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<h1>Project overview</h1>
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<p>We aim to create a defense system to fight against potato late blight. Our system consists of potatoes that is resistant to Phytophthora infestans, a fungus-like pathogen that causes potato late blight, by constructing a competitive inhibitor that can prevent the effector protein secreted by P. infestans from entering the potato cell. In case that the potato we genetically modified can’t fend off the P. infestans, we designed a soil based microbial fuel cell (SMFC) with engineered bacteria on the anode of the SMFC that can detect whether the potato tuber is infected or not. If the potato is infected and detected by our SMFC, we will spread defensin obtained from maca. The defensin can weaken and inhibit the growth of P. infestans while doing no harm to the environment since it doesn’t contain heavy metal like most fungicide.</p>
 
  
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<h1>Project overview</h1>
  
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<h2>Protection</h2>
 
  
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<h3>Design</h3>
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<h2>Background</h2>
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<p>After P. infestans penetrates the cell wall of potato, it will exploit the potato and in turn infect potatoes nearby within 3 days. Since it will infect other potatoes in such a short time that there is no effective biological method to react and inhibit the development of the disease, we decide to prevent the disease at the very beginning by rendering the potatoes the ability to prevent the invasion of P. infestans. Under certain conditions, the zoospores of P. infestans will attach to the surface of potato leaves, penetrate the cell wall by high turgor pressure and some enzymes, and secrete some effector protein, such as Avr3, into potato cells. The effector protein needs to bind to a transmembrane receptor called PI3P, which can mediate its entry into potato cell to translocate into host cell. It will suppress plant resistance gene-based immunity so that P. infestans can enter potato cells without any resistance. To stop the effector protein from entering potato cell, we found through literature research that reduction in translocated effector is a promising way to decrease the virulence of pathogens and improve disease resistance in potatoes. In research done by other scientists, FYVE protein domain from Hrs or EEA1 can also bind to PI3P receptor strongly in animal cells. We then decided to construct a FYVE protein domain with high affinity that can compete with the effector protein to inhibit the entry of P. infestans.</p>
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<h3>FYVE protein domain</h3>
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<p>Potatoes all over the world are in danger and this is not the first time in history. In the late 1840s, potato late blight epidemics caused the Great Famine in Ireland and one million people were starved to death. This devastating disease is caused by the plant pathogenic oomycetes, <i>Phytophthora infestans</i>. Even now, when potatoes are widely grown in 135 countries and consumed by 1 billion people, late blight is still causing serious problems, including food insecurity, economic losses, and environmental damages.</p>
  
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<p>FYVE protein domain is well conserved PI3P binding domain in various organisms with only 141 amino acids. FYVE protein domain originally existed in EEA1 and Hrs proteins in human and mouse, respectively. However, EEA1 and Hrs protein are too large and may take long time for the plant to degrade, we then decided to extract the protein domain from Hrs protein. On the other hand, monomeric FYVE has far lower affinity to PI3P than Hrs and it is not so stable. Therefore, we decided to construct a dimeric FYVE which has a higher affinity and is much more stable than monomeric FYVE. </p>
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<div style="margin-left:12.5%;width:75%;"><img src="https://static.igem.org/mediawiki/2015/e/eb/Nymu-overview-world.png" style="margin-left:13;padding-top:2%;padding-bottom:2%;width:90%;"><p style="font-weight:bold;font-size:14px;">Fig 1. Worldwide distribution of potato late blight caused by <i>P.infestans</i></p></div>
  
  
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<h3>Circuit design</h3>
 
  
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<img src="https://static.igem.org/mediawiki/2015/1/10/Circuit_design_dimeric_FYVE.png" style="padding-left:15%;">
 
  
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<p>Our goal in this part is to inhibit the entry of P. infestans effector protein, so the dimeric FYVE will be constitutively expressed in plant cell. The circuit will have a viral constitutive promoter CAMV35S following the coding sequence of dimeric FYVE.
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The circuit is shown on the right.</p>
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<p>Potato late blight costs an annual loss of 6.7 billion USD. Although late blight have only little impact on the food supply in some areas, crop loss can still force farmers out of business. To control late blight, fungicides are frequently used, up to once every 3 days. These fungicides have enormous costs financially, at $200 per acre of farmland. Moreover, these chemicals often seep underground or escape to nearby streams and contaminate water sources. Water samples from over the U.S. show that 75% of surface waters and 58% of groundwater wells contain at least one of the 33 potato fungicides.</p>
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<p>In modern agriculture, the use of fungicides and genetically modified potatoes are inefficient in fighting against potato late blight. Most strains of <i>P. infestans</i> have developed resistance against fungicides used nowadays. <i>P. infestans</i> secretes some enzymes and form high turgor pressure inside its cell to penetrate and colonize in potato cells. <i>P. infestans</i> infect potato leaves and tubers; eventually the entire plant rots and dies. The 2015 NYMU-Taipei iGEM team aims to prevent potatoes from being infected by this devastating disease and ensure global food security.</p>
  
  
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<h3>Experiment</h3>
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<p>To check whether dimeric FYVE, a protein domain in mice, will work well in plant, we fused GFP with dimeric FYVE in vector pSAT1-Venus-C and transfect the plasmid into a tobacco cell, BY-2. If the dimeric FYVE works well, we can see green fluorescence on the endosome of BY-2.</p>
 
  
  

Latest revision as of 12:11, 20 November 2015

Project overview

Background

Potatoes all over the world are in danger and this is not the first time in history. In the late 1840s, potato late blight epidemics caused the Great Famine in Ireland and one million people were starved to death. This devastating disease is caused by the plant pathogenic oomycetes, Phytophthora infestans. Even now, when potatoes are widely grown in 135 countries and consumed by 1 billion people, late blight is still causing serious problems, including food insecurity, economic losses, and environmental damages.

Fig 1. Worldwide distribution of potato late blight caused by P.infestans



Potato late blight costs an annual loss of 6.7 billion USD. Although late blight have only little impact on the food supply in some areas, crop loss can still force farmers out of business. To control late blight, fungicides are frequently used, up to once every 3 days. These fungicides have enormous costs financially, at $200 per acre of farmland. Moreover, these chemicals often seep underground or escape to nearby streams and contaminate water sources. Water samples from over the U.S. show that 75% of surface waters and 58% of groundwater wells contain at least one of the 33 potato fungicides.



In modern agriculture, the use of fungicides and genetically modified potatoes are inefficient in fighting against potato late blight. Most strains of P. infestans have developed resistance against fungicides used nowadays. P. infestans secretes some enzymes and form high turgor pressure inside its cell to penetrate and colonize in potato cells. P. infestans infect potato leaves and tubers; eventually the entire plant rots and dies. The 2015 NYMU-Taipei iGEM team aims to prevent potatoes from being infected by this devastating disease and ensure global food security.