Team:NCTU Formosa/project

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<li><span>Overview</span></li>
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<li><span>PBAN</span></li>
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{{:Team:NCTU Formosa/source/introo}}
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<li><span>Biobrick</span></li>
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<p class="tab_title">Overview </p>
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<p>Insect damage has been a serious problem for a long time over the whole world and it is difficult to be solved. People have took several methods to kill these harmful insects, but the methods have caused other problems to the environment. In order to solve these troublesome problems and not to cause side effect, we established an alternative way to solve it in an eco-friendly manner.</p>
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<p class="tab_title">PBAN  </p>
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<p>PBAN (Pheromone Biosynthesis Activating Neuropeptide) is a simple, short peptide. When it binds with a receptor on the pheromone gland of an insect, it will activates pheromone synthesis. Just like the pheromones, PBAN is species-specific, so we can use one kind of PBAN to enhance pheromone biosynthesis on the kind of insect that we are targeting.</p>
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<p class="tab_title">Brobrick</p>
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<p>In our basic design, we want to express PBAN in an easier way than the original complex process. To make it more convenient to observe and model, we ligate the reporter gene BFP into our biobrick.    </p>
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===Overview===
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===Motivation===
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====A Serious Problem====
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<p>From ancient times to the present day, harmful insects always greatly affect our daily life, and we don’t have any appropriate solution to these troubling insects. Every year, harmful insects bring significant economic loss to agriculture, which seriously disturb people's livelihood and economy.  </p><p>Take Brazil as an example, where the '''agricultural loss''' can reach up to''' 11 billion US dollars''' because most economic crops here such as coconut, coffee, and sugarcane are under serious insects attack. To control these obnoxious insects, '''1.4 billion''' has to be invested to purchase insecticides annually. Terrible as it may seem, Brazil is just one of the many cases in the world! The truth is that farmers worldwide are doing the same thing! To ensure good harvest, they use '''pesticides''' without much consideration about its side effects. In other words, a great amount of insecticides containing '''toxic substances''' is discharged to the environment every day. Obviously, it’s never a good solution. 
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</p>
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[[File:2014NCTU Formosa project Fig 1-1-1.png|center|thumb|1200px| Fig.1-1-1 Losses caused by pests in Brazil. This chart demonstrates the losses of various crops caused by pests.]]
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<h4>Impact of pest</h4>
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<p>Also, there is a data about the '''IPM (Integrated pest management)''' in USA. Integrated pest management, which is defined as integration of pest control based on predicted economic, ecological and sociological consequences, makes maximum use of naturally occurring control agents, including weather, disease organisms, predators and parasites.In brief,IPM means the ability to control harmful insects.The lower the number is, the more efficient we are able to control the pests in the evaluated area. In the data, we can see that crops are planted in lots of areas in USA.There are several kinds of plants such as corn and soybean which suffer from insect damage. Since we can only control the insect damage efficiently within a small, limited area. Therefore, as the production areas expand to an extent, the effect of control declines exponentially. Even though the production areas become larger, the total production might be lower.</p>
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<p>From ancient times to the present, harmful insects always play roles in our life, and we don’t have any appropriate solution to these annoying pests. Every year, harmful insects cause a great amount of loss to our agriculture, which seriously disturbs our people livelihood and economy.  </p><p>In order to make our discuss more trustworthy, we found some data to support our opinion. Firstly, we take Brazil for example. We can find several crucial economic crops such as coconut, coffee, and sugarcane are damaged by harmful insects. The total loss of crops in Brazil is about 11 billion US dollars. In addition, we can also find that people spent nearly 1.4 billion controlling these annoying pests. What's more? They have used a great number of insecticides  containing some toxic substances. However, this method will make human beings exposed to horrible situation.
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<p>
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</p>losses caused by pests in Brazil
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Therefore, what we need is a brand new method to solve this problem. </p>
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[[File:Pest data6.png|center|1200px| fig.1-1-1  This chart demonstrates the losses of various crops caused by pests.]]
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[[File:2014NCTU Formosa project Fig 1-1-2.png|center|thumb|1200px| Fig.1-1-2  This chart demonstrates IPMs of the various areas of different crops planted in the USA.]]
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<div><p></p></div>
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<p></p>
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<p>Secondly, there is also a data about the IPM (Integrated pest management) of USA. IPM means the ability to control pests. The IPM is reality number in global, and the lower of the number is, the more efficiently we control. In the data, we can see that lots of areas in USA are used to plant crops. There are several kinds of plants which are still under attack by harmful insects. We can only control the insect damage fairly well in a small limited production area. Therefore, as the production areas expand to an extent, the effect of control declines to a point. Even though the production areas become larger, the total production is lower.
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====Common Solutions====
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Therefore, what we need is invent a brand new method to solve this problem.</p>
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=====Chemical Control Method=====
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<p>
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Because of insects damage, human tried lots of ways to kill these harmful insects. In the 15<sup>th</sup> century, people used pesticide containing '''heavy metal''' such as Arsenic, Mercury and Plumbum to kill harmful insects, which was a catastrophe to the environment. Pesticides became more powerful along with the development of modern technology. In the 20<sup>th</sup> century, the agriculture prospered rapidly owing to the evolution of pesticides. But the pesticides are not only fatal to the insects but also harmful to the human body. People found this problem after several decades. The '''toxin''' of the pesticides will remain in creatures through the '''food chain''' and enter the '''human body''' eventually. </p>
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[[File:PIC.png|center|1200px| fig.1-1-2  This chart demonstrates the area we plant of crops IPM of various crops in USA.]]
 
