Team:Toulouse/ethics

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<div id="innercontenthome">
       <div class="centering" style="padding-top: 85px; padding-bottom:40px;">
       <div class="centering" style="padding-top: 85px; padding-bottom:40px;">
-
      <p class="texte">The ethical questioning turned out to be one of the major starting points of our project.  Acting on an established environment and modifying it is no mean feat and our thinking combines technical and philosophical point of views. The actual purpose of our project also leads us to undertake an ethical questioning about the role of the scientist regarding “useless” things such as the trees lining the Canal du Midi. </p>
+
<div id="column-left">
 +
<h3 class="title2" style="margin-top:10px; color:#333;">Summary :</h3>
 +
<ul class="menuleft">
 +
  <li style="margin-top:25px;"><a href="#select1">Protection of the beauty</a></li>
 +
  <li><a href="#select2">Human intervention in the nature</a></li>
 +
  <li><a href="#select3">SubtiTree</a></li>
 +
</ul>
 +
</div>
-
<p class="title1">Protection of the beauty</p>
+
<div class="column-right" style="width:75%; float:right;">
-
<p class="texte" style="text-align:center"><B>Is it the scientists’ role to protect beauty?</B>
+
 
 +
<!--CITATION-->
 +
<p class="citation">
 +
"Knowledge without conscience is but the ruin of the soul."</br>
 +
"Science sans conscience, n'est que ruine de l'âme", François Rabelais, 16<sup>th</sup> century.
</p>
</p>
-
<p class="texte"> Beauty is a feeling of satisfaction and is selfless. It is more a feeling than the property of a thing, this is not a notion we can clearly understand. Indeed, we can find something beautiful even when we don’t know the purpose of the object. There is always a distinction between natural beauty and artistic beauty according to Hegel, the famous author. The artistic beauty is born from our mind and our spirit: it is an element of signification of the work of art whereas the natural beauty of the object is external. In a way, the Canal du Midi combines both types of beauty: a natural one regarding the nature, the centenary plane trees but also an artistic one since the Canal was built by the human hands.
+
      <p class="texte">The ethical
-
Usually, scientists judge beauty as a superficial feature not deserving to undertake any kind of scientific efforts to maintain it. The traditional role of scientists is to solve global issues and to elaborate complex strategies in order to find useful solutions for everyone’s life.
+
questioning turned out to be one of the major starting points of our project.
-
Once made this observation, one may wonder why synthetic biology would be used only to protect the useless beauty of a local heritage such as the trees lining the Canal du Midi.  
+
Acting on an established environment and modifying it is by no mean trivial and
-
</p>
+
our combined technical and philosophical points of view. The actual purpose of our project also leads us to undertake an
 +
ethical questioning about the role of the scientist regarding “useless” things
 +
such as the trees lining along the Canal du Midi. </p>
-
<p class="texte">
+
<p class="title1" id="select1">Protection of the beauty</p>
-
This crucial interrogation leads us to consider science and synthetic biology in another way. <B> What if the role of scientists was also to make people rediscovering the beauty of nature?  What if the bases of new scientific challenges resulted from a more local scale? </B> Science does not have to be elitist, it has so much to gain opening itself to these challenges. First scientifically, as research is never useless and as we never know the impact and the scope of our results. Then, socially as we could measure the deep interest raised by our project within the population and the media. Adopting a new vision of synthetic biology, we probably make people change their mind about this innovative discipline.
+
<p class="title2">Is it the scientists’ role to protect beauty?
-
The traditional cold objectivity of science distances itself from the society. Scientists are also beings capable of feeling the beauty, sensitive to the charm of landscapes and <B>able to understand the usefulness of useless trees </B>…
+
-
The design of a strategy to protect useless beauty may seem senseless but we believe that it is also the scientist’s duty. Thus, it becomes essential to protect the beauty of this site.
+
</p>
</p>
 +
<p class="texte"> Beauty is a
 +
feeling of satisfaction and is selfless. It is more a feeling than the property
