Team:UNITN-Trento/Project/Fruit ripening

From 2013.igem.org

(Difference between revisions)
Line 1: Line 1:
-
<!--NEW PAGE-->
+
.tn-title {
-
{{:Team:UNITN-Trento/Templates/Default|<html><!--start content-->
+
    display: block;
-
<!--CSS-->
+
    margin-bottom: 0;
-
<link rel="stylesheet" href="https://2013.igem.org/wiki/index.php?title=Team:UNITN-Trento/CSS/Project_pages&action=raw&ctype=text/css" type="text/css" />
+
-
<link rel="stylesheet" href="https://2013.igem.org/wiki/index.php?title=Team:UNITN-Trento/CSS/Project/Fruit_ripening&action=raw&ctype=text/css" type="text/css" />
+
 +
    color: #bf1b39;
-
<!--JS-->
+
    font-family: "Boogaloo",Helvetica,sans-serif;
-
<script src="https://2013.igem.org/wiki/index.php?title=Team:UNITN-Trento/JS/Project/Fruit_ripening&action=raw&ctype=text/javascript"></script>
+
    font-variant: small-caps;
 +
    font-size: 3em;
 +
   
 +
    text-align: center;
 +
}
-
<!--PAGE-->
+
/*jmslideshow*/
-
<div class="container">
+
-
<div class="sheet">
+
-
<div>
+
-
<h1>Fruit ripening</h1>
+
-
+
-
<p>
+
.jms-slideshow {
-
We have tested several type of fruit to verify that our engineered bacteria are able to produce <b>ethylene</b> (cell transformed with <a href="http://parts.igem.org/Part:BBa_K1065001">BBa_K1065001</a>) and <b>methyl salicylate</b> (cell transformed with <a href="http://parts.igem.org/Part:BBa_K1065102">BBa_K1065102</a> and <a href="http://parts.igem.org/Part:BBa_K1065106">BBa_K1065106</a>) for enhancing or blocking the ripening of fruit, respectively.
+
    display: none;
-
</p>
+
    margin: 2em auto !important;
-
<p>
+
}
-
The flasks containing the induced culture were kept at 37 degree in agitation and connected to a sealed jar (or a desiccator), where the fruit was exposed to either ethylene or methyl salicylate (MeSA). Negative controls were either a fruit kept in airtight jar or a fruit placed in a jar connected to a flask with non-induced cells.
+
-
</p>
+
-
<p>
+
-
Within the same experiment we used fruit assumed to be at the same ripening stage (i.e.: the same bunch of bananas).
+
-
</p>
+
 +
.jms-slideshow,
 +
.jms-wrapper {
 +
    width: 100%;
 +
    height: 500px;
-
</div>
+
    font-family: arial;
 +
    font-size: 16px;
 +
    background-image: url(https://static.igem.org/mediawiki/2013/c/c3/Tn-2013-mainbg-Wood_pattern_2.jpg);
 +
    border: 0;
 +
    border-radius: 1em;
 +
    box-shadow: 2px 2px 4px #323232 !important;
 +
}
-
    <div id="ex1" class="step">
+
.step {
-
<table>
+
    height: 700px;
-
<tr>
+
    text-align: center;
-
<td>
+
    opacity: 1 !important;
 +
}
-
<p>
+
.step table {
-
<span class="title">Type of fruit</span>
+
    width: 100%;
-
Cavendish banana
+
    height: 100%;
-
</p>
+
    background-color: transparent;
-
<p>
+
    color: white;
-
<span class="title">Duration of experiment</span>
+
    text-align: justify;
-
6 days
+
}
-
</p>
+
-
<p>
+
-
<span class="title">Exposition to Ethylene</span>
+
-
6 days
+
-
</p>
+
-
<p>
+
-
<span class="title">Exposition to pure MeSA (LB-dissolved)</span>
+
-
6 days
+
-
</p>
+
-
<p>
+
-
<span class="title">Container</span>
+
-
Essicator (10.3L)
+
-
</p>
+
-
<p>
+
-
<span class="title">Comment</span>
+
-
Maturation started 4 days after exposition to Ethylene.  Methyl Salicylate in this experiment seems to ripen the fruit.
+
-
</p>
+
-
</td>
+
.step table img {
-
<td>
+
    margin-left: 2em;
-
<img src="https://static.igem.org/mediawiki/2013/3/39/Tn-2013-fruit-banana1.png" />
+
    max-width: 600px;
-
</td>
+
    max-height: 500px;
-
</tr>
+
}
-
</table>
+
-
    </div>
+
-
    <div id="ex2" class="step">
+
.step table .title {
-
<table>
+
    display: block;
-
<tr>
+
    margin-bottom: 5px;
-
<td>
+
    font-size: 1.3em;
 +
    font-weight: bold;
 +
    text-align: left;
 +
    text-shadow: 0 0 2px black;
 +
    border-bottom: 2px solid white;
 +
}
-
<p>
+
.jms-slideshow .jms-dots > span {
-
<span class="title">Type of fruit</span>
+
    background-color: #f3d19a !important;
-
Cherry plum
+
}
-
</p>
+
-
<p>
+
-
<span class="title">Duration of experiment</span>
+
-
4 days
+
-
</p>
+
-
<p>
+
-
<span class="title">Exposition to Ethylene</span>
+
-
4 days
+
-
</p>
+
-
<p>
+
-
<span class="title">Exposition to pure MeSA (LB-dissolved)</span>
+
-
4 days
+
-
</p>
+
-
<p>
+
-
<span class="title">Container</span>
+
-
Fruit in 1 L jar
+
-
</p>
+
-
<p>
+
-
<span class="title">Comment</span>
+
-
Ethylene successfully ripened plums after 4 days. Methyl salicylate in this experiment seems to ripen the fruit. We performed the experiment in duplicate, in a different jar (1.5L) and we obtained the same results.
+
-
</p>
+
-
</td>
+
.jms-slideshow .jms-dots .jms-dots-current > span {
-
<td>
+
    background-color: #ac5b3d !important;
-
<img src="https://static.igem.org/mediawiki/2013/5/52/Tn-2013-fruit_susine1.png" />
+
}
-
</td>
+
-
</tr>
+
-
</table>
+
-
    </div>
+
-
 
