Development and characterization of new polymorphic microsatellite markers in four sea anemones: Entacmaea quadricolor, Heteractis magnifica, Stichodactyla gigantea, and Stichodactyla mertensii

  • Remy Gatins
  • Pablo Saenz-Agudelo
  • Anna Scott
  • Michael L. Berumen
Original Paper

Abstract

Relatively few studies have investigated the genetic population structure of sea anemones. This is particularly true for sea anemones that host some of the most iconic fishes on coral reefs, the anemonefishes. One of the main reasons for this knowledge gap is the lack of appropriate genetic markers. We developed and characterized a total of 47 novel polymorphic microsatellite markers for four host sea anemone species from the Indo-Pacific: Entacmaea quadricolor (n = 16 microsatellite markers), Heteractis magnifica (n = 8), Stichodactyla mertensii (n = 13), and Stichodactyla gigantea (n = 10). Here, we report genetic diversity statistics from two different sampling locations for each anemone species. Overall, we found that most markers were highly polymorphic. On average, we found a mean of seven alleles per locus. Observed and expected heterozygosities displayed high variation among loci, ranging from 0.033 to 0.980 and from 0.038 to 0.927, respectively. Only four loci showed deviations of Hardy–Weinberg equilibrium in both populations and were identified as having null alleles. Additionally, two pairs of loci were identified to be in linkage disequilibrium in only one population. Host anemones are highly sought after in the marine aquarium trade and are susceptible to thermal bleaching. Although most studies focus on their obligate symbionts (the anemonefish), genetic analyses of host sea anemones can expand our understanding of the biology, connectivity, and population structure of these organisms and potentially help develop conservation strategies that will aid both the host and its symbionts.

