Journal Article

A molecular toolbox to estimate the number and diversity of <i>Variovorax</i> in the environment: application in soils treated with the phenylurea herbicide linuron

Karolien Bers, Kristel Sniegowski, Pieter Albers, Philip Breugelmans, Larissa Hendrickx, René De Mot and Dirk Springael

in FEMS Microbiology Ecology

Volume 76, issue 1, pages 14-25
Published in print April 2011 |
Published online January 2011 | e-ISSN: 1574-6941 | DOI: https://dx.doi.org/10.1111/j.1574-6941.2010.01028.x

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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.

Keywords: Variovorax; DGGE; real-time PCR; linuron; agricultural soil

Journal Article.  7112 words.  Illustrated.

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