丹顶鹤(Grus japonensis)性别鉴定的分子生物学方法

性别鉴定对鸟类特别是珍稀濒危鸟类的饲养管理、人工繁育、遗传疾病的防治和进化研究等方面都有非常重要的意义。丹顶鹤是我国一级保护动物，为单态性鸟，很难通过外观和形态来进行性别鉴别，给配对和人工繁殖造成了很大困难。本试验利用非伤害性取样方法，成功从丹顶鹤羽毛中提取了基因组DNA，并筛选出3对引物组合对丹顶鹤EE0.6序列的相关片段进行了特异性扩增。结果显示，这3对引物组合从雄鹤中扩增出一条片段，从雌鹤中扩增出两条片段，均可以对丹顶鹤的性别作出准确鉴定，解决了丹顶鹤人工繁育中的关键问题，并为其他濒危鸟类的准确性别鉴定提供了参考和借鉴。

The Molecular Method of Sex identification in Red-Crowned Cranes (Grus japonensis)

Sex identification was essential for the management, human-assisted breeding, prevention and cure of genetic disease and evolutionary studies of birds, especially for endangered birds. Red-Crowned Crane is a first class protected animal in China and a kind of monomorphic bird, so it’s difficult to identify the sex with morphological method, this situation challenged the couple and human-assisted breeding of Red-Crowned Crane. Genome DNA was extracted from feathers by using nondestructive sampling method and three combinations of primers were selected to amplify the EE0.6 sequence of Red-Crowned Crane specifically. The results showed that male birds displayed only one distinct amplification band, whereas two bands were amplified for female ones. With this molecular method in the article, the sex of Red-Crowned Crane could be identified successfully and accurately and then solved the key problem during the human-assisted breeding of Red-Crowned Crane. This method also gave a guide and reference for the accurate sex identification of other endangered birds.