Marine Biology

, Volume 156, Issue 8, pp 1681–1690 | Cite as

Multiple paternity and extra-group fertilizations in a natural population of California grunion (Leuresthes tenuis), a beach-spawning marine fish

  • Rosemary J. ByrneEmail author
  • John C. Avise
Original Paper


Although individuals in many fish species move to shallow waters to spawn, the California grunion (Leuresthes tenuis) is almost unique in its constitutive display of synchronous full-emergence beach spawning. During a spawning event, fish ride large waves onshore to spawn on beach land, where their eggs incubate terrestrially. Here, we employ molecular markers to ascertain how this unusual reproductive behavior impacts genetic parentage. We developed and utilized four highly polymorphic microsatellite markers to assess maternal and paternal contributions in a total of 682 progeny from 17 nests of a natural population of L. tenuis. Alleles deduced to be of paternal origin in progeny were used to determine the minimum number of sires per nest and to estimate the true number of sires per nest via Bayesian analysis. We document the following: (a) no instances of multiple maternity for progeny within a nest; (b) a high frequency of nests (88%) with multiple paternity; and (c) an appreciable fraction of nests (18%) in which the estimated number of genetic sires (as many as nine) proved to be greater than the observed number of male attendants, thus implicating occasional extra-group fertilization events. From these and other observations, we also conclude that spawning behavior in grunions may involve site choice but not explicit mate choice. In addition to providing the first analysis of molecular parentage in a beach-spawning fish, we compare our findings to those reported previously for a beach-spawning arthropod, and we discuss the forces that may be maintaining this peculiar reproductive behavior.


Beach Mate Choice Mating Behavior Sperm Competition Horseshoe Crab 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



This work was supported by the National Science Foundation (NSF Grant DGE-0638751) and the University of California, Irvine. Animals were collected with permission from the California Department of Fish and Game granted to R.J.B. with Scientific Collecting Permit ID Number SC-008834. We thank Felipe Barreto and Molly Burke for field assistance, and Felipe Barreto, Vimoksalehi Lukoschek, Andrey Tatarenkov, and two anonymous reviewers for thoughtful comments on the manuscript.

Open Access

This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.


