No abstract

Studies of reproductive biology in insects often require quantification of sperm production, transfer or storage. Here, we develop a quantitative real‐time PCR‐based assay using a Y‐specific marker for quantification of sperm from spermathecae of female Queensland fruit fly ('Q‐fly'), overcoming constraints typical of traditional sperm quantification methods. The assay enables accurate and reliable quantification of as few as 50 sperms and provides a means to analyse large numbers of samples with flexible timing. The real‐time PCR method enables revised understanding of how many sperms are stored by female Q‐flies, the distribution of storage between the two spermathecae and the relationship between copula duration and sperm storage. Real‐time PCR assays based on Y‐specific markers provide an effective solution for sperm quantification in tephritid flies, as well as in other insects and potentially other animals with sperm storage organs.

No abstract

Multiple mating by females, polyandry, is common in insects, including in tephritid fruit flies. Female insects that remate commonly store sperm of multiple males. How the sperm of different males contribute to paternity is an important element of sexual selection. Sexual behavior and reproduction of the Queensland fruit fly (Qfly), Bactrocera tryoni, has been extensively investigated both in relation to understanding this economically important species’ reproductive biology and in relation to implications for Sterile Insect Technique (SIT), whereby sterile flies are released to constrain reproduction of pest populations. Despite numerous studies of pre‐ and postcopulatory sexual selection in Qfly, there have been no direct studies of paternity patterns in polyandrous female Qflies. We used two morphologically distinguishable lines to investigate patterns of sperm use in Qfly. The two lines showed comparable mating performance evidenced by similar mating and remating frequency, copula duration, and proportion of second mate paternity (P2) between reciprocal crosses. The mechanism of sperm usage, with P2 close to 0.5 immediately after the second mating followed by gradual decrease of P2 as females aged, is most consistent with stratification or repositioning of sperm. Patterns observed in the present study are compared with the available information from other tephritid fruit flies, and are discussed in relation to this species’ reproductive biology, known patterns of sperm storage, and SIT.