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<div><p></p></div>
 
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[[File:2014NCTU Formosa project Fig 1-1-3.png|center|400px|thumb|Fig.1-2-1 Human health and environmental cost from pesticides in the United States is estimated at $9.6 billion. ]]
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====chemical control method====
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'''Damage to Human Body'''
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    <p> Pesticides includes herbicides, insecticides, fungicides and rodenticides, which will kill weeds, insects, fungus, rodents and others. In our project, we refer to insecticides as pesticides. The use of toxic pesticides to manage insect problems has become a common practice around the world. However, pesticides cause serious '''human health hazards''', ranging from short-term impacts such as headaches and nausea to chronic impacts like cancer and reproductive harm. There are also many researches revealing that exposure to pesticides will disrupt the endocrine system, the reproductive system and embryonic development.</p>
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'''Damage to the Environment'''
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    <p>Pesticides pollute the environment such as threatening '''biodiversity''' and '''weakening the natural systems'''. Pesticides have led to abnormally high mortality of America's honeybees. The population of bees has dropped by 29 % to 36 % each year since 2006. To our amazement, approximately 1/3 of the food we eat depends on bees for pollination!</p>
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    <p>Some scientists believe that amphibians and bats have become more susceptible to deadly diseases because their immune systems are weakened by pesticides. Pesticides also contaminate waterways and endangered fish and birds, causing '''ecological damage'''.</p>
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<p>
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=====Biological Control Method=====
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Because of the hazard of insects, human beings have come up with lots of ideas to kill these harmful insects. In the 15<sup>th</sup> century, people used heavy metals such as Arsenic, Mercury and Plumbum to kill harmful insects, which caused a catastrophe to the environment. Pesticides became more powerful along with the technology, in the 20<sup>th</sup> century, the agriculture developed rapidly just because of the evolution of pesticides. But the pesticides are not only fatal to the insects but also harmful to the human beings. People found this problem after several decades. The toxin of the pesticides will be kept in creatures by the food chain, and finally, goes into human's body. However, it's not too late to improve this situation. We can create an evolution of agriculture by a new method, and that is what we do!</p>
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<p>Since most chemical control methods are poisonous to the environment, humans began to look for alternative solutions. One of the  best known example is bacillus thuringiensis (Bt), which can efficiently kill insects with its crystal proteins. However, recent researches have shown that more and more insects have been '''resistant''' to this kind of protein.</p>
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[[File:NCTU pesticide.JPG|center|350px|thumb|fig.1-1-3 Human health and environmental cost from pesticides in the United States is estimated at $9.6 billion. ]]
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<p>Chemical control methods have caused fatal environmental damage and insects have grown resistant to both the chemicals substances and Bt. It is, however, not too late to improve this situation. We can create an agricultural revolution with our new method! </p>
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[[File:2014NCTU Formosa project Fig 1-1-4.png|center|500px|thumb|Fig.1-2-2  Comparison of both solutions. ]]
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<p>Pheromone trap is currently the most novel method to resolve insect damage problems. What's more, pheromone is a '''pollution-free''' substance that is produced by the insects, meaning that insects '''cannot resist''' it.  That's why we take biological synthesis of pheromone in ''E.coli'' to solve the problem. </p>
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=====Damage to human body=====
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====The Pheromone Trap====
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    <p> Pesticides includes substances that kill weeds (herbicides), insects (insecticides), fungus (fungicides), rodents (rodenticides), and others. The use of toxic pesticides to manage pest problems has become a common practice around the world. Pesticides are used almost everywhere, causing human health hazards, ranging from short-term impacts such as headaches and nausea to chronic impacts like cancer and reproductive harm. There is also mounting evidence that exposure to pesticides disrupts the endocrine system, wreaking havoc with the complex regulation of hormones, the reproductive system, and embryonic development.</p>
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=====Damage to the environment=====
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=====Introduction of Pheromones=====
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    <p>Pesticides wreak havoc on the environment, threatening biodiversity and weakening the natural systems upon which human survival depends. Pesticides have led to abnormal mass mortality of America's honeybees. The populations of bees have dropped by 29% to 36% each year since 2006. To our astonishment, fully 1/3 of the food we eat depends on bees for pollination!</p>
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<p>A pheromone is a secreted or excreted chemical factor that triggers a '''social response''' among members of the '''same species'''. Pheromones are capable of acting outside the body of the secreting individual to impact the behavior of the receiving individuals. </p>
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    <p>Secondly, some scientists believe that amphibians and bats have become more susceptible to deadly disease because their immune systems are weakened by pesticides. What's more, one kind of pesticides called herbicide, which can give a male frog a sex change. Genetically, the frogs are still males, but morphologically they are completely female and they can even mate successfully with other males and lay viable eggs. Pesticides have also contaminated waterways and endangered fish and birds, causing ecological damage.</p>
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    <p>By the way, pesticide resistance makes insecticides ineffective. Other factors in the speed with which a species evolves resistance are generation time and fecundity, that is, causing shorter generations and more offspring lead to resistance more quickly.</p>
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====Biological control method====
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<p>There are many kinds of pheromones such as alarm pheromones, food trail pheromones, sex pheromones, and many others that affect behavior or physiology. </p>
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　<p>Since most chemical control methods are poisonous to the environment, humans began to look for other alternative ways. One of the most well known example is bacillus thuringiensis (Bt), which can efficiently kill insects with its crystal proteins. However, recent researches have shown that more and more insects have been resistant to this kind of protein. It's time to find other better solutions.  
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<p>Sex pheromones are an important factor in finding a mating partner. When a female releases chemicals, the mating search is initiated, and the male moths begin their upwind motion toward their potential partner. Pheromones have the ability to propel '''long-distance''' attraction and are emitted by the  abdominal glands of female insects in most cases.</p>
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[[File:NCTU Overview.png|center|500px|thumb|fig.1-1-4  Comparison of both solutions. ]]</p>
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=====How to Produce Pheromones=====
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<p>
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There are 2 main ways to produce pheromones. One through chemical synthesis, and the other through biosynthesis approach.
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</p>
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====Solution to both questions====
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<p>
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'''Chemical Synthesis Approach Used in Factory for Massive Production'''
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</p>
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<p>
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It is '''difficult''' to make pheromone with a chemical synthesis approach. This approach requires many kinds of chemical compound which may cause '''pollution''' to the environment. Furthermore, the control of reacting condition may consume a lot of '''energy'''. An example of chemical synthesis approach: Leafroller moth, ''Bonagota cranaodes''.</p>
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[[File:Chemical synthesis approach.png|500px|thumb|center|Fig.1-3-1 If we want to produce the pheromone of Leafroller moth (''Bonagota cranaodes'') by chemical synthesis approach, we have to purchase expensive equipment to control the conditions. And it may produce toxic byproducts.<sup>(34)</sup>(Paulo H. G. Zarbin et al. 2007) ]]
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People invented pesticides to eliminate the pests, but the use of pesticides hurt the environment and cause harm to humans. Thus there have been conflicting issues, and have remained to be settled.
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<p>
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In order to solve the above-mentioned problems, we came up with a practical, inexpensive way.
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'''Simulating the Biosynthetic Pathway in Insects with ''E.coli'''''</p>
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<p>Biosynthesis approach requires the cooperation of different enzymes, which is '''too complicated''' for ''E.coli'' to carry out. In addition, ''E.coli'' lacks Glycoproteins for posttranslational modification. Therefore, it is nearly impossible for ''E.coli'' to produce pheromones.</p>
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[[File:2014NCTUPyramid.JPG|300px|thumb|center|fig.1-1-5 This is a diagram of our device, we will mention how this device can bring better results for our plan and how we improve its efficiency later.]]
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[[File:Gypsy moth pheromone biosynthesis.png|600px|thumb|center|Fig.1-3-2 If we want to produce the pheromone of Gypsy moth, ''Lymantria dispar''. We have to pass at least four complicated pathways. That will be highly stressful for ''E.coli'', and it may also cause low efficiency of producing pheromone.<sup>(35)</sup>(Russell Jurenka, 2004)]]
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====Our target insect====
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<p>
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<p class="bigtxt">Check photo to see introduce</p>
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'''Difficulty and Disadvantage of Pheromone Production'''
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<div class="center">
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</p>
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{{:Team:NCTU Formosa/source/project/test}}
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<p>As mentioned above, pheromone is '''too complicated''' to be produced either artificially or bio-synthetically. Although the cost of artificial chemical synthesis can be lowered through massive production, the '''environmental damage''' it causes is fatal. With that said, we decided to find an easier and better way to reach our goal.</p>
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{{:Team:NCTU Formosa/source/project/test3}}
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====PBAN- The Solution====
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To solve the impossibility of biologically synthesizing pheromone with ''E.coli'', we found a feasible solution. As mentioned above, our method should be '''pollution-free''', '''efficient''', '''insect resistance-free''', and '''cheap'''. Combining all these advantageous elements, we introduce you '''PBAN'''. Click the photos to learn more! 
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{{:Team:NCTU Formosa/source/project/test3}}
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<div class="ref">
<ol start="1">
<ol start="1">
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1.Pogue, Michael. "A review of selected species of Lymantria Huber [1819]". Forest Health Technology Enterprise Team. Retrieved September 14, 2012.<br>
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2.Eliminate Cutworms Using Natural Pest Control By Susan Glaese
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<li>Cabbage Moth - Caterpillar". Habitas.org.uk. Retrieved 2014-08-11.</li>
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March/April 1987
+
<li>Interesting (To Us) Photos From The Garden". Meades.org. Retrieved 2011-08-11.</li>
-
<br>
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<li>RXwildlife Sightings » Blog Archive » More Emergence". Rxwildlife.org.uk. 2009-06-03. Retrieved 2011-08-11.</li>
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3.  Laurence Mousson, Catherine Dauga, Thomas Garrigues, Francis Schaffner, Marie Vazeille & Anna-Bella Failloux (August 2005). "Phylogeography of Aedes (Stegomyia) aegypti (L.) and Aedes (Stegomyia) albopictus (Skuse) (Diptera: Culicidae) based on mitochondrial DNA variations". Genetics Research <br>
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<li>Practice Organic Cabbage Worm Control for a Chemical-Free Garden, by Barbara Pleasant, March 25, 2013</li>
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4. NPIC is a cooperative agreement between Oregon State University<br>
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<li>2003-2009 Project «Interactive Agricultural Ecological Atlas of Russia and Neighboring Countries. Economic Plants and their Diseases, Pests and Weeds</li>
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5.M. S. Ascunce et al., “Global Invasion History of the Fire Ant Solenopsis invicta”, Science, vol. 331, no. 6020, pp. 1066 - 1068, 2011<br>
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<li>Report of a Pest Risk Analysis Helicoverpa armigera, Hübner, 1808</li>
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6.organicgardening.com/learn-and-grow/fire-ant-control<br>
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<li>UC IPM Pest Management Guidelines:
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7.Scientists suggest fighting fire ants with ice By Chiu Yu-Tzu  /  STAFF REPORTER<br>
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Cotton, http://ucanr.edu/sites/ipm/pdf/pmg/pmgcotton.pdf</li>
-
8.Capinera, J.L. 1999. Fall armyworm Spodoptera frugiperda (J.E. Smith) (Insecta: Lepidoptera: Noctuidae<br>
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<li>Michigan State University’s invasive species factsheets(Oriental leafworm Spodoptera litura)
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9.Helicoverpa Diapause Induction and Emergence Tool
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, http://www.ipm.msu.edu/uploads/files/Forecasting_invasion_risks/orientalLeafworm.pdf</li>
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<li>Effect of tetra hydroxyl-ρ-benzoquinone on growth and
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metamorphosis of Spodoptera litura Fabr.
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(Lepidoptera: Noctuidae) larvae, Sujata Magdum
 +
, Seema Gupta
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</li>
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<li> Espinosa, A. and A.C. Hodges University of Florida, Spodoptera litura, BUGWOODWiki, http://wiki.bugwood.org/Spodoptera_litura.</li>
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<li>OROKIN, Fire Ant Anatomy, http://www.orkin.com/ants/fire-ant/fire-ant-anatomy</li>
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<li>Wikipedia, Red imported fire ant
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http://en.wikipedia.org/wiki/Red_imported_fire_ant</li>
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<li>
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Organic Gardening.com/learn-and-grow/fire-ant-control</li>
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<li>Cram101 Textbook Reviews, Life: The Science of Biology 8th Edition</li>
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<li>
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Wikipedia, Aedes aegypti
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, http://en.wikipedia.org/wiki/Aedes_aegypti</li>
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<li>National Pesticide Information Center, Mosquito Control Methods</li>
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<li>
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BugwoodWiki
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, Cutworms, by Dr. Steve L. Brown, Dr. Will Hudson</li>
 +
<li>Eliminate Cutworms Using Natural Pest Control By Susan Glaese
 +
March/April 1987</li>
 +
<li>
 +
Wikipedia, Lymantria dispar dispar
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, http://en.wikipedia.org/wiki/Lymantria_dispar_dispar</li>
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<li>
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FACT SHEET :
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GYPSY MOTH, http://www.metro-forestry.com/wp-content/uploads/2010/02/FactSheet_Gypsy-Moth.pdf</li>
 +
<li>
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Planet Natural, GYPSY MOTH</li>
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<li>
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Larry P. Pedigo, G. David Buntin, Handbook of Sampling Methods for Arthropods in Agriculture</li>
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<li>L. A. Hull, D. G. Pfeiffer & D. J. Biddinger, Apple Direct Pests</li>
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 +
<li>
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IPM, obliquebanded leafroller, http://www.nysipm.cornell.edu/factsheets/treefruit/pests/oblr/oblr.asp</li>
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<li>
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UC IPM Online, Leafrollers on Ornamental and Fruit Trees, Publication 7473 September 2010</li>
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<li>Tan Koon San, Dynastic China: An Elementary History</li>
 +
 
 +
<li>Pogue, Michael. "A review of selected species of Lymantria Huber [1819]". Forest Health Technology Enterprise Team. Retrieved September 14, 2012.</li>
 +
 
 +
<li>Laurence Mousson, Catherine Dauga, Thomas Garrigues, Francis Schaffner, Marie Vazeille & Anna-Bella Failloux (August 2005). "Phylogeography of Aedes (Stegomyia) aegypti (L.) and Aedes (Stegomyia) albopictus (Skuse) (Diptera: Culicidae) based on mitochondrial DNA variations". Genetics Research </li>
 +
<li>M. S. Ascunce et al., “Global Invasion History of the Fire Ant Solenopsis invicta”, Science, vol. 331, no. 6020, pp. 1066 - 1068, 2011</li>
 +
 
 +
<li>Scientists suggest fighting fire ants with ice By Chiu Yu-Tzu  /  STAFF REPORTER</li>
 +
<li>Capinera, J.L. 1999. Fall armyworm Spodoptera frugiperda (J.E. Smith) (Insecta: Lepidoptera: Noctuidae)</li>
 +
<li>Helicoverpa Diapause Induction and Emergence Tool
Introduction to the Helicoverpa armigera Genome Project
Introduction to the Helicoverpa armigera Genome Project
Helicoverpa armigera Genome Project updates on InsectaCentral
Helicoverpa armigera Genome Project updates on InsectaCentral
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Lepiforum
Lepiforum
Funet Taxonomy
Funet Taxonomy
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Fauna Europaea<br>
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Fauna Europaea</li>
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UC IPM Pest Management Guidelines: Cotton
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UC ANR Publication 3444
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<br>
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10. "Interesting (To Us) Photos From The Garden". Meades.org. Retrieved 2011-08-11.RXwildlife Sightings » Blog Archive » More Emergence". Rxwildlife.org.uk. 2009-06-03. Retrieved 2011-08-11.Cabbage Moth - Caterpillar". Habitas.org.uk. Retrieved 2014-08-11.<br>
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11.Authored by W. H. Reissig.
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Published by the New York State Agricultural Experiment Station, Geneva, A Division of the New York State College of Agriculture and Life Sciences, A Statutory College of the State University, Cornell University, Ithaca. Funded in part by an Extension Service-USDA, IPM Grant.<br>
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<li>Authored by W. H. Reissig.
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Published by the New York State Agricultural Experiment Station, Geneva, A Division of the New York State College of Agriculture and Life Sciences, A Statutory College of the State University, Cornell University, Ithaca. Funded in part by an Extension Service-USDA, IPM Grant.</li>
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<li>Paulo H. G. Zarbin; José A. F. P. Villar; Arlene G. Corrêa, Insect pheromone synthesis in Brazil: an overview, Journal of the Brazilian Chemical Society, vol.18 no.6 São Paulo  2007</li>
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===PBAN(Pheromone Biosynthesis Activating Neuropeptide)===
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===PBAN ( Pheromone Biosynthesis Activating Neuropeptide )===
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<p>PBAN (Pheromone Biosynthesis Activating Neuropeptide) is a kind of peptide that can activate biosynthesis of pheromones of specific insects. Once a PBAN binds with the''' G-protein coupled receptor''' located at an insect’s pheromone gland, it will send a signal to activate kinase and phosphatase, which in turn activates other enzymes that participate in the biosynthesis of insect sex pheromones. These pheromones are eventually emitted<sup>(35)</sup>.</p>
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====Introduction of Pheromones====
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<p>In nature, '''female insects''' such as '''moths release PBAN''' to '''stimulate the synthesis of pheromones''' in order to attract male moths during mating. PBAN can also facilitate the release of non-sex pheromones such as trail pheromones for ants. Overall, this kind of natural substance, PBAN, has many advantages in the following, which can complete our goals perfectly.
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<p>A pheromone is a secreted or excreted chemical factor that triggers a social response in members of the same species. Pheromones are chemicals capable of acting outside the body of the secreting individual to impact the behavior of the receiving individual. </p>
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<p>There are many kinds of pheromones such as alarm pheromones, food trail pheromones, sex pheromones, and many others that affect behavior or physiology. </p>
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<p>Our project - Operation Debug aims to use different PBAN from different moths and other insects to induce them to make their own sex pheromone. </p>
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<p>Sex pheromones, which are an important factor in finding a mating partner. When a female releases chemicals, the mate search is initiated, and the male moths begin their upwind motion toward their potential partner. Sex pheromones in particular are associated with long-range chemical communication of sex substances used in signaling a mating partner. Mate finding in moths involve sex pheromones that have the ability to propel long-distances and are emitted by the females abdominal glands in most cases.</p>
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<br>
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<br>
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=====Ways to make pheromones=====
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<p>
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There are 2 main ways to make pheromones. One is chemical synthesis, the other one is biosynthesis approach.
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</p>
</p>
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<p>
 
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'''1. Chemical synthesis approach：'''
 
-
<br>
 
-
  It is difficult to make pheromone by chemical synthesis approach. This approach needs to use many kinds of chemical compound which may cause some pollution to the environment. Furthermore, chemical approach needs to control the condition of reaction that may consume a lot of energy. An example of chemical synthesis approach: leafroller moth, Bonagota cranaodes.　
 
