RESUMEN
Vetiver is the only grass cultivated worldwide for the root essential oil, which is a mixture of sesquiterpene alcohols and hydrocarbons, used extensively in perfumery and cosmetics. Light and transmission electron microscopy demonstrated the presence of bacteria in the cortical parenchymatous essential oil-producing cells and in the lysigen lacunae in close association with the essential oil. This finding and the evidence that axenic Vetiver produces in vitro only trace amounts of oil with a strikingly different composition compared with the oils from in vivo Vetiver plants stimulated the hypothesis of an involvement of these bacteria in the oil metabolism. We used culture-based and culture-independent approaches to analyse the microbial community of the Vetiver root. Results demonstrate a broad phylogenetic spectrum of bacteria, including alpha-, beta- and gamma-Proteobacteria, high-G+C-content Gram-positive bacteria, and microbes belonging to the Fibrobacteres/Acidobacteria group. We isolated root-associated bacteria and showed that most of them are able to grow by using oil sesquiterpenes as a carbon source and to metabolize them releasing into the medium a large number of compounds typically found in commercial Vetiver oils. Several bacteria were also able to induce gene expression of a Vetiver sesquiterpene synthase. These results support the intriguing hypothesis that bacteria may have a role in essential oil biosynthesis opening the possibility to use them to manoeuvre the Vetiver oil molecular structure.
Asunto(s)
Bacterias/clasificación , Bacterias/metabolismo , Chrysopogon/microbiología , Aceites Volátiles/metabolismo , Bacterias/genética , Bacterias/aislamiento & purificación , Carbono/metabolismo , Chrysopogon/citología , Chrysopogon/ultraestructura , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Perfilación de la Expresión Génica , Genes de ARNr , Microscopía , Microscopía Electrónica , Datos de Secuencia Molecular , Filogenia , Proteínas de Plantas/biosíntesis , Raíces de Plantas/citología , Raíces de Plantas/microbiología , Raíces de Plantas/ultraestructura , ARN Bacteriano/genética , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Homología de Secuencia de Ácido Nucleico , Sesquiterpenos/metabolismoRESUMEN
Localization of Hg in root tissues of vetivergrass (Chrysopogon zizanioides) was investigated by micro-Proton Induced X-ray Emission (PIXE) spectrometry to gain a better understanding of Hg uptake and its translocation to the aerial plant parts. Tillers of C. zizanioides were grown in a hydroponic culture for 3 weeks under controlled conditions and then exposed to Hg for 10 days with or without the addition of the chelators (NH(4))(2)S(2)O(3) or KI. These treatments were used to study the effects of these chelators on localization of Hg in the root tissues to allow better understanding of Hg uptake during its assisted-phytoextraction. Qualitative elemental micro-PIXE analysis revealed that Hg was mainly localized in the root epidermis and exodermis, tissues containing suberin in all Hg treatments. Hg at trace levels was localized in the vascular bundle when plants were treated with a mercury solution only. However, higher Hg concentrations were found when the solution also contained (NH(4))(2)S(2)O(3) or KI. This finding is consistent with the observed increase in Hg translocation to the aerial parts of the plants in the case of chemically induced Hg phytoextraction.