ABSTRACT
Heavy metals are pollutants which affect all organisms. Since a small number of eukaryotes have been investigated with respect to metal resistance, we hypothesize that many genes that control this phenomenon remain to be identified. This was tested by screening soil eukaryotic metatranscriptomes which encompass RNA from organisms belonging to the main eukaryotic phyla. Soil-extracted polyadenylated mRNAs were converted into cDNAs and 35 of them were selected for their ability to rescue the metal (Cd or Zn) sensitive phenotype of yeast mutants. Few of the genes belonged to families known to confer metal resistance when overexpressed in yeast. Several of them were homologous to genes that had not been studied in the context of metal resistance. For instance, the BOLA ones, which conferred cross metal (Zn, Co, Cd, Mn) resistance may act by interfering with Fe homeostasis. Other genes, such as those encoding 110- to 130-amino-acid-long, cysteine-rich polypeptides, had no homologues in databases. This study confirms that functional metatranscriptomics represents a powerful approach to address basic biological processes in eukaryotes. The selected genes can be used to probe new pathways involved in metal homeostasis and to manipulate the resistance level of selected organisms.
Subject(s)
Drug Resistance/genetics , Eukaryota/drug effects , Eukaryota/genetics , Metals, Heavy/pharmacology , Soil Microbiology , Soil Pollutants/pharmacology , Yeasts/genetics , Gene Expression Profiling , Gene Library , Genetic Variation , Metals, Heavy/metabolism , Molecular Sequence Data , Soil Pollutants/metabolism , Yeasts/drug effectsABSTRACT
A quorum-quenching approach was exploited in order to identify functions regulated by quorum-sensing (QS) in the plant growth-promoting bacterium Azospirillum lipoferum. The AttM lactonase from Agrobacterium tumefaciens was shown to enzymatically inactivate N-acyl homoserine lactones (AHLs) produced by two A. lipoferum strains. The targeted analysis of several phenotypes revealed that in strain B518, a rice endophyte, AHL inactivation abolished pectinase activity, increased siderophore synthesis and reduced indoleacetic acid production (in stationary phase) but no effect was observed on cellulase activity or on swimming and swarming motilities. None of the tested phenotypes appeared to be under QS regulation in strain TVV3 isolated from the rice rhizosphere. Moreover, AHL inactivation had no deleterious effect on the phytostimulatory effect of the two strains in vitro. A global proteomic approach revealed little modification of protein patterns when comparing attM-expressing TVV3 and the wild-type strain, but numerous proteins appeared to be regulated by the AHL-mediated QS system in strain B518. Several proteins identified by MS-MS analysis were revealed to be implicated in transport (such as OmaA) and chemotaxis (ChvE). Altogether, the results indicate that in A. lipoferum, QS regulation is strain-specific and is dedicated to regulating functions linked to rhizosphere competence and adaptation to plant roots.