Team:Valencia UPV/Project/modules/methodology/EAG

EAG consists of a circuit and two electrodes connected to an amplifier which is closed by means of the moth antenna. A continuous clear air flow is blown over the antenna at a constant rate and the samples to be analysed are introduced inside the air stream. Once the stimulus is exerted to the antenna, for example the pheromone, it is locked on the antenna receptors, a signal transduction cascade is initiated and it is converted to an electrical impulse, which is registered [4]. As a result, base line undergoes an up and down or derivation, similar to those shown in an electrocardiogram. Different compounds elicit different electrophysiological responses but concentration could vary the registry, too. The characterization of DNA parts involved in pheromone biosynthesis was made by transient gene expression in N. benthamiana coupled to Electroantennography (NBTE-AEG). The volatiles synthesized by the Sexy Plant were puffed into the odour delivery system and the electrical response was registred.

Our moths were immobilized and the antennas were cut or detached from the male. Then, terminal ends of the antenna were cut and the resulting antenna fragment was placed in the electrodes. A small amount of each sample (1 ug) was loaded onto small pieces of filter paper inside the wide section of a Pasteur pipette.

The characterization of DNA parts involved in pheromone biosynthesis was made by transient gene expression in N. benthamiana coupled to Electroantennography (NBTE-AEG). The volatiles synthesized by the Sexy Plant were puffed into a device containing two small electrodes connected by a moth ́s antenna. Specific receptors in the antenna respond to the pheromones producing a measurable electric output. The volatiles in our Sexy Plants induced detectable electric pulses that could indicate a pheromone response, although further testing will be required for confirmation.

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