Difference between revisions of "Team:Tec-Chihuahua"

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            <h1>Description</h1></div>
 
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            <h3>ABSTRACT</h3>
 
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<p align="justify"><i>Erwinia amylovora</i> causes fire blight disease in some of the most important crops such as apple, pear and most of the members of the Rosaceae. Here the team Tec-Chihuahua presents its proposal to address this environmental issue by using synthetic biology techniques to synthesize three different enzymes that might inhibit most, if not all, the virulence factors of <i>E. amylovora</i>. The use of N-Acyl homoserine lactonase, encoded by the aiiA gene, would directly affect the AHLs by hydrolyzing and inhibiting the main quorum sensing molecule. Then, the Cyclic-di-GMP phosphodiesterase from the yhjH gene would linearize the c-di-GMP avoiding the formation of biofilm and encouraging the motility. For this last virulence factor, the team proposes to arrest flagellar rotation with a Putative Glycosyltransferase encoded by epsE. Also, we had already designed a biocontrol product that goes by the name of "Erwnions" which main value proposition it's our modified <i>Erwinia</i>. We had considered all the entrepreneurship, social and legal aspects of how this project will affect locally, nationally and worldwide.</p>
 
  
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<h3>INTRODUCTION</h3>
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<p align="justify">The Rosaceae, commonly called the rose family, is a family of flowering plants; it consists of about one hundred genera and three thousand species. It's distributed throughout the world, being especially common in North America, Europe, and Asia. </p>
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<h1> iGEM Tec-Chihuahua </h1>
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<p>Your team has been approved and you are ready to start the iGEM season! </p>
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<p align="justify"> One of the most conspicuous characteristics of Rosaceae is the variety of fruits produced by its species, which much of them are eaten by people in temperate regions; for example apples, pears, strawberries, and cherries. Also, many members of the family are important as ornamentals because of their foliage and flowers plus with the vast variety of species they are large enough to be sources of wood (Folta, 2009). Also, it is essential to highlight the economic impact of the Rosaceae. For example, the total world production of edible rosaceous fruits in 2005 was about 113 million tonnes. If this amount is converted to money considering a very conservative farm gate value of US$400 per tonne, this represents an economic value of $45 billion (Hummer, 2009).</p></div>
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<p align="justify">This family commonly suffers from different diseases, the majority caused by pathogenic bacteria. One of the most harmful diseases is known as Fire Blight, which can be developed in the flowers, fruit, branches, trunks, scaffold limbs, root systems and practically the whole plant (UCANR, 2011). Fire Blight was the first disease proven to be caused by bacteria, it was originally found in North America, but through time it has vastly expanded all around the globe, including many countries from Europe, Asia and other American countries like Canada and Mexico. Although it's been many years since its discovery, nowadays there's still not a highly effective way to counterattack this problem (Johnson, 2000).
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<p align="justify">Plague control and loss costs are estimated in approximately $100 million a year in the USA. Specifically, in Michigan in the year 2000, $42 million in losses were estimated because of the removal of about 400,000 apple trees, while approximately $68 million is estimated in losses in Washington and northern Oregon. The decline of apple and pear trees from their landscape can be expensive to replace and could have a negative effect in many areas. Although there are no recent investigations in Mexico about the epidemy, it was estimated that in 1994, Chihuahua, one of the most important producing regions of apples in Latin America, had 10% of its crops presenting the usual symptoms of the disease. In the long-run, the disease of the fire blight is a critical factor for economy and society (Johnson, 2016; Ramirez, 1994). </p>
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<p align="justify"> It's important to highlight that Fire blight is caused by a phytopathogenic bacteria; <i>Erwinia amylovora</i>. It is a Gram-negative, facultative anaerobic, rod-shaped bacteria widely distributed in forty-six countries in every continent except Antarctica. The main characteristics that define this plant pathogen as a significant threat are: </p>
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<p> American and European Foulbrood are diseases found in honey bee (Apis mellifera) bee hives all around the world that affect larva in their prepupal stage. The causal agents of these are two gram-positive bacteria, Paenibacillus larvae, and Melissococcus plutonius. Nowadays, two techniques for the treatment of AFB and EFB are used: antibiotics (chloramphenicol and oxytetracycline) and incineration of affected hives. The first technique promotes the development of antibiotic resistance in the bacteria and the second one causes great monetary losses for beekeepers. The production of heterologous proteins in Escherichia coli is proposed for P. larvae and M. plutonius inhibition. An expression vector with native bee antimicrobial peptide genes (defensin 1/abaecin/defensin 2/apidaecin) will be inserted in the bacteria. Interactions between these peptides will be confirmed through a factorial experiment with a factor for each peptide. The final product, purified proteins with a specific packaging that can be released in the presence of P. larvae and M. plutonius will be available for beekeepers to apply in their beehives and inhibit the proliferation of pathogenic bacteria.
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<h3>Before you start</h3>
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<p> Please read the following pages:</p>
 
