title: The W- and Z-linked EE0.6 sequences used for molecular sexing of captive Japanese crested ibis on Sado Island.
abstract: The Japanese crested ibis Nipponia nippon is a critically threatened bird. Accurate sexing is necessary to perform effective management of captive breeding toward a national project for a tentative release of the Japanese crested ibis on Sado Island. A PCR-based sexing method targeting a 0.6 kb EcoRI fragment (EE0.6) sequence on W chromosome with AWS03 and USP3 primers has been developed for the Japanese crested ibis. However, the primers were selected from the EE0.6 sequences from bird species other than the Japanese crested ibis. In this study, we determined the W- and Z-linked EE0.6 sequences in the Japanese crested ibis, and clarified Japanese crested ibis sequence mismatch in the binding sites of the primers. Further, we found no polymorphism in the primer binding sites among five founder birds for the Sado captive Japanese crested ibis population. These findings validated the PCR-based sexing method with the AWS03 and USP3 as accurate molecular sexing methods of captive Japanese crested ibis on the Sado Island. Additionally, we designed a primer set for a novel PCR-based sexing, based on the EE0.6 sequences obtained in this study. This novel sexing method may be useful for future ecological research following the release of Japanese crested ibis on Sado Island. This is the first report to show the EE0.6 sequences in Japanese crested ibis.
authors.affiliation: Department of Agrobiology, Faculty of Agriculture, Niigata University, Nishi, Niigata, Japan.
authors.forename: Kaori
authors.initials: K
authors.lastname: Kasuga
authors.forename: Maho
authors.initials: M
authors.lastname: Higashi
authors.forename: Takahisa
authors.initials: T
authors.lastname: Yamada
authors.forename: Toshie
authors.initials: T
authors.lastname: Sugiyama
authors.forename: Yukio
authors.initials: Y
authors.lastname: Taniguchi
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Iwaisaki
authors.forename: Kosuke
authors.initials: K
authors.lastname: Homma
authors.forename: Yuuichi
authors.initials: Y
authors.lastname: Wajiki
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Kaneko
authors.forename: Satoshi
authors.initials: S
authors.lastname: Yamagishi
mesh_terms: D000818:Animals; D001483:Base Sequence / Q000235:genetics; D001717:Birds / Q000235:genetics* / Q000502:physiology; D001947:Breeding; D002875:Chromosomes / Q000235:genetics; D005260:Female; D007564:Japan; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D017421:Sequence Analysis; D012732:Sex Determination Analysis / Q000379:methods*