Team:KU Leuven/Project/HoneydewSystem

From 2013.igem.org

iGem

Secret garden

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  • A video shows that two of our team members are having great fun at our favourite company. Do you know the name of the second member that appears in the video?
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  • We organised a symposium with a debate, some seminars and 2 iGEM project presentations. An iGEM team came all the way from the Netherlands to present their project. What is the name of their city?

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tree ladybugcartoon

Aphid Background

Crashcourse in aphid biology

Sticker System

BanAphid population oscillates!

E. coligy

Validating the BanAphids in vivo

Parts

BioBrick 'm all!

Data Page

All our achievements on one small page!

We aim to achieve a sustainable way to reduce the damage caused by aphid pests, and offer an effective alternative for insecticides. Our modified E. coli, the BanAphid (meaning ‘to ban aphids’ as well as with ‘benefits’), would imitate the effect of insecticides by using the aphid’s own alarm pheromone, E-β-farnesene, (EBF) to repel them off the plant. On top of that we want to attract aphid predators such as the ladybug by using Methyl Salicylate(MeS), a phytohormone. This way we make sure the aphids are thoroughly removed from the plant. MeS also activates plant defense mechanisms and EBF also attracts predators.

We have established possible hurdles in introducing this system in the agricultural industry and took steps to tackle them.
First we have to make sure that the plant cell’s metabolism is not overburdened .
Secondly we have to take into account that aphids might habituate to constitutive expression of EBF (De Vos et al., 2010, Kunert et al., 2010).
Finally, we do not want to attract the aphid’s natural predators when they are not needed. These factors along with input from potential end users all played a role in the design of the Honeydew system.

Designing the Honeydew System

In this Honeydew System we opt to spray our BanAphids onto the plants. If aphids do happen to escape the EBF repellent signal, they activate the MeS BioBrick due to the presence of honeydew. This direct contact with honeydew is why we call this our Honeydew System. MeS acts as a second line of defence and attracts natural predators of the aphids such as ladybugs and green lacewings.

E-β-Farnesene construction

EBF is the most universal aphid alarm pheromone. It is released from the cornicles of the aphids to warn itself and others against upcoming danger, such as the natural predators of aphids. The aphids undergo a change in gene expression that motivates them to mobilise and leave the plant but because EBF is highly susceptible to oxidation, we our BanAphids to produce EBF regularly.

Methyl Salicylate construction

Methyl Salicylate is a phytohormone released by plants when they are under attack, for instance, upon aphid infestation. It activates plant defence systems, as well as attract aphid predators, such as the ladybug or the green lacewing. We have focussed on increasing the MeS production of an existing brick, by increasing the production of its precursor, chorismate.

Methyl Salicylate - qPCR

To check whether our genes of the MeS brick are properly transcribed, we performed a qPCR. On top of this, the amount of mRNA we can measure serves as input data for the Methyl Salicylate model.