 +
of a thing, this is not a notion we can clearly understand. Indeed, we can find
 +
something beautiful even when we don’t know the purpose of the object...</p>
 +
<center>
 +
<img src="https://static.igem.org/mediawiki/2014/f/fa/Fontaine_Duchamp.jpg" width="400px">
 +
<p class="legend">Figure 1: Fontaine (Marcel Duchamp). Yes this is also art...</p></center>
-
<p class="title1">Human intervention in the nature</p>
+
<p class="texte">There is always
-
<p class="texte">The main question would be to understand the purpose of taking actions in nature. Does the mankind have the proper right to operate in nature? </p>
+
a distinction between natural beauty and artistic beauty according to Hegel,
-
<p class="texte" style="text-align:center"><B> The nature </B>
+
a German philosopher. The artistic beauty is born from our mind and our
 +
spirit: it is an element of signification of the work of art whereas the
 +
natural beauty of the object is external. In a way, the Canal du Midi combines
 +
both types of beauty: a natural one regarding the Nature, the centenary plane
 +
trees but also an artistic one since the Canal was built by the human hands.
 +
Usually, science judges beauty as a superficial feature not deserving to
 +
undertake any kind of scientific efforts to maintain it. The traditional role
 +
of science is to solve global issues and to elaborate complex strategies in
 +
order to find useful solutions for everyone’s life. Once made this observation,
 +
one may wonder why synthetic biology would be used only to protect the useless
 +
beauty of a local heritage such as the trees lining the Canal du Midi.
</p>
</p>
-
<p class="texte"> The nature is known as a creation of God. Thus, human is linked to the nature and for that reason the nature deserves to be respected and loved. The nature is also a source of life: indeed a man needs to breath, drink, eat. If he is deprived of these elements, he would die. Therefore, human survival depends on the nature. Everything we eat comes for the environment: vegetables, cereals, meat, fish. Even if everything has an origin in the nature, men have always taken action in the improvement of the natural species of food.</p>
+
<p class="texte">
 +
This crucial
 +
interrogation leads us to consider science and synthetic biology from a
 +
different point of view. <b>What if the role of scientists was also to make
 +
people rediscovering the beauty of Nature? What if the bases of new scientific
 +
challenges resulted from a more local scale? </b>Science does not have to be it has so much to gain opening itself to these
 +
challenges. First scientifically, as research is never
 +
useless and as we never know the impact and the scope of our results.
 +
Then socially, as we could measure the deep interest raised by our
 +
project within the population and the media. Adopting a new vision of synthetic
 +
biology, we will probably make people change their mind about this innovative
 +
discipline. <br>
 +
The traditional cold objectivity of science distances itself from the society.
 +
However, scientists are also being capable of feeling the beauty, sensitive to
 +
the charm of landscapes and <b>able to understand the usefulness of
 +
"useless" trees</b>…<br>
 +
The design of a strategy to protect useless beauty may seem senseless but we
 +
believe that it is also the scientist’s duty. We have to remember that thinking
 +
is what distinguish <i>Homo sapiens</i> from other species on earth and this
 +
"thinking" feature allows us to understand the world and be conscious
 +
of our human Nature (Descartes: <i>Cogito ergo sum</i>). The art is an object
 +
of philosophical thought. Consciousness raises humans above all others living
 +
creatures. Thus, it is necessary to respect and protect art. And thus, it
 +
becomes essential to preserve the beauty of the Canal du Midi.  
 +
</p>
-
<p class="texte">The nature is a source of wealth for the mankind. It ensures survival and development by giving men the wood, the rocks, the soil to build shelters. All these resources allow human to develop their civilization.
 
-
Besides that, the nature as resources and has more relationship with men. One functional relationship: the nature is a source of education. By observing it, the mankind has made major discoveries: animals show us some examples of social life attitudes and successful technics. And an emotional relationship: being in contact with nature can allow men to feel strong emotion, as describe by poets like Hugo and Lamartine.</p>
 
