Abstract
Male horseshoe crabs have two mating tactics. Some males come ashore attached to a female (clasping the posterior margin of the females' carapace with their modified pedipalps) and nest with her on the intertidal portion of the beach during the high tide. Other males come ashore unattached and crowd around nesting couples. Fertilization is external and unattached males that are in contact with a pair, i.e. ‘satellite’ males, release sperm, so the assumption has been that they are fertilizing eggs. We conducted a paternity analysis to determine the proportion of eggs fertilized by attached and satellite males. Pairs with one satellite were observed during nesting on beaches in Florida and Delaware and their eggs were collected and reared to the late trilobite or first instar horseshoe crab stage. DNA was extracted from these offspring and from each adult (female, attached and satellite male) for use in paternity analysis. A Limulus-specific hypervariable microsatellite locus was identified and primers were constructed to amplify this locus via the polymerase chain reaction (PCR). Genotypes of putative parents and offspring were determined by resolving length variants of these PCR products on acrylamide gels. This allowed us to determine parentage of the offspring. We demonstrate that satellite males fertilized 40% of the eggs on average, attached males fertilized 51% and 4% of the eggs that were laid by the female were fathered neither by the attached male nor by the satellite (and 5% could not be determined unambiguously). There is high variability in the success of satellite males, ranging from 0 to 88%. Part of this variability can be explained by the position of the satellite relative to the attached male. We discuss the mechanics of fertilization and the possible advantages for multiple mating in this species.
Similar content being viewed by others
References
Alcock J, Eickwort GC, Eickwort KR (1977) The reproductive behavior ofAnthidium maculosum (Hymenoptera: Megachilidae) and the evolutionary significance of multiple copulations by females. Behav Ecol Sociobiol 2:385–396
Arak A (1983) Male-male competition and mate choice in anuran amphibians. In: Bateson BP (ed) Mate choice. Cambridge University Press, Cambridge, pp 181–210
Assem J van den (1967) Territory in the three-spined sticklebackGasterosteus aculeatus: an experimental study in intra-specific competition (Behav Suppl 16). EJ Brill, Leiden
Barlow RB Jr, Powers MK, Howard H, Kass L (1986) Migration ofLimulus for mating: relation to lunar phase, tide height and sunlight. Biol Bull 171:310–329
Barthel KW (1974)Limulus: a living fossil. Naturwissenschaften 61:428–433
Botton ML, Loveland RE (1989) Reproductive risk: high mortality associated with spawning by horseshoe crabs (Limulus polyphemus) in Delaware Bay, USA. Mar Biol 101:143–151
Botton ML, Loveland RE (1992) Body size, morphological constraints, and mated pair formation in four populations of horseshoe crabs (Limulus polyphemus) along a geographic cline. Mar Biol 112:409–415
Brockmann HJ (1990) Mating behavior of horseshoe crabs,Limulus polyphemus. Behaviour 114:206–220
Brockmann HJ, Penn D (1992) Male mating tactics in the horseshoe crab,Limulus polyphemus. Anim Behav 44:653–665
Brown GG (1976) Scanning electron-microscopical and other observations of sperm fertilization reactions inLimulus polyphemus L. (Merostomata: Xiphosura). J Cell Sci 22:547–562
Brown GG, Clapper DL (1981) Procedures for maintaining adults, collecting gametes, and culturing embryos and juveniles of the horseshoe crab,Limulus polyphemus L. In: Hinegardner R, Atz J, Fay R, Fingerman M, Josephson R, Meinkoth N (eds) Laboratory animal management, marine invertebrates. National Academy Press, Washington DC, pp 268–290
Brown GG, Humphreys WJ (1971) Sperm-egg interactions ofLimulus polyphemus with scanning electron microscopy. J Cell Biol 51:904–907
Brown GG, Knouse JR (1973) Effects of sperm concentration, sperm aging, and other variables on fertilization in the horseshoe crab,Limulus polyphemus L. Biol Bull 144:462–470
Cavanaugh CM (1975) Observations on mating behavior inLimulus polyphemus. Biol Bull 149:422
