exhibited preferential growth on the plant root exudates,good survival in the rhizosphere and enhanced pollutant-degrading ability. To a similar end, Narasimhan
engineered a bacterium to utilize the predominant rootexudates of
for the remediation of polychlori-nated biphenyl (PCB) in soil. This nutritional bias for theinoculum provides a selective advantage for the strainwhile in the
rhizosphere. The inoculum, aPCB-degrading
PML2, was thencapable of degrading PCBs upon augmentation into the
rhizosphere, while growing on phenylpropa-noids, the main root exudates, as a ‘preferential’ growthsubstrate. This study highlights one of the mostpromising approaches for the future of bioaugmentation:the combination of a sustainable, semiselective nutrientsource in plant exudates with the rhizosphere competenceof a pollutant-degrading ‘superbug’. Additional techniques can be employed
toachieve superior strains for bioaugmentation. de Weert
demonstrated a novel method of enhancingcompetitive root-tip colonization in a
Pseudomonas ﬂuor- escens
strain through repeated cycling of the strain andmutant derivatives on the plant root
. The authorsobserved a 100- to 1000-fold increase in the strain’s root-tip-colonizing ability in tomato and grass plants. Theauthors concluded that enhanced competitive root-tipcolonization was a result of the accumulation of nonlethalmutations, which enabled the strain to rapidly adapt tothe plant rhizosphere. This approach to strain selection isparticularly attractive because it is rapid, inexpensive,does not require genetic engineering and is readilyapplicable to Heirloom strains.
Non-traditional strain selection
Uptothispoint,wehaveprovidedourvisionforthefutureof bioaugmentation: Heirloom species and
-directed strain selection. However, we recognize thatsigniﬁcant potential rests in a less traditional approach,termed ‘priming’. Priming is generally described aspredisposing an isolate or population of microorganismsto future conditions in which they are designed to performa function. Priming is routinely used in probiotic yogurtculture preparation, where
cultures are subjected to the mildly acidic environ-ment of the yogurt, ‘priming’ the inoculum for the harsh,acidic conditions within the stomach[21,22]. This pre-treatment, albeit unnecessary for acid-tolerant strains, isessential for the viable transit of nonacid-tolerant strainsthrough the stomach into the intestines.The priming approach has also been effectively demon-strated in environmental systems, whereby a portion of clean soil is enriched for pollutant-degrading microorgan-isms by repeated biostimulation with the relevant pollu-tant(s). The resulting soil itself, with its highly competentdegrader microbial community, is then used as theinoculum for the target polluted soil.This approach
has severaladvantages:(i)theinoculumisa consortiumof indigenous microorganisms which is potentially moreresilient to stress than a single isolate; (ii) the ‘primed’consortium is maintained within its native soil, poten-tially enhancing its survival in the target soil; and(iii) inclusion of unculturable microorganisms, whichmight contain one or more highly competent pollutantdegraders. In addition, the approach is theoreticallyexpandable to solving the issue of co-contaminated soil(i.e. one with more than one pollutant), which mightnecessitate the combination of singly primed soils or onesoil primed to degrade multiple contaminants.Takeuchi et al.were among the ﬁrst to applypriming to contaminated groundwater. The authorsinitially demonstrated cometabolism of trichloroethylene(TCE) by methanotrophs from a borehole in a contami-nated aquifer. They subsequently amended a poorlyperforming borehole with groundwater from the TCE-degrading borehole, resulting in a rapid decline in theTCE groundwater concentration. Treatment success wasattributed to the activity of the bioaugmented indigenousmethanotrophs within the ‘primed’ TCE-degradinggroundwater. Albeit a return to the ‘black box’ method of remediation, instinctively avoided by scientists, thisapproach is attractive from a practical perspective; it istechnically accessible, easily implemented, inexpensiveand environmentally friendly. It remains to be seen howeffective different matrices respond to priming (e.g. clay-primed soil augmented into sandy target soil, or high pHprimed soil augmented into netural or low pH target soil).Moreover, the approach has not been suitably tested forpotential scale-up to address large contaminated sites;however, priming clearly warrants a more thoroughinvestigation to answer these and other practicalquestions.
In time, it is hoped that bioaugmentation can fulﬁll itspotential to remediate contaminated soils and water,
. Many of the limitations to its applicability in theﬁeld are the result of an uncoordinated effort to addressthe social concerns of bioremediation along with thescientiﬁc interests. To this end, we propose that exploita-tion of Heirloom species should be coordinated betweenresearch laboratories, providing both scientists andregulators with a predictable, (globally) tested selectionof microorganisms, which would facilitate pollutantremediation as well as its public acceptance. In addition,we highlight two non-traditional approaches to strainselection, termed
-directed strain selection andpriming, which demonstrate great promise for remedia-tion of polluted matrices but remain largely on theperiphery of the discipline.
WewishtothanktheRefereesfortheirvaluablefeedbackandsuggestionsin preparing this manuscript, as well as Stephanie Hunter and PennyCarter for their assistance.
1 van Veen, J.A.
. (1997) Fate and activity of microorganismsintroduced into soil.
Microbiol. Mol. Biol. Rev.
61, 121–1352 Gilbert, E.S. and Crowley, D.E. (1998) Repeated application of carvone-induced bacteria to enhance biodegradation of poly-chlorinated biphenyls in soil.
TRENDS in Biotechnology
Vol.23 No.2 February 200576