Conservation Genetics

, Volume 13, Issue 1, pp 21–37 | Cite as

Combining multiple analytical approaches for the identification of population structure and genetic delineation of two subspecies of the endemic Arabian burnet moth Reissita simonyi (Zygaenidae; Lepidoptera)

  • C. F. C. Klütsch
  • R. J. Dyer
  • B. Misof
Research Article


Habitat fragmentation and landscape topology may influence the genetic structure and connectivity between natural populations. Six microsatellite loci were used to infer the population structure of 35 populations (N = 788) of the alpine Arabian burnet moth Reissita simonyi (Lepidoptera, Zygaenidae) in Yemen and Oman. Due to the patchy distribution of larval food plants, R. simonyi is not continuously distributed throughout the studied area and the two recognized subspecies of this endemic species (Reissita s. simonyi/R. s. yemenicola) are apparently discretely distributed. All microsatellites showed prevalence of null alleles and therefore a thorough investigation of the impact of null alleles on different population genetic parameters (F ST, inbreeding coefficients, and Population Graph topologies) is given. In general, null alleles reduced genetic covariance and independence of allele frequencies resulting in a more connected genetic topology in Population Graphs and an overestimation of pairwise F ST values and inbreeding coefficients. Despite the presence of null alleles, Population Graphs also showed a much higher genetic connectivity within subspecies (and lower genetic differentiation (via F ST)) than between; supporting existing taxonomic distinction. Partial Mantel tests showed that both geographical distance and altitude were highly correlated with the observed distribution of genetic structure within R. simonyi. In conclusion, we identified geographical and altitudinal distances in R. simonyi as well as an intervening desert area to be the main factors for spatial genetic structure in this species and show that the taxonomic division into two subspecies is confirmed by genetic analysis.


Reissita simonyi Microsatellites Isolation-by-distance Isolation-by-altitude Landscape connectivity Null alleles 



We would like to thank especially Abdul Karim Nasher and Masaá Al-Jumaily (Sanaá) for their hospitality and invaluable help in Yemen. We thank the Yemeni Scientific Research Foundation (YSRF) for providing visa invitations. We are very grateful to Dr. Mohamed Ali Hubaishan (AREA Mukalla), Abdul Gadr Al-Haimi, Dr. Gabr Al-Kirshi (GTZ office, Taiz), Peter Hein, Norbert Kilian, Harald Kürschner (all FU Berlin), Christoph Oberprieler and Jörg Meister (Regensburg) Bruno Mies (Essen), Matthias Schultz (Hamburg), Tony van Haarten (Sanaá) and all master students, who worked within the BIOTA-EAST project. We also thank Marie-Pierre Chapuis for answering our questions regarding FreeNA. Special thanks to all lab members and technicians of the ZFMK in Bonn. This is a publication of the ZFMK Molecular Systematic Unit. We thank the German Federal Ministry of Education and Research (BMBF, Germany) for supporting this study, grant no. 01LC0025/E16 within the frame of the BIOLOG program, subprogram BIOTA-East. The simulation work was supported in part by National Science Foundation grants DEB-0543102 and DEB-0640803 to RJD.

Supplementary material

10592_2011_259_MOESM1_ESM.pdf (205 kb)
Online Resource 1: a) Pairwise FST values among 35 populations examined in this study. Corrected pairwise FST values using the ENA method (lower triangle matrix: Chapuis & Estoup 2007) to account for the presence of null alleles and uncorrected pairwise FST values (upper triangle matrix). b) Geographical and altitudinal distances among 35 populations. Geographical distances between localities (in km; lower triangle matrix) and altitudinal distances (in m; upper triangle matrix). (PDF 204 kb)
10592_2011_259_MOESM2_ESM.pdf (43 kb)
Online Resource 2: Average null allele frequencies for all populations and each microsatellite locus separately using the IIM algorithm implemented in INEST (Chybicki & Burczyk 2009). In parentheses, (RSY) and (RSS) indicate the two respective subspecies, R. s. yemenicola and R. s. simonyi. (PDF 43 kb)
10592_2011_259_MOESM3_ESM.pdf (27 kb)
Online Resource 3: Average null allele frequencies for populations and each microsatellite locus separately using the EM algorithm (Dempster et al. 1977) implemented in FREENA (Chapuis & Estoup 2007). In parentheses, (RSY) and (RSS) indicate the respective subspecies, R. s. yemenicola and R. s. simonyi. (PDF 27 kb)
10592_2011_259_MOESM4_ESM.pdf (58 kb)
Online Resource 4: Pairwise genetic distances (Dc; Cavalli-Sforza and Edwards 1967) among 35 populations using the INA method (including null alleles) implemented in FreeNA (Chapuis & Estoup 2007). (PDF 58 kb)
10592_2011_259_MOESM5_ESM.pdf (54 kb)
Online Resource 5: Density distribution of the number of edges in population graphs with simulated null alleles (control = no null alleles, rare null = lowest frequency treated as null, common null = most frequent allele treated as null). Mean differences are marginally significant (Kruskal–Wallis Test; χ2 = 7.1683, df = 2, P = 0.028) but were not different in variance (modified Kruskal–Wallis; χ2 = .375, df = 2, P = 0.8292). (PDF 54 kb)


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Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  1. 1.Zoologisches Forschungsmuseum Alexander KoenigBonnGermany
  2. 2.Department of BiologyVirginia Commonwealth UniversityRichmond VirginiaUSA
  3. 3.Biozentrum Grindel & Zoologisches MuseumHamburgGermany
  4. 4.Biology DepartmentTrent UniversityPeterboroughCanada

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