MOLECULAR SEXING OF GREEN-CHEEKED CONURE (PYRRHURA MOLINAE) USING CHD1 AND NIPBL GENES
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Date
2014
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COLLEGE OF VETERINARY AND ANIMAL SCIENCES, POOKODE WAYANAD
Abstract
Sexing in birds is important for behavioral and ecological studies and also
to improveex situ captive breeding programme. More than 50% of the world’s
avian population are monomorphic and are difficult to sex phenotypically. Some
avian species exhibit marked sexual dimorphism as adults, but lack dimorphism as
juveniles. Monomorphic nature of many pet birds creates problems for pet
breeders in identifying the sex. Traditional methods of avian sexing were based on
behavioral observations, difference in size and shape, differences in the sounds
produced, cloacal examination, surgical techniques and cytogenetic analysis. Most
of them are time consuming, invasive and often generate ambiguous results. In the
present study, PCR based techniques were used to identify the sex of a popular pet
bird, Pyrrhura molinae (Green-cheeked conure), which is monomorphic in nature.
The gene CHD1 (encoding chromo helicase- DNA-binding protein 1) is located in
both the sex chromosomes, Z and W of birds.Intron 9 of CHDI gene was
amplified by PCR technique in DNA isolated from the feather. Retroposon
insertion in the region of amplification in Z chromosome generated a larger
fragment. As female birds are heterogametic (ZW) PCR amplification generated
fragments of two different sizes- 1100 bp and 600 bp, whereas in male birds (ZZ),
only a single fragment of 1100 bp was observed. It revealed two different patterns
in male and female birds which help to identify the sex accurately. Intron 16
region of CHDI gene was also amplified to see for sex specific difference in
amplicon pattern between male and female birds. Though five out of thirty birds
generated distinct amplification pattern rest of the birds did not generate
unambiguous results. The result was similar for NIPBL intron 16 also. It appears
that amplification of CHD1 intron 9 region is the most reliable method in sexing
of green cheeked conure. As the method is non-invasive, it produces minimum
stress to the bird and can be employed for easy and accurate sexing of the bird.
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