Detection of candidate gene LsACOS5 and development of InDel marker for male sterility by ddRAD-seq and resequencing analysis in lettuce.

A new breeding method of F hybrid using male sterility would open an exciting frontier in lettuce breeding, a self-pollinating crop. Male sterility is a crucial trait in F hybrid breeding. It is essential to map the causative gene for using male sterility. The ms-S, male-sterile (MS) gene of 'CGN17397', was mapped to linkage group (LG) 8 by ddRAD-seq and narrowed down between two markers using two F populations. This region spans approximately 10.16 Mb, where 94 genes were annotated according to the lettuce reference genome sequence (version8 from 'Salinas'). The whole-genome sequencing of the MS lines 'CGN17397-MS' and male-fertile (MF) lines 'CGN17397-MF' revealed that only one gene differed in the area of Lsat_1_v5_gn_8_148221.1, a homolog of acyl-CoA synthetase5 (ACOS5), and was deleted in the MS lines. It was reported that ACOS5 was needed for pollen wall formation and that the null mutants of ACOS5 were entirely male sterility in some plants. Thus, I concluded that Lsat_1_v5_gn_8_148221.1 designated as LsACOS5 was a biologically plausible candidate gene for the ms-S locus. By using the structural polymorphism of LsACOS5, an InDel marker was developed to select the MS trait. The results obtained here provide valuable information for the genic male-sterility in lettuce.