-
<p class="texte">The major important aspect is to be aware of the importance of the nature in the mankind’s life. By destroying and hammering the nature, we jeopardize our lives. We need the nature, we come from the nature and we depend on nature for our survival, our food, our discoveries and our civilisation. Thus, respecting, loving and preserving the environment is a question of survival.</p>
+
<p class="title1" id="select2">Human intervention on  Nature</p>
 +
<p class="texte">Our main question is to understand the complicated relationship between man and Nature.
 +
Does mankind have the proper right to change the Nature? Is modified Nature considered as artificial?</p>
 +
<p class="title2"> Mankind & Nature</p>
-
<p class="texte" style="text-align:center"><B> Impact of mankind </B>
+
<p class="texte"> Nature deserves
-
</p>
+
to be respected and loved. Mankind has always been linked to Nature as its
 +
survival depends on what comes out of the ground, the trees, the oceans… The
 +
Nature is a source of wealth for mankind. It ensures survival and development
 +
by giving men the wood, the rocks, the soil to build shelters. Being in contact
 +
with Nature can allow men to feel strong emotion, as describe by poets like
 +
Hugo and Lamartine.</p>
-
<p class="texte">Since the birth of humanity, men modified their environment, changing it for their own comfort and profit depending on their own desire.By increasing the spatial development, humans modify the natural environment. However, they become aware of the risks for the planet and start restoring what they previously transformed. But how have men modified the environment? How can they preserved the natural balance?</p>
 
-
<p class="texte">It is true mankind can have harmful actions for the nature and the environment such as the waste discharge, oil slicks, intensive fishing but also the introduction of devastating species such as the pathogen, Ceratocystis platani.</p>
+
<p class="texte">Since the birth
-
 
+
of humanity, man himself understood the importance of studying and mastering
-
<p class="texte">For decades, mankind realized that all its modifications treat humanity and the nature as a resource. Thanks to this understanding, a new desire was born, a wish to protect the nature and the wildness. Men want to protect and preserve the nature: they want to limit the results of human actions.
+
Nature to develop the civilization. Still today the most advanced technologies
-
Man fits with his position: he takes advantage of the environment and the environment takes advantage of the reasoned human interventions. There is an adaptation of the mankind toward the nature.
+
often try to mimic natural phenomena. With the development of civilization, men
 +
modified their environment, changing it for their own comfort depending on
 +
their own desire. With the increase of human activity, the natural environment
 +
is modified profundly. With industrialization, the
 +
natural environment suffered from waste discharges, oil slicks, intensive
 +
fishing (and many others...) but also the introduction of devastating species
 +
such as the pathogen,<i> Ceratocystis
 +
platani</i>. However, despite these negative aspects, men
 +
are capable of favorable actions to help the environment and fix their
 +
mistakes. The current trend is to limit the impact of human interventions on
 +
Nature, and hopefully this trend is not transient and will not vanish. A new
 +
desire is born, a wish to protect Nature and wilderness. Humanity can adhere to
 +
this position: human take advantage of the
 +
environment and the environment takes advantage of the reasoned human
 +
interventions. There is an adaptation of mankind to Nature. Moreover, humans
 +
can have empathy: people are capable of understanding emotions and cognitive
 +
states of other organisms. To respond to these feelings, humans have
 +
technological tools allowing them to fight against enemies. This is the case
 +
with our project: fighting <i>Ceratocystis
 +
platani</i>.  
</p>
</p>
-
<p class="texte">Moreover, humans have the capacity of empathy: people are able to understand the emotions and cognitive states of other organisms and to identify to them. To respond to these feelings, humans have technological tools allowing them to fight against enemies such as Ceratocystis platani.</p>
+
<p class="texte">In conclusion,
 +
by destroying and hammering the Nature, we jeopardize our lives. We need
 +
Nature, we come from Nature and we depend on Nature for survival, food,
 +
discoveries and civilisation. Respecting, loving and
 +
preserving the beauty of it is also a question of
 +
survival.</p>
 +
<p class="title2">Nature and artifice
 +
</p>
-
<p class="texte" style="text-align:center"><B> Natural and artificial</B>
+
<p class="texte">Talking about
 +
the Nature refers to the whole world with an exception: all the transformations
 +
made by mankind. Nature exists regardless of men and his interventions whereas
 +
artificial is everything that exists because to humans.
</p>
</p>
 +
<p class="texte">However,
 +
pretending that natural and artificial are opposite does not seem to be true.
 +
Man cannot create without the various elements provided by Nature, he then justs transform Nature. Thus we may wonder if there is a
 +
true difference between natural and artificial. The border between these two
 +
notions is not as obvious as it seems. The landscapes are shaped by the hand of
 +
man, animals are domesticated, and now bacteria are considered as cell
 +
factories. A natural reserve is artificially preserved as a result of human
 +
actions. Is there still something natural since the birth of mankind? Actually,
 +
the artifice is a slight modification of Nature and couldn’t exist by itself.
 +
The distinction between natural and artificial seems sterile and we clearly
 +
understand that these notions are inextricably linked and need each other to
 +
exist. </p>
-
<p class="texte">Talking about the nature refers to the whole world with an exception: all the transformations made by mankind. Thus, the nature consists in the real without all the artificial elements created by humans. The nature is existing regardless of men and his interventions whereas artificial is everything that exists thanks to humans.
+
<p class="texte">In conclusion,
-
However, pretending that natural and artificial are opposite does not seem to be true. Indeed, when a man creates something it becomes possible to discover naturel elements. Man cannot create without elements provided by the nature, he is just transforming the nature, changing the shape.</p>
+
isn't it our duty to use our unique position in the history of life and our human
 +
approach to try to replace the evolutive processes?</p>
-
<p class="texte"><I><CENTER>Thus we may wonder: is there a true difference between natural and artificial?</I></CENTER></p>
 
