The 6The 6thth Annual AnnualIslip High SchoolIslip High School

ScienceScience

SymposiumSymposium

http://www.reefkorea.org/member/board/data/data/Limulus_polyphemus.jpg

HISTORY

• For 445 million years L. polyphemus has remained unchanged.

• They often used in the fishing industry as inexpensive bait.

• A component of their blood is essential to the development of vaccines and implanted medical devices.

• In the past 20-30 years, scientists believe their numbers have dropped drastically. The causes of this drop are still a mystery.

HYPOTHESIS

H1: The initial hypothesis was that the presence of brown tide in the Limulus polyphemus’ habitat would cause a decline in the Limulus polyphemus population.

H2: The hypothesis for the second experiment was that due to the increasing ratio of male to female Limulus polyphemus, there would be an overall population decline.

PROCEDURESThe procedure for both experiments were similar. The goal was to use past surveys to compare and determine whether or not there was a trend or relationship between the variables.

In the first experiment this would be to observe the amount of brown tide cells as compared to the number of spawning females in both Delaware and New Jersey.

The procedure for the second experiment was to observe the increasing ratio of male to female Limulus polyphemus’ and the overall population of the Limulus polyphemus.

Year Yearly ISA Brown Tide Cells (per ml)

1999 .93204 1.96 x 10^5

2000 1.01971 6.10 x 10^5

2001 .82268 4.20 x 10^5

2002 .76155 1.70 x 10^3

2003 .80909 5.30 x 10^5

2004 .76031 2.2 x 10^6

Delaware

Year Yearly ISA Brown Tide Cells (per ml)

1999 .6149 2.89 x 10^6

2000 .80267 2.20 x 10^6

2001 .63964 1.90 x 10^6

2002 1.09376 1.75 x 10^6

2003 .82703 1.70 x 10^3

2004 .77643 1.70 x 10^3

New Jersey

CONCLUSION

• After analyzing the data, no correlation between brown tide blooms and L. polyphemus populations was found.

• Surveys from both New Jersey and Delaware refuted the hypothesis that brown tide played a direct role in the drop of L.polyphemus populations

Sex Ratio Study

• Because fisherman often take the larger females, a study was begun to examine the possibility of a link between male to female ratios and the L. polyphemus populations.

Year Males / Female Estimated Population

2001 3.38 5,604, 000

2002 3.48 4,480,000

2003 3.61 4,102,900

2004 3.85 3,395,000

2005 3.89 2,689,500

2006 4.53 2,654,400

2007 4.90 2,360,000

CONCLUSION

In every year since 2001, the male to female ratio has been increasing. In the last 3 years alone the record has been eclipsed the following year.

This overall shortage of females in comparison to the males may indicate an overall drop in fertilized eggs laid each year.

This trend would also go support the observation that the overall population has been declining in recent years.

FUTURE RESEARCH

In conclusion, the brown tide was found to have a small if any effect on the Limulus polyphemus populations.

In the second experiment, the male to female ratio increased as the overall population decreased.

Future studies of more complete survey counts may confirm suspicions that the overall North American L. polyphemus populations are in decline.

If so, male to female ratios present a viable cause for this decline, and may aid conservation efforts. By preserving females, the overall population may recover.

•Limulus Spawning Ability on Delaware Bay Shores 1990, Sea Grant, 1990•Horseshoe Crabs Imperiled? The Fate of a Species 350 Million Years in the Making- By John T. Tanacredi-Limulus in the Limelight-1996•The Horseshoe Crab: The Battle for a True Multiple-use Resource- By Jim Berkson and Carl N. Shuster, Jr.-Fisheries Management, Vol.24 No.11, 1999•Two Perspectives: Horseshoe Crabs During 420 Million Years, Worldwide, and the Past 150 Years in the Delaware Bay Area- Carl N. Shuster Jr.-Limulus in the Limelight-2000•A Unique Medial Product(LAL from the Horseshoe Crab and Monitoring the Delaware Bay Horseshoe Crab Population-By Benjie Lynn Swan-Limuli Laboratories, Inc.-Limulus in the Limelight-2001•Biomedical Products from the Horseshoe Crab: Lessons for Sound Resource Management and Conservation-By Thomas J. Novitsky-President/CEO, Cape Cod,Inc.-2001•Issues and Approaches in Regulation of the Horseshoe Crab Fishery-By Josh Eagle-Staff Attorney-National Audubon Society-2001•The Delaware Bay Horseshoe Crab Spawning Survey- By Benjie Lynn Swan, William R. Hall, Carl N. Shuster, Jr.-2001•Brown Tide blooms in Long Island's coastal waters linked to interannual variability in groundwater flow-By Julie Laroche , Robert Nuzzi , Robert Waters, Kevin Wyman , Paul Falkowski & Douglas Wallace- Brookhaven National Laboratory-Global Change Biology, Volume 3 Issue 5, pgs.397-410, 2003•The 2007 Delaware Bay Horseshoe Crab Spawning Survey-By Benjie Lynn Swan, William R. Hall and Carl N. Shuster, Jr.-2007

