Genetic relationships among isolates of Colletotrichum gloeosporioides from Stylosanthes spp. in Africa and Australia using RAPD and ribosomal DNA markers

Thirty-three isolates of Colletotrichum gloeosporioides from various Stylosanthes species collected in Africa and Australia and associated with restricted (type A), extensive (type B) or nontypical anthracnose lesions (type C) were first compared by random amplified polymorphic DNA (RAPD) analysis. A phylogenetic tree was constructed based on 118 reproducible polymorphic bands generated with 16 random primers, using the upgma method. Twenty-nine isolates were grouped in two main clusters, corresponding to types A and B, within which polymorphic subgroups were partially related to geographical origin. Strong similarities were observed among isolates of distant origin. Four isolates presented profiles completely different from the A and B types and were grouped in two additional clusters. To assess the phylogenetic relationship among isolates of various types and origins at the species level, the lnternal Transcribed Spacer region (ITS 1) of the ribosomal DNA was sequenced. Type A isolates and a restricted number of type B isolates selected in the RAPD clusters showed an homology of 99.4–100%. When compared with published sequence data, the isolates that were clustered separately in the phylogenetic tree, had the exact sequence of a C. gloeosporioides strain associated with the rotting of coffee berries, or of C. kahawae , the causal agent of coffee berry disease.