A PHYLOGENETIC ANALYSIS OF THE TRYPANOSOMA-CRUZI GENOME PROJECT CL BRENER REFERENCE STRAIN BY MULTILOCUS ENZYME ELECTROPHORESIS AND MULTIPRIMER RANDOM AMPLIFIED POLYMORPHIC DNA-FINGERPRINTING

We have assessed the phylogenetic status of the Trypanosoma cruzi Geno me Project CL Brener reference strain by multilocus enzyme electrophor esis (MLEE) and multiprimer random amplified polymorphic DNA (RAPD) in cluding a set of cloned stocks representative of the whole genetic div ersity of T. cruzi. MLEE and RAPD data gave congruent phylogenetic res ults. The CL Brener reference strain fell into the second major phylog enetic subdivision of T. cruzi, and was genetically very close to the Tulahuen reference strain. No reliable RAPD character and only one MLE E character permitted us to distinguish between the CL Brener and Tula huen reference strains. In contrast, many RAPD and MLEE characters wer e able to distinguish between the CL Brener reference strain and the o ther T. cruzi genotypes analyzed here, in particular the formerly desc ribed principal zymodemes I, II and III. It is suspected that both CL Brener and Tulahuen are hybrid genotypes, a fact that should be taken into account when interpreting sequence data. Moreover, our study conf irms that the species T. cruzi is genetically very heterogeneous. We r ecommend future comparison of sequencing data from the CL Brener refer ence strain with those of at least one radically distinct T. cruzi gen otype, belonging to the other major phylogenetic subdivision of this s pecies. (C) 1998 Elsevier Science B.V. All rights reserved.