Keywords

Cnidarian Connectivity Host anemone Papua New Guinea Population genetics Red Sea 

References

  1. Almany GR, Berumen ML, Thorrold SR, Planes S, Jones GP (2007) Local replenishment of coral reef fish populations in a marine reserve. Science 316:742–744. doi:10.1126/science.1140597 CrossRefPubMedGoogle Scholar
  2. Andras JP, Kirk NL, Drew Harvell C (2011) Range-wide population genetic structure of Symbiodinium associated with the Caribbean sea fan coral, Gorgonia ventalina. Mol Ecol 20:2525–2542. doi:10.1111/j.1365-294X.2011.05115.x CrossRefPubMedGoogle Scholar
  3. Ben-Horin T, Iacchei M, Selkoe KA, Mai TT, Toonen RJ (2009) Characterization of eight polymorphic microsatellite loci for the California spiny lobster, Panulirus interruptus and cross-amplification in other achelate lobsters. Conserv Genet Resour 1:193–197. doi:10.1007/s12686-009-9047-2 CrossRefGoogle Scholar
  4. Benjamini Y, Hochberg Y (1995) Controlling the false discovery rate: a practical and powerful approach to multiple testing. J R Stat Soc Ser B 57:289–300Google Scholar
  5. Berumen ML, Almany GR, Planes S, Jones GP, Saenz-Agudelo P, Thorrold SR (2012) Persistence of self-recruitment and patterns of larval connectivity in a marine protected area network. Ecol Evol 2:444–452. doi:10.1002/ece3.208 CrossRefPubMedPubMedCentralGoogle Scholar
  6. Billingham M, Ayre DJ (1996) Genetic subdivision in the subtidal, clonal sea anemone Anthothoe albocincta. Mar Biol 125:153–163. doi:10.1007/BF00350769 CrossRefGoogle Scholar
  7. Bocharova ES, Kozevich IA (2011) Modes of reproduction in sea anemones (Cnidaria, Anthozoa). Biol Bull Russ Acad Sci 38:849–860. doi:10.1134/S1062359011090020 CrossRefGoogle Scholar
  8. Bonin MC, Saenz-Agudelo P, Harrison HB, Nanninga GB, van der Meer MH, Mansour H, Perumal S, Jones GP, Berumen ML (2016) Characterization and cross-amplification of microsatellite markers in four species of anemonefish (Pomacentridae, Amphiprion spp.). Mar Biodivers 46:135–140. doi:10.1007/s12526-015-0336-6 CrossRefGoogle Scholar
  9. Brolund TM, Tychsen A, Nielsen LE, Arvedlund M (2004) An assemblage of the host anemone Heteractis magnifica in the northern Red Sea, and distribution of the resident anemonefish. J Mar Biol Assoc UK 84:671–674. doi:10.1017/S0025315404009737h CrossRefGoogle Scholar
  10. Buston PM, Jones GP, Planes S, Thorrold SR (2012) Probability of successful larval dispersal declines fivefold over 1 km in a coral reef fish. Proc R Soc B Biol Sci 279:1883–1888. doi:10.1098/rspb.2011.2041 CrossRefGoogle Scholar
  11. Chomsky O, Douek J, Chadwick NE, Dubinsky Z, Rinkevich B (2009) Biological and population-genetic aspects of the sea anemone Actiniaequina (Cnidaria: Anthozoa) along the Mediterranean coast of Israel. J Exp Mar Biol Ecol 375:16–20. doi:10.1016/j.jembe.2009.04.017 CrossRefGoogle Scholar
  12. Darling JA, Reitzel AM, Finnerty JR (2004) Regional population structure of a widely introduced estuarine invertebrate: Nematostella vectensis Stephenson in New England. Mol Ecol 13:2969–2981. doi:10.1111/j.1365-294X.2004.02313.x CrossRefPubMedGoogle Scholar
  13. Darling JA, Reitzel AM, Finnerty JR (2006) Characterization of microsatellite loci in the widely introduced estuarine anemone Nematostella vectensis. Mol Ecol Notes 6:803–805. doi:10.1111/j.1471-8286.2006.01350.x CrossRefGoogle Scholar
  14. Dunn DF (1981) The clownfish sea anemones: Stichodactylidae (Coelenterata: Actiniaria) and other sea anemones symbiotic with pomacentrid fishes. Trans Am Philos Soc 71:3–115CrossRefGoogle Scholar
  15. Edmands S, Potts DC (1997) Population genetic structure in brooding sea anemones (Epiactis spp.) with contrasting reproductive modes. Mar Biol 127:485–498. doi:10.1007/s002270050037 CrossRefGoogle Scholar
  16. Faircloth BC (2008) MSATCOMMANDER: detection of microsatellite repeat arrays and automated, locus-specific primer design. Mol Ecol Resour 8:92–94. doi:10.1111/j.1471-8286.2007.01884.x CrossRefPubMedGoogle Scholar
  17. Fautin DG, Allen GR (1997) Anemonefishes and their host sea anemones: a guide for aquarists and divers. Western Australian Museum, PerthGoogle Scholar
  18. Fautin DG, Guo CC, Hwang JS (1995) Costs and benefits of the symbiosis between the anemoneshrimp Periclimenes brevicarpalis and its host Entacmaea quadricolor. Mar Ecol Prog Ser 129:77–84. doi:10.3354/meps129077 CrossRefGoogle Scholar