-
<p class="texte">Men already changed nature: they are surrounded by animals and plants which are not wild anymore. So can we still consider them as natural? They come from human wishes and not from natural selection, so what is the limit between natural and artificial? An important aspect for us is the ability to think about the involvements of their choice regarding changes in environment for example in our project. </p>
+
<p class="title2">Back to our project</p>
-
<p class="texte"><I><CENTER>One question can be asked: isn’t it our duty to use our unique position in the history of life and our human approach to try to replace the evolutive processes?</I></CENTER></p>
 
-
<p class="texte">By all means, the natural world and its organization would be drastically changed to reduce the different kind of pain that the organisms can feel such as the infection of Ceratocystis platani with the plane trees. Indeed, promoting the idea that even non-human organisms pain in the nature is a serious ethical problem becomes a major goal to be taken care of. Furthermore, cancer, depression, malaria are natural results of evolutive processes that we consider to be life threatening and we need to fight. Why would it not be the same for our plane trees? It becomes ethicaly intolerable to realize the cruelty of the nature for the plants and our purpose is to encourage people to realize this.</p>
+
<p class="texte">These
 +
inextricable links are obviously the basis of our project. We aim to
 +
artificially preserve a natural heritage shaped by Pierre Paul Riquet hundreds years ago. Fighting a
 +
naturally occurring form of life that threatens it maybe just an imitation of
 +
the natural evolution process. What is considered today as ‘non-natural’
 +
may be one day regarded differently. To the extent that everything is done not
 +
to unbalance the ecosystem, our intervention can be judged rightful, even more
 +
than the use of chemicals.</p>
-
<p class="texte">Of course, men are responsible for their mistakes such as the introduction of Ceratocytis platani and should fix it by any way. But is it too stupid to think that nature will respond to this threat? Since ages nature was able to keep its balance, if one specie dies another takes the place. So if all plane-trees die by this disease, there will be another species which will be able to take up this ecological niche. This project just shows the wish of humanity to interfere in the nature. </p>
 
-
<p class="title1">SubtiTree</p>
+
<p class="title1" id="select3">SubtiTree</p>
-
<p class="texte"><I><CENTER>Survival in the environment</I></CENTER></p>
+
 +
<p class="title2"> Potential strategies discussed
 +
<br> (See more details in the <a href="https://2014.igem.org/Team:Toulouse/Project/Spreading">Spreading</a> dedicated page)
 +
</p>
 +
 +
<p class="texte">To be sure that
 +
SubtiTree will not survive and spread in the
 +
environment, many strategies were discussed to improve our bacterium:
-
<p class="texte">SubtiTree will live in sap tree, that is why we use one endophyte Bacillus subtilis strain. In order to contain our bacteria in this area during a short period of time, we modified some of its survival characteristics. 
+
<br>- Avoid the survival in the natural environment (outside the tree) thanks to a proline auxotrophy system
-
First, the control of synthetic bacterium location concerned us,that is why we border it inside the plane tree. Sap tree is known to contain a certain quantity of proline which could allow bacterial growth whereas the soil is composed by only trace of this amino acids. Thus we decide to delete the proline production pathway to make an auxotrophic bacterium for this amino acid.
+
<br>- Prevent the sporulation of <i>Bacillus subtilis</i> to make it annual 
 +
<br>- Avoid gene transfers between SubtiTree and a wild
 +
type bacterium thanks to a toxin-antitoxin system
 +
<br>- Use an integrative plasmid to improve the genetic stability
</p>
</p>
-
<p class="texte">Secondly, SubtiTree is an annual bacterium to avoid any kind of mutations. It is known that endophyte bacteria must sporulate to survive to winter. Thus, we chose a B.subtilis strain without late genes of sporulation. So SubtiTree is a non-sporulation bacterium, which means that in stress conditions (cold temperature, food lack …) or outside the sap, the bacterium will not resist and die.</p>
 
