Spiroplasma strain MQ-lT (T = type strain) from the hemolymph of the vespid wasp Monobia quadridens differed serologically from other spiroplasma species, groups, and subgroups. Cells of strain MQ-lT were helical and motile and possessed a single cytoplasmic membrane, with no evidence of a cell wall. The organism grew in conventional mycoplasma medium, in serum fraction, SM-1, MlD, and SP-4 liquid media, and on SP-4 solid medium in either aerobic or anaerobic environments. The optimum temperature for growth was 32"C, but multiplication occurred over a wide temperature range (10 to 37°C). The doubling time at 32°C in M1D medium was 1.9 h. Strain MQ-lT catabolized glucose but hydrolyzed neither arginine nor urea. Previous work showed that strain MQ-lT has a unique methylase, previously known only in eucaryotes. Also, strain MQ-lT induces production of tumor necrosis factor in bone marrow macrophages. The guanine-plus-cytosine content of the DNA was 28 f 1 mol%. The genome size of strain MQ-lT was 940 kb (627 MDa); a similar strain, MQ-8, had a genome size of 985 kb (657 MDa). Strain MQ-lT and its allies have the smallest genomes known in the genus Spiroplasma. Strain MQ-1 (= ATCC 33825) is designated the type strain of a new species, Spirophsma monobiae.The genus Spiroplasma (36), originally thought to consist of plant pathogens (22), was later shown, largely through the efforts of T. B. Clark and his associates (7,15), to be associated with arthropods. As the immense diversity of this genus unfolded, numerous interesting properties emerged. For example, group I organisms (2) were the only spiroplasmas found to be associated with plant disease or with plant-sucking bugs. Yet, the subgroup 1-2 spiroplasma, Spiroplasma melliferum (8), which was closely related to the plant spiroplasmas, was primarily associated with bees. One subgroup of this cluster (subgroup I-6), which appears to occur in the insect-plant surface habitat (15), had a guanineplus-cytosine (G+C) content that was higher than the G+C contents of its cohort subgroups (5). Another spiroplasma (subgroup I-4), isolated only once, appeared to be associated with ticks. Group I1 spiroplasmas (34) produced sex ratio abnormalities in Drosophila flies. Group 111 spiroplasmas (9) (Spiroplasma jloricola) proved to be associated with magnoliaceous flowers and their beetle and fly visitors and to have idiosyncratic metabolic pathways (6) that were readily studied because of the rapid growth of the species. Group IV appeared to be a widely distributed cluster of spiroplasmas, some of whose members, including type strain B31 (Spiroplasma apis), were pathogenic (19, 20) for honeybees @pis mellifera). Groups V (Spiroplasma mirum [30]) and VI (29) were associated with ticks. Thus, each of the first six established spiroplasma groups appeared to possess unusual biological and/or ecological properties.For some time, the group VII spiroplasmas, strain MQ-lT (T = type strain) and its allies, appeared to be exceptions to the emerging uniqueness of newly discovered strains. H...