  1. Alcock J, Eickwort GC, Eickwort KR (1977) The reproductive behavior of Anthidium maculosum (Hymenoptera: Megachilidae) and the evolutionary significance of multiple copulations by females. Behav Ecol Sociobiol 2:385–396. doi: CrossRefGoogle Scholar
  2. Avise JC (2004) Molecular markers, natural history, and evolution, 2nd edn. Sinauer Associates, Sunderland, MSGoogle Scholar
  3. Avise JC, Jones AG, Walker D, DeWoody JA et al (2002) Genetic mating systems and reproductive natural histories of fishes: lessons for ecology and evolution. Annu Rev Genet 36:19–45. doi: CrossRefGoogle Scholar
  4. Brantley RK, Bass AH (1994) Alternative male spawning tactics and acoustic signals in the plainfin midshipman fish Porichthys notatus Girard (Teleostei, Batrachoididae). Ethology 96:213–232CrossRefGoogle Scholar
  5. Breder CM Jr, Rosen DE (1966) Modes of reproduction in fishes. Natural History Press, Garden City, NYGoogle Scholar
  6. Brockmann HJ (1996) Satellite male groups in horseshoe crabs, Limulus polyphemus. Ethology 102:1–21CrossRefGoogle Scholar
  7. Brockmann HJ, Colson T, Potts W (1994) Sperm competition in horseshoe crabs (Limulus polyphemus). Behav Ecol Sociobiol 3:153–160. doi: CrossRefGoogle Scholar
  8. Brockmann HJ, Nguyen C, Potts W (2000) Paternity in horseshoe crabs when spawning in multiple-male groups. Anim Behav 60:837–849. doi: CrossRefGoogle Scholar
  9. Brookfield JFY (1996) A simple new method for estimating null allele frequency from heterozygote deficiency. Mol Ecol 5:453–455. doi: CrossRefGoogle Scholar
  10. Chesser RK, Baker RJ (1996) Effective sizes and dynamics of uniparentally and diparentally inherited genes. Genetics 144:1225–1235PubMedPubMedCentralGoogle Scholar
  11. Clark FN (1925) The life history of Leuresthes tenuis, an atherine fish with tide controlled spawning habits. Calif Dep Fish Game Fish Bull 10:1–51Google Scholar
  12. Dakin EE, Avise JC (2004) Microsatellite null alleles in parentage analysis. Heredity 93:504–509. doi: CrossRefGoogle Scholar
  13. David LR (1939) Embryonic and early larval stages of the grunion, Leuresthes tenuis, and of the sculpin, Scorpaena guttata. Copeia 1939:75–81. doi: CrossRefGoogle Scholar
  14. DeWoody JA, DeWoody YD, Fiumera AC, Avise JC (2000) On the number of reproductives contributing to a half-sib progeny array. Genet Res 75:95–105. doi: CrossRefGoogle Scholar
  15. Emery AM, Wilson IJ, Craig S, Boyle PR, Noble LR (2001) Assignment of paternity groups without access to parental genotypes: multiple mating and developmental plasticity in squid. Mol Ecol 10:1265–1278. doi: CrossRefGoogle Scholar
  16. Gomendio M, Harcourt AH, Roldán ERS (1998) Sperm competition in mammals. In: Birkhead TR, Møller AP (eds) Sperm competition and sexual selection. Academic Press, London, pp 667–756CrossRefGoogle Scholar
  17. Gronell AM (1989) Visiting behaviour by females of the sexually dichromatic damselfish, Chrysiptera cyanea (Teleostei: Pomacentridae): a probable method of assessing male quality. Ethology 81:89–122CrossRefGoogle Scholar
  18. Gross MR, Sargent RC (1985) The evolution of male and female parental care in fishes. Am Zool 25:807–822CrossRefGoogle Scholar
  19. Halliday TR (1983) The study of mate choice. In: Bateson P (ed) Mate choice. Cambridge University Press, Cambridge, pp 3–32Google Scholar
  20. Halliday T, Arnold SJ (1987) Multiple mating by females: a perspective from quantitative genetics. Anim Behav 35:939–941. doi: CrossRefGoogle Scholar
  21. Hamilton MB, Pincus EL, Di Fiore A, Flescher RC (1999) Universal linker and ligation procedures for construction of genomic DNA libraries enriched for microsatellites. Biotechniques 27:500–507CrossRefGoogle Scholar
  22. Hancock JM (1999) Microsatellites and other simple sequences: genomic context and mutational mechanisms. In: Goldstein DB, Schlötterer C (eds) Microsatellites: evolution and applications. Oxford University Press, New York, pp 1–9Google Scholar
