Team:NCTU Formosa/project
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Revision as of 12:08, 25 September 2014
- Overview
- PBAN
- Brobrick
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Contents
Overview
Insect damage have been a serious problem for a long time over the whole world and it is difficult to be solved. People take several methods to kill these harmful insects, but the methods have cause other problems to the environment. In order to solve the troublesome problem and not to have the bad side effect, we established an alternative way to solve it in an eco-friendly manner.
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PBAN
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.
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Brobrick
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.
Overview
Impact of pest
In agriculture, insect problems have existed for a long time, and have been difficult to be solved. In addition, all agricultural products on earth every year suffer from harm and damage caused by these insects. Therefore, it goes without saying that insect problem can be regard as a global economic problem.
Losses caused by pests on crops include direct damage, drug costs and the experimental costs, The loss which is most difficult to estimate is the consumption of medical resources on human health hazards caused by pesticides.
We can see this credible chart, insect damage recorded in Brazil region, pests generally reduce crop yields by 10 percent, which is each crop have reduced the value of hundreds of millions. Our goal is committed to reducing agricultural losses, safeguarding human health, and maintaining ecological balance.
Pesticide hazards
Because of the hazard of insects, human beings to make a lot of ideas to kill these harmful insects. In 15th century,people used heavy metals to kill harmful insects, such as Arsenic, Mercury and Plumbum, which caused a catastrophe to the environment.
Pesticide becomes more powerful along with the technology, in 20th century,the agriculture develops rapidly just because of the evolution of pesticide. But the pesticide is not only harmful to the insects but also harmful to the human beings. People found this problem after several decades. The toxin of the pesticide will be kept in creatures by the food chain,and finally go 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 it is what we do!
Solution to both questions
People invent pesticides to eliminate the pests, but the use of pesticides will hurt environments and cause harm to humans. Thus there have been conflicting issues, and have been difficult to be solved. In order to solve the above-mentioned problems, we came up with a practical, inexpensive way.
Reference
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http://en.wikipedia.org/wiki/Lymantria_dispar_dispar
http://www.motherearthnews.com/homesteading-and-livestock/natural-pest-control-zmaz87mazgoe.aspx#axzz3EEpOIwz2
http://en.wikipedia.org/wiki/Aedes_aegypti
http://npic.orst.edu/pest/mosquito/control.html
http://en.wikipedia.org/wiki/Red_imported_fire_ant
http://www.organicgardening.com/learn-and-grow/fire-ant-control
http://www.taipeitimes.com/News/taiwan/archives/2004/10/31/2003209074
http://wiki.bugwood.org/Spodoptera_litura
http://en.wikipedia.org/wiki/Helicoverpa_armigera
http://www.ipm.ucdavis.edu/PMG/r114301511.html
http://en.wikipedia.org/wiki/Cabbage_Moth
http://nysipm.cornell.edu/factsheets/treefruit/pests/oblr/oblr.asp
http://en.wikipedia.org/wiki/Pesticide
PBAN(Pheromone Biosynthesis Activating Neuropeptide)
Introduction
PBAN(Pheromone Biosynthesis Activating Neuropeptide) is one kind of peptides that can activate biosynthesis of pheromones of many kinds of insects. 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.
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.
Features of PBAN
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.
2. The coding sequence for a PBAN is usually around 100 basepairs. Thus, it is easy for E.coli to express. We can even combine several different PBAN sequences into one BioBrick assembly. (Check this out at our BioBrick page)
How are we going to use PBAN?
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.
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.
Owing to the first feature mentioned above, PBANs are species-specific, which means that it doesn't matter if other kind of insect fly into our device and eat PBANs, because the insects we don't want to catch will not be stimulated by PBANs to produce pheromone; our PBANs are only for what we want to catch, we are sure that our method won't affect other kinds of insects.
Peptidase-resistant pyrokinin/PBAN
Some peptidase-resistant pyrokinin/PBAN with Pro,Oic or Hyp residues can enhance resistance to peptidase hydrolysis. From the figure down , we can see that peptidase-resistant pyrokinin/PBAN can maintain its original concentration , however , the natural PBAN will be degraded by peptidases bound to corn earworm (H. zea) Malpighian tubule tissue. So, if we want to maintain the concentration of the natural PBAN in insects’ body , we must offer them food with PBAN constantly.
Application of PBAN
Reference
- Miriam Altstein, Role of neuropeptides in sex pheromone production in moths,Peptides 25 (2004) 1491–1501.
- Ada Rafaeli, Pheromone biosynthesis activating neuropeptide (PBAN): Regulatory role and mode of action, General and Comparative Endocrinology 162 (2009) 69–78
- 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 +
- 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
- 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
- Man-Yeon Choi and Robert K. Vander Meer, Ant Trail Pheromone Biosynthesis Is Triggered by a Neuropeptide Hormone, PLoS Onev.7(11); 2012PMC3511524
Design
Biobrick Design
We searched the DNA sequences of the PBANs from many kinds of moths on NCBI, then contrasted to the amino sequences from papers so that we can selected the DNA fragments which directly correspond to gland-stimulating function, By ligating the Ribosome binding site(B0034) and PBAN DNA sequence, we were able to make E.coli directly produce these PBANs instead of the original complex process of PBAN biosynthesis in insects. We had gotten nine kinds of PBANs, each of which is from one kind of moth ,after we constructed the PBAN biobricks, the B0034+BFP+J61048 biobrick was ligated behind the PBAN biobrick in order to make modeling each PBAN biobrick respectively more easy in the future and observing the production of PBANs more convenient for us.
Reference
- 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
- part BBa_K115002;TUDelft Registry of Standard Biological Parts
Device
Introduction
Since insects react 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 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.
Mechanism
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 PBANs and bag for pests. When the harmful insects we want to catch eat our PBANs, 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.
To get more professional suggestions, we went to the National Chung Hsing university,and visited the professor Hau You Tzeng who major in mosquito. This tour benefited us very much. We got some knowledge about how to design breeding cage. With this experience, we finally designed our own breeding cage.
Design
For the materials of our device, We use Acrylic Sheet or balsa. The fomer is transparent and safer than glass. The latter is cheap, light, and easy to cut. For the process , We google some informations of our factories, and use Colddraw to hand out the design chart. We also go everywhere to buy what we need for our device. For example, the tool for cutting wood and LED light were just named a few.
Table below shows the exploded view of a complete device:
Introduction | Function | schematic | |
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Outer shell | We shaped the device into a pyramid. Its special layout also enriches the device with mysterious colors. |
1. Outer shell is composed of 4 triangular acrylic planes which has a trapezoid entrance to let bugs in and release the smell of PBAN. 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. |
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Inner shell | 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. |
1. The inner space can contain a purse-string bag to collect insects we want. 2. 4 rods are assembled like a pound sign on the top to sustain the bag. 3. There would be a culture dish placed on the bottom. 4. A blue LED light bulb will be installed around the top of the inner shell plane to attract the first female insect. |
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Tenon | 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 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.
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.
3. Inner shell is removable so it’s easier to add new PBAN solution.
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.
5. Purse-string bag is cheap and easy to switch.
6. The PBAN system can run day and night. Its function won’t be affected by sunlight.