title: Diversity of thermophilic and non-thermophilic Crenarchaeota at 80 degrees C.
abstract: A hot spring in the solfataric field of Pisciarelli (Naples-Italy) was analysed for Archaeal diversity. Total DNA was extracted from the environment, archaeal 16S rRNA genes were amplified with Archaea specific primers, and a clone library consisting of 201 clones was established. The clones were grouped in 10 different groups each representing a specific band pattern using restriction fragment length polymorphism (RFLP). Members of all 10 groups were sequenced and phylogenetically analyzed. Surprisingly, a high abundance of clones belonging to non-thermophilic Crenarchaeal clusters were detected together with the thermophilic archaeon Acidianus infernus in this thermophilic environment. Neither Sulfolobus species nor other hyperthermophilic Crenarchaeota were detected in the clone library. The relative abundance of the sequenced clones was confirmed by terminal restriction fragment analyses. Amplification of 16S rRNA genes from Archaea transferred from the surrounding environment was considered negligible because DNA from non-thermophilic Crenarchaeota incubated under conditions similar to the solfatara could not be PCR amplified after 5 min.
authors.affiliation: Environmental Microbiology & Biotechnology Group, BioCentrum-Danmarks Tekniske Universitet Søltofts Plads, Building 227, DK-2800 Lyngby, Denmark.
authors.forename: Thomas
authors.initials: T
authors.lastname: Kvist
authors.forename: Anett
authors.initials: A
authors.lastname: Mengewein
authors.forename: Stefanie
authors.initials: S
authors.lastname: Manzei
authors.forename: Birgitte K
authors.initials: BK
authors.lastname: Ahring
authors.forename: Peter
authors.initials: P
authors.lastname: Westermann
mesh_terms: D001483:Base Sequence; D003001:Cloning, Molecular; D019615:Crenarchaeota / Q000235:genetics* / Q000302:isolation & purification; D019641:DNA, Archaeal / Q000235:genetics / Q000302:isolation & purification; D017753:Ecosystem; D005618:Fresh Water / Q000382:microbiology; D015723:Gene Library; D019766:Genes, Archaeal; D014644:Genetic Variation; D006358:Hot Temperature; D017404:In Situ Hybridization, Fluorescence; D007558:Italy; D010802:Phylogeny; D012150:Polymorphism, Restriction Fragment Length; D019642:RNA, Archaeal / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D016944:Sulfolobus / Q000235:genetics / Q000302:isolation & purification; D013696:Temperature
chemical_list: D019641:DNA, Archaeal; D019642:RNA, Archaeal; D012336:RNA, Ribosomal, 16S