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Real‐time PCR ‐based Y‐specific sperm quantification assay in Queensland fruit fly: Insights to patterns of sperm storage
Author(s) -
Shadmany J.,
Lee S. F.,
Taylor P. W.
Publication year - 2021
Publication title -
insect molecular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.955
H-Index - 93
eISSN - 1365-2583
pISSN - 0962-1075
DOI - 10.1111/imb.12697
Subject(s) - sperm , biology , spermatheca , real time polymerase chain reaction , andrology , botany , genetics , gene , medicine
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.