Team:Valencia UPV/Project/eag

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<p><h3 class="hook" align="left"><a>Project</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results">Project Results</a> > <a>Electroantennography</a></h3></p><br/><br/>
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<p><h3 class="hook" align="left"><a>Project</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results">Results</a> > <a>Electroantennography</a></h3></p><br/><br/>
<div align="center"><span class="coda"><roja>E</roja>lectroantennography</span> </div><br/><br/>
<div align="center"><span class="coda"><roja>E</roja>lectroantennography</span> </div><br/><br/>
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<img width="300px" style="float:right; margin-left: 15px;" src="https://static.igem.org/mediawiki/2014/f/f3/VUPVIMG_20141006_135403.jpg" alt="EAG_1"></img>
<img width="300px" style="float:right; margin-left: 15px;" src="https://static.igem.org/mediawiki/2014/f/f3/VUPVIMG_20141006_135403.jpg" alt="EAG_1"></img>
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<p>As explained in the methodology section (<a href="#" class="normal-link-page">see Methodology: Electroantennography</a>) we performed an  electroantennography (EAG) to test the moth response to pheromones. Insects can detect pheromones through their antennae, then an electrical impulse is transmitted from them to the brain in order to trigger moth response to the pheromones. The EAG allows us to detect these electrical impulses by connecting one insect antenna to two electrodes that will amplify this impulse in order to be detected.</p><br/>
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<p>As explained in the methodology section (<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/EAG" class="normal-link-page">see Methodology: Electroantennography</a>) we performed an  electroantennography (EAG) to test the moth response to pheromones. Insects can detect pheromones through their antennae, then an electrical impulse is transmitted from them to the brain in order to trigger moth response to the pheromones. The EAG allows us to detect these electrical impulses by connecting one insect antenna to two electrodes that will amplify this impulse in order to be detected.</p><br/><br/>
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<br/><p style="text-align: justify; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 1</span>. Healthy crop field.</p></div>
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<p>We connected one antenna from a male moth, Sesamia nonagrioides , with the two electrodes. Then , an air current with a leaf extract containing our pheromones was applied (Figure 3. Signal 1). As it can be appreciated, as the extract was applied the antenna transmitted an electrical impulse. This was the moth response to our insect pheromones produced in plant.</p><br/>
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<br/><p style="text-align: right; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 1</span>. Side view of an insect antenna connected to EAG electrodes.</p><br/>
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<img width="400px" style="float:left; margin-right: 15px;" src="https://static.igem.org/mediawiki/2014/6/69/VUPVIMG_4156.JPG" alt="EAG_2"></img>
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<p>We connected one antenna from a male moth, Sesamia nonagrioides , with the two electrodes. Then , an air current with a leaf extract containing our pheromones was applied (Figure 1. Signal 1). As it can be appreciated, as the extract was applied the antenna transmitted an electrical impulse. This was the moth response to our insect pheromones produced in plant.</p><br/>
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<br/><p style="text-align: justify; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 1</span>. Healthy crop field.</p></div>
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<div align="center"><img width="400px" src="https://static.igem.org/mediawiki/2014/6/69/VUPVIMG_4156.JPG" alt="EAG_2"></img></div><br/>
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<div align="center"><p style="text-align: center; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 1</span>. Genetic construct of the Glandular trichomes specific promoter PCPS2 and GFP.</p></div><br/>
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<p>As explained in the methodology section (<a href="#" class="normal-link-page">see Methodology: Electroantennography</a>) we performed an electroantennography (EAG) to test the moth response to pheromones. Insects can detect pheromones through their antennae, then an electrical impulse is transmitted from them to the brain in order to trigger moth response to the pheromones. The EAG allows us to detect these electrical impulses by connecting one insect antenna to two electrodes that will amplify this impulse in order to be detected.</p><br/><br/>
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</div>
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<div><img style="float:left; margin-right: 15px;" width="300px" src="https://static.igem.org/mediawiki/2014/6/69/VUPVIMG_4156.JPG" alt="EAG_2"></img></div><br/>
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<div align="center"><img width="50%" src="https://static.igem.org/mediawiki/2014/2/28/VUPVPCPS2-GFP.png" alt="GFP"></img></div><br/>
 
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<div align="center"><p style="text-align: center; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 1</span>. Genetic construct of the Glandular trichomes specific promoter PCPS2 and GFP.</p></div><br/>
 