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  <li>Fast plant propagation that could induce the disappearance of the affected crop in a single vegetative period.</li>
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<li> <a href="https://2018.igem.org/Competition">Competition Hub</a> </li>
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  <li>Great capacity to disseminate by different means.</li>
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<li> <a href="https://2018.igem.org/Competition/Deliverables/Wiki">Wiki Requirements page</a></li>
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  <li>Its ability to survive in the tissues of host plants.</li>
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<li> <a href="https://2018.igem.org/Resources/Template_Documentation">Template documentation</a></li>
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<li>Devastating effects with high economic impact.</li>
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<li>No effective control methods.</li>
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<p align="justify">Also, it is relevant to mention that <i>E. amylovora</i> counts with four virulence factors, which play an important role in its pathogenicity. The first two are the injectisome and the secretion of exopolysaccharides (EPS), both produced via quorum sensing. The other two factors, biofilm formation, and motility are regulated by the concentration of C-di-GMP. As a solution to this problem, we propose the use of synthetic biology to attack these virulence factors, this proposal will be thoroughly explained in the detailed project description.</p><br>
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  <h3>Detailed Project Description</h3>
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<p align="justify">The four virulence factors of <i>Erwinia amylovora</i> mentioned before play an important role in its phytopathogenicity, giving it the necessary qualities to become an important threat to the agricultural sector. Our project objective is to dismiss the Fire Blight problem by inhibiting the quorum sensing, biofilm formation and motility of <i>E. amylovora</i>, consequently stopping its virulence. With the help of genetic engineering, synthetic biology, and the iGEM registry, our proposal involves the use of three genes: aiiA, yhjH, and epsE. These genes encode for different enzymes that will inhibit the virulence factors.</p>
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                        <p align="justify">During the quorum sensing process of some Gram-negative bacteria (as is the case for <i>E. amylovora</i>), communicating molecules called AHLs (acyl-homoserine lactones) are used. These molecules regulate the expression and synthesis of two key EPS, amylovoran and levan (Molina <i>et al</i>., 2005). The aiiA gene encodes for the autoinducer inactivation enzyme A, better known as N-Acyl homoserine lactonase, an enzyme that hydrolyzes and inactivates the AHLs, thus provoking the degradation of the communication signals. When the quorum sensing is interrupted, inhibition of the injectisome and EPS expression is expected as a consequence. At the same time, the AHLs inhibition is expected to impact directly in the T3SS. This supposition is based on the recurrent reports that had established a relation between this virulent system and the quorum sensing in <i>Vibrio fischeri</i>. Nevertheless, it is still unknown how it will affect on <i>E. amylovora</i> but this can be considered as an opportunity for better characterization of the aiiA gene</p>     
 
