Characterising microsatellite loci in the blue crane (Grus paradisea)

Abstract

The blue crane (Grus paradisea) is endemic to southern Africa and has the smallest geographical range of the 15 crane species occurring world-wide. Although this species is still found throughout most of its historic range, it has experienced a significant and rapid decline in numbers over the last 20 years. One factor causing this decline is the illegal removal of chicks from the wild. Permits are required to keep, trade in and breed cranes in captivity. However, birds must be captive bred in order to obtain a permit. Therefore, chicks taken illegally from the wild are fraudulently incorporated into an existing captive population under the pretence that they offspring of a legal captive pair. This study describes the development of a set of microsatellite markers to assist the identification of illegal trade in the blue crane. These markers can ultimately be used to verify the relationship between the offspring and its claimed parents by performing parentage analyses. Forty microsatellite loci were obtained from genomic libraries previously developed in two other crane species and tested for cross-species utility in the blue crane. In addition, 42 loci were developed for this study from a blue crane species-specific genomic microsatellite library, of which 19 were tested for polymorphism in this species. The microsatellite markers characterised here were also tested for their utility in two other crane species: wattled crane (G. carunculatus) and grey-crowned crane (Balearica regulorum). One locus, Gamu007, was found to be sex-linked and therefore excluded from the set of markers. A total of 28 polymorphic loci were tested for the suitability in parentage analysis in the blue crane. Of these, a set of 16 loci were determined to be as suitable for this purpose. These loci were shown to be inherited in a Mendelian fashion in a single blue crane family. In addition, statistical analysis of the loci were identified as exhibiting linkage equilibrium, this was supported by their distant association on a predicted Grus microsatellite map based on the chicken genome. The selected loci were also identified as having a low frequency of null alleles as well as a total first and second parent exclusion power of 0.9999 and 1.0000, respectively. These loci provide a valuable tool for parentage testing in blue crane, and may also be valuable in population genetic studies to assist conservation strategies. In addition, this set may be used to assist legal cases involving the illegal trade in blue cranes upon completion of additional microsatellite marker validation procedures. Twenty-seven loci were polymorphic in the wattled and grey-crowned crane. These could provide a valuable source of micro satellite loci in these species, and could potentially eliminate the need for the development of a species-specific microsatellite library.