  19. Giles EC, Saenz-Agudelo P, Berumen ML, Ravasi T (2013) Novel polymorphic microsatellite markers developed for a common reef sponge, Stylissa carteri. Mar Biodivers 43:237–241. doi:10.1007/s12526-013-0151-x CrossRefGoogle Scholar
  20. Guo CC, Hwang JS, Fautin DG (1996) Host selection by shrimps symbiotic with sea anemones: a field survey and experimental laboratory analysis. J Exp Mar Biol Ecol 202:165–176. doi:10.1016/0022-0981(96)00020-2 CrossRefGoogle Scholar
  21. Hattori A (2002) Small and large anemonefishes can coexist using the same patchy resources on a coral reef, before habitat destruction. J Anim Ecol 71:824–831. doi:10.1046/j.1365-2656.2002.00649.x CrossRefGoogle Scholar
  22. Hill R, Scott A (2012) The influence of irradiance on the severity of thermal bleaching in sea anemones that host anemonefish. Coral Reefs 31:273–284. doi:10.1007/s00338-011-0848-x CrossRefGoogle Scholar
  23. Hobbs JPA, Frisch AJ, Ford BM, Thums M, Saenz-Agudelo P, Furby KA, Berumen ML (2013) Taxonomic, spatial and temporal patterns of bleaching in anemones inhabited by anemonefishes. PLoS One 8:e70966. doi:10.1371/journal.pone.0070966 CrossRefPubMedPubMedCentralGoogle Scholar
  24. Hoeksema BW, Crowther AL (2011) Masquerade, mimicry and crypsis of the polymorphic sea anemone Phyllodiscus semoni and its aggregations in South Sulawesi. Contrib Zool 80:251–268Google Scholar
  25. Holbrook SJ, Schmitt RJ (2004) Growth, reproduction and survival of a tropical sea anemone (Actiniaria): benefits of hosting anemonefish. Coral Reefs 24:67–73. doi:10.1007/s00338-004-0432-8 CrossRefGoogle Scholar
  26. Howells EJ, Van Oppen MJH, Willis BL (2009) High genetic differentiation and cross-shelf patterns of genetic diversity among Great Barrier Reef populations of Symbiodinium. Coral Reefs 28:215–225. doi:10.1007/s00338-008-0450-z CrossRefGoogle Scholar
  27. Kearse M, Moir R, Wilson A, Stones-Havas S, Cheung M, Sturrock S, Buxton S, Cooper A, Markowitz S, Duran C, Thierer T, Ashton B, Meintjes P, Drummond A (2012) Geneious Basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics 28:1647–1649. doi:10.1093/bioinformatics/bts199 CrossRefPubMedPubMedCentralGoogle Scholar
  28. Madduppa HH, von Juterzenka K, Syakir M, Kochzius M (2014) Socio-economy of marine ornamental fishery and its impact on the population structure of the clown anemonefish Amphiprion ocellaris and its host anemones in Spermonde Archipelago, Indonesia. Ocean Coast Manag 100:41–50. doi:10.1016/j.ocecoaman.2014.07.013 CrossRefGoogle Scholar
  29. Mariscal RN (1970) The nature of the symbiosis between Indo-Pacific anemone fishes and sea anemones. Mar Biol 6:58–65. doi:10.1007/BF00352608 CrossRefGoogle Scholar
  30. Nanninga GB, Saenz-Agudelo P, Manica A, Berumen ML (2014) Environmental gradients predict the genetic population structure of a coral reef fish in the Red Sea. Mol Ecol 23:591–602. doi:10.1111/mec.12623 CrossRefPubMedGoogle Scholar
  31. Peakall R, Smouse PE (2012) GenALEx 6.5: genetic analysis in excel. Population genetic software for teaching and research—an update. Bioinformatics 28:2537–2539. doi:10.1093/bioinformatics/bts460 CrossRefPubMedPubMedCentralGoogle Scholar
  32. Pinsky ML, Montes HR Jr, Palumbi SR (2010) Using isolation by distance and effective density to estimate dispersal scales in anemonefish. Evolution 64:2688–2700. doi:10.1111/j.1558-5646.2010.01003.x CrossRefPubMedGoogle Scholar
  33. Planes S, Jones GP, Thorrold SR (2009) Larval dispersal connects fish populations in a network of marine protected areas. Proc Natl Acad Sci U S A 106:5693–5697. doi:10.1073/pnas.0808007106 CrossRefPubMedPubMedCentralGoogle Scholar
  34. Priest MA, Almany GR, Braun CD, Hamilton RJ, Lozano-Cortés DF, Saenz-Agudelo P, Berumen ML (2015) Isolation and characterization of 29 microsatellite markers for the bumphead parrotfish, Bolbometopon muricatum, and cross amplification in 12 related species. Mar Biodivers 45:861–866. doi:10.1007/s12526-014-0278-4 CrossRefGoogle Scholar
  35. Randall JE, Fautin DG (2002) Fishes other than anemonefishes that associate with sea anemones. Coral Reefs 21:188–190. doi:10.1007/s00338-002-0234-9 Google Scholar
  36. Raymond M, Rousset F (1995) GENEPOP (version 1.2): population genetics software for exact tests and ecumenicism. J Hered 86:248–249Google Scholar
  37. Reitzel AM, Herrera S, Layden MJ, Martindale MQ, Shank TM (2013) Going where traditional markers have not gone before: utility of and promise for RAD sequencing in marine invertebrate phylogeography and population genomics. Mol Ecol 22:2953–2970. doi:10.1111/mec.12228 CrossRefPubMedPubMedCentralGoogle Scholar