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<p align="justify">Concerning C-di-GMP there we found that it is a key component of the regulatory networks that govern the expression of virulence traits in <i>E. amylovora</i>. This secondary intracellular messenger molecule is involved in the regulation of many cellular processes in numerous bacterial species, including the transition from a motile to sessile lifestyle, virulence, biosynthesis of exopolysaccharides and adhesion structures. Thus it is demonstrated that at high concentration of c-di-GMP induces a positive regulation of biofilm formation and represses motility. The gene yhjH encodes for Cyclic-di-GMP phosphodiesterase, which makes di-GMP linear. As consequence of this molecule disruption, the biofilm formation would be inhibited, but it will promote an increase in motility due to a low or nonexisting c-di-GMP concentration (Jonas,2009).</p>
 
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<p align="justify">At this point it is necessary to inhibit the fourth and last virulence factor for this pathogen, motility. Here we include the third gene epsE which encodes to a Putative Glycosyltransferase, arresting flagellar rotation, by disengaging motor force-generating elements in cells embedded in the biofilm matrix. According to the bibliography, it is believed that the espE enzyme interacts with FliG interrupting the MotA-FliG interaction, stopping the flagellar spinning resembling a clutch. The specific reaction and region that interacts with FliG are still unknown (Jonas, 2009)</p>
 
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  <h3>Applications and Implications </h3>
 
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<p align="justify">As described in the past section, the project will involve the synthesis and expression of four biobricks. Three of them will express each of the genes described above with the objective of making an individual characterization against the pathogen. On the final stage of the project, the fourth biobrick would be created as a combination of them to establish the efficiency these enzymes would have over <i>E. amylovora</i> at the same time and medium. The team expects a complete or partial inhibition of the virulence from the phytopathogen, if this happens to be true, then the project could be extrapolated to several applications. In this section, the process of selection of a specific application.</p>
 
  
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                                <p align="justify">For example, the team could create a GMO that serves as a biocontrol in the Rosaceae crops competing and inhibiting the most important pathogen. Nevertheless, this would implicate a more complex genetic circuit which takes into consideration things like the environment, inducible promoters, and secretion of the enzymes through proteic capsules. Also, it would be important to analyze which bacteria would be the ideal considering that it would be in direct contact with crops for human consumption.</p><p align="justify"> A more simple application could be the creation of a spray product that contains the enzymes in the solution. In this case, it would be necessary to consider facts like the concentration of the proteins in the product, storage, proteic capsules, the protein's lifetime, as well as economic viability. </p> <p align="justify">At last, the team’s vision of the project was the creation of a transgenic plant able to defend itself against <i>E. amylovora</i>. This vision could mean that there wouldn't be a need to use antibiotics anymore having a positive impact on the economy of the agriculture sector. However, creating a transgenic plant for the human consumption would imply several issues to consider like the genetic circuit, briefly described above, as well as the possible effects that the enzymes could have inside a human body (toxicity, bioavailability, etc.).</p>
 
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<h3> Styling your wiki </h3>
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<p>You may style this page as you like or you can simply leave the style as it is. You can easily keep the styling and edit the content of these default wiki pages with your project information and completely fulfill the requirement to document your project.</p>
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<p>While you may not win Best Wiki with this styling, your team is still eligible for all other awards. This default wiki meets the requirements, it improves navigability and ease of use for visitors, and you should not feel it is necessary to style beyond what has been provided.</p>
  
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<p align="justify">To be honest, at the beginning of the project, the whole team was really lost on the final application of the project due to several ideas that emerged along the project development. It wasn't until we started our Integrated Human Practices that we were able to establish this edition project as a biocontrol which the main component is modified/sterile <i>Erwinia</i>. We took the Sterile Insect Technique as an idea where the sterilization means that <i>Erwinia</i> has lost all of his virulence factors. It is important to highlight the fact that all of the genetic requirements to be a biosecure product and the implications were reviewed with gorvernamental entities (also as part of our <a href="https://2017.igem.org/Team:Tec-Chihuahua/HP/Gold_Integrated">Integrated HP </a> and implemented directly into our <a href="https://2017.igem.org/Team:Tec-Chihuahua/Applied_Design">design</a>).
 
