title: A molecular toolbox to estimate the number and diversity of Variovorax in the environment: application in soils treated with the phenylurea herbicide linuron.
abstract: Real-time PCR and PCR-denaturing gradient gel electrophoresis (DGGE) approaches that specifically target the Variovorax 16S rRNA gene were developed to estimate the number and diversity of Variovorax in environmental ecosystems. PCR primers suitable for both methods were selected as such that the enclosed sequence showed maximum polymorphism. PCR specificity was maximized by combining PCR with a targeted endonuclease treatment of template DNA to eliminate 16S rRNA genes of the closely related Acidovorax. DGGE allowed the grouping of PCR amplicons according to the phylogenetic grouping within the genus Variovorax. The toolbox was used to assess the Variovorax community dynamics in agricultural soil microcosms (SMs) exposed to the phenylurea herbicide linuron. Exposure to linuron resulted in an increased abundance within the Variovorax community of a subgroup previously linked to linuron degradation through cultivation-dependent isolation. SMs that were treated only once with linuron reverted to the initial community composition 70 days after linuron exposure. In contrast, SMs irrigated with linuron on a long-term base showed a significant increase in Variovorax number after 70 days. Our data support the hypothesis that the genus Variovorax is involved in linuron degradation in linuron-treated agricultural soils.
authors.affiliation: Division of Soil and Water Management, Katholieke Universiteit Leuven, Leuven, Belgium. karolien.bers@ees.kuleuven.be
authors.forename: Karolien
authors.initials: K
authors.lastname: Bers
authors.forename: Kristel
authors.initials: K
authors.lastname: Sniegowski
authors.forename: Pieter
authors.initials: P
authors.lastname: Albers
authors.forename: Philip
authors.initials: P
authors.lastname: Breugelmans
authors.forename: Larissa
authors.initials: L
authors.lastname: Hendrickx
authors.forename: René
authors.initials: R
authors.lastname: De Mot
authors.forename: Dirk
authors.initials: D
authors.lastname: Springael
mesh_terms: D000383:Agriculture; D001483:Base Sequence; D042621:Comamonadaceae / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D058645:Denaturing Gradient Gel Electrophoresis; D017753:Ecosystem; D006540:Herbicides / Q000378:metabolism*; D008044:Linuron / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D012987:Soil / Q000737:chemistry; D012988:Soil Microbiology; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D006540:Herbicides; D012336:RNA, Ribosomal, 16S; D012987:Soil; D008044:Linuron