title: Phylogenetic diversity of Acidobacteria in a former agricultural soil.
abstract: Although Acidobacteria represent the most abundant bacterial phylum in many soils, knowledge of acidobacterial diversity is still rather incomplete. We, therefore, examined the diversity of 16S rRNA genes affiliated with this phylum in a former arable soil via three independent approaches: (1) screening of a fosmid metagenome library for inserts containing Acidobacteria-like 16S rRNA genes; (2) PCR-cloning using general bacterial primers; and (3) PCR-cloning with acidobacterial-specific primers. Bacterial-specific libraries compared rhizosphere versus bulk soil samples, revealing a higher proportion of acidobacterial sequences in bulk soil libraries (P<0.001). Bacterial libraries recovered the greatest diversity, and sequence examination suggested that sequence mismatches with the Acidobacteria-specific primers limited the coverage of the metagenome library screening and specific library approaches. Together, these results expand knowledge of the distribution and diversity of Acidobacteria in soil environments and highlight important technical considerations in the molecular analysis of Acidobacteria diversity.
authors.affiliation: Department of Terrestrial Microbial Ecology, Netherlands Institute of Ecology (NIOO-KNAW), Heteren, The Netherlands.
authors.forename: Anna
authors.initials: A
authors.lastname: Kielak
authors.forename: Agata S
authors.initials: AS
authors.lastname: Pijl
authors.forename: Johannes A
authors.initials: JA
authors.lastname: van Veen
authors.forename: George A
authors.initials: GA
authors.lastname: Kowalchuk
mesh_terms: D001419:Bacteria / Q000145:classification* / Q000302:isolation & purification*; D044822:Biodiversity; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D015723:Gene Library; D020459:Genes, rRNA; D008969:Molecular Sequence Data; D010802:Phylogeny; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D012988:Soil Microbiology
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S