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Conservation Genetics

, Volume 13, Issue 1, pp 89–98 | Cite as

Genetic diversity, compatibility patterns and seed quality in isolated populations of Cypripedium calceolus (Orchidaceae)

  • Henrik Æ. Pedersen
  • Hanne N. Rasmussen
  • Imalka M. Kahandawala
  • Michael F. Fay
Research Article

Abstract

Cypripedium calceolus has suffered an alarming decline, and today mainly occurs in small and isolated populations. In Denmark there are only two populations, close to each other and situated far from other European stands. One population is stagnant or in slow decline, whereas the other is in rapid increase. We examined the levels of genetic diversity and compatibility and seed quality following experimental crosses. No genetic variation could be detected in plastid and nuclear markers within or between the two populations—in contrast to results previously reported from other European populations of C. calceolus. This may indicate a founder effect in both populations, but it could also be the outcome of prolonged inbreeding or reflect a genetic bottleneck after the populations were established. According to fruit dimensions and frequency of fully developed seeds there was full self-compatibility in the stagnant population, and partial late-acting self-incompatibility in the proliferating population. In combination with previous reports from other countries, this suggests that several self-incompatibility systems may occur in C. calceolus. Seeds from the older and stagnant population performed more poorly in germination tests in vitro than seeds from the thriving population. The difference needs not be genetically based, but could be due to environmental differences during seed maturation, producing different seed quality or dormancy characteristics. However, low level of genetic diversity within the populations may affect their ability to adapt and the possibility of inbreeding depression should be investigated.

Keywords

Asymbiotic germination Conservation Inbreeding depression Microsatellites Seed quality Self-incompatibility 

Notes

Acknowledgments

This project was supported financially by the Spar Nord Foundation and “Det Obelske Familiefond”, and public and private land owners kindly allowed us to work on their premises. The genetic study was in part funded by Natural England. We gratefully thank the Danish nature conservation authorities for readily issuing all necessary permits, Aage Pedersen and Kurt Æ. Pedersen for assistance in the field and Tiiu Kull for providing Estonian literature.

