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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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Abstract

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.

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Acknowledgments

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.

Author Contributions

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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Authors and Affiliations

  1. Department of Natural Resources, Tromsø Museum, UiT The Arctic University of Norway, 9037, Tromsø, Norway

    Dilli P. Rijal, Torbjørn Alm & Inger G. Alsos

  2. School of Biology and Center of Excellence in Phylogeny of Living Organisms, University of Tehran, 14155-6455, Tehran, Iran

    Mohsen Falahati-Anbaran

  3. Department of Biology, Norwegian University of Science and Technology, 7491, Trondheim, Norway

    Mohsen Falahati-Anbaran

Authors
  1. Dilli P. Rijal
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  2. Mohsen Falahati-Anbaran
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  3. Torbjørn Alm
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  4. Inger G. Alsos
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Corresponding author

Correspondence to Dilli P. Rijal.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Online resource 1

Microsatellite library of H. persicum with 3904 sequences containing microsatellite motifs (TXT 1173 kb)

Online resource 2

List of 30 biologically and 134 bioinformatically validated microsatellite markers (XLSX 47 kb)

Fig. S1

Screen shot of some of the markers amplified in H. persicum in a singleplex. PET labeled marker Hp_02 with two alleles at 131.5 and 135.6, Hp_08 with two alleles at 170.6 and 173.5, and Hp_18 with single allele at 256.8 bp (base pair); HEX labeled marker Hp_19 with two alleles at 170 and 180.1, and Hp_05 with one allele at 125 bp; 6-FAM labeled marker Hp_09 with single allele at 159 bp; and NED labeled marker Hp_10 and Hp_16 with single allele each at 187.6 and 135.6 bp respectively (see Tables 3 and 4 of the paper for details) (PDF 81 kb)

Fig. S2

Screen shot of 10 co-amplified markers of multiplex I in H. persicum (see Tables 3 and 4 of the paper for details) (PDF 56 kb)

Fig. S3

Screen shot of 6 co-amplified markers of multiplex II in H. persicum (see Tables 3 and 4 of the paper for details) (PDF 59 kb)

Fig. S4

Screen shot of 9 co-amplified markers of multiplex III in H. persicum (see Tables 3 and 4 of the paper for details) (PDF 64 kb)

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Rijal, D.P., Falahati-Anbaran, M., Alm, T. et al. Microsatellite Markers for Heracleum persicum (Apiaceae) and Allied Taxa: Application of Next-Generation Sequencing to Develop Genetic Resources for Invasive Species Management. Plant Mol Biol Rep 33, 1381–1390 (2015). https://doi.org/10.1007/s11105-014-0841-y

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