The phylogenetic positions of 22 isolates that morphologically resemble members of the family Planctomycetaceae were determined by sequence analysis of genes coding for 16S rRNA. While nine and eight isolates could be assigned to the genera Planctomyces and Pirellula, respectively, three strains grouped near Isosphaera pallida and one strain was closely related to Gemmata obscuriglobus. No isolate was found to be related to a previously described species of any of the four genera at the species level. Morphological characters and sequence idiosyncrasies of genes coding for 16S rRNA of the isolates generally correlated with features described for the four genera to which the isolates could be assigned. One strain stands phylogenetically isolated and may be representative of a novel genus of the family. Comparison with environmental clone sequences representing planctomycetes in soil and water revealed that three of the novel isolates were related to one clone of soil origin, but no close relationships between clones and the other new strains were found. The study reveals that the biodiversity of planctomycetes is significantly greater than was previously determined.During the 1920s, Gimesi (9) described planktonic, colonyforming microorganisms consisting of threadlike forms which bore spherical structures. These features were interpreted as fungal hyphae and exospores, respectively; therefore, this organism was described as a fungus, Planctomyces bekefii. A decade later, Henrici and Johnson (11), unaware of Gimesi's work, observed morphologically similar organisms which they interpreted as bacteria. The name Blastocaulis sphaerica was proposed for these stalked forms. The authors also found spore-forming, drop-shaped, colony-forming, budding bacteria lacking a stalk, which they believed to be identical to Pasteuria ramosa (29). These bacteria were later renamed Blastobacter henricii (62).In the following decades, P. bekefii-like bacteria from middle and northern Europe (12,18,23,30,45,55), Java (34), India, and Vietnam (19) were repeatedly reported. Besides P. bekefii, other budding bacteria were observed in diverse aquatic habitats such as eutrophic freshwater lakes (17,23,31,32), fish ponds (18), brackish water (13, 17), aquarium water and marine sediments (17), and forest brook sediments (61). Stalkless organisms have been found in seawater (17) and freshwater lakes (15). A thermophilic form was found at 60ЊC in hot springs in Tiberias, Israel (21). Several Planctomyces species, such as P. condensatus (46), P. stranskae and P. subulatus (55), P. ferrimorula (56), P. gracilis, P. crassus, and P. guttaeformis (18), and Blastocaulis kljasmensis (62), were described. The identity of P. bekefii and B. sphaerica was described by Hirsch (14), who concluded that the name Planctomyces should have priority over Blastocaulis. The genus was restricted to ''spherical, oblong or pear-shaped bacteria with long and slender stalks when in the mature state.'' Consequently, organisms that lacked a stalk were excluded from the gen...