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Revision as of 21:50, 21 September 2011

Module 1: Phyto-Route

Chemotaxis is the movement of bacteria based on attraction or repulsion of chemicals. Roots secrete a variety of compounds that E. coli are not attracted to naturally. Accordingly, we engineered a chemoreceptor into our chassis that can sense malate, a common root exudate, so that it can swim towards the root. Additionally, E. coli are actively taken up by plant roots, which will allow targeted IAA delivery into roots by our system.

Specifications

1. The bacteria should actively move towards roots.

• For this, the bacteria need to be able to sense a common root exudate. We have chosen E. coli chemotaxis to be rewired towards L (-) malic acid (also referred to as malate), a compound found in the Citric acid cycle. It is secreted by the roots at low concentrations.

2. Uptake of bacteria into roots.

• Uptake of bacteria into the roots followed by secretion of chemicals presents a novel platform for modifying plants without genetically modifying the plant genomes.

3. Efficient expression of foreign genes in our chassis.

• Due to introduction of genes from soil bacteria into E. coli effect of the codon bias can play role in expression of our constructs. Therefore we have to ensure that expression of our construct is not constrained by this phenomenon.

3. The construct must be as modular as possible.

• Early on in our project we modelled that different concentrations of chemoreceptors on the surface of the cell allow the chemotaxis pathway to be saturated at different concentrations of malate. This means that we might be able to control the exact distance at which the chassis will stop performing biased walk towards the plant. Testing this construct with different promoters might allow us to study different saturation frequencies.