-
[[File:Chemical synthesis approach.png|500px|thumb|center|fig.1-2-1 If we want to produce the pheromone of leafroller moth and Bonagota cranaodes by Chemical synthesis approach ,we have to buy some expensive equipments to control the conditions.And it may produce some byproduct which is toxic. ]]
 
-
</p>
 
-
<br>
 
-
<p>
 
-
'''2.Biosynthesis approach：'''
 
-
<br>
 
-
  Biosynthesis approach needs lots of enzymes to catalyzed, which is too complicated for E.coli to carry out. In addition, E.coli is lack of Glycoproteins that can do posttranslational modification. Therefore, it is nearly impossible for E.coli to produce pheromones in a normal way. An example of Biosynthesis approach: Gypsy moth, Lymantria dispar.
 
-
[[File:Gypsy moth pheromone biosynthesis.png|600px|thumb|center|fig.1-2-2 If we want to produce the pheromone of Gypsy moth, Lymantria dispar. We have to pass at least four complicated pathways. That will be high loading for E.coli,and it may also cause low efficiency of producing pheromone.
+
[[File:2014NCTUGprotein.jpg|800px|thumb|center|Fig.2-1-1 This is the biosynthetic pathway of pheromones. Once PBAN comes in contact with the PBAN receptor, the receptor will send a signal to activate other enzymes that participate in the biosynthesis of insect pheromones.]]
-
]]
+
-
</p>
+
====Features of PBAN====
-
====Introduction of PBAN====
+
<p>1. '''Species-specific''': PBAN is species-specific just like pheromones, meaning that every kind of insect produces specific PBAN that only binds with its specific receptor, resulting in the production of a particular pheromone. </p>
-
<p>PBAN (Pheromone Biosynthesis Activating Neuropeptide) is one kind of peptides that can activate biosynthesis of pheromones of insects we target. Once a PBAN binds with the G-protein coupled receptor on an insect’s pheromone gland, the signal send by the G-protein coupled receptor activates the kinase and phosphatase, and then kinase and phosphatase can activate enzymes that participate in the biosynthesis of insect pheromone, which will be emitted.</p>
+
-
<p>In nature, female insects such as moths release PBAN during mating to stimulate the synthesis of pheromones in order to attract their male counterparts. PBAN can also facilitate the release of non-sex pheromones such as trail pheromones for ants.
+
<p>2. '''Small and simple''': The coding sequence for a PBAN is only around 100 base pairs. For ''E.coli'', 100 base pairs is totally within its working capacity. Therefore, ''E.coli'' can be a low-cost PBAN factory. By transforming the DNA sequences for different PBAN into the ''E.coli'', we can even gain a variety of PBANs. 　</p>
-
  </p>
+
<p>3. '''Secreted directly''': Because PBAN can be synthesized by the insect itself, the insect would not form a resistance to it compare to use pesticide. </p>
-
[[File:2014NCTUGprotein.jpg|400px|thumb|center|fig.1-2-3 Working mechanism of PBAN ]]
+
<p>In conclusion, using PBAN is totally a environmental friendly way for solving harmful insects problems with easily triggering the production of pheromone by contacting with PBAN receptors. </p>
 +
</div></div>
 +
<div class="li"><div class="card">
-
====Features of PBAN====  
+
===Main Idea===
 +
<p>We plan to produce PBANs through ''E.coli'' and make the PBAN come in contact with the target insect. The target insect will then start producing pheromones and attract more target insects.</p>
-
<p>1. PBAN is species-specific just like pheromones, that means every kinds of insects which can produce pheromone have it's specific PBAN， which can only bind with it's specific receptor and only stimulate the biosynthesis of a specific pheromone. </p>
+
====Pheromone Production====
-
<p>2. The coding sequence for a PBAN is usually around 100 base pairs. Thus, it is easy for E.coli to express. We can even combine several different PBAN sequences into one BioBrick assembly or even make different regulation for different PBANs.　</p>
+
<p>Before we could employ PBAN to accomplish our goals, we need to obtain enough background knowledge to evaluate the workability of our plan through paper research. First, we had to confirm whether an insect can take in vitro PBAN and allow it to function. From “Pheromonotropic Activity of Naturally Occurring Pyrokinin Insect Neuropeptides (FXPR-Lamide) in ''Helicoverpa zea''”<sup>(37)</sup>, this method has been proven possible. In this experiment, 0.05 pmol to 200 pmol of PBAN had been injected in to corn earworm and the amount of pheromone produced had been measured. The result (Fig.1-2-4) showed that PBAN produced by insects itself can effectively increase the production of pheromone. The maximized amount of pheromone production was reached with only "10 pmol" of PBAN. This experiment means as long as '''PBAN enters''' '''the body of the insect''', it highly possible that''' PBAN''' can '''stimulate the production of pheromone'''.</p>
-
====Effect Testing of PBAN from Reference====
+
[[File:PBAN圖表.jpg|thumb|950px|center|Fig.3-1-1 Effect of dose of Hez-PBAN on stimulation in vivo pheromonotropic activity in vitrugin females of H. zea (bars indicate SEM. n=8). This figure shows that PBAN can efficiently stimulate pheromone production in just a small amount (10 pmol).(Rellal.Abernathy et al, 1995)]]
-
<p>Because we regard PBAN as a leading character in our project, we have to realize what kind of problem we would meet with if we want to make use of PBAN. First, we had to confirm that whether PBAN can work when getting in the moth’s body from outside, we got a paper titled “Pheromonotropic Activity of Naturally Occurring Pyrokinin Insect Neuropeptides (FXPRLamide) in Helicoverpa zea” from ELSEVIER, experiments of the research were conducted with Helicoverpa zea (corn earworm) in this research, researchers injected different amount (from 0.05 pmol to 200 pmol) of PBAN of Helicoverpa zea (named Hez-PBAN) into their bodies directly, and then measured the amount of pheromone produced, the result (fig.1-2-4) showed that PBAN has function as long as it gets in moth’s body, and the 2 pmol dose of Hez-PBAN was the minimum dose required to stimulate production of the maximum amount of pheromone, that revealed we are able to stimulate the production of pheromone with only little amount of PBAN which gets in moth’s body, whether we fed or injected PBAN.</p>
+
====A Problem of Degradation by Peptidase====
 +
<p> The only problem for us to use PBAN is that PBAN would be '''degraded''' quickly in an insect’s body. In order to understand this problem, we searched some former papers and we found another paper from Peptides, “Enhanced oral availability/pheromonotropic activity of peptidase-resistant topical amphiphilic analogs of pyrokinin/PBAN insect neuropeptides<sup>(36)</sup> .” In this experiment, ''Heliothis virescens'' was the experimental insect and amphiphilic analogs of pyrokinin/PBAN were used instead of natural PBAN. This way, the analog can resist peptidase, as shown in the graph below. The amount of the natural PBAN decreases quickly while the analogs degrades slowly (Fig.3-2-1). However, we found the amphiphilic analogs of '''pyrokinin/PBAN peptide''' requires many special artificial modification, which means ''E.coli'' absolutely cannot produce this artificial PBAN that can easily resist peptidase. Thus, we need more brain storming to think of a way to use PBAN even if it may be degraded by peptidase.</p>
-
[[File:PBAN圖表.jpg|800px|thumb|center|fig.1-2-4 Effect of dose of Hez-PBAN on stimulation of in vivo pheromonotropic activity in vitrugin females of H. zea (bars indicate SEM. n=8).]]
+
[[File:PBAN圖表3-2.jpg|450px|thumb|center|Fig.3-2-1  Stability of the natural pyrokinin/PBAN analog LPK, and the peptidase-resistant pyrokinin/PBAN analogs Hex-Phe-Thr-Hyp-Trp-Gly-NH2 and Hex-Phe-Thr-Oic-Trp-Gly-NH2 to hydrolysis by peptidases bound to corn earworm (H. zea) Malpighian tubule tissue. Measurement of the amount of remaining peptide was made by HPLC at 30, 60, and 120 min. The data points represent the means of at least three replicates.(Ronald J. Nachmana, Peter E.A. Teal, Allison Strey, 2002)]]
-
====Difficulty of Natural PBAN about Peptidase Degradation====  
+
====Our Solution====  
-
<p>But we still had the second problem, we had known that peptidase in the insect’s body can degrade peptides including PBAN, so the effect of PBAN can’t continue for a long time. In order to solve this problem, we search some former papers and we found another paper from ELSEVIER, it was titled “Enhanced oral availability/pheromonotropic activity of peptidase-resistanttopical amphiphilic analogs of pyrokinin/PBAN insect neuropeptides”, Heliothis virescens was used in this research, researchers produced amphiphilic analogs of pyrokinin/PBAN with chemical synthesis method, they used hydroxyproline (Hyp) and octahydroindole-2-carboxyl  (Oic) to replace the Proline (Pro) of the PBAN C-terminal penpapeptides (Phe-Thr-Pro-Arg-Leu-NH2), that made PBAN be able to resist peptidase, they compared the stability of the natural pyrokinin/PBAN analog LPK, and the peptidase-resistant pyrokinin/PBAN analogs (5 nmol was used), we can see that natural PBAN will be reduced over time in the moth’s body (fig.1-2-5).</p>
+
<p>The results above did not sound good to us, but there was another experiment that interested us in the same research. Researchers also conducted '''oral test''' with '''artificial PBAN'''/pyrokinin analogs (Fig.3-3-1), in this experiment, they fed moths with '''sugar solution''' which contained their PBAN analogs, and measured the amount of pheromone produced over time. (Blank: natural PBAN in low concentration) This result inspired us. Although natural PBANs can’t be maintained for a long time in moth’s body, we could solve this problem by simply '''feeding moths''' with '''high concentration PBANs''' continually.</p>
-
[[File:PBAN圖表3-2.jpg|450px|thumb|center|fig.1-2-5  Stability of the natural pyrokinin/PBAN analog LPK (_), and the peptidase-resistant pyrokinin/PBAN analogs Hex-Phe-Thr-Hyp-Trp-Gly-NH2 (901)(_) and Hex-Phe-Thr-Oic-Trp-Gly-NH2 (904) (_) to hydrolysis by peptidases bound to corn earworm (H. zea) Malpighian tubule tissue. Measurement of the amount of remaining peptide was made by HPLC at 30, 60, and 120 min. The data points represent the means of at least three replicates]]
+
[[File:WIKI 3-3.jpg|450px|thumb|center|Fig.3-3-1 Amount of pheromone, relative to the maximum pheromone amount induced by injected PBAN, produced by Hyp-analog and Oic-analog, 1.5, 3, 4, and 6h following oral administration. Dotted line at 100% denotes maximal pheromone production of injected PBAN (positive control). This figure shows a very important information that blank (natural PBAN in low concentration：50 pmol, the blank was measured when it was 2 hr after feeding with nature PBAN) still has some ability to stimulate pheromone production. Thus, we decided to try to feed the female target insects with high concentration PBAN continuously.(Ronald J. Nachmana, Peter E.A. Teal, Allison Strey, 2002) ]]
-
====Our New Idea of using PBAN====
+
<p> Thus, as long as '''female moths''' suck the high concentration''' PBAN''' solution''' continuously''', the PBAN in the moths’ body will be replenished continually even if peptidase degradation occurs. Then, there is a high possibility of PBAN being absorbed by the moths’ body from the digestive system and succeed in stimulating the pheromone gland. Thus, if we feed the female insects with high concentration PBAN solution continuously, the female insects will produce pheromones for us, which theoretically solves the problem of PBAN's inability to maintain for a long time in the insects’ body. As to how we produce PBAN and apply our concept to capture wanted harmful insects, we will explain in the following.</p>
-
<p>This results above sounded not good for us, but there was other experiment interested us in the same research, researchers also conducted oral test with their PBAN (fig.1-2-6), in this experiment, they fed moths with sugar solution which contained their PBAN analogs, and measure the amount of pheromone produced over time, that gave us inspiration, although natural PBAN can’t be maintained for a long time in moth’s body, we could solve this problem by feeding moths with PBAN continually, after combining this thought with the method of the oral experiment, we figured out an idea, instead of killing all of the moths.</p>
+
-
<p>It is very possible that the PBAN in the food will penetrate into moths’ body from the digestive system. Thus, if female moths suck the food, PBAN in the moths’ body will be replenished, which solves the problem that PBAN can not maintain for a long time in the moths’body. As to how we produce PBAN and apply our concept to capture wanted harmful insects, we will explain in the following.</p>
+
====How We Are Going to Use PBAN?====
 +
[[file:How_we_are_going_to_use_PBAN.jpg|center|thumb|700px| Fig.3-4-1  Overview of our project.]]
 +
<p>In our project, we will biologically synthesize PBAN with the ''E.coli''. We store the PBAN inside a trapping device [https://2014.igem.org/Team:NCTU_Formosa/project#Device (check this out at our Device page)]. In the device, there will be appropriate lighting and nutrient sources that will attract insects. </p>  
-
[[File:WIKI 3-3.jpg|450px|thumb|center|fig.1-2-6 Amount of pheromone, relative to injected PBAN, produced by Hyp-analog 901 and Oic-analog 904, 1.5, 3, 4, and 6 h following oral administration.Dotted line at 100% denotes maximal production of injected PBAN (positive control).]]
+
<p>Once an insect is attracted into our device and ingests the nutrient sources we provide, it will also inevitably come in contact with our PBAN. As the PBAN works and activates the pheromone synthesis of the attracted insect, more of this species of insect’s counterparts will be attracted and later captured. </p>
-
====How are we going to use PBAN?====
+
<p>Owing to the first feature mentioned above, PBAN is species-specific, which means that it doesn't matter if other kinds of insects fly into our device and eat the PBAN. This is because the insects we don't want to catch will not be stimulated by PBAN to produce pheromone. Therefore, the PBAN is only for what we want to catch, and we are sure that our method won't affect other kinds of insects. </p>
-
In our project, we will biologically synthesize PBAN with our E.coli. We store the PBAN inside a trapping device (check this out at our Device page). In the device, there will be appropriate lighting and nutrient sources that will attract insects.  
+
-
[[File:PBAN_mechanism-2.jpg|450px|thumb|center||fig.1-2-7 In this picture,different kinds of insects are attracted by blue light,and they accidentally eat our PBAN when they enjoy the syrup.]]
 
