Summary: The group of planctomycetes represents a separate line of descent within the domain Bacteria. Two phylum-specific 16S rRNA-targeted oligonucleotide probes for planctomycetes have been designed, optimized for in situ hybridization and used in different habitats to detect members of the group in situ. The probes, named PLA46 and PLA886, are targeting all or nearly all members of the planctomycete line of descent. Planctomycetes could be detected in almost all samples examined, e.g. a brackish water lagoon, activated sludge, and other wastewater habitats. In situ probing revealed quite uniform morphology and spatial arrangement of the detected cells but profound differences in abundance ranging from less than 01% to several percentage of the total cells. Single coccoid cells with diameters between 1 and 25 m were dominating in most samples with the exception of the lagoon, in which rosettes of pear-shaped cells were abundant. The planctomycetes showed generally no hybridization signals with the bacterial probe EUB338, which is in accordance with base changes in their 16S rRNA sequences. A discrete ultrastructure of planctomycete cells was suggested by double staining with rRNA-targeted probes and the DNA-binding dye 4',6-diamidino-2-phenylindole (DAPI). The probe-conferred fluorescence was distributed in a ring-shaped manner around a central DAPI spot. The two probes developed extend the existing set of group-specific rRNA-targeted probes and help to elucidate the basic composition of bacterial communities in a first step of differential analysis. In situ hybridization of environmental samples indicated widespread presence of planctomycetes in different ecosystems.
AmannR.,
LudwigW.,
SchleiferK.H.1995; Phylogenetic identification and in situ detection of individual microbial cells without cultivation.. Microbiol Rev 59:143–169
AznarR.,
AmaroC.,
GarayE.,
AlcaideE.1991; Physicochemical and bacteriological parameters in a hypereutrophic lagoon (Albufera lake, Valencia, Spain).. Zentbl Mikrobiol 146:311–321
BondP.L.,
HugenholtzP.,
KellerJ.,
BlackallL.L.1995; Bacterial community structures of phosphate-removing and nonphosphate-removing activated sludges from sequencing batch reactors.. Appl Environ Microbiol 61:1910–1916
FuerstJ.A.,
GwilliamH.G.,
LindsayM.,
LichanskaA.,
BelcherC.,
VickersJ.E.,
HugenholtzP.1997; Isolation and molecular identification of planctomycete bacteria from postlarvae of the giant tiger prawn, Penaeus monodon.
. Appl Environ Microbiol 63:254–262
GiovannoniS.J.,
SchabtachE.,
CastenholzR.W.1987; lsosphaera pallida gen. nov. and comb.nov. a gliding, budding eubacterium from hot springs.. Arch Microbiol 147:276–284
GodonJ.-J.,
ZumsteinE.,
DabertP.,
HabouzitF.,
MolettaR.1997; Molecular microbial diversity of an anaerobic digestor as determined by small-subunit rDNA sequence analysis.. Appl Environ Microbiol 63:2802–2813
LeeS.-Y.,
BollingerJ.,
BezdicekD.,
OgramA.1996; Estimation of the abundance of an uncultured soil bacterial strain by a competitive quantitative PCR method.. Appl Environ Microbiol 62:3787–3793
LiesackW.,
StackebrandtE.1992; Occurrence of novel groups of the domain Bacteria as revealed by analysis of genetic material isolated from an Australian terrestrial environment.. J Bacteriol 174:5072–5078
LindsayM.R.,
WebbR.I.,
FuerstJ.A.1997; Pirellulosomes: a new type of membrane-bounded cell compartment in planctomycete bacteria of the genus Pirellula.
. Microbiology 143:739–748
NeefA.,
ZaglauerA.,
MeierH.,
AmannR.,
LemmerH.,
SchleiferK.H.1996; Population analysis in a denitrifying sand filter: conventional and in situ identification of Paracoccus spp. in methanol-fed biofilms.. Appl Environ Microbiol 62:4329–4339
SchlesnerH.1994; The development of media suitable for the microorganisms morphologically resembling Planctomyces spp., Pirellula spp., and other Planctomycetales from various aquatic habitats using dilute media.. Syst Appl Microbiol 17:135–145
SchlesnerH.,
StackebrandtE.1986; Assignment of the genera Planctomyces and Pirella to a new family Planctomycetaceae fam. nov. and description of the order Planctomycetales ord. nov.. Syst Appl Microbiol 8:174–176
StaleyJ.T.,
FuerstJ.A.,
GiovannoniS.,
SchlesnerH.1992; The order Planctomycetales and the genera Planctomyces, Pirellula, Gemmata, and lsosphaera.
. In The Prokaryotes 2nd edn IV: pp. 3710–3731BalowsA., TrüperM., DworkinW.,
HarderW.,
SchleiferK.H.
Edited by New York: Springer;
WagnerM.,
AmannR.,
LemmerH.,
SchleiferK.H.1993; Probing activated sludge with proteobacteria-specific oligonucleotides: inadequacy of culture-dependent methods for describing microbial community structure.. Appl Environ Microbiol 59:1520–1525
WardN.,
RaineyF.A.,
StackebrandtE.,
SchlesnerH.1995; Unraveling the extent of diversity within the order Plancto- mycetales.
. Appl Environ Microbiol 61:2270–2275
Ward-RaineyN.,
RaineyF.A.,
WellingtonE.M.H.,
StackebrandtE.1996; Physical map of the genome of Planctomyces limnopbilus, a representative of the phylogenetically distinct planctomycete lineage.. J Bacteriol 178:1908–1913
ZardaB.,
HahnD.,
ChatzinotasA.,
SchönhuberW.,
NeefA.,
AmannR.,
ZeyerJ.1997; Analysis of bacterial community structure in bulk soil by in situ hybridization.. Arch Microbiol 168:185–192