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<p>We connected one antenna from a male moth, Sesamia nonagrioides , with the two electrodes. Then , an air current with a leaf extract containing our pheromones was applied (Figure 1. Signal 1). As it can be appreciated, as the extract was applied the antenna transmitted an electrical impulse. This was the moth response to our insect pheromones produced in plant.</p><br/><br/>
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<p><br/><br/>As a control, we also applied an air current with no pheromones in suspension. (Figure 3. Signal 2) The antena did not transmit any electrical signal.</p><br/>
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<p>As a control, we also applied an air current with no pheromones in suspension. (Figure 1. Signal 2) The antena did not transmit any electrical signal.</p><br/><br/>
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<br/><p style="text-align: right; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 2</span>. Upper view of an insect antenna connected to EAG electrodes .</p><br/>
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<br/><br/><br/><br/><br/><br/><br/>
<div align="center"><img width="500px" src="https://static.igem.org/mediawiki/2014/5/53/VUPVEAGi.png" alt="EAG"></img></div><br/>
<div align="center"><img width="500px" src="https://static.igem.org/mediawiki/2014/5/53/VUPVEAGi.png" alt="EAG"></img></div><br/>
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<div align="center"><p style="text-align: center; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 1</span>. Electroantennography analysis of Sesamia nonagroides response to sexual pheromones produced in genetically engineered Nicotiana Benthamiana plants.</p></div><br/>
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<div align="center"><p style="text-align: center; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 3</span>. Electroantennography analysis of Sesamia nonagroides response to sexual pheromones produced in genetically engineered Nicotiana Benthamiana plants. Signal1: Antennal response to the Sexy Plant leaf extract. Signal 2: Antennal response to an air puff.</p></div><br/>
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<p>With these results, we can positively say that moths respond to our pheromones produced in genetically engineered Nicotiana benthamiana plants.</p><br/><br/>
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<p>The volatiles in our Sexy Plants induced detectable electric pulses that could indicate a pheromone response, although further testing will be required for confirmation.</p><br/><br/>
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<a class="button-content" id="goto-left" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/results/constructs"><strong>&larr; Go to Constructs</strong></a>
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<a class="button-content" id="goto-left" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/results/pheromone_analysis"><strong>&larr; Go to Pheromone Analysis</strong></a>
<a class="button-content" id="goto-middle" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/results"><strong>Go to Results</strong></a>
<a class="button-content" id="goto-middle" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/results"><strong>Go to Results</strong></a>
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<a class="button-content" id="goto-right" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/results/pheromone_analysis"><strong>Go to Pheromone Analysis &rarr;</strong></a></div></br></br></br>
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<a class="button-content" id="goto-right" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/results/biosafety"><strong>Go to Biosafety &rarr;</strong></a></div></br></br></br>
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Latest revision as of 03:54, 18 October 2014

Project > Results > Electroantennography



Electroantennography


EAG_1

As explained in the methodology section (see Methodology: Electroantennography) we performed an electroantennography (EAG) to test the moth response to pheromones. Insects can detect pheromones through their antennae, then an electrical impulse is transmitted from them to the brain in order to trigger moth response to the pheromones. The EAG allows us to detect these electrical impulses by connecting one insect antenna to two electrodes that will amplify this impulse in order to be detected.



We connected one antenna from a male moth, Sesamia nonagrioides , with the two electrodes. Then , an air current with a leaf extract containing our pheromones was applied (Figure 3. Signal 1). As it can be appreciated, as the extract was applied the antenna transmitted an electrical impulse. This was the moth response to our insect pheromones produced in plant.



Figure 1. Side view of an insect antenna connected to EAG electrodes.


EAG_2



As a control, we also applied an air current with no pheromones in suspension. (Figure 3. Signal 2) The antena did not transmit any electrical signal.



Figure 2. Upper view of an insect antenna connected to EAG electrodes .









EAG

Figure 3. Electroantennography analysis of Sesamia nonagroides response to sexual pheromones produced in genetically engineered Nicotiana Benthamiana plants. Signal1: Antennal response to the Sexy Plant leaf extract. Signal 2: Antennal response to an air puff.


The volatiles in our Sexy Plants induced detectable electric pulses that could indicate a pheromone response, although further testing will be required for confirmation.