  38. Rousset F (2008) GENEPOP’007: a complete re-implementation of the GENEPOP software for Windows and Linux. Mol Ecol Resour 8:103–106. doi:10.1111/j.1471-8286.2007.01931.x CrossRefPubMedGoogle Scholar
  39. Rozen S, Skaletsky H (1999) Primer3 on the WWW for general users and for biologist programmers. Bioinform Methods Protoc 132:365–386CrossRefGoogle Scholar
  40. Russo CAM, Solé-Cava AM, Thorpe JP (1994) Population structure and genetic variation in two tropical sea anemones (Cnidaria, Actinidae) with different reproductive strategies. Mar Biol 119:267–276. doi:10.1007/BF00349566 CrossRefGoogle Scholar
  41. Saenz-Agudelo P, Jones GP, Thorrold SR, Planes S (2011a) Detrimental effects of host anemone bleaching on anemonefish populations. Coral Reefs 30:497–506. doi:10.1007/s00338-010-0716-0 CrossRefGoogle Scholar
  42. Saenz-Agudelo P, Jones GP, Thorrold SR, Planes S (2011b) Connectivity dominates larval replenishment in a coastal reef fish metapopulation. Proc Biol Sci 278:2954–2961. doi:10.1098/rspb.2010.2780 CrossRefPubMedPubMedCentralGoogle Scholar
  43. Scribner KT, Gust JR, Fields RL (1996) Isolation and characterization of novel salmon microsatellite loci: cross-species amplification and population genetic applications. Can J Fish Aquat Sci 53:833–841CrossRefGoogle Scholar
  44. Sherman CDH, Ayre DJ (2008) Fine-scale adaptation in a clonal sea anemone. Evolution 62:1373–1380. doi:10.1111/j.1558-5646.2008.00375.x CrossRefPubMedGoogle Scholar
  45. Sherman CDH, Peucker AJ, Ayre DJ (2007) Do reproductive tactics vary with habitat heterogeneity in the intertidal sea anemone Actinia tenebrosa? J Exp Mar Biol Ecol 340:259–267. doi:10.1016/j.jembe.2006.09.016 CrossRefGoogle Scholar
  46. Shick JM, Lamb AN (1977) Asexual reproduction and genetic population structure in the colonizing sea anemone Haliplanella luciae. Biol Bull 153:604–617CrossRefGoogle Scholar
  47. Shick JM, Hoffmann RJ, Lamb AN (1979) Asexual reproduction, population structure, and genotype–environment interactions in sea anemones. Am Zool 19:699–713. doi:10.1093/icb/19.3.699 CrossRefGoogle Scholar
  48. van der Meer MH, Hobbs JP, Jones GP, van Herwerden L (2012) Genetic connectivity among and self-replenishment within island populations of a restricted range subtropical reef fish. PLoS One 7:e49660. doi:10.1371/journal.pone.0049660 CrossRefPubMedPubMedCentralGoogle Scholar
  49. van Oosterhout C, Hutchinson WF, Wills DPM, Shipley P (2004) MICRO-CHECKER: software for identifying and correcting genotyping errors in microsatellite data. Mol Ecol Notes 4:535–538. doi:10.1111/j.1471-8286.2004.00684.x CrossRefGoogle Scholar
  50. Veale AJ, Lavery SD (2012) The population genetic structure of the waratah anemone (Actinia tenebrosa) around New Zealand. New Zeal J Mar Freshw Res 46:523–536. doi:10.1080/00288330.2012.730053 CrossRefGoogle Scholar
  51. Wabnitz C, Taylor M, Green EP, Razak T (2003) From ocean to aquarium: the global trade in marine ornamental species. UNEP-WCMC, CambridgeGoogle Scholar
  52. Waits LP, Luikart G, Taberlet P (2001) Estimating the probability of identity among genotypes in natural populations: cautions and guidelines. Mol Ecol 10:249–256. doi:10.1046/j.1365-294X.2001.01185.x CrossRefPubMedGoogle Scholar
  53. Weir BS, Cockerham CC (1984) Estimating F-statistics for the analysis of population structure. Evolution 38:1358–1370CrossRefGoogle Scholar
  54. Wood EM (1985) Exploitation of coral reef fishes for the aquarium trade. Report to the Marine Conservation Society. Marine Conservation Society, UKGoogle Scholar

Copyright information

© Senckenberg Gesellschaft für Naturforschung and Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  1. 1.Red Sea Research Center, Division of Biological and Environmental Science and EngineeringKing Abdullah University of Science and TechnologyThuwalSaudi Arabia
  2. 2.Department of Ecology and Evolutionary BiologyUniversity of California Santa CruzSanta CruzUSA
  3. 3.Facultad de Ciencias, Instituto de Ciencias Ambientales y EvolutivasUniversidad Austral de ChileValdiviaChile
  4. 4.National Marine Science Centre and Marine Ecology Research Centre, School of Environmental Science and ManagementSouthern Cross UniversityCoffs HarbourAustralia

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