  23. Hassler C, Brockmann HJ (2001) Evidence for use of chemical cues by male horseshoe crabs when locating nesting females (Limulus polyphemus). J Chem Ecol 27:2319–2335. doi: CrossRefGoogle Scholar
  24. Hauswaldt JS, Glenn TC (2003) Microsatellite DNA loci from the diamondback terrapin (Malaclemys terrapin). Mol Ecol Notes 3:174–176. doi: CrossRefGoogle Scholar
  25. Hoelzel AR, Green A (1998) PCR protocols and population analysis by direct DNA sequencing and PCR-based DNA fingerprinting. In: Hoelzel AR (ed) Molecular genetic analysis of populations: a practical approach, 2nd edn. Oxford University Press, New York, pp 201–235Google Scholar
  26. Jamieson A, Taylor SCS (1997) Comparisons of three probability formulae for parentage exclusion. Anim Genet 28:397–400. doi: CrossRefGoogle Scholar
  27. Jennions MD, Petrie M (2000) Why do females mate multiply? A review of the genetic benefits. Biol Rev Camb Philos Soc 75:21–64. doi: CrossRefGoogle Scholar
  28. Jones GP (1981) Spawning-site choice by female Pseudolabrus celidotus (Pisces: Labridae) and its influence on the mating system. Behav Ecol Sociobiol 8:129–142. doi: CrossRefGoogle Scholar
  29. Leggett WC, Frank KT (1990) The spawning of the capelin. Sci Am 262:102–107CrossRefGoogle Scholar
  30. Levitan DR (1991) Influence of body size and population density on fertilization success and reproductive output in a free-spawning invertebrate. Biol Bull 181:261–268. doi: CrossRefGoogle Scholar
  31. Levitan DR (1998) Sperm limitation, gamete competition, and sexual selection in external fertilizers. In: Birkhead TR, Møller AP (eds) Sperm competition and sexual selection. Academic Press, London, pp 175–217CrossRefGoogle Scholar
  32. Mackiewicz M, Fletcher DE, Wilkins SD, DeWoody JA, Avise JC (2002) A genetic assessment of parentage in a natural population of dollar sunfish (Lepomis marginatus) based on microsatellite markers. Mol Ecol 11:1877–1883. doi: CrossRefGoogle Scholar
  33. Martin KLM, Van Winkle RC, Drais JE, Lakisic H (2004) Beach-spawning fishes, terrestrial eggs, and air breathing. Physiol Biochem Zool 77:750–759. doi: CrossRefGoogle Scholar
  34. Middaugh DP (1981) Reproductive ecology and spawning periodicity of the Atlantic silverside, Menidia menidia (Pisces, Atherinidae). Copeia 1981:766–776. doi: CrossRefGoogle Scholar
  35. Middaugh DP, Kohl HW, Burnett LE (1983) Concurrent measurement of intertidal environmental variables and embryo survival for the California grunion, Leuresthes tenuis, and Atlantic silverside, Menidia menidia (Pisces: Atherinidae). Calif Fish Game 69:89–96Google Scholar
  36. Miller DJ, Lea RN (1972) Guide to the coastal marine fishes of California. Calif Depart Fish Game Fish Bull 157:1–235Google Scholar
  37. Neff BD (2001) Genetic paternity analysis and breeding success in bluegill sunfish (Lepomis macrochirus). J Hered 92:111–119. doi: CrossRefGoogle Scholar
  38. Parker GA (1970) Sperm competition and its evolutionary consequences in the insects. Biol Rev Camb Philos Soc 45:525–567. doi: CrossRefGoogle Scholar
  39. Parker GA (1990) Sperm competition games: raffles and roles. Proc R Soc Lond B Biol Sci 242:120–126. doi: CrossRefGoogle Scholar
  40. Pemberton JM, Slate J, Bancroft DR, Barrett JA (1995) Nonamplifying alleles at microsatellite loci: a caution for parentage and population studies. Mol Ecol 4:249–252. doi: CrossRefGoogle Scholar
  41. Pennington JT (1985) The ecology of fertilization of echinoid eggs: the consequences of sperm dilution, adult aggregation, and synchronous spawning. Biol Bull 169:417–430. doi: CrossRefGoogle Scholar
  42. Petersen CW, Warner RR (1998) Sperm competition in fishes. In: Birkhead TR, Møller AP (eds) Sperm competition and sexual selection. Academic Press, London, pp 435–463CrossRefGoogle Scholar