-
<p>Once an insect is attracted into our device and ingests the nutrient sources we provide, it will also inevitably come in contact with our PBAN, which is evenly mixed with the nutrient sources. As the PBAN works its magic and activates the pheromone synthesis of the attracted insect, more of this species of insect’s counterparts will be attracted and later captured. </p>
 
-
[[File:PBAN_mechanism-3.jpg|450px|thumb|center||fig.1-2-8 Because the E.coli in the picture only produce moth PBAN. Therefore,only the pheromone of moth is produced,and a great number of moths will be attracted.]]
 
-
 
-
Owing to the first feature mentioned above, PBAN are species-specific, which means that it doesn't matter if other kind of insect fly into our device and eat PBAN, because the insects we don't want to catch will not be stimulated by PBANs to produce pheromone; our PBAN are only for what we want to catch, and we are sure that our method won't affect other kinds of insects.
 
-
 
-
====PBAN we use====
 
-
Check photo to see petite sequence.
 
-
<div class="center">
 
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{{:Team:NCTU Formosa/source/project/test2}}</div>
 
======Reference======
======Reference======
<div class="ref">
<div class="ref">
-
<ol start="1">
+
<ol start="35">
-
<li>Miriam Altstein， Role of neuropeptides in sex pheromone production in moths, ELSEVIER, Peptides 25 (2004) 1491–1501.</li>
+
<li>Russell Jurenka, Insect Pheromone Biosynthesis, Topics in Current Chemistry (2004) 239: 97– 132
-
<li>Ada Rafaeli, Pheromone biosynthesis activating neuropeptide (PBAN): Regulatory role and mode of action, General and Comparative Endocrinology 162 (2009) 69–78</li>
+
DOI 10.1007/b95450, 2004</li>
-
<li>Ronald J. Nachmana, Peter E.A. Teal, Allison Strey, Enhanced oral availability/pheromonotropic activity of peptidase-resistant topical amphiphilic analogs of pyrokinin/PBAN insect neuropeptides, ELSEVIER, Peptides 23 (2002) 2035–2043</li>+
+
<li>Ronald J. Nachmana, Peter E.A. Teal, Allison Strey, Enhanced oral availability/pheromonotropic activity of peptidase-resistant
-
<li>Russell Jurenka1 and Ada Rafaeli, Regulatory role of PBAN in sex pheromone biosynthesis of heliothine moths, frontiers in ENDOCRINOLOGY, published: 10 October 2011 doi: 10.3389/fendo.2011.00046</li>
+
topical amphiphilic analogs of pyrokinin/PBAN insect neuropeptides, Peptides 23 (2002) 2035–2043.</li>
-
<li>Dr. Ashok K. Raina andJulius J. Menn, Pheromone biosynthesis activating neuropeptide: From discovery to current status, Issue Archives of Insect Biochemistry and Physiology, Article first published online: 7 FEB 2005 DOI: 10.1002/arch.940220112</li>
+
<li>RELLA L. ABERNATHY, RONALD J. , PETER E. A. TEAL, OKITSUGU YAMASHITAS
-
<li>Man-Yeon Choi and Robert K. Vander Meer, Ant Trail Pheromone Biosynthesis Is Triggered by a Neuropeptide Hormone, PLoS Onev.7(11); 2012PMC3511524</li>
+
-
<li>RELLA L. ABERNATHY, RONALD J. NACHMAN, PETER E. A. TEAL, OKITSUGU YAMASHITAS
+
AND JAMES H. TUMLINSON, Pheromonotropic Activity of Naturally Occurring
AND JAMES H. TUMLINSON, Pheromonotropic Activity of Naturally Occurring
Pyrokinin Insect Neuropeptides (FXPRLamide)
Pyrokinin Insect Neuropeptides (FXPRLamide)
-
in Helicoverpa zea, ELSEVIER, Peptides Volume 16, Issue 2, 1995, Pages 215–219</li>
+
in Helicoverpa zea, Peptides, Vol. 16, No. 2, pp. 215-219, 1995.</li>
-
 
+
</ol></div>
-
</ol>
+
-
</div>
+
</div></div>
</div></div>
-
 
<div class="li"><div class="card">
<div class="li"><div class="card">
-
===Design===
+
===Biobrick Design===
 +
====Basic Biobrick Design====
 +
[[File:HALFPBAN.png|400px|thumb|center|Fig.4-1-1 This is our basic design of biobrick.]]
 +
We searched the DNA sequences of the PBANs of many kinds of insects on NCBI, then compared them to the amino acid sequences from papers so that we can select the DNA fragments that directly correspond to gland-stimulating function. By ligating the constitutive promoter (BBa_J23101), ribosome binding site (BBa_B0034) and PBAN DNA sequence with a terminator (BBa_J61048) at last (we delimit this sequence as basic part), we were able to make ''E.coli'' '''directly''' produce these PBANs '''continuously''' instead of the original complex process of PBAN biosynthesis in insects.
-
====Biobrick Design====
+
====Multifunctionality====
-
[[File:HALFPBAN.png|457px|thumb|center||fig.1-3-1 This is our basic design of biobrick.]]
+
[[File:LONGPBAN.png|750px|thumb|center|Fig.4-2-1 Best Potential of Our PBAN Biobrick - Multifunctional.]]
-
<p></p>
+
-
<p>
+
-
We searched the DNA sequences of the PBANs from many kinds of insects on NCBI, then contrasted to the amino sequences from papers so that we can select the DNA fragments which directly correspond to gland-stimulating function. By ligating the constitutive promoter (J23101), ribosome binding site (B0034) and PBAN DNA sequence with a terminator (J61048) at the last (we delimit this sequence as basic part), we were able to make E.coli directly produce these PBANs continusely instead of the original complex process of PBAN biosynthesis in insects. </p>
+
-
<p>
+
-
Our biobrick design can be customized to different pest problems. For instance, there is a farm damaged by 3 kinds of moths : Lymantria dispar, Spodoptera litura and Mamestra brassicae. What we only have to do is change the PBAN DNA sequence of 3 basic parts into the PBAN DNA sequence of Lymantria dispar, Spodoptera litura and Mamestra brassicae. Then we assemble these 3 basic parts together. After these pests are attracted into our device, they will ingest these three kinds of PBANs. Subsequently, these 3 species of moths will produce their own pheromones to attract the same species. </p>
+
-
<p>
+
-
It allows us to assemble these basic parts together because the base pairs of these basic parts are small. Therefore, we can assemble different basic parts which contain different PBAN DNA sequences to resolve different pest problems. </p>   
+
 +
We can assemble these basic parts together easily because the number of base pairs of these basic parts are '''small'''. We can assemble different basic parts that contain different PBAN DNA sequences to resolve different insect problems. Therefore, our biobrick design can be '''customized''' according to the users' needs. For instance, if there is a farm harmed by 3 kinds of moths: ''Lymantria dispar'', ''Spodoptera litura'' and ''Mamestra brassicae'', what we have to do is to ligate the PBAN DNA sequence of 3 basic parts into one plasmid and let the ''E.coli'' express these PBANs. After these three kinds of moth ingest these three kinds of PBANs, subsequently, these 3 species of moths will produce their own pheromones to attract their same-species counterparts. To put it '''simply''', our PBAN basic parts can be assembled together with any combination in infinite possibility.
-
 
+
For even more creative ideas, each of the PBAN basic parts can use different promoters, RBS and terminators to make many '''different regulation'''. Thus, not only can we produce many different PBANs with just one strain of ''E. coli'', but these parts can be regulated to our desire. Thus, our PBAN biobricks really have infinite potential!
-
 
+
   
-
 
+
-
 
+
-
 
+
-
======Reference======
+
-
<div class="ref">
+
-
<ol start="9">
+
-
<li>Torsten Waldminghaus, Nadja Heidrich, Sabine Brantl and Franz Narberhaus .(2007). FourU: a novel type of RNA thermometer in Salmonella . Molecular Microbiology , 65(2): 413–424 DOI:10.1111/j.1365-2958.2007.05794.x</li>
+
-
<li>part BBa_K115002;TUDelft Registry of Standard Biological Parts</li>
+
-
</li>
+
-
</ol>
+
-
</div>
+
</div></div>
</div></div>
-
 
