Fine mapping of complex traits in non-model species: using next generation sequencing and advanced intercross lines in Japanese quail.
BACKGROUND
As for other non-model species, genetic analyses in quail will benefit greatly from a higher marker density, now attainable thanks to the evolution of sequencing and genotyping technologies. Our objective was to obtain the first genome wide panel of Japanese quail SNP (Single Nucleotide Polymorphism) and to use it for the fine mapping of a QTL for a fear-related behaviour, namely tonic immobility, previously localized on Coturnix japonica chromosome 1. To this aim, two reduced representations of the genome were analysed through high-throughput 454 sequencing: AFLP (Amplified Fragment Length Polymorphism) fragments as representatives of genomic DNA, and EST (Expressed Sequence Tag) as representatives of the transcriptome.


RESULTS
The sequencing runs produced 399,189 and 1,106,762 sequence reads from cDNA and genomic fragments, respectively. They covered over 434 Mb of sequence in total and allowed us to detect 17,433 putative SNP. Among them, 384 were used to genotype two Advanced Intercross Lines (AIL) obtained from three quail lines differing for duration of tonic immobility. Despite the absence of genotyping for founder individuals in the analysis, the previously identified candidate region on chromosome 1 was refined and led to the identification of a candidate gene.


CONCLUSIONS
These data confirm the efficiency of transcript and AFLP-sequencing for SNP discovery in a non-model species, and its application to the fine mapping of a complex trait. Our results reveal a significant association of duration of tonic immobility with a genomic region comprising the DMD (dystrophin) gene. Further characterization of this candidate gene is needed to decipher its putative role in tonic immobility in Coturnix.
INRA, UMR444 Laboratoire de Génétique Cellulaire, Castanet-Tolosan, F-31326, France.
Laure
L
Frésard
Sophie
S
Leroux
Patrice
P
Dehais
Bertrand
B
Servin
Hélène
H
Gilbert
Olivier
O
Bouchez
Christophe
C
Klopp
Cédric
C
Cabau
Florence
F
Vignoles
Katia
K
Feve
Amélie
A
Ricros
David
D
Gourichon
Christian
C
Diot
Sabine
S
Richard
Christine
C
Leterrier
Catherine
C
Beaumont
Alain
A
Vignal
Francis
F
Minvielle
Frédérique
F
Pitel
D054458:Amplified Fragment Length Polymorphism Analysis; D000818:Animals; D030161:Avian Proteins / Q000235:genetics*; D002645:Chickens / Q000235:genetics; D002874:Chromosome Mapping; D002875:Chromosomes; D003370:Coturnix / Q000235:genetics*; D003433:Crosses, Genetic; D016189:Dystrophin / Q000235:genetics*; D020224:Expressed Sequence Tags; D005260:Female; D056726:Genetic Association Studies; D016678:Genome; D005838:Genotype; D059014:High-Throughput Nucleotide Sequencing; D046353:Immobility Response, Tonic; D008297:Male; D010641:Phenotype; D020641:Polymorphism, Single Nucleotide; D040641:Quantitative Trait Loci; D059467:Transcriptome
D030161:Avian Proteins; D016189:Dystrophin