Difference between revisions of "Team:Goettingen/Demonstrate"

Line 8: Line 8:
 
             </div>
 
             </div>
  
−
             <p>Until now our detection method worked perfectly under laboratory conditions, so the next step was inevitably to test it under “real life conditions”, or more precisely, we put our method under trial. Hence, we purchased the product Roundup<sup>&reg;</sup> from our local home improvement store. It has to be considered, that even though glyphosate is the active ingredient in Roundup<sup>&reg;</sup>, it contains additional substances that could pose negative side effects.
+
             <p>Until now our detection method worked perfectly under laboratory conditions, so the next step was inevitably to test it under “real life conditions”, or more precisely, we put our method under trial. Hence, we purchased the product Roundup<sup>&reg;</sup> from our local home improvement store. It has to be considered, that even though glyphosate is the active ingredient in Roundup<sup>&reg;</sup>, it contains additional substances (actually it is a soap to facilitate the contact between glyphosate and the plant surface) that could pose negative side effects.
−
                 In a first trial phase, we wanted to investigate if Roundup<sup>&reg;</sup> affects our modified Bacillus strains in a similar way as pure glyphosate (<strong>Figure X</strong>).</p>
+
                 In a first trial phase, we wanted to investigate if Roundup<sup>&reg;</sup> affects our modified <i>B. subtilis</i> strains in a similar way as pure glyphosate (see Figure 1).</p>
 
             <div class="article_picture">
 
             <div class="article_picture">
 
                 <img src="https://static.igem.org/mediawiki/2018/9/9e/T--Goettingen--growth_with_roundup_roundup.png">
 
                 <img src="https://static.igem.org/mediawiki/2018/9/9e/T--Goettingen--growth_with_roundup_roundup.png">
−
                 <p>Figure X: Growth curves from the <em>Bacillus&nbsp;subtilis</em> wildtype strain 168 with increasing amounts of the herbicidal mixture Roundup<sup>&reg;</sup>. The optical density was measured at &lambda;=600&nbsp;nm.</p>
+
                 <p>Figure 1. Growth of the <em>B. subtilis</em> wild type strain 168 with increasing amounts of the herbicidal mixture Roundup<sup>&reg;</sup>. The optical density was measured at a wavelength of &lambda;=600&nbsp;nm.</p>
 
             </div>
 
             </div>
−
             <p>Indeed, the strain shows similar behaviour in the presence of Roundup<sup>&reg;</sup> like with pure glyphosate!</p>
+
             <p>As shown in Figure 1, growth of <em>B. subtilis</em> was severely inhibited in the presence of low amounts of Roundup<sup>&reg;</sup> as it was the case with pure glyphosate!</p>
 
             <p>
 
             <p>
−
                 Furthermore, we repeated our competition experiments with Roundup<sup>&reg;</sup> and compared it to our previous results. The signals from GFP and mOrange fit with the experiments where pure glyphosate was used.
+
                 Furthermore, we repeated our competition experiments with Roundup<sup>&reg;</sup> and compared it to our previous results (see Figure 2). The signals from GFP and mOrange fit with the experiments where pure glyphosate was used.
 
             </p>
 
             </p>
 
             <div class="article_picture">
 
             <div class="article_picture">
 
                 <img src="https://static.igem.org/mediawiki/2018/3/34/T--Goettingen--competitionassay_roundup.png">
 
                 <img src="https://static.igem.org/mediawiki/2018/3/34/T--Goettingen--competitionassay_roundup.png">
−
                 <p>Figure X: This figure shows the effects of Roundup<sup>&reg;</sup> on the competition assay we performed previously. The fluorescence of GFP is shown on the ordinate, the abscissa depicts the time. GFP was excited at &lambda;=488&nbsp;nm and emission was measured at 509&nbsp;nm. mOrange was excited at &lambda;=529&nbsp;nm, emission was recorded at &lambda;=562&nbsp;nm.<br />A:&nbsp;Fluorescence of GFP. With increasing amounts of Roundup<sup>&reg;</sup>, the fluorescence of GFP decreases.<br />B:&nbsp;Fluorescence of mOrange. With increasing amounts of Roundup<sup>&reg;</sup>, the fluorescence increases.</p>
+
                 <p>Figure 2. The effect of Roundup<sup>&reg;</sup> on the competition assay that we have performed previously. GFP was excited at &lambda;=488 nm and emission was measured at 509 nm. mOrange was excited at &lambda;=529 nm, emission was recorded at &lambda;=562 nm. (A) &nbsp;Fluorescence of GFP. With increasing amounts of Roundup<sup>&reg;</sup>, the fluorescence of GFP decreases. (B) &nbsp;Fluorescence of mOrange. With increasing amounts of Roundup<sup>&reg;</sup>, the fluorescence increases.</p>
 
             </div>
 
             </div>
−
             <p>This results show that our detection method works perfectly with the commercially available product Roundup<sup>&reg;</sup>!</p>
+
 
