Engineered endophytic bacteria improve phytoremediation of water-soluble, volatile, organic pollutants.

Phytoremediation of highly water soluble and volatile organic xenobiotics is often inefficient because plants do not completely degrade these compounds through their rhizospheres. This results in phytotoxicity and/or volatilization of chemicals through the leaves, which can cause additional environmental problems. We demonstrate that endophytic bacteria equipped with the appropriate degradation pathway improve the in planta degradation of toluene. We introduced the pTOM toluene-degradation plasmid of Burkholderia cepacia G4 into B. cepacia L.S.2.4, a natural endophyte of yellow lupine. After surface-sterilized lupine seeds were successfully inoculated with the recombinant strain, the engineered endophytic bacteria strongly degraded toluene, resulting in a marked decrease in its phytotoxicity, and a 50-70% reduction of its evapotranspiration through the leaves. This strategy promises to improve the efficiency of phytoremediating volatile organic contaminants.

Endophytic bacterial diversity in poplar trees growing on a BTEX-contaminated site: the characterisation of isolates with potential to enhance phytoremediation.

The diversity of endophytic bacteria found in association with poplar was investigated as part of a larger study to assess the possibility and practicality of using endophytic bacteria to enhance in situ phytoremediation. Endophytic bacteria were isolated from the root, stem and leaf of two cultivars of poplar tree growing on a site contaminated with BTEX compounds. They were further characterised genotypically by comparative sequence analysis of partial 16S rRNA genes and BOX-PCR genomic DNA fingerprinting, and phenotypically by their tolerance to a range of target pollutants, heavy metals and antibiotics. One hundred and 21 stable, morphologically distinct isolates were obtained, belonging to 21 genera, although six isolates could not be identified with confidence to a genus. The endophytic bacteria exhibited marked spatial compartmentalisation within the plant, suggesting there are likely to be species-specific and non-specific associations between bacteria and plants. A number of isolates demonstrated the ability to degrade BTEX compounds or to grow in the presence of TCE. This study demonstrates that within the diverse bacterial communities found in poplar several endophytic strains are present that have the potential to enhance phytoremediation strategies.

Field note: hydraulic containment of a BTEX plume using poplar trees.

In 1999, 275 poplar trees were planted on a field site near a car factory in order to install a bioscreen. The aim was to combine the biodegradation activities of poplar and its associated rhizosphere and endophytic microorganisms for containing a BTEX contaminated groundwater plume. This BTEX plume occurred as the result of leaking solvents and fuel storage tanks. Monitoring, conducted overa 6-year period (1999-2005) after the planting of the trees suggested that the poplar trees and their associated microorganisms had, once the tree roots reached the contaminated groundwater zone, an active role in the remediation of the BTEX plume, resulting in full containment of the contamination. Analysis of the microbial communities associated with poplar demonstrated that, once the poplar roots got in contact with the BTEX contaminated groundwater, enrichment occurred of both rhizosphere and endophytic bacteria that were able to degrade toluene. Interestingly, once the BTEX plume was remediated, the numbers of toluene degrading rhizosphere and endophytic bacteria decreased below the detection limit, indicating that their population resulted from selective enrichment by the presence of the contaminants.

Colonisation of poplar trees by gfp expressing bacterial endophytes.

With the exception of nitrogen fixing bacteria, there is little known about the colonisation patterns or population sizes of bacterial endophytes in deciduous trees. This study describes the isolation, identification, construction and re-colonisation patterns of three green fluorescent protein(gfp):kanamycin(R) labelled bacterial endophytes when re-introduced into poplar trees, their original host plant. Two of these endophytes showed considerable colonisation in the roots and stems of inoculated plants. gfp expressing cells of all three strains were observed to colonise the xylem tissue of the root. All three strains proved to be efficient rhizosphere colonisers, supporting the theory that the rhizosphere can serve as a source of bacterial endophytes.