<div class="li"><div class="card">
<div class="li"><div class="card">
===Device===
===Device===
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[[File:NCTU_Formosa_2014_Project_Device1.jpg|thumb|center|700px|fig1-4-1 Our blue light pyramidal device]]
+
In order to actually attract and capture the insect, we take the different weather conditions such as '''light''' and '''temperature''' into consideration. The different conditions will change the efficiency of insect attraction. Therefore, we design a device that can be used in different conditions, which would allow farmers to choose the best way for their local condition.
 +
[[File:NCTU_Formosa_2014_Project_Device1.jpg|thumb|center|700px|Fig.5-1-1 Our blue light pyramidal device.]]
====Introduction====
====Introduction====
<p>
<p>
-
   Since insects behave widely different to the gravity force, we design a device which could catch specific kind of insect species that we want. For example, Agrotis ypsilon Rottemberg and Spodoptera litura falls into the kind of moth that have negative geotaxis. So we made a trap with accessible pathway in the bottom. Once an insect enters the device, it could only goes up and be trapped inside the pyramid. However, after field investigation, we found some insects still escape from the device. Then we came out with a new version trap with doubled layers, inner shell and outer shell.  
+
   Since insects behave widely different to the gravity force, we design a device which could catch specific kinds of insect species that we want. For example, ''Agrotis ypsilon'' (Rottemberg) and ''Spodoptera litura'' fall into the kind of moth that have negative geotaxis '''(antigravity, tendency to fly upwards)'''. Thus, we made a trap with accessible pathway at the bottom. Once an insect enters the device, it could only go up and be trapped inside the pyramid. However, after field investigation, we found some insects still escape from the device. Then we came up with a new version trap with doubled layers, inner shell and outer shell.  
<br><br><br><br>
<br><br><br><br>
</p>
</p>
-
====Mechanism====
+
====Assembling Process====
-
<div>
+
<p>
 +
Before the detailed description of how we design our device, we can show you a simple animation of the '''assembling process''' of our device.
 +
As you can see, our device is very easy to use. Just assemble the outer shell, inner shell (containing PBAN) and tenons together to complete the device. Then, you can put the device at where you want to attract the harmful insects. </p><div style="margin:0pt auto; width:50%;"></div>
 +
　　　　                   
 +
<div style="margin:0pt auto; width:50%;">{{:Team:NCTU Formosa/source/project/Device Assembling Process}}</div>
 +
<br><br><br><br><br>
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<div class="left">
+
====Mechanism of Attraction====
-
[[File:NCTU Formosa 2014 Project Device2.jpg|200px|thumb|center|ssss]]
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</div>
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<div class="right">
+
　　　　　　　　　　　　{{:Team:NCTU Formosa/source/mechanism}}
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First, we divide our design chart into two parts－exterior and interior. The exterior is just like the appearance of pyramid, and the interior is used to equip PBAN and bag for pests. When the harmful insects we want to catch eat our PBAN, they will release pheromone, and attract the same species. We can use blue light or the smell of pheromone to attract insect at first. After they go into our device, the design of our device will take advantages of their characteristic that insects always fly high to escape and make them stuck in our device. When we take away the outer shield, the hock on the outer shield will close the bag, and the insects will be caught. In addition, the four tenons at the corner can firm up our device.
+
-
</div>
+
-
</div>
+
<p>
 +
As you can see, we divide our design into two parts－'''exterior and interior'''. The exterior is just like the appearance of a pyramid, and the interior is designed to accommodate PBAN. At the beginning of using the device, we turn on the blue light LEDs to attract the target harmful insects. When the target female insects are stuck in the interior and eat the food mixed with PBAN solution, they will release pheromones, and attract same-species counterparts. Even if we turn off the blue light, our device can attract many target male insects because the female insects inside our device is still in rut and releasing sex pheromone to attract their mates.
 +
After the harmful insects go into our device, the design of our device will take advantage of their habit. Insects always '''fly high to escape''' so they will be '''stuck in the top of our device'''. </p>
Line 294: Line 287:
====Device Design====
====Device Design====
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<p>For the materials of our device, Acrylic is a polymer created when giant carbon molecules combine chemically. Finished acrylic sheet exhibits glass-like qualities – clarity, brilliance, transparency, translucence – but at half the weight and up to 10 times the impact resistance. It can be tinted or colored, mirrored or made opaque. A number of coatings can be applied to a sheet or finished part for performance enhancing characteristics such as scratch resistance, anti-fogging, glare reduction and solar reflective.
+
<p>More detail information about how we design our pyramidal device can downloaded in the file below.</p>
-
Because it's a thermoplastic and softens under extremely high temperatures, acrylic can be formed to virtually any shape. Incredibly durable, acrylic is a suitable solution over a broad temperature range, and has superior weathering properties compared to other plastics.</p>
+
<ul class="download">
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[[File:Acrylic.jpg|300px|thumb|center|fig.1-4-3 Acrylic Sheet]]
+
[https://drive.google.com/file/d/0ByIV-UvWEH5cLXJrbnFtc014bHM/view Device_Design_Download]
 +
</ul>
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<br><br><br>
+
====Process to Assemble Our Device & Main Idea of Our Device Design====
-
<p>
+
<br>
-
　For the process, we searched for some information of our factories, and handed out the design chart. We also went everywhere to buy what we need for our device.</p>
+
'''Outer Shell：'''
-
<p>
+
<table width="100%;">
-
　First we use PowerPoint to draw the draft. Because PowerPoint can offer us a drawing tools, we chose it to draw our draft. It did not need a lot of technique. We only have to pay attention to present the brain idea perfectly. In the figure below you can see some of the draft.</p>
+
<td>[[File:Outer_Shell.JPG|350px|thumb|center|Fig.5-5-1 Outer shell real product.]]</td>
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[[File:NCTU_Formosa_2014_Project_Device3.jpg|400px|thumb|left|fig1-4-5 In this picture you can see the inner design.]]
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<td>[[File:Outer_Shell_1.JPG|270px|thumb|center|Fig.5-5-2 Outer shell schematic diagram.]]</td>
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[[File:NCTU_Formosa_2014_Project_Device4.jpg|400px|thumb|right|fig.1-4-6 In this picture you can see the overall appearance of roughly.]]
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</table>
-
<br><br><br><br><br>
+
-
<p>
+
-
　Also we use PowerPoint to double check the design. Because if the software has the problems, the manufacturers can open this file to see the right industry with map. It is very important to prevent this thing from happening. In the figure below you can see some of the draft.</p>
+
-
[[File:NCTU_Formosa_2014_Project_Device5.jpg|400px|thumb|left|fig1-4-7 In this picture, we can see the bottom front view and overlooking view.]]
+
-
[[File:NCTU_Formosa_2014_Project_Device6.jpg|400px|thumb|right|fig.1-4-8 In this picture, we can see the outer shell.]]
+
-
<br><br><br><br><br>
+
-
<p>
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-
　Second we use one kind of graphics software ‘‘CorelDRAW’’ to draw the formal design picture. In this software, we can learn how to draw industry with map to cooperate with manufacturers. They usually use it to make design. So we have to learn how to use it software. In the figure below you can see some of the draft.</p>
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[[File:NCTU_Formosa_2014_Project_Device7.jpg|400px|thumb|left|fig1-4-9 In this picture, we can see our bottom and latch.]]
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[[File:NCTU_Formosa_2014_Project_Device8.jpg|400px|thumb|right|fig.1-4-10 In this picture, we can see the outer shell.]]
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<br><br><br><br><br>
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<p>
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-
　Now we get the acrylic models finished. We could begin to do our best to assemble and install the inner shell, bottom and outer shell. Although the latch was very expensive, we did it ourselves. The blue light bulb were also assembled by ourselves. Accomplishing our device is very interesting and exciting. Be fun in welding.</p>
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[[File:NCTU_Formosa_2014_Project_Device9.jpg|350px|thumb|left|fig1-4-11]]
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[[File:NCTU_Formosa_2014_Project_Device10.jpg|350px|thumb|right|fig.1-4-12]]
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[[File:NCTU_Formosa_2014_Project_Device11.jpg|350px|thumb|center|fig.1-4-13]]
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====Process to assemble our device & main idea of our device design====
 
-
'''Outer Shell：'''
 
<p>
<p>
　We shaped the device into a pyramid. Its special layout also enriches the device with mysterious colors. </p>
　We shaped the device into a pyramid. Its special layout also enriches the device with mysterious colors. </p>
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<p>
<p>
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1. Outer shell is composed of 4 triangular acrylic planes which has a trapezoid entrance to let bugs in and release the smell of pheromone produced by the insects which eat our PBAN. </p>
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1. The outer shell is composed of 4 triangular acrylic planes which has a trapezoid entrance. When insects come inside and ingest PBAN solution, they will release pheromone. With the pheromone scent, our device can trap and collect more and more insects of the same species. </p>
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[[File:NCTU_Formosa_2014_ProductD1.jpg|350px|thumb|center|fig.1-4-14]]
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<br>
<br>
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<p>
 
-
2. There is a hook installed on the outer plane which attaches to the collecting bag inside. When we pull up the outer shell, the bag inside would also be sealed simultaneously so the bugs won’t escape.</p>
 
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[[File:NCTU_Formosa_2014_Project_Device12.jpg|350px|thumb|center|fig.1-4-15]]
 
-
<br><br><br><br><br>
 
'''Inner Shell：'''
'''Inner Shell：'''
 +
<table width="100%;">
 +
<td>[[File:Inner_Shell.JPG|350px|thumb|center|Fig.5-5-3 Inner shell real product.]]</td>
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<td>[[File:Inner_Shell_1.JPG|270px|thumb|center|Fig.5-5-4 Inner shell schematic diagram.]]</td>
 +
</table>
 +
 +
<p>
<p>
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　Similar to the outer one, inner shell are also composed of 4 trapezoid planes and removable from the base. The only different part is that its top is not sealed in order to contain the collecting bag.</p>
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　Similar to the outer one, the inner shell is also composed of 4 trapezoid planes and is removable from the base. The only difference  is that its top is not sealed so to allow entry of the captured insects.</p>
 +
 
<p>
<p>
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1. The inner space can contain a purse-string bag to collect insects we want.</p>
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1. There would be a container of '''PBAN''' solution placed at the bottom. </p>
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[[File:NCTU_Formosa_2014_ProductD3.jpg|350px|thumb|center|fig.1-4-16]]
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<br><br><br><br><br>
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<p>
<p>
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2. 4 rods are assembled like a pound sign on the top to sustain the bag. </P>
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2. '''Blue LED light bulbs''' will be installed around the top of the inner shell plane to attract the first female insect. </p>
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<p>
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<br>
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3. There would be a PBAN solution placed on the bottom. </p>
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'''Latch：'''
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<p>
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<table width="100%;">
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4. A blue LED light bulb will be installed around the top of the inner shell plane to attract the first female insect. </p>
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<td>[[File:Tenon.JPG|350px|thumb|center|Fig.5-5-5 Latch real product.]]</td>
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[[File:NCTU_Formosa_2014_ProductD4.jpg|350px|thumb|center|fig.1-4-17]]
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<td>[[File:Tenon_1.JPG|270px|thumb|center|Fig.5-5-6 Latch schematic diagram.]]</td>
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<br><br><br><br><br>
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</table>
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'''Tenon：'''
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 +
 
<p>
<p>
A part to stabilize the pyramid.</p>
A part to stabilize the pyramid.</p>
Line 358: Line 337:
<p>
<p>
2. To make sure the outer shell can combine with the inner one tightly.</p>
2. To make sure the outer shell can combine with the inner one tightly.</p>
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[[File:NCTU_Formosa_2014_ProductD5.jpg|350px|thumb|center|fig.1-4-18]]
 
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<br><br><br><br><br>
 
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====Assembling Process====
 
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<p>
 
-
We can assemble the outer shell、inner shell and tenon together to get our completed pyramid device.</p>
 
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[[File:NCTU_Formosa_2014_Project_Device13.jpg|500px|thumb|center|fig.1-4-19 Device Assembling]]
 
<br><br><br><br><br>
<br><br><br><br><br>
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====Advantages====
====Advantages====
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<p>
 
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<br>
 
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1.We successfully apply the geotaxis of targeted female moth to trap them in our device, forcing them releasing sex pheromone by our PBAN to attract more same-kind insects.<br>
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<p>1. We successfully trap the targeted female moth in our device, forcing them to release sex  
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2.The hook attached to the purse-string bag can seal it simultaneously when we want to remove the outer shell. By doing so, no insects would flee from the bag and safety problems can also be solved.<br>
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      pheromone by ingesting PBAN, which results in attracting more insects of the same kind.<br><br></p>
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3.Inner shell is removable so it’s easier to add new PBAN solution.<br>
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<p></p>
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4.Compared to conventional light bulb, LED bulb is much brighter and conserves more energy. It could powered by battery so it’s also easier to establish.<br>
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5.Purse-string bag is cheap and easy to switch. <br>
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<p>2. The inner shell is removable so it’s easier to replenish new PBAN solution and the food for the insect.<br><br></p>
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6.The PBAN system can run day and night. Its function won’t be affected by sunlight.<br>
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<p></p>
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7.Pyramid is good at looking and can enriches the entire device with a technological feeling.<br>
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</p>
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<p>3. Compared to conventional light bulbs, LED is much''' brighter''' and '''conserves more energy'''. It could powered by battery so it’s also easier in practical use.<br><br></p>
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<p></p>
 +
 
 +
<p>4. The PBAN system can run '''day and night'''. Its function won’t be affected by sunlight.<br><br></p>
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<p></p>
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<p>5. Pyramid is a good-looking form and can enrich the entire device with a technological feeling.<br><br></p>
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Latest revision as of 03:59, 18 October 2014

Project

Change the font size right here



Contents

Motivation

A Serious Problem

From ancient times to the present day, harmful insects always greatly affect our daily life, and we don’t have any appropriate solution to these troubling insects. Every year, harmful insects bring significant economic loss to agriculture, which seriously disturb people's livelihood and economy.