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<b>REFERENCES</b><br>
 
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<p align="justify"><b>Hummer.</b> (2009). Rosaceae: Taxonomy, Economic Importance, Genomics. may 31, 2017, de US
 
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Department of Agriculture Sitio web: https://hort.purdue.edu/newcrop/janick-papers/rosaceae.pdf </p>
 
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<p align="justify"><b>Johnson, Kenneth B.</b> "Fire Blight of Apple and Pear." The American Phytopathological Society. The Plant
 
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Health Instructor, 2000. Web. 15 Nov. 2016.</p>
 
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<p align="justify"><b>Jonas, K.</b> (2009). Regulation of c-di-GMP metabolism in Biofilms. 2017, de Future Medicine Sitio web:
 
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http://www.futuremedicine.com/doi/abs/10.2217/fmb.09.7</p>
 
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<p align="justify"><b>K.M. Folta, S.E. Gardiner</b> (2009), Genetics and Genomics of Rosaceae, Plant Genetics and Genomics:
 
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Crops and Models 6, DOI 10.1007/978-0-387-77491-6 1</p>
 
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<p align="justify"><b>Molina, L et al.. </b>(2005). Autoinduction in <i>Erwinia amylovora</i>: Evidence of an Acyl-Homoserine Lactone
 
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Signal in the Fire Blight Pathogen. JOURNAL OF BACTERIOLOGY, 187, p. 3206–3213. 25/05/201, De
 
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NCBI Base de Datos.</p>
 
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<p align="justify"><b>Ramirez.</b> (1994). Manejo del tizón de Fuego del Manzano en la Sierra de Chihuahua. 31 may, 20017, de
 
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INIFAP Sitio web:
 
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http://biblioteca.inifap.gob.mx:8080/jspui/bitstream/handle/123456789/2521/Manejo%20del%20tizon%20d
 
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e%20fuego%20del%20manzano%20en%20la%20sierra%20de%20chihuahua.pdf?sequence=1</p>
 
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<p align="justify"><b>UCANR.</b> (2011). Fire Blight. 23 may, 2017, de UCANR Sitio web:
 
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http://ipm.ucanr.edu/PMG/PESTNOTES/pn7414.html</p><br>
 
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<h3> Uploading pictures and files </h3>
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<p> You must upload any pictures and files to the iGEM 2018 server. Remember to keep all your pictures and files within your team's namespace or at least include your team's name in the file name. </p>
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<p>When you upload, set the "Destination Filename" to <b> T--YourOfficialTeamName--NameOfFile.jpg</b>. (If you don't do this, someone else might upload a different file with the same "Destination Filename", and your file would be erased!)</p>
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<h3> Wiki template information </h3>
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<p>We have created these wiki template pages to help you get started and to help you think about how your team will be evaluated. You can find a list of all the pages tied to awards here at the <a href="https://2018.igem.org/Judging/Pages_for_Awards">Pages for awards</a> link. You must edit these pages to be evaluated for medals and awards, but ultimately the design, layout, style and all other elements of your team wiki is up to you!</p>
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<h3> Editing your wiki </h3>
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<p>Use WikiTools - Edit in the black menu bar to edit this page</p>
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<h3>Tips</h3>
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<p>This wiki will be your team’s first interaction with the rest of the world, so here are a few tips to help you get started: </p>
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<li>State your accomplishments! Tell people what you have achieved from the start. </li>
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<li>You have a global audience! Consider the different backgrounds that your users come from.</li>
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<li>Make sure information is easy to find; nothing should be more than 3 clicks away.  </li>
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<li>Avoid using very small fonts and low contrast colors; information should be easy to read.  </li>
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<li>Start documenting your project as early as possible; don’t leave anything to the last minute before the Wiki Freeze. For a complete list of deadlines visit the <a href="https://2018.igem.org/Calendar">iGEM 2018 calendar</a> </li>
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<li>Have lots of fun! </li>
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<h3>Inspiration</h3>
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<p> You can also view other team wikis for inspiration! Here are some examples:</p>
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<ul>
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<li> <a href="https://2014.igem.org/Team:SDU-Denmark/"> 2014 SDU Denmark </a> </li>
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<li> <a href="https://2014.igem.org/Team:Aalto-Helsinki">2014 Aalto-Helsinki</a> </li>
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<li> <a href="https://2014.igem.org/Team:LMU-Munich">2014 LMU-Munich</a> </li>
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<li> <a href="https://2014.igem.org/Team:Michigan"> 2014 Michigan</a></li>
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<li> <a href="https://2014.igem.org/Team:ITESM-Guadalajara">2014 ITESM-Guadalajara </a></li>
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<li> <a href="https://2014.igem.org/Team:SCU-China"> 2014 SCU-China </a></li>
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</ul>
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Revision as of 16:48, 11 June 2018

