Cryptoendolithic microorganisms from stratified communities in Antarctic sandstone were studied for physiological diversity and possible interactions. Cultures of 25 bacteria, five fungi, and two green algae from one boulder grew with a wide variety of organic carbon or nitrogen sources, they exhibited varied exoenzymatic activities and were psychrophilic or psychrotrophic. Many isolates excreted vitamins into the medium and were stimulated by other vitamins. Organic acid excretion and siderophore formation were common, but antibiotic activity was rare. Plasmids were found in 24% of the bacteria, and many of these strains showed resistance to antibiotics and heavy metals. A small plasmid (2.9 kb) from strain AA-341 was electrotransferred into sensitive isolates, thereby rendering these resistant to ampicillin and Cr 3÷. Bacterial cultures in spent algal medium and coculture with algae demonstrated beneficial (rarely inhibitory) interactions. A search for free organic compounds in zones of the sandstone community revealed sugars, sugar alcohols, organic acids and amino acids -in many cases the same compounds that were excreted into the laboratory medium. Data presented here indicate low taxonomic but high physiological diversity among these heterotrophic cryptoendoliths. This physiological diversity, as well as the spatial separation in layers with distinct activities, allows coexistence within the community and contributes to the stability of this ecosystem.
I nst it ut fur Al lg emei ne Mi kro biologie, U n iversitat Kiel, Am Botanischen Garten 1-9, 0-241 18 Kiel, Germany Almost complete 16s rDNA sequences from the type strains of seven species of the genus Hyphomicrobium and of Filomicrobium fusiforme have been determined. The Hyphomicrobium species form two phylogenetic clusters that are only moderately related to each other. While cluster I contains the type species Hyphomicrobium vulgare, Hyphomicrobium aestuarii, Hyphomicrobium hollandicum and Hyphomicrobium zavarzinii, cluster II comprises Hyphomicrobium facilis, Hyphomicrobium denitrificans and Hyphomicmbium methylovomm. Within the two species clusters, the species are highly related. Phylogenetically, Filomicmbium fusiforme clusters moderately with Hyphomicrobium species. The lack of distinguishing phenotypical properties presently excludes the possibility of describing cluster II as a new genus.
Abstract. The aim of this study was to compare structural differences in the nirS-type denitrifying microbial communities along the environmental gradients observed in the water column and coastal sediments of the Baltic Sea. To link community structure and environmental gradients, denitrifier communities were analyzed by terminal restriction fragment length polymorphism (T-RFLP) based on nirS as a functional marker gene for denitrification. nirS-type denitrifier community composition was further evaluated by phylogenetic analysis of nirS sequences from clone libraries. T-RFLP analysis indicated some overlap but also major differences between communities from the water column and the sediment. Shifts in community composition along the biogeochemical gradients were observed only in the water column while denitrifier communities were rather uniform within the upper 30 mm of the sediment. Specific terminal restriction fragments (T-RFs) indicative of the sulfidic zone suggest the presence of nitrate-reducing and sulfide-oxidizing microorganisms that were previously shown to be important at the suboxic-sulfidic interface in the water column of the Baltic Sea. Phylogenetic analysis of nirS genes from the Baltic Sea and of sequences from marine habitats all over the world indicated distinct denitrifier communities that grouped mostly according to their habitats. We suggest that these subgroups of denitrifiers had developed after selection through several factors, i.e. their habitats (water column or sediment), impact by prevalent environmental conditions and isolation by large geographic distances between habitats.
Budding methylotrophic bacteria resembling Hyphomicrobium spp. were counted for 12 months in a German sewage treatment plant by most-probable-number (MPN) methods. Influent samples contained up to 2 ؋ 10 4 cells ml ؊1 , activated sludge consistently contained 1 ؋ 10 5 to 5 ؋ 10 5 cells ml ؊1 , and the effluent contained 1 ؋ 10 3 to 4 ؋ 10 3 cells ml ؊1 . The receiving lake had only 2 to 12 cells ml ؊1 . Six morphological groups with different growth requirements could be observed among 1,199 pure cultures that had been isolated from MPN dilutions. With dot blot DNA hybridizations, 671 isolates were assigned to 30 hybridization groups (HGs) and 84 could not be classified. Only HG 22 hybridized with a known species, Hyphomicrobium facilis IFAM B-522. Fourteen HGs (HGs 8 to 20 and HG 22) were specific for the lake; most others occurred only in the treatment plant. HGs 1, 3, and 26 were found in the activated sludge tank throughout the year, and HGs 27 and 28 were found for most of the year. In summary, it was demonstrated that bacteria with nearly identical and specific morphologies and nutritional types showed a high level of genetic diversity, although they were isolated under the same conditions and from the same treatment plant or its receiving lake. A directional exchange of these genetically different populations was possible but less significant, as was shown by the establishment of distinct populations in specific stations.
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