Streptococci and acholeplasmas have a close evolutionary relationship. We examined their genomes to determine what chromosomal losses occurred to produce the smaller acholeplasma genomes and found by RNA-DNA hybridization that Streptococcus cremoris and S. lactis possess at least five or at least six ribosomal RNA gene sets, respectively, while acholeplasmas have only two rRNA gene sets. Other important deficiencies identified in mycoplasmas are associated with envelope or RNA genes, and analysis of these chromosomal deletions may lead to an understanding of how mycoplasmas evolved from walled bacteria.