+
-
<div id="ex3" class="step">
+
-
<table>
+
-
<tr>
+
-
<td>
+
-
 
+
-
<p>
+
-
<span class="title">Type of fruit</span>
+
-
Cherry tomatoes
+
-
</p>
+
-
<p>
+
-
<span class="title">Duration of experiment</span>
+
-
7 days
+
-
</p>
+
-
<p>
+
-
<span class="title">Exposition to Ethylene</span>
+
-
1 day
+
-
</p>
+
-
<p>
+
-
<span class="title">Exposition to MeSA (BBa_K1065106)</span>
+
-
7 days
+
-
</p>
+
-
<p>
+
-
<span class="title">Container</span>
+
-
Fruit in 0.5 L jar
+
-
</p>
+
-
<p>
+
-
<span class="title">Comment</span>
+
-
Ethylene successfully ripened the Cherry tomatoes.
+
-
MeSA present the same coloration and consistence as the two control.
+
-
One of the tomatoes in the control ripened, probably due at different stage of maturation of the fruit when it was placed in the jar.
+
-
 
+
-
</p>
+
-
 
+
-
</td>
+
-
<td>
+
-
<img src="https://static.igem.org/mediawiki/2013/2/24/Tn-2013-fruit_pomodorini2.png" />
+
-
</td>
+
-
</tr>
+
-
</table>
+
-
    </div>
+
-
 
+
-
 
+
-
    <div id="ex4" class="step">
+
-
<table>
+
-
<tr>
+
-
<td>
+
-
 
+
-
<p>
+
-
<span class="title">Type of fruit</span>
+
-
Heirloom tomatoes
+
-
</p>
+
-
<p>
+
-
<span class="title">Duration of experiment</span>
+
-
9 days
+
-
</p>
+
-
<p>
+
-
<span class="title">Exposition to Ethylene</span>
+
-
2 days
+
-
</p>
+
-
<p>
+
-
<span class="title">Container</span>
+
-
Fruit in 1 L jar
+
-
</p>
+
-
<p>
+
-
<span class="title">Comment</span>
+
-
Ethylene successfully ripened the tomatoes.
+
-
</p>
+
-
 