References

  1. Antonelli A, Dahlberg CJ, Carlgren KHI, Appelqvist T (2009) Pollination of the lady’s slipper orchid (Cypripedium calceolus) in Scandinavia—taxonomic and conservational aspects. Nord J Bot 27:266–273CrossRefGoogle Scholar
  2. Borba EL, Semir J, Shepherd GJ (2001) Self-incompatibility, inbreeding depression and crossing potential in five Brazilian Pleurothallis (Orchidaceae) species. Ann Bot 88:89–99CrossRefGoogle Scholar
  3. Brachet S, Jubier MF, Richard M, Jung-Muller B, Frascaria-Lacoste N (1999) Rapid identification of microsatellite loci using 5′ anchored PCR in the common ash Fraxinus excelsior. Mol Ecol 8:160–163Google Scholar
  4. Brigham CA (2003) Factors affecting persistence in formerly common and historically rare plants. In: Brigham CA, Schwartz MW (eds) Population viability in plants: conservation, management, and modeling of rare plants. Springer, Berlin, pp 59–97Google Scholar
  5. Brzosko E, Ratkiewicz M, Wróblewska A (2002) Allozyme differentiation and genetic structure of the lady’s slipper (Cypripedium calceolus) island populations in north-east Poland. Bot J Linn Soc 138:433–440CrossRefGoogle Scholar
  6. Brzosko E, Wróblewska A, Ratkiewicz M, Till-Bottraud I, Nicole F, Baranowska U (2009) Genetic diversity of Cypripedium calceolus at the edge and in the centre of its range in Europe. Ann Bot Fenn 46:201–214Google Scholar
  7. Charlesworth D, Charlesworth B (1987) Inbreeding depression and its evolutionary consequences. Annu Rev Ecol Syst 18:237–268CrossRefGoogle Scholar
  8. Charlesworth D, Charlesworth B (1990) Inbreeding depression with heterozygote advantage and its effect on selection for modifiers changing the outcrossing rate. Evolution 44:870–888CrossRefGoogle Scholar
  9. Cribb P (1997) The genus Cypripedium. Timber Press, Portland, The Royal Botanic Gardens, KewGoogle Scholar
  10. Darwin C (1877) The various contrivances by which orchids are fertilised by insects, 2nd edn. John Murray, LondonGoogle Scholar
  11. Daumann E (1968) Zur Bestäubungsökologie von Cypripedium calceolus L. Österr Bot Z 115:434–446CrossRefGoogle Scholar
  12. DeMauro MM (1993) Relationship of breeding system to rarity in the lakeside daisy (Hymenoxys acaulis var. glabra). Conserv Biol 7:542–550CrossRefGoogle Scholar
  13. Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull Bot Soc Am 19:11–15Google Scholar
  14. Dudash MR, Fenster CB (2000) Inbreeding and outbreeding depression in fragmented populations. In: Young AG, Clarke GM (eds) Genetics, demography and viability of fragmented populations. Cambridge University Press, Cambridge, pp 35–53CrossRefGoogle Scholar
  15. Ellstrand NC, Elam DR (1993) Population genetic consequences of small population size: implications for plant conservation. Annu Rev Ecol Syst 24:217–242CrossRefGoogle Scholar
  16. Erneberg M, Holm B (1999) Bee size and pollen transfer in Cypripedium calceolus (Orchidaceae). Nord J Bot 19:363–367CrossRefGoogle Scholar
  17. Fay MF, Cowan RS (2001) Plastid microsatellites in Cypripedium calceolus (Orchidaceae): genetic fingerprints from herbarium specimens. Lindleyana 16:151–156Google Scholar
  18. Fay MF, Bone R, Cook P, Kahandawala I, Greensmith J, Harris S, Pedersen HÆ, Ingrouille MJ, Lexer C (2009) Genetic diversity in Cypripedium calceolus (Orchidaceae) with a focus on northwestern Europe, as revealed by plastid DNA length polymorphisms. Ann Bot 104:517–525PubMedCrossRefGoogle Scholar
  19. Ferdy JB, Loriot S, Sandmeier M, Lefranc M, Raquin C (2001) Inbreeding depression in a rare deceptive orchid. Can J Bot 79:1181–1188CrossRefGoogle Scholar
  20. Ferrer MM, Good-Avila SV, Montaña C, Domínguez CA, Eguiarte LE (2009) Effect of variation in self-incompatibility on pollen limitation and inbreeding depression in Flourensia cernua (Asteraceae) scrubs of contrasting density. Ann Bot 103:1077–1089PubMedCrossRefGoogle Scholar
  21. Fischer M, Matthies D (1998) RAPD variation in relation to population size and plant fitness in the rare Gentianella germanica (Gentianaceae). Am J Bot 85:811–819PubMedCrossRefGoogle Scholar
  22. Fischer M, Matthies D, Schmid B (1997) Responses of rare calcareous grassland plants to elevated CO2: a field experiment with Gentianella germanica and Gentiana cruciata. J Ecol 85:681–691CrossRefGoogle Scholar
  23. Fischer M, van Kleunen M, Schmid B (2000) Genetic Allee effects on performance, plasticity and developmental stability in a clonal plant. Ecol Lett 3:530–539CrossRefGoogle Scholar
  24. Grøntved J (1948) Danmarks Topografisk-Botaniske Undersøgelse iværksat af Dansk Botanisk Forening. Nr. 15. Orchidéernes Udbredelse i Danmark. Bot Tidsskr 47:277–370Google Scholar
  25. Hiscock SJ, McInnis SM (2003) The diversity of self-incompatibility systems in flowering plants. Plant Biol 5:23–32CrossRefGoogle Scholar
  26. Honnay O, Bossuyt B (2005) Prolonged clonal growth: escape route or route to extinction? Oikos 108:427–432CrossRefGoogle Scholar
  27. Husband BC, Schemske DW (1996) Evolution of the magnitude and timing of inbreeding depression in plants. Evolution 50:54–70CrossRefGoogle Scholar
  28. Kahandawala IM (2009) Genome size evolution and conservation genetics in Cypripedium (Orchidaceae). PhD thesis, Birkbeck, University of LondonGoogle Scholar
  29. Kahn AP, Morse DH (1991) Pollinium germination and putative ovule penetration in self- and cross-pollinated common milkweed Asclepias syriaca. Am Midl Nat 126:61–67CrossRefGoogle Scholar
