Microsatellite Markers for Heracleum persicum (Apiaceae) and Allied Taxa: Application of Next-Generation Sequencing to Develop Genetic Resources for Invasive Species Management
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Invasive giant hogweeds are infamous in Europe for causing ecological and economic damage, but genetic resources for their study are limited. We used next-generation sequencing to develop a microsatellite library for Heracleum persicum, a widespread invasive in Nordic countries. In addition, these markers were cross amplified with the closely related Heracleum mantegazzianum, Heracleum sosnowskyi, Heracleum sphondylium, and the putative hybrid H. persicum × H. sphondylium, as well as the more distantly related Anthriscus sylvestris. We designed and validated 164 primer pairs. A cost-effective PCR approach with modified forward primer, reverse primer, and fluorescently labeled universal tail was used to test the functionality of each marker. Twenty-five of thirty markers screened on eight geographically distant samples of H. persicum were polymorphic. The number of alleles was 2–4 whereas the expected and observed heterozygosity varied from 0.06 to 0.84 and 0.0 to 1.0 respectively. The cross-species amplification efficiency was 84–100 %, in which 60–76 % of the cross-species amplified markers were polymorphic for Heracleum taxa including H. persicum × sphondylium. Three out of eight of the cross-amplified markers were polymorphic in Anthriscus sylvestris. Ordination revealed a clear genetic structure of Heracleum taxa. Thus, these markers can serve as important genetic resources for understanding taxonomy, population genetics, and phylogeny of giant hogweeds and their hybrids, which in turn, is expected to contribute to invasive species management.
KeywordsInvasive species Giant hogweeds Hybridization Microsatellites Universal tails Biodiversity conservation
We would like to thank Atefeh Pirany for collecting samples from Iran. We are grateful to Jens-Petter Kjemprud, Embassy of Norway in Iran, for delivering samples from Iran. We would also like to thank Sarka Jahodova for sample contribution. The authors are thankful to all the sample contributors listed in Tables 1 and 2. We would like to thank two anonymous reviewers for their constructive comments which were invaluable while preparing this manuscript. Finally, we would like to thank Matthew Perisin from Department of Ecology and Evolution, University of Chicago, for language correction. This project was funded by Tromsø Museum, UiT—the Arctic University of Norway.
DPR, TA, and IGA designed the project, obtained funding, and performed the fieldwork. MFA instructed DPR during primer optimization and laboratory work. DPR performed the laboratory work and wrote the manuscript. All coauthors commented on the manuscript.
Conflict of Interest
The authors declare no conflict of interest.
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