The efficacy of Cot-based gene enrichment in wheat (Triticum aestivum L.).
We report the results of a study on the effectiveness of Cot filtration (CF) in the characterization of the gene space of bread wheat (Triticum aestivum L.), a large genome species (1C = 16,700 Mb) of tremendous agronomic importance. Using published Cot data as a guide, 2 genomic libraries for hexaploid wheat were constructed from the single-stranded DNA collected at Cot values > 1188 and 1639 M x s. Compared with sequences from a whole genome shotgun library from Aegilops tauschii (the D genome donor of bread wheat), the CF libraries exhibited 13.7-fold enrichment in genes, 5.8-fold enrichment in unknown low-copy sequences, and a 3-fold reduction in repetitive DNA. CF is twice as efficient as methylation filtration at enriching wheat genes. This research suggests that, with improvements, CF will be a highly useful tool in sequencing the gene space of wheat.
Lamoureux
Didier
D
Wheat Genetics Resource Center, Department of Plant Pathology, Kansas State University, Manhattan 66506, USA.
Peterson
Daniel G
DG
Li
Wanlong
W
Fellers
John P
JP
Gill
Bikram S
BS
D003001:Cloning, Molecular; D017343:Genes, Plant; D018745:Genome, Plant; D011237:Predictive Value of Tests; D012091:Repetitive Sequences, Nucleic Acid; D017422:Sequence Analysis, DNA / Q000639:trends; D014908:Triticum / Q000235:genetics*