Clapper DL, Brown GG (1980) Sperm motility in the horseshoe crab,Limulus polyphemus L. I. Sperm behavior near eggs and motility initiation by egg extracts. Develop Biol 76:341–349
Clapper DL, Epel D (1985) TheLimulus sperm motility-initiating peptide initiates acrosome reactions in sea water lacking potassium. J Exp Zool 236:211–217
Cohen JA, Brockmann HJ (1983) Breeding activity and mate selection in the horseshoe crab,Limulus polyphemus. Bull Mar Sci 33:274–281
Dominey WJ (1980) Female mimicry in male bluegill sunfish — a genetic polymorphism? Nature 284:546–548
Dominey WJ (1981) Maintenance of female mimicry as a reproductive strategy in bluegill sunfish (Lepomis macrochirus) Environ Biol Fish 6:59–64
Fahrenbach WH (1973) Spermiogenesis in the horseshoe crab,Limulus polyphemus. J Morphol 140:31–52
Fairchild L (1984) Male reproductive tactics in an explosive breeding toad population. Am Zool 24:407–418
Feng AS, Narins PM (1991) Unusual mating behavior of Malaysian treefrogs,Polypedates leucomystax. Naturwissenschaften 78:362–365
Feng AS, Narins PM (1992) Reply [to Letters to the editor responding to: unusual mating behavior of Malaysian treefrogs,Polypedates leucomystax]. Naturwissenschaften 79:138–139
Fukuyama K (1991) Spawning behaviour and male mating tactics of a foam-nesting treefrog,Rhacophorus schlegehi. Anim Behav 42:193–199
Georges M, Laquarre AS, Castelli M, Hanset R, Vassart G (1988) DNA fingerprinting in domestic animals using four different minisatellite probes. Cytogenetics and Cell Genetics 47:127–131
Gross MR (1984) Sunfish, salmon, and the evolution of alternative reproductive strategies and tactics in fishes. In: Potts GW, Wootton RJ (eds) Fish reproduction: strategies and tactics. Academic Press, London, pp 55–75
Gross MR (1991a) Evolution of alternative reproductive strategies: frequency-dependent sexual selection in male bluegill sunfish. Philos Trans R. Soc Lond B 332:59–66
Gross MR (1991b) Salmon breeding behavior and life history evolution in changing environments. Ecology 72:1180–1186
Gross MR, Charnov EL (1980) Alternative male life histories in bluegill sunfish. Proc Natl Acad Sci USA 77:6937–6940
Halliday TR (1983) The study of mate choice. In: Bateson P (ed) Mate choice. Cambridge University Press, Cambridge, pp. 3–32
Halliday TR, Arnold SJ (1987) Multiple mating by females: a perspective from quantitative genetics. Anim Behav 35:939–941
Halliday TR, Verrell PA (1984) Sperm competition in amphibians. In: Smith RL (ed) Sperm competition and the evolution of animal mating systems. Academic Press, New York, pp 487–508
Höglund J, Säterberg S (1989) Sexual selection in common toads: correlates with age and body size. J Evol Biol 2:367–372
Hutchings JA, Myers RA (1988) Mating success of alternative maturation phenotypes in male Atlantic salmon,Salmo salar. Oecologia 75:169–174
Jeffreys AJ, Wilson V, Thein SL (1985) Hypervariable ‘minisatellite’ regions in human DNA. Nature 314:67–73
Jennions, MD, Backwell PRY, Passmore NI (1992) Breeding behaviour of the African frog,Chiromantis xerampelina: multiple spawning and polyandry. Anim Behav 44:1091–1100
Jordan WC, Youngson AF (1992) The use of genetic marking to assess the reproductive success of mature male Atlantic salmon parr (Salmo salar, L.) under natural spawning conditions. J Fish Biol 41:613–618
Kasuya E (1992) Letters to the editor responding to: unusual mating behavior of Malaysian treefrogs,Polypedates leucomystax. Naturwissenschaften 79:138
Katano O (1992) Spawning tactics of paired males of the dark chub,Zacco temmincki, reflect potential fitness costs of satellites. Environ Biol Fishes 35:343–350
Keenleyside MHA (1972) Intraspecific intrusions into nests of spawning longear sunfish (Pisces: Centrarchidae). Copeia 1972:272–278
Knowlton N, Greenwell SR (1984) Male sperm competition avoidance mechanisms: the influence of female interests. In: Smith RL (ed) Sperm competition and the evolution of animal mating systems. Academic Press, Inc, New York, pp 61–84
Kodric-Brown A (1986) Satellites and sneakers: opportunistic male breeding tactics in pupfish (Cyprinodon pecosensis) Behav Ecol Sociobiol 19:425–432
