Methyl Filtration As a Tool For Enrichment Of Sugarcane Euchromatic Regions

Modern sugarcane cultivars are hybrids of Saccharum officinarum and Saccharum spontaneous species. Hybrids have complex polyploidy genomes that difficult the studies of structure and organization of the genome. Here, we have used methyl filtration with McrBC digestion to enrich for euchromatic regions of the sugarcane genome. Total genomic DNA of the sugarcane cultivar SP70-1143 was extracted from purified nuclei isolated from leaves, and digested with McrBc. Three DNA libraries were prepared: in the first, DNA was sheared to 200 bp; then two jumping libraries of 2kb and 5kb, respectively, were prepared. The three fractions were then sequenced by Illumina, yielding approximately 24 Gb of sequences. The reads were then mapped to the sorghum genome to estimate the coverage of the coding regions. Initial analyzes indicate a significant representation of genes. Additional analyzes are in progress to determine the representation of regulatory portions of the genome and of DNA sequences representing noncoding RNA.  This study is an initial step towards the discovering of structure and organization of sugarcane genome.

Supported by CNPq, FINEP/Genoprot, CAPES and INCT