LAUSR

Green peafowl ( Pavo muticus ), a representative of the Southeast Asia Phasianidae, is assessed as globally endangered with populations that are declining. It is highly sensitive to human disturbance and habitat change, two threats that are unlikely to be removed soon in present range countries, and that increase the need for long-term continuous effort for in situ conservation. Captive breeding for reintroduction is potentially an important measure for localized population restoration. But green peafowl can hybridize with blue peafowl ( P. cristatus ) and produce fertile offspring that can backcross with ancestral species. Hybrids backcrossed to green peafowl for a few generations are indistinguishable from pure green peafowl in appearance. Such hybrids must be excluded from captive breeding programs to avoid anthropogenic genetic contamination. In this study, we developed technology to identify hybrids using mitochondrial Cyt b and COI gene sequences, and 16 microsatellites. We studied maternal species assignment for excluding individuals of the blue peafowl maternal line and genetic clustering analysis to exclude individuals containing blue peafowl nuclear genes. Maternal species assignment was accomplished using K2P pairwise genetic distance computed using transition nucleotide substitution. Nuclear genetic clustering analysis was performed using STRUCTURE (assuming K = 2) and NewHybrids. The method was validated using reference pure green (n = 18) and blue (n = 21) peafowl and known hybrids (n = 12). Two criteria were set up to discriminate hybrids based on the probability that an individual is either pure green or pure blue peafowl at 95% confidence. This method was applied to identification of 59 peafowl of ambiguous genetic composition. Our study provides an example for establishing a purity test workflow for other species that can hybridize and has potential for use in reintroduction of genetically pure green peafowl.