Use of [13C]/[12C] ratios as an indicator of the role of microorganisms in protection of plants from the phytotoxic action of naphthalene

Using molecular and isotopic mass spectrometry, we investigated the toxic effect of naphthalene as a representative of polycyclic aromatic hydrocarbons (PAHs) on plants growing under sterile conditions and plants inoculated with microorganisms capable and incapable of naphthalene degradation. Tobacco plants of the Samsun variety were grown in a closed gas-nutrient system on a mineral medium with sucrose as a carbon source. Naphthalene used as a toxicant at a concentration of 5.2 × 10−4% contained 13C isotope whose amount was characterized by the value δ13C = +281.4 ± 0.6‰ relative to the PDB standard and differed from that of sucrose, the main source of carbon (δ13C = −12.0 ± 0.1‰). Degradation of naphthalene was determined by the inclusion of its carbon in metabolic CO2 and plant tissues (the root, stem, leaves). The effect of naphthalene on plants was indicated by the rates of O2 production and CO2 uptake during the light period as compared with the dark period of exposure. A decrease of the toxic effect of naphthalene on plants was observed only at the inoculation of plants with Pseudomonas aureofaciens BS1393 rhizosphere bacteria bearing plasmid pBS216, which controls the naphthalene biodegradation ability. The occurrence of other heterotrophic microorganisms incapable of naphthalene degradation had no similar protective effect.