Journal Article

Genetic diversity in <i>Cypripedium calceolus</i> (Orchidaceae) with a focus on north-western Europe, as revealed by plastid DNA length polymorphisms

Michael F. Fay, Ruth Bone, Peter Cook, Imalka Kahandawala, Jennifer Greensmith, Stacey Harris, Henrik Æ. Pedersen, Martin J. Ingrouille and Christian Lexer

in Annals of Botany

Published on behalf of The Annals of Botany Company

Volume 104, issue 3, pages 517-525
Published in print August 2009 | ISSN: 0305-7364
Published online May 2009 | e-ISSN: 1095-8290 | DOI: http://dx.doi.org/10.1093/aob/mcp116
Genetic diversity in Cypripedium calceolus (Orchidaceae) with a focus on north-western Europe, as revealed by plastid DNA length polymorphisms

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Background and Aims

Cypripedium calceolus, although widespread in Eurasia, is rare in many countries in which it occurs. Population genetics studies with nuclear DNA markers on this species have been hampered by its large nuclear genome size. Plastid DNA markers are used here to gain an understanding of variation within and between populations and of biogeographical patterns.

Methods

Thirteen length-variable regions (microsatellites and insertions/deletions) were identified in non-coding plastid DNA. These and a previously identified complex microsatellite in the trnL-trnF intergenic spacer were used to identify plastid DNA haplotypes for European samples, with sampling focused on England, Denmark and Sweden.

Key Results

The 13 additional length-variable regions identified were two homopolymer (polyA) repeats in the rps16 intron and a homopolymer (polyA) repeat and ten indels in the accD-psa1 intergenic spacer. In accD-psa1, most of these were in an extremely AT-rich region, and it was not possible to design primers in the flanking regions; therefore, the whole intergenic spacer was sequenced. Together, these new regions and the trnL-trnF complex microsatellite allowed 23 haplotypes to be characterized. Many were found in only one or a few samples (probably due to low sampling density), but some commoner haplotypes were widespread. Most of the genetic variation was found within rather than between populations (83 vs. 18%, respectively). Two haplotypes occurred from the Spanish Pyrenees to Sweden.

Conclusions

Plastid DNA data can be used to gain an understanding of patterns of genetic variation and seed-mediated gene flow in orchids. Although these data are less information-rich than those for nuclear DNA, they present a useful option for studying species with large genomes. Here they support the hypothesis of long-distance seed dispersal often proposed for orchids.

Keywords: Biogeography; Cypripedium calceolus; genome size; plastid microsatellites; population genetics; seed dispersal

Journal Article.  5626 words.  Illustrated.

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

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