Take Brazil as an example, where the agricultural loss can reach up to 11 billion US dollars because most economic crops here such as coconut, coffee, and sugarcane are under serious insects attack. To control these obnoxious insects, 1.4 billion has to be invested to purchase insecticides annually. Terrible as it may seem, Brazil is just one of the many cases in the world! The truth is that farmers worldwide are doing the same thing! To ensure good harvest, they use pesticides without much consideration about its side effects. In other words, a great amount of insecticides containing toxic substances is discharged to the environment every day. Obviously, it’s never a good solution.

Fig.1-1-1 Losses caused by pests in Brazil. This chart demonstrates the losses of various crops caused by pests.


Also, there is a data about the IPM (Integrated pest management) in USA. Integrated pest management, which is defined as integration of pest control based on predicted economic, ecological and sociological consequences, makes maximum use of naturally occurring control agents, including weather, disease organisms, predators and parasites.In brief,IPM means the ability to control harmful insects.The lower the number is, the more efficient we are able to control the pests in the evaluated area. In the data, we can see that crops are planted in lots of areas in USA.There are several kinds of plants such as corn and soybean which suffer from insect damage. Since we can only control the insect damage efficiently within a small, limited area. Therefore, as the production areas expand to an extent, the effect of control declines exponentially. Even though the production areas become larger, the total production might be lower.

Therefore, what we need is a brand new method to solve this problem.

Fig.1-1-2 This chart demonstrates IPMs of the various areas of different crops planted in the USA.

Common Solutions

Chemical Control Method

Because of insects damage, human tried lots of ways to kill these harmful insects. In the 15th century, people used pesticide containing heavy metal such as Arsenic, Mercury and Plumbum to kill harmful insects, which was a catastrophe to the environment. Pesticides became more powerful along with the development of modern technology. In the 20th century, the agriculture prospered rapidly owing to the evolution of pesticides. But the pesticides are not only fatal to the insects but also harmful to the human body. People found this problem after several decades. The toxin of the pesticides will remain in creatures through the food chain and enter the human body eventually.


Fig.1-2-1 Human health and environmental cost from pesticides in the United States is estimated at $9.6 billion.

Damage to Human Body

Pesticides includes herbicides, insecticides, fungicides and rodenticides, which will kill weeds, insects, fungus, rodents and others. In our project, we refer to insecticides as pesticides. The use of toxic pesticides to manage insect problems has become a common practice around the world. However, pesticides cause serious human health hazards, ranging from short-term impacts such as headaches and nausea to chronic impacts like cancer and reproductive harm. There are also many researches revealing that exposure to pesticides will disrupt the endocrine system, the reproductive system and embryonic development.

Damage to the Environment

Pesticides pollute the environment such as threatening biodiversity and weakening the natural systems. Pesticides have led to abnormally high mortality of America's honeybees. The population of bees has dropped by 29 % to 36 % each year since 2006. To our amazement, approximately 1/3 of the food we eat depends on bees for pollination!

Some scientists believe that amphibians and bats have become more susceptible to deadly diseases because their immune systems are weakened by pesticides. Pesticides also contaminate waterways and endangered fish and birds, causing ecological damage.

Biological Control Method

Since most chemical control methods are poisonous to the environment, humans began to look for alternative solutions. One of the best known example is bacillus thuringiensis (Bt), which can efficiently kill insects with its crystal proteins. However, recent researches have shown that more and more insects have been resistant to this kind of protein.

Chemical control methods have caused fatal environmental damage and insects have grown resistant to both the chemicals substances and Bt. It is, however, not too late to improve this situation. We can create an agricultural revolution with our new method!

Fig.1-2-2 Comparison of both solutions.

Pheromone trap is currently the most novel method to resolve insect damage problems. What's more, pheromone is a pollution-free substance that is produced by the insects, meaning that insects cannot resist it. That's why we take biological synthesis of pheromone in E.coli to solve the problem.

The Pheromone Trap

Introduction of Pheromones

A pheromone is a secreted or excreted chemical factor that triggers a social response among members of the same species. Pheromones are capable of acting outside the body of the secreting individual to impact the behavior of the receiving individuals.

There are many kinds of pheromones such as alarm pheromones, food trail pheromones, sex pheromones, and many others that affect behavior or physiology.

Sex pheromones are an important factor in finding a mating partner. When a female releases chemicals, the mating search is initiated, and the male moths begin their upwind motion toward their potential partner. Pheromones have the ability to propel long-distance attraction and are emitted by the abdominal glands of female insects in most cases.



How to Produce Pheromones

There are 2 main ways to produce pheromones. One through chemical synthesis, and the other through biosynthesis approach.

Chemical Synthesis Approach Used in Factory for Massive Production

It is difficult to make pheromone with a chemical synthesis approach. This approach requires many kinds of chemical compound which may cause pollution to the environment. Furthermore, the control of reacting condition may consume a lot of energy. An example of chemical synthesis approach: Leafroller moth, Bonagota cranaodes.

Fig.1-3-1 If we want to produce the pheromone of Leafroller moth (Bonagota cranaodes) by chemical synthesis approach, we have to purchase expensive equipment to control the conditions. And it may produce toxic byproducts.(34)(Paulo H. G. Zarbin et al. 2007)

Simulating the Biosynthetic Pathway in Insects with E.coli

Biosynthesis approach requires the cooperation of different enzymes, which is too complicated for E.coli to carry out. In addition, E.coli lacks Glycoproteins for posttranslational modification. Therefore, it is nearly impossible for E.coli to produce pheromones.

Fig.1-3-2 If we want to produce the pheromone of Gypsy moth, Lymantria dispar. We have to pass at least four complicated pathways. That will be highly stressful for E.coli, and it may also cause low efficiency of producing pheromone.(35)(Russell Jurenka, 2004)

Difficulty and Disadvantage of Pheromone Production

As mentioned above, pheromone is too complicated to be produced either artificially or bio-synthetically. Although the cost of artificial chemical synthesis can be lowered through massive production, the environmental damage it causes is fatal. With that said, we decided to find an easier and better way to reach our goal.

PBAN- The Solution

To solve the impossibility of biologically synthesizing pheromone with E.coli, we found a feasible solution. As mentioned above, our method should be pollution-free, efficient, insect resistance-free, and cheap. Combining all these advantageous elements, we introduce you PBAN. Click the photos to learn more!


  • Cabbage Moth
    Mamestra brassicae
  • Cotton bollworm
    Helicoverpa armigera （Hubner）
  • Oriental Leafworm Moth
    Spodoptera litura
  • Red imported fire ant
    Solenopsis invicta
  • Yellow fever mosquito
    Aedes (Stegomyia) aegypyi
  • Black cutworm
    Agrotis ipsilon
  • Gypsy Moth
    Lymantria dispar
  • Leafrollers
    Statherotis leucaspis Meyrick
  • Silkworm
    Bombyx mori
Mamestra brassicae

Spread:This moths has a natural range across Europe, Asia, and North Africa.

Characteristics:The larva is green, khaki, grey-brown or brown with dark spots(1)(2)(3). The topside is darker than the bottom side and a yellow or light brown stripe goes round the middle portion by the spots.

Damage:The caterpillar of this species is seen as a pest for commercial agriculture. Often referred to as the "imported cabbageworm" they are a serious pest to cabbage and other mustard family crops. It can also be a pest of cultivated brassicas and sweet peas, but it feeds on a wide range of other plants.

Control: Organic controls(4) for cabbage worms include handpicking, excluding them with row cover barriers, or treating with a Bt pesticide. Cabbage worms are found throughout North America, and more than one species may be found in the same garden.


Helicoverpa armigera （Hubner）

Spread:The pink bollworm has spread to cotton-growing regions throughout the world.

Characteristics: The larva is green, khaki, grey-brown or brown with dark spots(5). The topside is darker than the bottom side and a yellow or light brown stripe goes round the middle portion by the spots(32).

Damage: The cotton bollworm is a highly polyphagous species.[6] The most important crop hosts are tomato, cotton, pigeon pea, chickpea, sorghum and cowpea. Other hosts include groundnut, okra, peas, field beans, soybeans, lucerne, Phaseolus spp., other Leguminosae, tobacco, potatoes, maize, flax, Dianthus, Rosa, Pelargonium, Chrysanthemum, Lavandula angustifolia, a number of fruit trees, forest trees and a range of vegetable crops(6).

Control: Cultural controls, with the exception of the use of Bt cotton and the use of mating disruption and sprays of the Entrust formulation of spinosad are acceptable to use on organically grown cotton(7).


Spodoptera litura

Spread:Widely distributed in Asia and Oceania.

Asia: Afghanistan, Bangladesh, Cambodia, China, Hong Kong, Indonesia, India, Iran, Japan, Laos, Malaysia, Myanmar, Nepal, North Korea, Oman, Pakistan, Philippines, Singapore, South Korea, Sri Lanka, Taiwan, Thailand, Vietnam. Oceania: Australia, Guam, New Caledonia, New Zealand, Micronesia, Papua New Guinea,

Samoa, other Pacific islands. United States: Hawaii(8).

Characteristics: Adult moths measure between 15-20 mm (0.59-0.79 inches) in length and have a wingspan of 30-38 mm (1.18-1.5 inches). Forewings are gray to reddish-brown, with a complex pattern of creamy streaks and paler lines along the veins. Hind wings are grayish-white with grayish-brown margins. Males have a blue-grey band from the upper corner (apex) to the inner margin of each forewing. Larvae have bright yellow stripes along the back and the sides. Larval color varies from pale green to dark green(9)(31).

Damage: Oriental Leafworm Moth Spodoptera litura is a Noctuid moth which is considered as an agricultural pest. It is also known as the Cluster caterpillar, Cotton leafworm, Tobacco cutworm, and Tropical armyworm. It has a very wide host range of over 120 plant species, including: lettuce, cabbage, beetroot, peanuts, geranium, cotton, banana, fuchsias, acacia, African oil palm, amaranth, alfalfa, strawberry, sorghum, sugarcane, tomatoes, asparagus, apple, eggplant, beet, beans, broccoli, elephants ear, horsetail she oak, corn, flax, lantana, papaya, orange, mango, leek, among many others.

