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Chloroplast microsatellite markers reveal Zannichellia haplotypes across Europe using herbarium DNA

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The successful amplification of consensus chloroplast microsatellite markers (cpSSRs) from 22-year-old dried fruits of Zannichellia, originally sampled as a reference herbarium collection for an isozyme study, is reported. The DNA obtained from herbarium tissues was not degraded and revealed amplicons of the same length in old achenes and leaves than in fresh materials of both tissues from the same site. DNA extraction was also performed on single achenes of about 1 mg dry weight. Out of the ten cpSSRs tested, four gave reliable PCR products in both fresh and dried plant materials. Three polymorphic cpSSR loci allowed to distinguish five haplotypes, A, B, C, D, and E from the ancient DNA. Haplotype A is widespread in Z. palustris subsp. palustris and subsp. repens from West-European inland fresh water localities and in Z. major from the brackish waters of the Baltic Sea. Haplotype B is found in Z. palustris from the Baltic Sea, whereas haplotype C seems to be confined to Z. pedunculata, a taxon from northwestern Europe. Haplotypes D and E were observed in Z. palustris and Z. peltata, respectively, from the Mediterranean region. A haplotype network and neighbor-joining tree revealed the possibility of a cryptic species within Z. palustris. The chloroplast microsatellite primers show a potential in detecting haplotypes of Zannichellia, especially at taxon level, as well as in detecting geographical patterns across distant European fresh and brackish water habitats.


Document Type: Research Article

Publication date: 2007-06-01

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  • The Belgian Journal of Botany (now known as Plant Ecology and Evolution) is an international journal open to all fields of plant sciences. Please note, however, that papers restricted to purely nomenclatural matters or to floristical data of only local interest will not be accepted. The Journal appears in one volume of two issues per year. It publishes reviews, original research papers, short notes, letters to the editor, and book reviews. Click here for current issues of this journal
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