Phylogenetic analysis of sclerospora graminicola using internal transcribed spaced region-2

Abstract

The internal transcribed spacer region (ITS) from fourteen samples of Sclerospora graminicola was amplified using a nested PCR. These regions of the rDNA were amplified by a combination of conserved primers (ITS 1 and 6 and ITS 3 and 4). These products were cloned into a puc-4 vector and transformed into E.coli competent cells. The inserts from transformants were isolated, then sequenced by the Gene Technologies Lab, Texas A& M University. The sequences produced were and aligned with the ITS2 region of other known Oomycete fungi( Peronospora, Phytophthora and Pythium) using CLUSTALX. Phylogenetic relationship among sequences from Indian and African samples was examined using a consensus bootstrap parsimony tree. All the samples from India, Mali, Nigeria and Niger, with the exception of Niger 4, formed a monophyletic group with the Oomycetes ( Peronospora, Phytophthora and Pythium). The three samples Niger 4, BFaso 2 and BFaso 17 from Burkina Faso shared a close relationship with the outgroup Cladosporium herbareum.