-
<p class="texte">These two characteristics of SubtiTree show that it’s an annual bacterium, which can only grow in sap tree. Combined they prevent any long term colonization of an ecological niche by SubtiTree against wild bacteria. These prevent any long term effect.</p>
 
-
<p class="texte"> We try to respond to bacterial spread problem, at the same time we wonder about horizontal gene transfer. The transmission of some genes from our synthetic bacterium to wild bacteria could be block with two modifications. 
+
<p class="title2">Public perception
-
First, as used in genetic engineering, plasmids are a tremendous way to transfer some genic information. In this way, it’s not the best way to restrict the collecting transgenic DNA. That is why we insert our genes in the bacterium chromosome.
+
</p>
</p>
-
<p class="texte">Secondly, some bacteria are able to accept chromosomal DNA, like Bacillus subtilis. Thus, the previous strategy is enough not efficient to prevent transfer. We could include the toxin/anti-toxin system. The toxin gene is near to our synthetic gene, contrary to the anti-toxin gene. So if there is a transfer of our synthetic gene, the toxin gene will be included in the transfer. Receiver bacteria will die by the production of the toxic protein. </p>
+
<p class="title3">Political and public adhesion</p>
-
<p class="texte">Our synthetic genes are not the only problem in the design of SubtiTree. One of the side effects of our cloning method is the persistence of antibiotic resistance genes. This is incompatible with the introduction of SubtiTree in the environment. It is possible to delete this resistance in chromosome. To conclude, the spreading limitation as shown before makes the use of SubtiTree acceptable in the wild.  
+
<p class="texte">Due to our
-
</p>
+
strong implication in preserving this magnificent work of art, our project
 +
interested several governmental services. Indeed some municipalities and
 +
regional councils supported our local engagement. Beyond that, our project
 +
interests the highest level of the “Canal du Midi” administration: the national
 +
navigation authority (VNF) and the Ministry of agriculture. Both of them funded
 +
this project. They are now looking for the continuation of the project after
 +
the iGEM competition. This is clearly a sign that we targeted the right
 +
question. </p>
-
<p class="texte" style="text-align:center"><B> Public perception </B>
+
<p class="texte">This project
-
</p>
+
also received the attention of the public through several articles in
 +
newspapers, television, radio and internet. First we had just a local coverage,
 +
but days after days there were more and more media interested in SubtiTree. This mediatic coverage
 +
allowed us to contact concerned citizens who participated to the development of
 +
this project. This interaction with the public allowed us to explain and
 +
promote public knowledge of synthetic biology. </p>
-
<p class="texte">Thanks to the bolts we used in SubtiTree construction, its application interest civil services. Indeed some city halls and regional council support our local engagement. Beyond that, our project interests the highest level of the “Canal du Midi” administration: the national navigation authority and the ministry of agriculture fund this project. They are still interest in to continue this work after the iGEM competition context. </p>
 
-
<p class="texte">This project also received the attention of the public through several article in newspaper, television, radio and internet. First we have just a local coverage, but day after day there is some national media which come to us. This coverage media allows us to have contact with regular people and thanks to that they participate in the development of this project. This interaction with the public allows us to develop the safety matter of our project. The support they give to us is the result of the scientific popularization of our project.  </p>
+
<p class="title3">Safety principle</p>
-
<p class="texte">One single tree infected by the canker, that is a perimeter of a couple of hundred meters which is included in the prophylactic cut. So there a healthy population in this area, we want to protect this tree. There is a common principle in agronomics that said that you prefer to save the population to the subject. According to this concept the introduction of our GEM seems acceptable. A result of the subtitree’s introduction is the modification of endophytic microbial fauna, but without any treatment organisation of the entire population will be messier. As subtitree is an annual bacterium, its side effect will not last in the time, this disruption in this balance will return to the normal. </p>
+
<p class="texte">One single tree
 +
infected by Canker, and all the trees located in an area of a couple of hundred
 +
meters around are included in the prophylactic cut. We acted to preserve the
 +
surrounding trees. The modification of the endophytic
 +
microbial fauna generated by the introduction of the engineered bacterium has
 +
to be compared to the introduction of chemicals. They contain chlorine atom and
 +
aromatic hydrocarbon, so their remediation is complicated and they represent a
 +
source of pollution. By shortening the lifespan to one season and minimizing
 +
the risks of spreading, we plan a safe and environmental-friendly way to fight
 +
Canker.  
-
<p class="texte">Furthermore, usual fungicides used come from chemical industry. They contain chlorine atom and aromatic hydrocarbon, so their remediation is complicated and they represent a source of pollution. We propose to use in this project biofungicide, so they are eco-friendly and more specific. Thus they seem to be a good way to fight against the fungal infection.  </p>
 
