Abstract
To establish phylogenetic relationships and estimate the intra and interspecific divergence, the amplification and the sequencing of the internal transcribed spacers of ribosomal DNA (ITS = ITS1 + 5.8S + ITS2) were analyzed in Tunisian complex Lolium-Festuca DNA. These spacer regions have evolved mainly by point mutations. Results revealed a high level of polymorphism within studied species. Significant similarities were observed between these two species and showed the existence of an important phylogenetic relationship. Besides, this molecular approach has revealed two new clusterings, with a homologous ITS gene namely: Bromus hordeaceus and Hordeum murinum subsp. This could be explained by the conservation of an ancestral ITS gene in some fescue plants. Thus, Tunisian tall fescue and perennial ryegrass may derive from Bromus hordeaceus and Hordeum murinum subsp. Considerable morphology and bioclimatic distribution similarities were discovered in ITS sequences within the same species. This study can be of great help to identify suitable accessions that could be used in local fescue and ryegrass improvement program.