  43. Porter BA, Fiumera AC, Avise JC (2002) Egg mimicry and allopaternal care: two mate-attracting tactics by which nesting striped darter (Etheostoma virgatum) males enhance reproductive success. Behav Ecol Sociobiol 51:350–359. doi: CrossRefGoogle Scholar
  44. Rausher MD (1983) Ecology of host-selection behavior in phytophagous insects. In: Denno RF, McClure MS (eds) Variable plants and herbivores in natural and managed systems. Academic Press, New York, pp 223–257CrossRefGoogle Scholar
  45. Raymond M, Rousset F (1995) GENEPOP (version 1.2): population genetics software for exact tests and ecumenicism. J Hered 86:248–249CrossRefGoogle Scholar
  46. Rieger JF, Binckley CA, Resetarits WJ Jr (2004) Larval performance and oviposition site preference along a predation gradient. Ecology 85:2094–2099. doi: CrossRefGoogle Scholar
  47. Schwab RL, Brockmann HJ (2007) The role of visual and chemical cues in the mating decisions of satellite male horseshoe crabs, Limulus polyphemus. Anim Behav 74:837–846. doi: CrossRefGoogle Scholar
  48. Seymour RS, Bradford DF (1995) Respiration of amphibian eggs. Physiol Zool 68:1–25CrossRefGoogle Scholar
  49. Smyder EA, Martin KLM (2002) Temperature effects on egg survival and hatching during the extended incubation period of California grunion, Leuresthes tenuis. Copeia 2002:313–320. doi:[0313:TEOESA]2.0.CO;2 CrossRefGoogle Scholar
  50. Strathmann RR, Hess HC (1999) Two designs of marine egg masses and their divergent consequences for oxygen supply and desiccation in air. Am Zool 39:253–260CrossRefGoogle Scholar
  51. Sugg DW, Chesser RK (1994) Effective population sizes with multiple paternity. Genetics 137:1147–1155PubMedPubMedCentralGoogle Scholar
  52. Taborsky M (1994) Sneakers, satellites, and helpers–parasitic and cooperative behavior in fish reproduction. Adv Stud Behav 23:1–100. doi: CrossRefGoogle Scholar
  53. Taborsky M (2001) The evolution of bourgeois, parasitic, and cooperative reproductive behaviors in fishes. J Hered 92:100–110. doi: CrossRefGoogle Scholar
  54. Thomaz D, Beall E, Burke T (1997) Alternative reproductive tactics in Atlantic salmon: factors affecting mature parr success. Proc R Soc Lond B Biol Sci 264:219–226. doi: CrossRefGoogle Scholar
  55. Thompson WF, Thompson JB (1919) The spawning of the grunion (Leuresthes tenuis). Calif Depart Fish Game Fish Bull 3:1–29Google Scholar
  56. Tregenza T, Wedell N (2002) Polyandrous females avoid costs of inbreeding. Nature 415:71–73. doi: CrossRefGoogle Scholar
  57. Walker BW (1949) Periodicity of spawning by the Grunion, Louresthes tenuis, an Atherine Fish. PhD thesis, University of California, Los AngelesGoogle Scholar
  58. Walker BW (1952) A guide to the grunion. Calif Fish Game 38:409–420Google Scholar
  59. Warner RR (1988) Traditionality of mating-site preferences in a coral reef fish. Nature 335:719–721. doi: CrossRefGoogle Scholar
  60. Warner RR (1990) Male versus female influences on mating-site determination in a coral reef fish. Anim Behav 39:540–548. doi: CrossRefGoogle Scholar
  61. Weber JL, Wong C (1993) Mutation of human short tandem repeats. Hum Mol Genet 2:1123–1128. doi: CrossRefGoogle Scholar
  62. Yamahira K (1996) The role of intertidal egg deposition on survival of the puffer, Takifugu niphobles (Jordan et Snyder), embryos. J Exp Mar Biol Ecol 198:291–306. doi: CrossRefGoogle Scholar

Copyright information

© The Author(s) 2009

Open AccessThis is an open access article distributed under the terms of the Creative Commons Attribution Noncommercial License (, which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.

Authors and Affiliations

  1. 1.Department of Ecology and Evolutionary BiologyUniversity of California, IrvineIrvineUSA

Personalised recommendations