Kusano T, Toda M, Fukuyama K (1991) Testes size and breeding systems in Japanese anurans with special reference to large testes in the treefrog,Rhacophorus arboreus (Amphibia: Rhacophoridae). Behav Ecol Sociobiol 29:27–31
Loveland RE, Botton ML (1992) Size dimorphism and the mating system in horseshoe crabs,Limulus polyphemus L. Anim Behav 44:907–916
Maekawa K, Onozato H (1986) Reproductive tactics and fertilization success of mature male Miyabe charr,Salvelinus malma miyabei. Environ Biol Fishes 15:119–129
Maynard Smith J (1978) The evolution of sex. Cambridge University Press, Cambridge
Parker G (1990) Sperm competition games: raffles and roles. Proc R Soc Lond B 242:120–126
Penn D, Brockmann III (in press) Age-biased stranding and righting in male horseshoe crabs (Limulus polyphemus). Anim Behav
Perrill SA, Gerhardt HC, Daniel R (1978) Sexual parasitism in the green tree frog (Hyla cinerea). Science 200:1179–1180
Petrie, M, Hall M, Halliday T, Budgey H, Pierpoint C (1992) Multiple mating in a lekking bird: why do peahens mate with more than one male and with the same male more than once? Behav Ecol Sociobiol 31:349–358
Pyburn WF (1970) Breeding behavior of the leaf frogsPhyllomedusa callidryas andPachymedusa dacnicolor in Mexico. Copeia 1970:209–218
Queller DC, Strassmann JE, Hughes CR (1993) Microsatellites and kinship. Trends Ecol Evol 8:285–288
Rico C, Kuhnlein U, Fitzgerald GJ (1992) Male reproductive tactics in the threespine stickleback — an evaluation of DNA fingerprinting. Mol Ecol 1:79–87
Ridley M (1988) Mating frequency and fecundity in insects. Biol Rev 63:509–549
Riedl RJ, Machan R (1972) Hydrodynamic patterns in lotic intertidal sands and their bioclimatological implications. Mar Biol 13:179–209
Rowland WJ (1979) Stealing fertilizations in the fourspine stickle-back,Apeltes quadracus. Am Natl 114:602–604
Rudloe AE (1980) The breeding behavior and patterns of movement of horseshoe crabsLimulus polyphemus, in the vicinity of breeding beaches in Apalachee Bay, Florida. Estuaries 3:177–183
Rudloe AE, Rudloe J (1981) The changeless horseshoe crab. Natl Geogr Mag 159:562–572
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor
Shoger RL, Brown GG (1970) Ultrastructural study of sperm-egg interactions of the horseshoe crab,Limulus polyphemus L. (Merostomata: Xiphosura). J Submicr Cytol 2:167–179
Shuster CN Jr (1979) Distribution of the American horseshoe “crab”Limulus polyphemus. In: Cohen E (ed) Biomedical applications of the horseshoe crab (Limulidae). Liss, New York, pp 3–26
Shuster CN Jr (1982) A pictorial review of the natural history and ecology of the horseshoe crab,Limulus polyphemus, with reference to other Limulidae. In: Bonaventura J, Bonaventura C, Tesh S (eds) Physiology and biology of horseshoe crabs: studies on normal and environmentally stressed animals. Liss, New York, pp 1–52
Svärd L, Wiklund C (1986) Different ejaculate delivery strategies in first versus subsequent matings in the swallowtail butterflyPapilio macehaon L. Behav Ecol Sociobiol 18:325–330
Taborsky M, Hudde B, Wirtz P (1987) Reproductive behaviour and ecology ofSymphodus (Crenilabrus) ocellatus, a European wrasse with four types of male behaviour. Behaviour 102:82–118
Tautz D (1989) Hypervariability of simple sequences as a general source for polymorphic DNA markers. Nucleic Acids Res 17:6463–6471
Watson PJ (1993) Foraging advantage of polyandry for female sierra dome spiders (Linyphia litigiosa: Linyphiidae) and assessment of alternative direct benefit hypothesis. Am Nat 141:440–465
Wells KD (1979) Reproductive behavior and male mating success in a neotropical toad,Bufo typhonius, Biotropica 11:301–307
Westneat DF, Sherman PW Morton ML (1990) The ecology and evolution of extra-pair copulations in birds. In: Power DM (ed) Current Ornithology, vol 7. Plenum Press, New York, pp 331–369
Author information
Authors and Affiliations
Additional information
Communicated by R. Gibson
Rights and permissions
About this article
Cite this article
Brockmann, H.J., Colson, T. & Potts, W. Sperm competition in horseshoe crabs (Limulus polyphemus). Behav Ecol Sociobiol 35, 153–160 (1994). https://doi.org/10.1007/BF00167954
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00167954