Copulation in many sexually cannibalistic spiders is associated with a loss of function of the male reproductive organs and, as a consequence, males that survive sexual cannibalism may nevertheless be unable to subsequently copulate successfully. Sexual cannibalism is common in the Australian golden orb-web spider (Nephila plumipes), in which the tip of the conductor typically breaks during copulation. Thus, male mating frequency may be physiologically limited to two females, irrespective of the male's ability to avoid cannibalism or the opportunity to locate and court additional, receptive females. Laboratory experiments revealed that the likelihood of the conductor breaking depends upon the copulatory history of the female insemination duct: males were more likely to break their conductor if they inseminated a 'virgin' rather than 'mated' insemination duct. However, the choice of insemination duct did not influence the duration of copulation or quantity of sperm transferred. In field populations, the proportion of males with both conductors broken increased during the course of the mating season, but while males with broken conductors did not copulate successfully with virgin females, they were nevertheless observed on the webs of immature females. We suggest that male N. plumipes with broken conductors on the webs of females are most likely mate guarding, as this appears to be the most effective mechanism of securing paternity.