-
<p class="texte" style="text-align:center"><B> Feasability </B>
+
<p class="title2">Feasability
</p>
</p>
 +
<p class="texte">We wonder about
 +
the feasibility of tree’s treatment. As we used endophytic
 +
bacteria, we can count on the natural growth of SubtiTree
 +
inside the sap. So we can inject few bacteria to be sure to have enough
 +
bacteria to protect the tree. Some researchers (Xianling
 +
Ji<sup>1</sup> et al) already injected <i>B. subtilis</i> in plants and
 +
observed an increase of bacteria concentration to a maximum of 10<sup>5</sup>
 +
bacteria/mL</p>
 +
 +
<p class="texte">As we aim to
 +
inject a small quantity of bacteria, this treatment remains cheaper than the
 +
injection of several liters of chemical fungicides. In addition, this injection
 +
prevents the preventive tree cutting, which is very expensive. Cutting one tree
 +
cost around € 3000. The administration in charge of the protection of the
 +
“Canal du Midi” already plans to spend 220 million euros to cut and replant all
 +
trees along the Canal. Besides the important cost of cutting trees, it will
 +
destroy one of the symbols of south-western France. </p>
-
<p class="texte">We wonder about the feasibility of tree’s treatment. As we used endophytic bacteria, we can count on the natural growth of SubtiTree inside the sap. So we can inject few bacteria to be sure to have enough bacteria to protect the tree. Some researchers already inject bacillus subtilis strain in plants and observe an increase of bacteria concentration to a maximum of 105 bacteria/mL. </p>
+
<p class="texte">We know that SubtiTree could be improved in many ways, but in the  
 +
iGEM’s circumstances we could not have the time to go
 +
deeper. First, we can improve the fixation module. Using chitin as fixation
 +
anchor is simple but not enough specific to fix just one fungus type. That’s
 +
why we first think to fix SubtiTree to one protein
 +
included in the <i>Ceratocystis
 +
platani</i>’s
 +
membrane: CP. The bacterial prototype designed this summer can be optimized to
 +
trigger the fungicides production when the binding is completed, and to be more
 +
specific changing the peptides produced.</p>
 +
<p class="title1">References</p>
-
<p class="texte">As we inject a poor quantity of bacteria, the injection remains cheaper than the injection of several litter of fungicide. Plus this injection prevents the cutting of trees, which is very expensive. Cutting one tree cost around 3 K€. The administration in charge of the protection of the “Canal du Midi” already plans to spend 220 million euros to cut and replant all trees along the Canal. Besides the important cost of cutting tree, it will destroy one of the symbols of south-west France. </p>
+
<li class="tree"><p class="texte"> Xianling Ji, Guobing Lu, Yingping Gai, Chengchao Zheng,and Zhimei Mu.<b> Biological control against bacterialwilt and colonization of
 +
mulberry byan endophytic <i>Bacillus subtilis</i> strain </b>. FEMS Microbiol Ecol. 65 (2008) 565–573. </p></li>
 +
</div>
-
<p class="texte">We know that SubtiTree could be improve in many ways, but in the iGEM’s circumstances we could not have the time to go deeper. First, we can improve the fixation module. Using chitin as fixation anchor is simple but not enough specific to fix just one fungus type. That’s why we first think to fix SubtiTree to one protein included in the Ceratocystis platani’s membrane: CP. </p>
+
<div class="clear"></div>

Latest revision as of 03:26, 18 October 2014