A triple-primer PCR approach for the sex identification of endangered Phasianidae birds

摘要

To manage and conserve endangered Phasianidae species and to study their ecology, behavior, and population structure, sex identification is necessary. A search for the pheasant sex-linked marker (PSL) was carried out by comparing the sequences of chicken sex chromosomes. A triple-primer PCR approach was established in the present study based on a novel PSL for the sex identification of Phasianidae birds. The triple-primer system amplified a 202-bp PSL-W-specific fragment in female Phasianidae birds, whereas there is a 107-bp PSL-Z-specific fragment in the both sexes, thus making the female distinct. We also found a 102-bp AT-rich repeat sequence inserted into the PSL-W-specific region in Tetraophasis szechenyii generating a ∼300-bp PSL-W fragment in females. The sex of 11 Phasianidae species was distinguished clearly by the triple-primer PCR approach indicating that this method is both quick and reliable and can be easily applied to a wider range of Phasianidae species. Due to the relatively small size of the amplification products, the triple-primer PCR approach can also be applied to noninvasive samples such as feathers.