Erwinions

iGEM Tec-Chihuahua

Your team has been approved and you are ready to start the iGEM season!

Abstract

American and European Foulbrood are diseases found in honey bee (Apis mellifera) bee hives all around the world that affect larva in their prepupal stage. The causal agents of these are two gram-positive bacteria, Paenibacillus larvae, and Melissococcus plutonius. Nowadays, two techniques for the treatment of AFB and EFB are used: antibiotics (chloramphenicol and oxytetracycline) and incineration of affected hives. The first technique promotes the development of antibiotic resistance in the bacteria and the second one causes great monetary losses for beekeepers. The production of heterologous proteins in Escherichia coli is proposed for P. larvae and M. plutonius inhibition. An expression vector with native bee antimicrobial peptide genes (defensin 1/abaecin/defensin 2/apidaecin) will be inserted in the bacteria. Interactions between these peptides will be confirmed through a factorial experiment with a factor for each peptide. The final product, purified proteins with a specific packaging that can be released in the presence of P. larvae and M. plutonius will be available for beekeepers to apply in their beehives and inhibit the proliferation of pathogenic bacteria.

Before you start

Please read the following pages:

Styling your wiki

You may style this page as you like or you can simply leave the style as it is. You can easily keep the styling and edit the content of these default wiki pages with your project information and completely fulfill the requirement to document your project.

While you may not win Best Wiki with this styling, your team is still eligible for all other awards. This default wiki meets the requirements, it improves navigability and ease of use for visitors, and you should not feel it is necessary to style beyond what has been provided.

Uploading pictures and files

You must upload any pictures and files to the iGEM 2018 server. Remember to keep all your pictures and files within your team's namespace or at least include your team's name in the file name.

When you upload, set the "Destination Filename" to T--YourOfficialTeamName--NameOfFile.jpg. (If you don't do this, someone else might upload a different file with the same "Destination Filename", and your file would be erased!)

UPLOAD FILES

Wiki template information

We have created these wiki template pages to help you get started and to help you think about how your team will be evaluated. You can find a list of all the pages tied to awards here at the Pages for awards link. You must edit these pages to be evaluated for medals and awards, but ultimately the design, layout, style and all other elements of your team wiki is up to you!

Editing your wiki

On this page you can document your project, introduce your team members, document your progress and share your iGEM experience with the rest of the world!

Use WikiTools - Edit in the black menu bar to edit this page

EDIT PAGE

Tips

This wiki will be your team’s first interaction with the rest of the world, so here are a few tips to help you get started:

  • State your accomplishments! Tell people what you have achieved from the start.
  • Be clear about what you are doing and how you plan to do this.
  • You have a global audience! Consider the different backgrounds that your users come from.
  • Make sure information is easy to find; nothing should be more than 3 clicks away.
  • Avoid using very small fonts and low contrast colors; information should be easy to read.
  • Start documenting your project as early as possible; don’t leave anything to the last minute before the Wiki Freeze. For a complete list of deadlines visit the iGEM 2018 calendar
  • Have lots of fun!

Inspiration

You can also view other team wikis for inspiration! Here are some examples:

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