  30. Keller LF, Waller DM (2002) Inbreeding effects in wild populations. Trends Ecol Evol 17:230–241CrossRefGoogle Scholar
  31. Kéry M, Matthies D, Spillmann HH (2000) Reduced fecundity and offspring performance in small populations of the declining grassland plants Primula veris and Gentiana lutea. J Ecol 88:17–30CrossRefGoogle Scholar
  32. Kull T (1988) Identification of clones in Cypripedium calceolus (Orchidaceae). Proc Acad Sci Est SSR Biol 37:195–198 + 1 plGoogle Scholar
  33. Kull T (1998) Fruit-set and recruitment in populations of Cypripedium calceolus L. in Estonia. Bot J Linn Soc 126:27–38Google Scholar
  34. Kull T (1999) Biological flora of the British Isles no. 208. List Br. Vasc. Pl. (1958) 623, 1. Cypripedium calceolus L. J Ecol 87:913–924CrossRefGoogle Scholar
  35. Kull T, Paaver T (1997) Patterns of aspartate amino-transferase variation in relation to population size, founder effect and phytogeographic history in Cypripedium calceolus. Proc Est Acad Sci Biol Ecol 46:4–11Google Scholar
  36. Lange J (1885) Mødet d. 23e Oktober 1884. Medd Bot Foren Kjøbenhavn 1:147–149Google Scholar
  37. Les DH, Reinartz JA, Esselman EJ (1991) Genetic consequences of rarity in Aster furcatus (Asteraceae), a threatened, self-incompatible plant. Evolution 45:1641–1650CrossRefGoogle Scholar
  38. Light MHS, MacConaill M (2002) Climatic influences on flowering and fruiting of Cypripedium parviflorum var. pubescens. In: Kindlmann P, Willems JH, Whigham DF (eds) Trends and fluctuations in terrestrial orchid populations. Backhuys Publishers, Leiden, pp 85–97Google Scholar
  39. Malmgren S (1993) Asymbiontisk fröförökning i stor skala av Anacamptis, Ophrys, Orchis och andra orkideer med runda rotknöler. Sven Bot Tidskr 87:221–234Google Scholar
  40. McGowen MH, Vaillancourt RE, Pilbeam DJ, Potts BM (2010) Sources of variation in self-incompatibility in the Australian forest tree, Eucalyptus globulus. Ann Bot 105:737–745PubMedCrossRefGoogle Scholar
  41. Nicolé F, Brzosko E, Till-Bottraud I (2005) Population viability analysis of Cypripedium calceolus in a protected area: longevity, stability and persistence. J Ecol 93:716–726CrossRefGoogle Scholar
  42. Nilsson LA (1979) Anthecological studies on the lady’s slipper, Cypripedium calceolus (Orchidaceae). Bot Not 132:329–347Google Scholar
  43. Ramsay MM, Dixon KW (2003) Propagation science, recovery and translocation of terrestrial orchids. In: Dixon KW, Kell SP, Barrett RL, Cribb PJ (eds) Orchid conservation. Natural History Publications (Borneo), Kota Kinabalu, pp 259–288Google Scholar
  44. Ramsay MM, Stewart J (1998) Re-establishment of the lady’s slipper orchid (Cypripedium calceolus L.) in Britain. Bot J Linn Soc 126:173–181Google Scholar
  45. Rasmussen HN (1995) Terrestrial orchids from seed to mycotrophic plant. Cambridge University Press, CambridgeCrossRefGoogle Scholar
  46. Rasmussen HN, Whigham DF (1993) Seed ecology of dust seeds in situ: a new study technique and its application in terrestrial orchids. Am J Bot 80:1374–1378CrossRefGoogle Scholar
  47. Seavey SR, Bawa KS (1986) Late-acting self-incompatibility in angiosperms. Bot Rev 52:195–219CrossRefGoogle Scholar
  48. Smithson A (2006) Pollinator limitation and inbreeding depression in orchid species with and without nectar rewards. New Phytol 169:419–430PubMedCrossRefGoogle Scholar
  49. Sprunger S (2001) The cultivation of Cypripedium calceolus from seedlings. Orchid Rev 109:118–120Google Scholar
  50. Swofford DL (1998) PAUP* 4.0b: phylogenetic analysis using parsimony (and other methods). Sinauer, SunderlandGoogle Scholar
  51. Terschuren J (1999) Action plan for Cypripedium calceolus in Europe. Nature and environment no. 100. Council of Europe Publishing, StrasbourgGoogle Scholar
  52. Thomsen L, Ankersen H (1996) Fruesko (Cypripedium calceolus) er i reproduktionsbiologisk krise ved Buderupholm–men hvorfor? URT 20:35–39Google Scholar
  53. Torelli G (1989) Impollinazione del Cypripedium calceolus L. http://www.alexdepri.it/goto/orchids/orchidsfile/cypripcalceolus_imp1
  54. Vandepitte K, Roldán-Ruiz I, Jacquemyn H, Honnay O (2010) Extremely low genotypic diversity and sexual reproduction in isolated populations of the self-incompatible lily-of-the-valley (Convallaria majalis) and the role of the local forest environment. Ann Bot 105:769–776PubMedCrossRefGoogle Scholar
  55. Whitlock MC, Ingvarsson PK, Hatfield (2000) Local drift load and the heterosis of interconnected populations. Heredity 84:452–457PubMedCrossRefGoogle Scholar
  56. Wind P (1997) Overvågning af danske orkidéer 1987–1995. Flora Fauna (Aarhus) 103:49–71Google Scholar
  57. Wind P (2000) Naturovervågning. Overvågning af orkideer 2000. Arbejdsrapport fra DMU nr. 163. Danmarks Miljøundersøgelser, Miljøministeriet, KaløGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Henrik Æ. Pedersen
    • 1
  • Hanne N. Rasmussen
    • 2
  • Imalka M. Kahandawala
    • 3
  • Michael F. Fay
    • 3
  1. 1.Botanical Garden and Museum, Natural History Museum of DenmarkUniversity of CopenhagenCopenhagen KDenmark
  2. 2.Forest and Landscape, LIFE FacultyUniversity of CopenhagenFrederiksberg CDenmark
  3. 3.Jodrell Laboratory, Royal Botanic GardensKew, RichmondUK

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