Microsatellite Markers for Heracleum persicum (Apiaceae) and Allied Taxa: Application of Next-Generation Sequencing to Develop Genetic Resources for Invasive Species Management
- 383 Downloads
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.
- Anonymous (2009) Type-it Microsatellite PCR hand book: for reliable multiplex PCR-based analysis of microsatellites without the need for optimization. Qiagen, Hilden, Germany. http://www.qiagen.com/resources/resourcedetail?id=d6135896-0466-4d9d-aeef-1c4f23f8964e&lang=en. Accessed 15 December 2012
- Anonymous (2011) emPCR amplification method manual - Lib L LV. 454 Sequencing, Branford, Connecticut, USA. http://454.com/downloads/my454/documentation/gs-flx-plus/emPCR-Amp-Lib-L-LV-Method-Manual_XLPlus_May2011.pdf. Accessed 07 November 2014
- Butler J (2005) Constructing STR multiplex assays. In: Carracedo A (ed) Forensic DNA Typing Protocols, vol 297. Methods in Molecular Biology. Humana Press, Totowa, New Jersey, pp 53–65Google Scholar
- Elvebakk A (1992) Has Heracleum laciniatum auct. scand. been introduced to North Norway through England? Polarflokken 16:313–316Google Scholar
- Fading P, Watson MF (2005) Heracleum Linnaeus. In: Zhengyi W, Raven P (eds) Flora of China (Apiaceae through Ericaceae), vol 14. Missouri Botanical Garden Press, St. Louis, pp 194–202Google Scholar
- Fröberg L (2010) Heracleum L. In: Jonsell B, Karlsson T (eds) Flora Nordica (Thymelaeaceae to Apiaceae), vol 6. The Swedish Museum of Natural History, Stockholm, pp 224–234Google Scholar
- Henry P, Provan J, Goudet J, Guisan A, JahodovÁ Š, Besnard G (2008) A set of primers for plastid indels and nuclear microsatellites in the invasive plant Heracleum mantegazzianum (Apiaceae) and their transferability to Heracleum sphondylium. Mol Ecol Resour 8:161–163. doi: 10.1111/j.1471-8286.2007.01911.x CrossRefPubMedGoogle Scholar
- Nielsen C, Ravn HP, Nentwig W, Wade M (eds) (2005) The giant hogweed best practice manual. Guidelines for the management and control of an invasive weed in Europe. Forest and Landscape Denmark, Hoersholm, Denmark, p 44Google Scholar
- Sell P, Murrell G (2009) Flora of Great Britain and Ireland. Volume 3 (Mimosaceae-Lentibulariaceae) vol 3. Cambridge University Press, New YorkGoogle Scholar
- Stace C (2010) New flora of the British Isles, 3rd edn. Cambridge University Press, CambridgeGoogle Scholar
- Thiele J, Otte A (2007) Impact of Heracleum mantegazzianum on invaded vegetation and human activities. In: Pysek P, Cock MJW, Nentwig W, Ravn HP (eds) Ecology and management of giant hogweed (Heracleum mantegazzianum). CABI, Wallingford, pp 144–156Google Scholar