Exome sequencing in allotetraploid wheat genome
Targeted analysis of nucleotide and copy number variation by exon capture in allotetraploid wheat genome
A total of 3497 full length cDNA downloaded from the TriFlcDNA database were captured and sequenced in wild emmer wheat and cultivated durum wheat. Efficiency and performance of capture assay in polyploid genome was evaluated. This data was further used to assess the nucleotide variation and CNV between these two genomes.
Ability of grass species to adapt to various habitats is attributed to the dynamic nature of their genomes, which have been shaped by multiple rounds of ancient and recent polyploidization. To gain a better understanding of the nature and extent of variation in functionally relevant regions of a polyploid genome, we developed a sequence capture assay to compare exonic sequences of allotetraploid wheat accessions.   A sequence capture assay was designed for the targeted re-sequencing of 3.5 Mb exon regions which surveyed a total of 3,497 genes from allopolyploid wheat. This data was used to describe SNPs, copy number variation and homoeologous sequence divergence in coding regions. A procedure for variant discovery in the polyploid genome was developed and experimentally validated. About 1% and 24% of discovered SNPs were loss-of-function and non-synonymous mutations, respectively. Under-representation of replacement mutations was identified in several groups of genes involved in translation and metabolism. Gene duplications were predominant in cultivated wheat, while more gene deletions than duplications were identified in wild wheat.   We demonstrated that, even though the level of sequence similarity between targeted polyploidy genomes and capture baits can bias enrichment efficiency, exon capture is a powerful approach for variant discovery in polyploids. Our results suggest that allopolyploid wheat can accumulate new variation in coding regions at a high rate. This process has a potential to broaden functional diversity and generate new phenotypic variation that eventually can play critical role in the origin of new adaptations and important agronomic traits.