Sex-specific DNA markers for sexing in Orthopsittaca manilata, Neophema pulchella and Cacatua galerita elenora

• Main Authors: Kai-Hau Lian, 練凱豪
• Other Authors: Yan-Ming Horng
• Format: Others
• Language: zh-TW
• Online Access: http://ndltd.ncl.edu.tw/handle/33831532216156910411

碩士 === 國立嘉義大學 === 動物科學系研究所 === 103 === Red-bellied macaw (Orthopsittaca manilata) is native of northern South America, it has a bright red feather covering their abdomen as they named. Turquoise parrot (Neophema pulchella) is native of southeast Australia. Medium sulphur-crested cockatoo (Cacatua galerita elenora) is the one of a subspecies in four subspecies of sulphur-crested cockatoo (Cacatua galerita), it is native of Aru island in Indonesia. Red-bellied macaw, turquoise parrot and medium sulphur-crested cockatoo is hard to distinguish between males and females based on their external morphology. This study was performed to investigate the sex-specific fingerprinting by RAPD-PCR for sexing in red-bellied macaw (Orthopsittaca manilata), turquoise parrot (Neophema pulchella) and medium sulphur-crested cockatoo (Cacatua galerita elenora). The result showed that a female specific band in the length of 305 bp was found in the fingerprints of female red-bellied macaw amplified by random primer 1. The female-specific fragment was isolated and cloned into vector for nucleotide sequencing. The RbMsex F&;R primers were designed according to the cloned female-specific sequence for red-bellied macaw sexing by PCR. The PCR results showed that a 305 bp specific band can be observed in the females only, and a 400 bp band can be found in both of males and females. In addition, the RbMsex F&;R primers are also available for sexing of Ara ararauna, Ara chloroptera, Diopsittaca nobilis, Ara macao, Harlequin Macaw, Ara auricollis, Ruby Macaw, Camelot Macaw and Catalina Macaw. A female specific band in the length of 366 bp was found in the fingerprints of female turquoise parrot amplified by random primer 2. The female-specific fragment was isolated and cloned into vector for nucleotide sequencing. The Npulsex F&;R primers were designed according to the cloned female-specific sequence for turquoise parrot sexing by PCR, and the 18S ribosomal gene primers is added for internal control. The PCR results showed that a 366 bp specific band can be observed in the females only. In addition, the Npulsex F&;R primers are also available for sexing of Neopsephotus splendida, Platycercus eximius, Platycercus flaveolus and Platycercus bourkii. A female specific band in the length of 626 bp was found in the fingerprints of female medium sulphur-crested cockatoo amplified by random primer 3. The female-specific fragment was isolated and cloned into vector for nucleotide sequencing. The MCTsex F&;R primers were designed according to the cloned female-specific sequence for medium sulphur-crested cockatoo sexing by PCR, and the 18S ribosomal gene primers is added for internal control. The PCR results showed that a 626 bp specific band can be observed in the females only. In addition, the MCTsex F&;R primers are also available for sexing of Cacatua alba, Cacatua ophthalmica, Cacatua ducorpsii, Eolophus roseicapilla and Cacatua moluccensis. In conclusion, the primers RbMsex F&;R, Npulsex F&;R and MCTsex F&;R can be used easily and accurately for PCR sexing in red-bellied macaw, nine species of macaw, turquoise parrot, four species of Australian parrots, medium sulphur-crested cockatoo and five species of family Cacatuidae, respectively.