Team:Toulouse/Result/experimental-results

From 2014.igem.org

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However, no effect was detected for the strain expressing the GAFP-1 gene, supposing a synergistic effect between these two peptides.</br>
However, no effect was detected for the strain expressing the GAFP-1 gene, supposing a synergistic effect between these two peptides.</br>
Regarding EcAMP and the triple-fungicides operon, no effect has been detected on the fungal growth. Several factors can explain these results: a number of post-transcriptional modifications are required to have a functional EcAMP and in addition to that, sequencing results of these constructs showed some differences with the original designed sequence.
Regarding EcAMP and the triple-fungicides operon, no effect has been detected on the fungal growth. Several factors can explain these results: a number of post-transcriptional modifications are required to have a functional EcAMP and in addition to that, sequencing results of these constructs showed some differences with the original designed sequence.
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<p class="texte">Inhibition halos are not visible with supernatants, probably because of their low concentrations in the extracellular medium.
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Another effect was noted with the same strains expressing D4E1 and GAFP-1 + D4E1 on another fungus <i>Aspergillus brasiliansis</i>. This effect is comparable to the one previously noted with low concentration of sulfate copper. </br>
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<p class="texte">Inhibition halos are not visible with supernatants, probably because of their low concentrations in the extracellular medium.
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<p class="texte">
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Another effect was noted with the same strains expressing D4E1 and GAFP-1 + D4E1 on another fungus <i>Aspergillus brasiliansis</i>. This effect is comparable to the one previously noted with low concentration of sulfate copper. </br>
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The choice of our chassis appears to be optimal as we noted that wild type <i>Bacillus subtilis</i> disturbs the hyphae growth of the fungi. Some strains of <i>Bacillus subtilis</i> (qst 713) are already used as Biofungicides for use on several minor crops to treat a variety of plant diseases and fungal pathogens.</br>
The choice of our chassis appears to be optimal as we noted that wild type <i>Bacillus subtilis</i> disturbs the hyphae growth of the fungi. Some strains of <i>Bacillus subtilis</i> (qst 713) are already used as Biofungicides for use on several minor crops to treat a variety of plant diseases and fungal pathogens.</br>
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After this set of experiments, the strain expressing the operon GAFP-1 + D4E1 has shown to be the best candidate to play a major role in the fight against fungal diseases such as Canker stain. Our team decided to follow the experiments on a model plant.</br>
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After this set of experiments, the strains expressing D4E1 and expressing GAFP-1 + D4E1 have shown to be the best candidates to play a major role in the fight against fungal diseases such as Canker stain. Keeping in mind our objective, <b> we decided to tests these strains in a model plant </b> : <i>Nicotiana benthamiana</i>.</br> These tests were performed in the National Institute for the Agronomic Research by experts in this domain.  
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Thanks to the diversity of anti-fungal peptides, this strategy can be adapted to different types of diseases, with different degree of specifity, etc.
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<p class="title2">3. <i> In planta </i> tests with SubtiTree
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<div style="float:left; width:215px;">
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<img style="width:215px;" img src="http://parts.igem.org/wiki/images/a/af/In_planta.jpg" style="margin-top:5px" />
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</div> 
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<div style="float: right; width:700px; margin-left:44px;">
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<p class="texte">
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The tests were performed on a laboratory model plant : Nicotiana bentamiana. The plant was injected with 5.106 or 108 bacteria (either WT, D strain, or DG strain). Xh after the bacterial injection, Sclerotinia sclerotiorum, a plant pathogenic fungus infected the leaves. The percentage of sites showing necrosis and the size of the lesions are measured X hours post infection.
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A dramatic decrease of these two factors can be noted with the transformed strains.++
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PHOTO SUR PLANTE
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TABLEAU DE RESULTAT
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<p class="texte">
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Thanks to the diversity of anti-fungal peptides, this strategy can be adapted to different types of diseases, with different degree of specifity, etc.
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</p>
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</div>
    
    

Revision as of 08:36, 11 October 2014