| Lushi chicken is unique flakiness feather native breed which could lay green-shelleggs in China, there are both color of green and brown in Lushi chicken eggshell. Butthere are only about2～3%individuals that could lay green-shell eggs of the totalpopulations. As the Oocyan（O） which decides the color of green eggshell is located onthe chromosome1st, so two marker locus points from the1st chromosome of chickenwere selected in this study, which were closely linked to Oocyan. Microsatellite lociAY493302and Endogenous virus factor EV1loci were selected in this study. Bygathered blood of150green-shell chicken,50brown-shell chicken and other18white-shell native chicken、extracted genomic DNA and carried out PCR amplification, thegenotype differences among these three eggshell colors and the polymorphisms wereanalyzed and discussed, and the correlationship between these two marker locus andeggshell color were furthermore analyzed. The results of this study will be providedtheoretical basis and technical support for molecular marker-assisted selection of greenshell traits in future. The results were shown as bellow：1. The results showed that the genotype distributions of this MicrosatelliteAY493302marker among these three eggshell color chicken were significantlydifferent, the frequency of genotype AA（225bp/225bp） were0.3969in green-shellchicken. Obviously, this genotype existed only in green-shell chicken, while not inbrown-shell chicken and white-shell chicken respectively.The sequencing revealed that the base in the19th bp and217th bp were both G ingenetype AA, and the base in the19th bp was C and217th bp was G in genetype ABfor Lushi green-shell chicken. Base in the19th bp was C and217th bp was T ingenetype AB for Lushi brown-shell chicken and native white-shell chicken.By compared green-shell AA type sequence with homologous sequence of Gallusgallus in NCBI database, the results were showed that an extremely high level of identity （99%）, V00425.1, J02015.1and NW003763484.1which were located inchicken chromosome1st.2. The results showed that the genotype distributions of this EV1marker lociamong these three eggshell color chicken were significantly different, the frequency ofgenotype AA （251bp/251bp） were0.4348in green-shell chicken. Obviously, thisgenotype existed only in green-shell chicken, while not in brown-shell chicken andwhite-shell chicken respectively.The sequencing revealed that the base in the52th bp was T、54th bp was T and129th bp was A in genetype AA for green-shell chicken; and the base in the52th bpwas T、54th bp was T and129th bp A was changed T in genetype AB for green-shellchicken; but the base in the52th bp was G、54th bp was G and129th bp was T inLushi brown-shell chicken and native white-shell chicken.By compared green-shell AA type sequence with homologous sequence of Gallusgallus in NCBI database, the results were shown a high level of identity（89%）,NW003763686.1, which was located in chicken chromosome2st.3. The total populations PIC of of Lushi green-shell chicken,brown-shell chickenand native white-shell chicken on microsatellite AY493302and EV1marker locusrespectively was0.3325、0.3750、0.3750, and0.3233、0.3750、0.3750, which aremoderate polymorphism locus.The loci heterozygosity of Lushi green-shell chicken, brown-shell chicken andnative white-shell chicken on microsatellite AY493302and EV1marker locusrespectively was0.4212、0.5000、0.5000, and0.4055、0.5000、0.5000, whichshow that has a certain degree of genetic diversity among Lushi green-shellchicken,brown-shell chicken and native white-shell chicken.4. The results showed that the total populations of Lushi green-shellchicken,brown-shell chicken and other native white-shell chicken on microsatelliteAY493302and EV1marker locs respectively were not accorad with the Hardy-weinberg law by chisquare test.By chisquare test, the results showed that those different genotypes on themicrosatellite AY493302marker and EV1marker locus were significantly related withgenotype on the green shell gene loci. |
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