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Morphological variability, cytotype diversity, and cytogeography of populations traditionally called Dactylorhiza fuchsii in Central Europe

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Abstract

The morphological variation and cytotype diversity were investigated among Central European populations traditionally recognized as Dactylorhiza fuchsii, recently incorporated in D. maculata s.l. Flow cytometry was employed to assess the ploidy levels of 738 individuals from 77 localities and multivariate morphometrics for a total of 531 individuals from 27 localities. Three ploidy levels were found: diploid (2n = 2x = 40), DNA-triploid and tetraploid (2n = 4x = 80). Whereas diploids and tetraploids often occurred as pure-cytotype populations, individuals of DNA-triploids always co-occurred with at least one of the other cytotypes. Qualitative morphological traits were inferred to be the most important drivers of morphological variation among the investigated plants, with the most striking differences in flower colouration and leaf spotting. The combination of morphological and cytological characters enabled to delimit two separate groups of populations. The first corresponded to D. maculata subsp. fuchsii with morphologically indistinguishable diploid, DNA-triploid and tetraploid individuals, sometimes occurring in mixed-ploidy populations. A complex geographical pattern of cytotype distributions was observed, with diploids scatteredly occurring throughout Central Europe except for Bohemian Massif, which was dominated by tetraploids. The other group of populations represented newly described in this study D. maculata subsp. sooana, subsp. nova, morphologically well-defined and strictly diploid taxon with a restricted geographical range, occurring in the Western Carpathians. A new combination for a hybrid taxon D. × dinglensis nothosubsp. smitakii, comb. nova (= D. maculata subsp. sooana × D. majalis subsp. majalis), was also proposed.

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Acknowledgements

We are thankful to many botanists who guided us in the field, helped us with searching localities or samplings, namely, A. Čejková, I. Jongepierová, A. Kantor, N. Kantorová, J. Košnar, E. Pietorová, J. Plekanec, M. Popelářová, M. & R. Puscarciuc, P. Salenka, A. Schmotzer, G. Schneeweiss, R. Štencl, Z. Václavová, V. Vlačiha, M. & M. Wolanin and many others. We are grateful to M. Hroneš and M. Jandová for service of the FCM laboratory at the Palacký University and their methodological support, as well as J. Doležel and J. Vrána, who enabled us to work in the laboratory of the Institute of Experimental Botany in Olomouc. We furthermore appreciate M. Oulehlová, who scanned the type specimen, and P. Trávníček for valuable consultations. We are grateful to both reviewers for critical reading and improvement of the manuscript. The authorities in nature conservation are acknowledged for granting us with the necessary permits. Research was supported by internal grant of the Palacký University IGA_PrF_2021_001 and by Austrian Federal Ministry of Education, Science and Research (BMBWF) within program Aktion CZ-AT, administrated by OeAD-GmbH.

Funding

VT was supported by internal grant of the Palacký University IGA_PrF_2021_001 and by Austrian Federal Ministry of Education, Science and Research within program Aktion CZ-AT, administrated by OeAD-GmbH.

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Authors and Affiliations

  1. Plant Biosystematics & Ecology RG, Department of Botany, Faculty of Science, Palacký University, Šlechtitelů 27, 783 71, Olomouc, Czech Republic

    Vojtěch Taraška, Martin Duchoslav & Bohumil Trávníček

  2. Regional Museum in Jeseník, Zámecké náměstí 1, 790 01, Jeseník, Czech Republic

    Vojtěch Taraška

  3. U Trojáku 4644, 760 05, Zlín, Czech Republic

    Petr Batoušek

  4. Department of Botany and Biodiversity Research, Faculty of Life Sciences, University of Vienna, Rennweg 14, 1030, Wien, Austria

    Eva M. Temsch & Hanna Weiss-Schneeweiss

Authors
  1. Vojtěch Taraška
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  2. Petr Batoušek
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  3. Martin Duchoslav
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  4. Eva M. Temsch
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  5. Hanna Weiss-Schneeweiss
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  6. Bohumil Trávníček
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Contributions

VT, PB, and BT conceived the project and conducted the field work. VT, EMT and HWS performed the analysis of genome size and chromosome numbers. MD designed and conducted the statistical analysis. VT drafted the manuscript with significant contributions of MD and BT. All authors commented on and approved the manuscript. BT supervised the project.

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Correspondence to Vojtěch Taraška.

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Supplementary Information

Information on Electronic Supplementary Material

Information on Electronic Supplementary Material

Online Resource 1. Locality details of Dactylorhiza *fuchsii.

Online Resource 2. Details to (sub)populations of Dactylorhiza *fuchsii. Averaged relative DNA content (= fluorescence ratios between the positions of the sample and internal reference standard G0/G1 peaks) of investigated populations.

Online Resource 3. Explanations to quantitative characters used in the morphometrics.

Online Resource 4. Relative DNA content (= fluorescence ratios between the positions of the sample and internal reference standard G0/G1 peaks) of six plants with counted chromosome numbers; the stain was either DAPI or PI. All values are calculated relative to the Pisum sativum cv. ‘Ctirad’ as internal reference standard.

Online Resource 5. Absolute genome sizes (GS) of five individuals of Dactylorhiza *fuchsii estimated by flow cytometry.

Online Resource 6. Box plots of characters analysed for the fuchsii-2x, fuchsii-4x and sooana groups.

Online Resource 7. Holotype of Dactylorhiza maculata subsp. sooana Batoušek, Taraška & Trávn.

Online Resource 8. Images of Dactylorhiza maculata subsp. sooana, D. maculata subsp. fuchsii and D. ×dinglensis nothosubsp. smitakii.

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Taraška, V., Batoušek, P., Duchoslav, M. et al. Morphological variability, cytotype diversity, and cytogeography of populations traditionally called Dactylorhiza fuchsii in Central Europe. Plant Syst Evol 307, 51 (2021). https://doi.org/10.1007/s00606-021-01770-3

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