The W- and Z-linked EE0.6 sequences used for molecular sexing of captive Japanese crested ibis on Sado Island.
The W- and Z-linked EE0.6 sequences used for molecular sexing of captive Japanese crested ibis on Sado Island.
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DOI：
10.1111/j.1740-0929.2011.00971.x
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发表时间：
2012
期刊：
影响因子：
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通讯作者：
Kaori Kasuga;Maho Higashi;Takahisa Yamada;T. Sugiyama;Y. Taniguchi;H. Iwaisaki;Kosuke Homma;Y. Wajiki;Y. Kaneko;S. Yamagishi
中科院分区：
文献类型：
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作者：
Kaori Kasuga;Maho Higashi;Takahisa Yamada;T. Sugiyama;Y. Taniguchi;H. Iwaisaki;Kosuke Homma;Y. Wajiki;Y. Kaneko;S. Yamagishi
日本朱鹮是一种严重受威胁的鸟类。为了对佐渡岛朱鹮试放国家项目的圈养繁殖进行有效管理，必须进行准确的性别鉴定。针对日本朱鹮，开发了一种基于 PCR 的性别鉴定方法，该方法使用 AWS03 和 USP3 引物，针对 W 染色体上的 0.6 kb EcoRI 片段 (EE0.6) 序列。然而，引物选自朱鹮以外的鸟类的EE0.6序列。在本研究中，我们确定了朱鹮的W连锁和Z连锁EE0.6序列，并阐明了朱鹮引物结合位点的序列不匹配。此外，我们发现佐渡圈养朱鹮种群的五种始祖鸟的引物结合位点没有多态性。这些发现验证了基于 PCR 的 AWS03 和 USP3 性别鉴定方法是佐渡岛上圈养的朱鹮的准确分子性别鉴定方法。此外，我们根据本研究中获得的 EE0.6 序列，设计了用于新型 PCR 性别鉴定的引物组。这种新颖的性别鉴定方法可能对佐渡岛朱鹮放归后的未来生态研究有用。这是第一份显示日本朱鹮 EE0.6 序列的报告。
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.