Control: The use of Bacillus thuringiensis (BT) may effectively control this pest. Other forms of biological, horticultural, and cultural control that have been studied include: planting near derris and garlic plants, breeding resistant plants from wild plants for example groundnuts from wild groundnuts, breeding resistant plants using bacterium Bacillus thuringiensis genes, using a Baculovirus, using the nematode Steinernema carpocapsae, and using the fly Exorista japonica(10).

Solenopsis invicta

Spread:The red imported fire ant, a eusocial species, are far more aggressive than most ant species. Animals, including humans, often encounter them by inadvertently stepping on one of their mounds, which causes the ants to swarm up the legs, attacking en masse. The ants respond to pheromones released by the first ant that attacks, thereafter stinging in concert(12).

Characteristics: Fire ants are red and black in coloration and, like all insects, they are protected by a hard exoskeleton and have six legs. Worker ants have round heads with mandibles, an armored thorax midsection and an abdomen, made up of the pedicle and the gaster. The head is typically copper brown in color. In addition to their mandibles, fire ant workers also possess an abdominal stinger(11)(29).

Damage: They are considered to be a pest, not only because of the physical pain they can inflict, but also because their mound-building activity can damage plant roots, lead to loss of crops(12).

Control: Hot water Pouring hot water on the mounds is effective and environmentally friendly, but may require 3 or 4 applications to kill the colony. Water should be at least scalding hot, but does not need to be boiling. This works best when you use 3 to 4 gallons of water in each application. WARNING: Hot water kills grass and shrubbery and may cause severe burns if spilled. Liquid nitrogen: it could be the most effective and most environmental friendly method to eradicate the species(30)(13).

Aedes (Stegomyia) aegypti

Spread:The yellow fever mosquito, Aedes aegypti, is a mosquito that can spread the dengue fever, chikungunya, and yellow fever viruses, and other diseases. The mosquito can be recognized by white markings on its legs and a marking in the form of a lyre on the thorax. The mosquito originated in Africa, but is now found in tropical and subtropical regions throughout the world(14).

Characteristics: The mosquito can be recognized by white markings on its legs and a marking in the form of a lyre on the thorax.

Damage: The yellow fever mosquito, Aedes aegypti, is a mosquito that can spread the dengue fever, chikungunya, and yellow fever viruses, and other diseases(15).

Control: Empty water from containers such as flower pots, birdbaths, pet water dishes, cans, gutters, tires and buckets regularly to disrupt the mosquito breeding cycle. Consider using an insect repellent, be sure to follow the label directions for applying the repellent. For help selecting a mosquito repellent, try our Insect Repellent Locator(16).

Agrotis ipsilon

Spread: This Caterpillar can be found, as various species, through be serious foliage feeders on some crops such as peanuts.hout North America.

Characteristics: Cutworms common in Georgia fields are black (Agrotis ipsilon (Ashmed)), granulate (Agrotis subterranea (Fabricius)) and variegated cutworm (Peridroma saucia(Hubner)). These are moths in the family Noctuidae. Full-grown cutworm larvae are 1.5 to 2 inches long. Coloration will vary among species, but all tend to be stout-bodied caterpillars with four sets of prolegs. They have the tendency to curl into a ball when disturbed(17).

Damage: Almost any plant can be attacked in the seedling stage. Cotton and certain vegetables sometimes have stand reductions(17).

Control: Bacillus thuringiensis, a widely available caterpillar-killing bacterium,is a very effective control for climbing cutworms as well as for the surface feeders(18).

Lymantria dispar


Spread: It has a range which covers Europe, Africa, and North America(19).

Characteristics:Gypsy moth caterpillars change appearance as they grow. Young caterpillars are black or brown and about ¼ inch (0.6 cm) in length. As they grow, bumps develop along their backs along with coarse, black hairs. Each of the 11 sections of a developed caterpillar will have two coloured spots, the first five pairs, blue, and the last six, red. Mature caterpillars can be as long as 2 ½ inches (6.35 cm)(20).

Damage: It is classified as a pest, and its larvae consume the leaves of over 500 species of trees, shrubs and plants. The gypsy moth is one of the most destructive pests of hardwood trees in the eastern United States.he gypsy moth was considered a nuisance just ten years after their release. It included an account of all the trees being defoliated, caterpillars covering houses and sidewalks and that the caterpillars would rain down upon residents. The first outbreak occurred in 1889. An eradication program was begun in 1890.

Control: Tanglefoot Pest Barrier or Sticky Tree Bands can be placed around tree trunks to help curtail the caterpillars movement into and out of the tree canopy. Apply Bacillus thuringiensis, var. kurstaki or Monterey Garden Insect Spray (Spinosad) to the leaves of trees to kill gypsy moth caterpillars(21).

Statherotis leucaspis Meyrick

Spread: The obliquebanded leafroller (OBLR) is native to and widely distributed throughout temperate North America(22).

Characteristics: hatched larvae have a yellowish green body and a black head and thoracic shield. Mature larvae are 20 to 25 mm in length and the head and thoracic shield may be either black or various shades of brown(23).

Damage: Leafrollers, the larvae of certain tortricid moths, often feed and pupate within the protection of rolled-up leaves. Several species can cause problems on fruit and ornamental trees in California. The fruittree leafroller, Archips argyrospila, is the most common leafroller pest in landscapes throughout the state. It occurs on many ornamental trees—including ash, birch, California buckeye, box elder, elm, locust, maple, poplar, rose, and willow—and is particularly damaging to deciduous and live oaks. It also attacks numerous fruit and nut trees including almond, apple, apricot, caneberries, cherry, citrus, pear, plum, prune, quince, and walnut(24).

Control: Several parasites attack OBLR larvae but do not adequately control the pest. Apply sprays during June to kill the first summer brood adults and newly hatching larvae(24).

Bombyx mori

Spread: the place where farmers want to feed.

Characteristics : It is entirely dependent on humans for its reproduction and does not occur naturally in the wild(26).

Damage: None

Control: Not necessary


Reference
  1. Cabbage Moth - Caterpillar". Habitas.org.uk. Retrieved 2014-08-11.
  2. Interesting (To Us) Photos From The Garden". Meades.org. Retrieved 2011-08-11.
  3. RXwildlife Sightings » Blog Archive » More Emergence". Rxwildlife.org.uk. 2009-06-03. Retrieved 2011-08-11.
  4. Practice Organic Cabbage Worm Control for a Chemical-Free Garden, by Barbara Pleasant, March 25, 2013
  5. 2003-2009 Project «Interactive Agricultural Ecological Atlas of Russia and Neighboring Countries. Economic Plants and their Diseases, Pests and Weeds
  6. Report of a Pest Risk Analysis Helicoverpa armigera, Hübner, 1808
  7. UC IPM Pest Management Guidelines: Cotton, http://ucanr.edu/sites/ipm/pdf/pmg/pmgcotton.pdf
  8. Michigan State University’s invasive species factsheets(Oriental leafworm Spodoptera litura) , http://www.ipm.msu.edu/uploads/files/Forecasting_invasion_risks/orientalLeafworm.pdf
  9. Effect of tetra hydroxyl-ρ-benzoquinone on growth and metamorphosis of Spodoptera litura Fabr. (Lepidoptera: Noctuidae) larvae, Sujata Magdum , Seema Gupta
  10. Espinosa, A. and A.C. Hodges University of Florida, Spodoptera litura, BUGWOODWiki, http://wiki.bugwood.org/Spodoptera_litura.
  11. OROKIN, Fire Ant Anatomy, http://www.orkin.com/ants/fire-ant/fire-ant-anatomy
  12. Wikipedia, Red imported fire ant http://en.wikipedia.org/wiki/Red_imported_fire_ant
  13. Organic Gardening.com/learn-and-grow/fire-ant-control
  14. Cram101 Textbook Reviews, Life: The Science of Biology 8th Edition
  15. Wikipedia, Aedes aegypti , http://en.wikipedia.org/wiki/Aedes_aegypti
  16. National Pesticide Information Center, Mosquito Control Methods
  17. BugwoodWiki , Cutworms, by Dr. Steve L. Brown, Dr. Will Hudson
  18. Eliminate Cutworms Using Natural Pest Control By Susan Glaese March/April 1987
  19. Wikipedia, Lymantria dispar dispar , http://en.wikipedia.org/wiki/Lymantria_dispar_dispar
  20. FACT SHEET : GYPSY MOTH, http://www.metro-forestry.com/wp-content/uploads/2010/02/FactSheet_Gypsy-Moth.pdf
  21. Planet Natural, GYPSY MOTH
  22. Larry P. Pedigo, G. David Buntin, Handbook of Sampling Methods for Arthropods in Agriculture
  23. L. A. Hull, D. G. Pfeiffer & D. J. Biddinger, Apple Direct Pests
  24. IPM, obliquebanded leafroller, http://www.nysipm.cornell.edu/factsheets/treefruit/pests/oblr/oblr.asp
  25. UC IPM Online, Leafrollers on Ornamental and Fruit Trees, Publication 7473 September 2010
  26. Tan Koon San, Dynastic China: An Elementary History
  27. Pogue, Michael. "A review of selected species of Lymantria Huber [1819]". Forest Health Technology Enterprise Team. Retrieved September 14, 2012.
  28. Laurence Mousson, Catherine Dauga, Thomas Garrigues, Francis Schaffner, Marie Vazeille & Anna-Bella Failloux (August 2005). "Phylogeography of Aedes (Stegomyia) aegypti (L.) and Aedes (Stegomyia) albopictus (Skuse) (Diptera: Culicidae) based on mitochondrial DNA variations". Genetics Research
  29. M. S. Ascunce et al., “Global Invasion History of the Fire Ant Solenopsis invicta”, Science, vol. 331, no. 6020, pp. 1066 - 1068, 2011
  30. Scientists suggest fighting fire ants with ice By Chiu Yu-Tzu / STAFF REPORTER
  31. Capinera, J.L. 1999. Fall armyworm Spodoptera frugiperda (J.E. Smith) (Insecta: Lepidoptera: Noctuidae)
  32. Helicoverpa Diapause Induction and Emergence Tool Introduction to the Helicoverpa armigera Genome Project Helicoverpa armigera Genome Project updates on InsectaCentral Helicoverpa Genome Project database on-line Lepiforum Funet Taxonomy Fauna Europaea
  33. Authored by W. H. Reissig. Published by the New York State Agricultural Experiment Station, Geneva, A Division of the New York State College of Agriculture and Life Sciences, A Statutory College of the State University, Cornell University, Ithaca. Funded in part by an Extension Service-USDA, IPM Grant.
  34. Paulo H. G. Zarbin; José A. F. P. Villar; Arlene G. Corrêa, Insect pheromone synthesis in Brazil: an overview, Journal of the Brazilian Chemical Society, vol.18 no.6 São Paulo 2007


PBAN ( Pheromone Biosynthesis Activating Neuropeptide )

PBAN (Pheromone Biosynthesis Activating Neuropeptide) is a kind of peptide that can activate biosynthesis of pheromones of specific insects. Once a PBAN binds with the G-protein coupled receptor located at an insect’s pheromone gland, it will send a signal to activate kinase and phosphatase, which in turn activates other enzymes that participate in the biosynthesis of insect sex pheromones. These pheromones are eventually emitted(35).

In nature, female insects such as moths release PBAN to stimulate the synthesis of pheromones in order to attract male moths during mating. PBAN can also facilitate the release of non-sex pheromones such as trail pheromones for ants. Overall, this kind of natural substance, PBAN, has many advantages in the following, which can complete our goals perfectly.

Fig.2-1-1 This is the biosynthetic pathway of pheromones. Once PBAN comes in contact with the PBAN receptor, the receptor will send a signal to activate other enzymes that participate in the biosynthesis of insect pheromones.

Features of PBAN

1. Species-specific: PBAN is species-specific just like pheromones, meaning that every kind of insect produces specific PBAN that only binds with its specific receptor, resulting in the production of a particular pheromone.