+
-
</td>
+
-
<td>
+
-
<img src="https://static.igem.org/mediawiki/2013/1/1b/Tn-2013-fruit_pomodorigrandi2.png" />
+
-
</td>
+
-
</tr>
+
-
</table>
+
-
    </div>
+
-
 
+
-
 
+
-
<div id="ex5" class="step">
+
-
<table>
+
-
<tr>
+
-
<td>
+
-
 
+
-
<p>
+
-
<span class="title">Type of fruit</span>
+
-
Date tomatoes
+
-
</p>
+
-
<p>
+
-
<span class="title">Duration of experiment</span>
+
-
7 days
+
-
</p>
+
-
<p>
+
-
<span class="title">Exposition to Ethylene</span>
+
-
2 day w/, 3 days w/o, 2 days w/
+
-
</p>
+
-
<p>
+
-
<span class="title">Exposition to MeSA (BBa_K1065106)</span>
+
-
2 day w/, 3 days w/o, 2 days w/
+
-
</p>
+
-
<p>
+
-
<span class="title">Container</span>
+
-
Fruit in 0.5 L jar
+
-
</p>
+
-
<p>
+
-
<span class="title">Comment</span>
+
-
No difference showing compared with the control.
+
-
Given that the tomatoes exposed at 30°C in our lab, we confirmed that the hot temperature block the ripening of the tomatoes. [reference]
+
-
 
+
-
</p>
+
-
 
+
-
</td>
+
-
<td>
+
-
<img src="https://static.igem.org/mediawiki/2013/5/5a/Tn-2013-fruit_datteri1.png" />
+
-
</td>
+
-
</tr>
+
-
</table>
+
-
    </div>
+
-
 
+
-
 
+
-
    <div id="ex6" class="step">
+
-
<table>
+
-
<tr>
+
-
<td>
+
-
 
+
-
<p>
+
-
<span class="title">Type of fruit</span>
+
-
Cherry tomatoes
+
-
</p>
+
-
<p>
+
-
<span class="title">Duration of experiment</span>
+
-
8 days
+
-
</p>
+
-
<p>
+
-
<span class="title">Exposition to Ethylene</span>
+
-
1 days
+
-
</p>
+
-
<p>
+
-
<span class="title">Immersion in MeSA pure</span>
+
-
8 days
+
-
</p>
+
-
<p>
+
-
<span class="title">Container</span>
+
-
Fruit in 0.5 L jar
+
-
</p>
+
-
<p>
+
-
<span class="title">Comment</span>
+
-
In this experiment the fruit was immersed in pure MeSA and then left in the jar for the remaining time of the test. MeSA had a toxic effect on the fruit, as it can be observed by skin and pulp colour.
+
-
Ethylene work as aspected.
+
-
</p>
+
-
 
+
-
</td>
+
-
<td>
+
-
<img src="https://static.igem.org/mediawiki/2013/7/7d/Tn-2013-fruit_pomodorini1.png" />
+
-
</td>
+
-
</tr>
+
-
</table>
+
-
    </div>
+
-
 
+
-
   
+
-
    <div id="ex7" class="step">
+
-
<table>
+
-
<tr>
+
-
<td>
+
-
 
+
-
<p>
+
-
<span class="title">Type of fruit</span>
+
-
Blackberries
+
-
</p>
+
-
<p>
+
-
<span class="title">Duration of experiment</span>
+
-
7 days
+
-
</p>
+
-
<p>
+
-
<span class="title">Exposition to Ethylene</span>
+
-
1 day
+
-
</p>
+
-
<p>
+
-
<span class="title">Exposition to MeSA (BBa_K1065106)</span>
+
-
1 day
+
-
</p>
+
-
<p>
+
-
<span class="title">Container</span>
+
-
Fruit in 0.5 L jar
+
-
</p>
+
-
<p>
+
-
<span class="title">Comment</span>
+
-
All Blackberries ripened simultaneously (colour change), however fruit exposed to ethylene had a more intense and typical smell of blackberries.
+
-
</p>
+
-
 
+
-
</td>
+
-
<td>
+
-
<img src="https://static.igem.org/mediawiki/2013/7/73/Tn-2013-fruit_more1.png" />
+
-
</td>
+
-
</tr>
+
-
</table>
+
-
    </div>
+
-
 
