf of The Annals of Botany Company

Volume 111, issue 4, pages 663-673
Published in print April 2013 | ISSN: 0305-7364
Published online February 2013 | e-ISSN: 1095-8290 | DOI: http://dx.doi.org/10.1093/aob/mct004
trnL-F is a powerful marker for DNA identification of field vittarioid gametophytes (Pteridaceae)

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  • Ecology and Conservation
  • Evolutionary Biology
  • Plant Sciences and Forestry

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Background and Aims

The gametophyte phase of ferns plays an important role in habitat selection, dispersal, adaptation and evolution. However, ecological studies on fern gametophytes have been impeded due to the difficulty of species identification of free-living gametophytes. DNA barcoding provides an alternative approach to identifying fern gametophytes but is rarely applied to field studies. In this study, an example of field vittarioid gametophyte identification using DNA barcoding, which has not been done before, is given.

Methods

A combination of distance-based and tree-based approaches was performed to evaluate the discriminating power of three candidate barcodes (matK, rbcL and trnL-F) on 16 vittarioid sporophytes. Sequences of the trnL-F region were generated from 15 fern gametophyte populations by tissue-direct PCR and were compared against the sporophyte dataset, using BLAST.

Key Results trnL-F

earns highest primer universality and discriminatory ability scores, whereas PCR success rates were very low for matK and rbcL regions (10·8 % and 41·3 %, respectively). BLAST analyses showed that all the sampled field gametophytes could be successfully identified to species level. Three gametophyte populations were also discovered to be living beyond the known occurrence of their sporophyte counterparts.

Conclusions

This study demonstrates that DNA barcoding (i.e. reference databasing, tissue-direct PCR and molecular analysis), especially the trnL-F region, is an efficient tool to identify field gametophytes, and has considerable potential in exploring the ecology of fern gametophytes.

Keywords: DNA barcoding; field gametophyte; independent gametophyte; vittarioid fern

Journal Article.  5577 words.  Illustrated.

Subjects: Ecology and Conservation ; Evolutionary Biology ; Plant Sciences and Forestry

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