2. Small and simple: The coding sequence for a PBAN is only around 100 base pairs. For E.coli, 100 base pairs is totally within its working capacity. Therefore, E.coli can be a low-cost PBAN factory. By transforming the DNA sequences for different PBAN into the E.coli, we can even gain a variety of PBANs. 　

3. Secreted directly: Because PBAN can be synthesized by the insect itself, the insect would not form a resistance to it compare to use pesticide.

In conclusion, using PBAN is totally a environmental friendly way for solving harmful insects problems with easily triggering the production of pheromone by contacting with PBAN receptors.

Main Idea

We plan to produce PBANs through E.coli and make the PBAN come in contact with the target insect. The target insect will then start producing pheromones and attract more target insects.

Pheromone Production

Before we could employ PBAN to accomplish our goals, we need to obtain enough background knowledge to evaluate the workability of our plan through paper research. First, we had to confirm whether an insect can take in vitro PBAN and allow it to function. From “Pheromonotropic Activity of Naturally Occurring Pyrokinin Insect Neuropeptides (FXPR-Lamide) in Helicoverpa zea”(37), this method has been proven possible. In this experiment, 0.05 pmol to 200 pmol of PBAN had been injected in to corn earworm and the amount of pheromone produced had been measured. The result (Fig.1-2-4) showed that PBAN produced by insects itself can effectively increase the production of pheromone. The maximized amount of pheromone production was reached with only "10 pmol" of PBAN. This experiment means as long as PBAN enters the body of the insect, it highly possible that PBAN can stimulate the production of pheromone.

Fig.3-1-1 Effect of dose of Hez-PBAN on stimulation in vivo pheromonotropic activity in vitrugin females of H. zea (bars indicate SEM. n=8). This figure shows that PBAN can efficiently stimulate pheromone production in just a small amount (10 pmol).(Rellal.Abernathy et al, 1995)

A Problem of Degradation by Peptidase

The only problem for us to use PBAN is that PBAN would be degraded quickly in an insect’s body. In order to understand this problem, we searched some former papers and we found another paper from Peptides, “Enhanced oral availability/pheromonotropic activity of peptidase-resistant topical amphiphilic analogs of pyrokinin/PBAN insect neuropeptides(36) .” In this experiment, Heliothis virescens was the experimental insect and amphiphilic analogs of pyrokinin/PBAN were used instead of natural PBAN. This way, the analog can resist peptidase, as shown in the graph below. The amount of the natural PBAN decreases quickly while the analogs degrades slowly (Fig.3-2-1). However, we found the amphiphilic analogs of pyrokinin/PBAN peptide requires many special artificial modification, which means E.coli absolutely cannot produce this artificial PBAN that can easily resist peptidase. Thus, we need more brain storming to think of a way to use PBAN even if it may be degraded by peptidase.

Fig.3-2-1 Stability of the natural pyrokinin/PBAN analog LPK, and the peptidase-resistant pyrokinin/PBAN analogs Hex-Phe-Thr-Hyp-Trp-Gly-NH2 and Hex-Phe-Thr-Oic-Trp-Gly-NH2 to hydrolysis by peptidases bound to corn earworm (H. zea) Malpighian tubule tissue. Measurement of the amount of remaining peptide was made by HPLC at 30, 60, and 120 min. The data points represent the means of at least three replicates.(Ronald J. Nachmana, Peter E.A. Teal, Allison Strey, 2002)

Our Solution

The results above did not sound good to us, but there was another experiment that interested us in the same research. Researchers also conducted oral test with artificial PBAN/pyrokinin analogs (Fig.3-3-1), in this experiment, they fed moths with sugar solution which contained their PBAN analogs, and measured the amount of pheromone produced over time. (Blank: natural PBAN in low concentration) This result inspired us. Although natural PBANs can’t be maintained for a long time in moth’s body, we could solve this problem by simply feeding moths with high concentration PBANs continually.

Fig.3-3-1 Amount of pheromone, relative to the maximum pheromone amount induced by injected PBAN, produced by Hyp-analog and Oic-analog, 1.5, 3, 4, and 6h following oral administration. Dotted line at 100% denotes maximal pheromone production of injected PBAN (positive control). This figure shows a very important information that blank (natural PBAN in low concentration：50 pmol, the blank was measured when it was 2 hr after feeding with nature PBAN) still has some ability to stimulate pheromone production. Thus, we decided to try to feed the female target insects with high concentration PBAN continuously.(Ronald J. Nachmana, Peter E.A. Teal, Allison Strey, 2002)

Thus, as long as female moths suck the high concentration PBAN solution continuously, the PBAN in the moths’ body will be replenished continually even if peptidase degradation occurs. Then, there is a high possibility of PBAN being absorbed by the moths’ body from the digestive system and succeed in stimulating the pheromone gland. Thus, if we feed the female insects with high concentration PBAN solution continuously, the female insects will produce pheromones for us, which theoretically solves the problem of PBAN's inability to maintain for a long time in the insects’ body. As to how we produce PBAN and apply our concept to capture wanted harmful insects, we will explain in the following.

How We Are Going to Use PBAN?

Fig.3-4-1 Overview of our project.

In our project, we will biologically synthesize PBAN with the E.coli. We store the PBAN inside a trapping device (check this out at our Device page). In the device, there will be appropriate lighting and nutrient sources that will attract insects.

Once an insect is attracted into our device and ingests the nutrient sources we provide, it will also inevitably come in contact with our PBAN. As the PBAN works and activates the pheromone synthesis of the attracted insect, more of this species of insect’s counterparts will be attracted and later captured.

Owing to the first feature mentioned above, PBAN is species-specific, which means that it doesn't matter if other kinds of insects fly into our device and eat the PBAN. This is because the insects we don't want to catch will not be stimulated by PBAN to produce pheromone. Therefore, the PBAN is only for what we want to catch, and we are sure that our method won't affect other kinds of insects.


Reference
  1. Russell Jurenka, Insect Pheromone Biosynthesis, Topics in Current Chemistry (2004) 239: 97– 132 DOI 10.1007/b95450, 2004
  2. Ronald J. Nachmana, Peter E.A. Teal, Allison Strey, Enhanced oral availability/pheromonotropic activity of peptidase-resistant topical amphiphilic analogs of pyrokinin/PBAN insect neuropeptides, Peptides 23 (2002) 2035–2043.
  3. RELLA L. ABERNATHY, RONALD J. , PETER E. A. TEAL, OKITSUGU YAMASHITAS AND JAMES H. TUMLINSON, Pheromonotropic Activity of Naturally Occurring Pyrokinin Insect Neuropeptides (FXPRLamide) in Helicoverpa zea, Peptides, Vol. 16, No. 2, pp. 215-219, 1995.

Biobrick Design

Basic Biobrick Design

Fig.4-1-1 This is our basic design of biobrick.

We searched the DNA sequences of the PBANs of many kinds of insects on NCBI, then compared them to the amino acid sequences from papers so that we can select the DNA fragments that directly correspond to gland-stimulating function. By ligating the constitutive promoter (BBa_J23101), ribosome binding site (BBa_B0034) and PBAN DNA sequence with a terminator (BBa_J61048) at last (we delimit this sequence as basic part), we were able to make E.coli directly produce these PBANs continuously instead of the original complex process of PBAN biosynthesis in insects.

Multifunctionality

Fig.4-2-1 Best Potential of Our PBAN Biobrick - Multifunctional.

We can assemble these basic parts together easily because the number of base pairs of these basic parts are small. We can assemble different basic parts that contain different PBAN DNA sequences to resolve different insect problems. Therefore, our biobrick design can be customized according to the users' needs. For instance, if there is a farm harmed by 3 kinds of moths: Lymantria dispar, Spodoptera litura and Mamestra brassicae, what we have to do is to ligate the PBAN DNA sequence of 3 basic parts into one plasmid and let the E.coli express these PBANs. After these three kinds of moth ingest these three kinds of PBANs, subsequently, these 3 species of moths will produce their own pheromones to attract their same-species counterparts. To put it simply, our PBAN basic parts can be assembled together with any combination in infinite possibility.

For even more creative ideas, each of the PBAN basic parts can use different promoters, RBS and terminators to make many different regulation. Thus, not only can we produce many different PBANs with just one strain of E. coli, but these parts can be regulated to our desire. Thus, our PBAN biobricks really have infinite potential!

Device

In order to actually attract and capture the insect, we take the different weather conditions such as light and temperature into consideration. The different conditions will change the efficiency of insect attraction. Therefore, we design a device that can be used in different conditions, which would allow farmers to choose the best way for their local condition.

Fig.5-1-1 Our blue light pyramidal device.

Introduction

Since insects behave widely different to the gravity force, we design a device which could catch specific kinds of insect species that we want. For example, Agrotis ypsilon (Rottemberg) and Spodoptera litura fall into the kind of moth that have negative geotaxis (antigravity, tendency to fly upwards). Thus, we made a trap with accessible pathway at the bottom. Once an insect enters the device, it could only go up and be trapped inside the pyramid. However, after field investigation, we found some insects still escape from the device. Then we came up with a new version trap with doubled layers, inner shell and outer shell.



Assembling Process

Before the detailed description of how we design our device, we can show you a simple animation of the assembling process of our device. As you can see, our device is very easy to use. Just assemble the outer shell, inner shell (containing PBAN) and tenons together to complete the device. Then, you can put the device at where you want to attract the harmful insects.

　　　　






Mechanism of Attraction

　　　　　　　　　　　　

As you can see, we divide our design into two parts－exterior and interior. The exterior is just like the appearance of a pyramid, and the interior is designed to accommodate PBAN. At the beginning of using the device, we turn on the blue light LEDs to attract the target harmful insects. When the target female insects are stuck in the interior and eat the food mixed with PBAN solution, they will release pheromones, and attract same-species counterparts. Even if we turn off the blue light, our device can attract many target male insects because the female insects inside our device is still in rut and releasing sex pheromone to attract their mates. After the harmful insects go into our device, the design of our device will take advantage of their habit. Insects always fly high to escape so they will be stuck in the top of our device.




Device Design

More detail information about how we design our pyramidal device can downloaded in the file below.

Process to Assemble Our Device & Main Idea of Our Device Design


Outer Shell：

Fig.5-5-1 Outer shell real product.
Fig.5-5-2 Outer shell schematic diagram.


　We shaped the device into a pyramid. Its special layout also enriches the device with mysterious colors.

1. The outer shell is composed of 4 triangular acrylic planes which has a trapezoid entrance. When insects come inside and ingest PBAN solution, they will release pheromone. With the pheromone scent, our device can trap and collect more and more insects of the same species.


Inner Shell：

Fig.5-5-3 Inner shell real product.
Fig.5-5-4 Inner shell schematic diagram.


　Similar to the outer one, the inner shell is also composed of 4 trapezoid planes and is removable from the base. The only difference is that its top is not sealed so to allow entry of the captured insects.

1. There would be a container of PBAN solution placed at the bottom.

2. Blue LED light bulbs will be installed around the top of the inner shell plane to attract the first female insect.


Latch：

Fig.5-5-5 Latch real product.
Fig.5-5-6 Latch schematic diagram.


A part to stabilize the pyramid.

1. Stabilize 4 corners of the bottom.

2. To make sure the outer shell can combine with the inner one tightly.






Advantages

1. We successfully trap the targeted female moth in our device, forcing them to release sex pheromone by ingesting PBAN, which results in attracting more insects of the same kind.

2. The inner shell is removable so it’s easier to replenish new PBAN solution and the food for the insect.

3. Compared to conventional light bulbs, LED is much brighter and conserves more energy. It could powered by battery so it’s also easier in practical use.

4. The PBAN system can run day and night. Its function won’t be affected by sunlight.

5. Pyramid is a good-looking form and can enrich the entire device with a technological feeling.

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