−
             <p>The overall aim of our project was to devise a procedure that allows to determine if a sample is contaminated with glyphosate. Consequently we analysed soil samples that were previously treated with Roundup<sup>&reg;</sup>. Our product is based on the intraspecies competition of a sensitive strain expressing constitutively the &beta;&ndash;galactosidase and a glyphosate-resistant strain. In the following figure, the function of our product is depicted (<strong>Figure X</strong>).</p>
+
 
−
             <p>Roundup<sup>&reg;</sup> is sprayed on soil. Then, water is applied to solve the glyphosate from the sample. The solution is filtered through a filter device to avoid any other contaminations like other microorganisms. The sample drops onto solid media powder with all necessary ingredients and <em>Bacillus&nbsp;subtilis</em> spores from the two competing strains. The sample is incubated for 10&nbsp;h at 37°C with agitation. Now, the blue color can be compared to a test strip. The intensity of the blue color determines the concentration of glyphosate in the sample.</p>
+
 
 +
             <p>These results show that our detection method works perfectly with the commercially available product Roundup<sup>&reg;</sup>!</p>
 +
             <p>The overall aim of our project was to develop a procedure that allows to determine if a sample is contaminated with glyphosate. Consequently, we analyzed soil samples that were previously treated with Roundup<sup>&reg;</sup>. Our product is based on the intraspecies competition of a sensitive strain constitutively expressing the <em>lacZ</em> gene encoding the &beta;&ndash;galactosidase and a non-labelled glyphosate-resistant strain. The principle of our product is depicted in Figure 3.</p>
 +
 
 +
             <p>
 +
 
 +
 
 +
First, Roundup<sup>&reg;</sup> was sprayed on top of a soil sample. Second, water was applied to extract the glyphosate from the soil sample. Third, the solution was filtered through a filter device to remove soil fragments and microorganisms. Fourth, the sample is applied onto solid media powder with all necessary ingredients including spores of the two competing<em>B. subtilis</em> strains. Fifth, the sample was incubated for 10 h at 37°C with agitation. Finally, the blue color can be compared to a test strip. The intensity of the blue color roughly determines the concentration of glyphosate in the sample.</p>
 
             <p>The usage of our product is also shown in the following movie.</p>
 
             <p>The usage of our product is also shown in the following movie.</p>
 
             <p>
 
             <p>

Revision as of 11:25, 11 October 2018

Real life conditions

Until now our detection method worked perfectly under laboratory conditions, so the next step was inevitably to test it under “real life conditions”, or more precisely, we put our method under trial. Hence, we purchased the product Roundup® from our local home improvement store. It has to be considered, that even though glyphosate is the active ingredient in Roundup®, it contains additional substances (actually it is a soap to facilitate the contact between glyphosate and the plant surface) that could pose negative side effects. In a first trial phase, we wanted to investigate if Roundup® affects our modified B. subtilis strains in a similar way as pure glyphosate (see Figure 1).

Figure 1. Growth of the B. subtilis wild type strain 168 with increasing amounts of the herbicidal mixture Roundup®. The optical density was measured at a wavelength of λ=600 nm.

As shown in Figure 1, growth of B. subtilis was severely inhibited in the presence of low amounts of Roundup® as it was the case with pure glyphosate!

Furthermore, we repeated our competition experiments with Roundup® and compared it to our previous results (see Figure 2). The signals from GFP and mOrange fit with the experiments where pure glyphosate was used.

Figure 2. The effect of Roundup® on the competition assay that we have performed previously. GFP was excited at λ=488 nm and emission was measured at 509 nm. mOrange was excited at λ=529 nm, emission was recorded at λ=562 nm. (A)  Fluorescence of GFP. With increasing amounts of Roundup®, the fluorescence of GFP decreases. (B)  Fluorescence of mOrange. With increasing amounts of Roundup®, the fluorescence increases.

These results show that our detection method works perfectly with the commercially available product Roundup®!

The overall aim of our project was to develop a procedure that allows to determine if a sample is contaminated with glyphosate. Consequently, we analyzed soil samples that were previously treated with Roundup®. Our product is based on the intraspecies competition of a sensitive strain constitutively expressing the lacZ gene encoding the β–galactosidase and a non-labelled glyphosate-resistant strain. The principle of our product is depicted in Figure 3.

First, Roundup® was sprayed on top of a soil sample. Second, water was applied to extract the glyphosate from the soil sample. Third, the solution was filtered through a filter device to remove soil fragments and microorganisms. Fourth, the sample is applied onto solid media powder with all necessary ingredients including spores of the two competingB. subtilis strains. Fifth, the sample was incubated for 10 h at 37°C with agitation. Finally, the blue color can be compared to a test strip. The intensity of the blue color roughly determines the concentration of glyphosate in the sample.

The usage of our product is also shown in the following movie.

VIDEO!!!