Plant & Animal Genomes XVI Conference

January 12-16, 2008
Town & Country Convention Center
San Diego, CA

Cot-Filtration In Banana (Musa acuminata): A Promising Tool For Gene Discovery

Rita Aert¹ , Eva Hribová² , Jaroslav Dolezel² , Rony Swennen¹ , László Sági¹

¹  Laboratory of Tropical Crop Improvement, Division of Crop Biotechnics, Katholieke Universiteit Leuven, Kasteelpark Arenberg 13, B-3001 Leuven, Belgium

²  Laboratory of Molecular Cytogenetics and Cytometry, Institute of Experimental Botany, Sokolovská 6, CZ-77200 Olomouc, Czech Republic

The efficacy of Cot filtration and its application to gene discovery in the banana genome will be presented. Two Cot-libraries (Cot100 and Cot500) and an unfiltered library of Musa acuminata (var. ‘Calcutta 4’) were constructed. In total, 1317 clones (or 627,120 bp) were sequenced in these libraries, and 244 novel genic sequences, of which only four showed homology to genes previously annotated in banana, were identified. The expression of two coding sequences, allene oxide cyclase and peroxidase, was demonstrated to indicate genuity of Cot-filtered aequences. Cot-filtration efficiency was established by comparing sequences obtained at Cot100 and Cot500 to sequences acquired in an unfiltered (shotgun) library. Cot100 clones contained 1.8-fold gene enrichment, a 1.75-fold increase in unknown low-copy sequences and a 2.5-fold reduction of the repetitive DNA. Increasing the Cot value to Cot500 fortified gene enrichment and reduction of repetitive DNA fraction. These results indicate that Cot-filtration in banana is a useful means for the discovery of novel genes.