The Mysterious Case of the Microbe in the Soil
By Emm Fulk, graduate student, Rice University, Houston, TX, USA
Microbial biosensors sense and environmental signal and,
in response, produce a measurable reporter such as a gas.
Image by E. Fulk
Flagella? Check. Cell wall? Check. Cheesy music? Yes, that's right. What's a microbial detective without a melodramatic theme song?
Studies of soil microbiology have traditionally - and necessarily - been conducted from the outside in. Net fluxes of nutrients and gases from the soil environment can be chemically measured. High-throughput sequencing techniques can give metagenomic data, which provides a snapshot of the general composition of a soil microbiome. This is, essentially, a stakeout - we can get a general sense of a soil community by observing the surrounding environment and may be able to infer some activities by measuring who and what comes in and out. These strategies give us an overall picture of soil communities and their net interactions within the ecosystem but lack the spatial, temporal and chemical sensitivity to fully understand the internal dynamics of soil microbiomes.
We need a microbe on the inside.
The idea of using living microbes as biosensors is not especially new. To survive and adapt to new environmental stresses, such as nutrient or water deprivation, microbes have evolved networks to sense these changes and adapt their metabolism accordingly. Tying these naturally-evolved systems to a measurable reporter (for example, a fluorescent protein) is a logical step for understanding how microbes interact with their environment. Think of a light bulb and a light switch. The light switch senses whether it is on or off. The light bulb reports on the ON/OFF state of the light switch. Even if you can't see the switch, you can infer whether it is on or off by looking at the light bulb. Likewise, we can detect when a particular environmental condition elicits a microbial response by monitoring
the production of the reporter.
I know just the microbe for the job… he's sensitive, discreet and reports only to me.
For microbial biosensors to be useful in soil, their sensors and reporters must both be suitable for monitoring interesting
environmental conditions - for example, drought conditions, concentrations of nitrogen or carbon species, or cell-cell
communication signals. Sensors for -osmotic stress, nitrate, quorum sensing molecules and various heavy metals have been or are being
developed. These systems must also be sensitive to an environmentally relevant level of the signal. If too sensitive or not sensitive
enough, reporter production is not triggered at the right times to give useful information. Ideally, reporters need to be measurable in
situ. Traditional fluorescent or pigmented reports can't be used because, well, you can't see them in soil. Expanding our toolbox of
reporters would allow us to monitor cell growth as well as environmental response and to measure multiple signals.
The name's coli. Escherichia coli.
In standard biosensors without memory, microbes produce a reporter directly
in proportion to the level of signal. In biosensors with memory, the reporter is induced by the first instance of the environmental cue and remains on thereafter. The top cartoon shows how these two scenarios compare at the level of individual microbes. The bottom graph illustrates how reporter production differs at the whole-sample level. Image by E. Fulk
Even after engineering a useful biosensor, it remains to find suitable host microbe. Most synthetic biology is initially done in E. coli, because it is relatively well understood and easy to engineer. E. coli can also a good first organism to test biosensor function. Expanding our ability to engineer other microorganisms is a key challenge both for biosensors in soil ecology and for synthetic biology as a whole. Numerous other organisms - for example, several soil-dwelling Pseudomonas species - have successfully been engineered as biosensors. However expanding our repertoire of bacterial hosts will enable biosensors to be used in more microbial communities.
Where were the suspects on the night of the…high nitrate concentration?
Recent developments in synthetic biology have greatly expanded our ability to manipulate microbes and perform increasingly difficult computations. For example, microbes can now be programmed to produce a reporter only if both signal 1 AND signal 2 are present. Other logic functions (1 OR 2, 1 AND NOT 2, etc.) could allow for studying specific combinations of environmental conditions, such as in hot spots or during hot moments.