+
-
<p>
+
-
The results obtained varied according to the fruit tested. We tested: bananas, cherry plums, cherries and heirloom tomatoes, plums, blackberries, kiwifruit, pears and others. Results were evaluated qualitatively by observing skin color changes and pulp texture or firmness.
+
-
</p>
+
-
<p>
+
-
In some cases we were able to detect different levels of fragrance, however this results were too subjective and we discarded them.
+
-
</p>
+
-
<p>
+
-
We were also able to perform a iodine coloration test on starch-containing fruit.
+
-
</p>
+
-
<p>
+
-
We also performed quantitative evaluations of ethylene effects by fruit compression strength test.
+
-
</p>
+
-
<p>
+
-
In summary: cell transformed with <a href="http://parts.igem.org/Part:BBa_K1065001">BBa_K1065001</a> (our ethylene producing device) successfully ripen all the fruit tested when compared to the negative control, except for a few cases (Test with Date Tomatoes) where no significant effects were observed. We believe that in the first case the experiment did not work because the temperature in the laboratory (in those days) was over 30 degrees. We know the importance of temperature on ripening of vine-rape fruit (with external color). In our specific case the too high temperature (>25°C) caused inhibition of ethylene production which is needed for good lycopene synthesis. [reference]
+
-
</p>
+
-
<p>
+
-
Cells transformed with <a href="http://parts.igem.org/Part:BBa_K1065102">BBa_K1065102</a> and <a href="http://parts.igem.org/Part:BBa_K1065106">BBa_K1065106</a> (our methyl salicylate producing devices) did not show any difference when compared to the negative control.  We were not able to determine if MeSA actually inhibited fruit ripening because the experiments were not long enough to allow ripening of the control. We demonstrated by XX that YY ml of bacteria culture (induced at O.D.600 equal to 0.6) transformed with our device produce successfully MeSA at a concentration of xx ppm in the culture medium. However, the final concentration of the inhibitor in the gas phase in the jar could be lower than the needed concentration. We are now in the process of better estimating these values.
+
-
</p>
+
-
<p>
+
-
Exposure to pure MeSA instead caused an excessive ripening when added to the culture medium while it had a toxic effect when the fruit was drenched with it. MeSA was reported to slow down ripening at high concentration (XX) and to accelerate the process at lower concentration (YY). [ref] We think that under the used conditions its concentration was above the threshold required to inhibit ripening.
+
-
</p>
+
-
 
+
-
</div>
+
-
</div>
+
-
</div>
+
-
 
+
-
<!--end content--></html>|<html>https://static.igem.org/mediawiki/2013/1/1f/Tn-2013-headerbg-Sfondoch.jpg</html>|<html>https://static.igem.org/mediawiki/2013/c/c1/Tn-2013-headerbg-Sfondoch_or.jpg</html>}}
+

Revision as of 13:18, 16 September 2013

.tn-title { 

   display: block;

   margin-bottom: 0;

   color: #bf1b39;

   font-family: "Boogaloo",Helvetica,sans-serif;
   font-variant: small-caps;
   font-size: 3em;
   
   text-align: center;

}

/*jmslideshow*/

.jms-slideshow { 

   display: none;
   margin: 2em auto !important;

}

.jms-slideshow, .jms-wrapper { 

   width: 100%;
   height: 500px;

   font-family: arial;
   font-size: 16px;

   background-image: url(Tn-2013-mainbg-Wood_pattern_2.jpg);
   border: 0;
   border-radius: 1em;
   box-shadow: 2px 2px 4px #323232 !important;

}

.step { 

   height: 700px;
   text-align: center;
   opacity: 1 !important;

}

.step table { 

   width: 100%;
   height: 100%;
   background-color: transparent;
   color: white;
   text-align: justify;

}

.step table img { 

   margin-left: 2em;
   max-width: 600px;
   max-height: 500px;

}

.step table .title { 

   display: block;
   margin-bottom: 5px;
   font-size: 1.3em;
   font-weight: bold;
   text-align: left;
   text-shadow: 0 0 2px black;
   border-bottom: 2px solid white;

}

.jms-slideshow .jms-dots > span { 

   background-color: #f3d19a !important;

}

.jms-slideshow .jms-dots .jms-dots-current > span { 

   background-color: #ac5b3d !important;

}

Retrieved from "http://2013.igem.org/Team:UNITN-Trento/Project/Fruit_ripening"