http://www.reefkorea.org/member/board/data/data/Limulus_polyphemus.jpg

The 6The 6thth Annual AnnualIslip High SchoolIslip High School

ScienceScience

SymposiumSymposium

Amateur Guided Rocketry

Using the Sun as a Guidance

Design and construction of a computer Guided rocket

Experiment by: Michael Nelson

Background InformationBackground Information

Up until now rockets have been Up until now rockets have been guided by very expensive avionics guided by very expensive avionics systems.systems.

GMLRS United States Army

About the Basic StampThe Basic Stamp is a micro controller with a small specialized Basic interpreter. ( Built into ROM)It is made by Parallax, Inc. and has been quite popular with electronics hobbyist since the early 1990’s due to its low threshold of learning and ease of use, because of its simple language.

The basic stamp is in the form of a DIP chip. It has all of the elements of a microprocessor system.

ProposalProposal

To use a predetermined source of light, such To use a predetermined source of light, such as the sun; as a fixed point, so the rocket can as the sun; as a fixed point, so the rocket can stabilize itself.stabilize itself.

To create programming that will allow the To create programming that will allow the rocket to lock on to different sources of light.rocket to lock on to different sources of light.

What the Basic Stamp will seeWhat the Basic Stamp will see

SUN

Sun

What the Basic Stamp will seeWhat the Basic Stamp will see

SUN

Sun

What the Basic Stamp will seeWhat the Basic Stamp will see

SUN

Sun

Space CADSpace CAD Software that allows a user to design, Software that allows a user to design,

then simulate a rocket launch. then simulate a rocket launch.

Rocket DiagramRocket Diagram

Rocket SpecificationsWeight:Launch weight: 5.56 lb. Empty weight: 3.32 lb.

Engine configuration:

Stage 1 Engines: 1 * Aerotech G80 (3.76 oz.)

Flight Prediction results:

Max. height: 742.00 ft. Max. velocity: 249.23 ft/s Max. acceleration: 269.04 ft/s² Best delay: 6.15 s

Min. launch rod length: 6.00 ft.

Stability results:

Center Of Pressure (CP) : 2.89 ft. behind nose cone Center Of Gravity (CG) : 1.87 ft. behind nose coneRocket is stable (4.09 calibers)Other Rocket data:Rocket length: 4.58 ft.

BibliographyBibliography

Urbano, Leonardo, and Kam Moshe. Urbano, Leonardo, and Kam Moshe. Design And Construction of a Design And Construction of a computer Guided Rocketcomputer Guided Rocket. Drexel . Drexel University: 2006. University: 2006.

Verhage, Paul, Verhage, Paul, Near Space ProgramNear Space Program. . 2004 2004

Balkelock, John, Automatic Control of Balkelock, John, Automatic Control of Aircraft and Missiles. 1991. Aircraft and Missiles. 1991.

Amateur Guided Rocketry

Using the Sun as a Guidance

Design and construction of a computer Guided rocket

The 6The 6thth Annual AnnualIslip High SchoolIslip High School

ScienceScience

SymposiumSymposium

The Biological Sterilization of Petunia xhybrida Using the

Excessmicrosporcytes1 Gene

By Michael Fahner

Abstract/HypothesisThe EMS1 gene is a male sterile mutant identified in the

Arabidopsis plant. The mutant gene results in minimal phenotypic effects, aside from abnormal growth of

cells in the tapetum layer of the anther. If the EMS1 gene can be successfully amplified in the Petunia

xhybrida then the sterile EMS1 Petunia would not be able to spread pollen to nearby fields. To amplify the EMS1 in the Petunia plant, the gene first needed to be

isolated. The gene was to be isolated from Petunia cDNA using a Polymerase Chain Reaction procedure. The EMS1 gene is a relatively small gene, consisting of about 500 base pairs. Various temperatures in the

PCR program, primer concentrations, template dilutions and other variables were tested to develop an

effective Polymerase Chain Reaction procedure, to isolate the EMS1 Gene.

Brief History Of The Arabidopsis• Arabidopsis is a small flowering weed.

• It belongs to the Brassicaceae family. (along with other crops such as radish and cabbage)

• It has a relatively small genome, making mutations such as the EMS1 gene easier to identify.

•The EMS1 gene is a male sterile mutant.

• The gene’s expression results in the production of extra microspores that later form sterile pollen.

• The gene prohibits the growth of the tapetum cells which are located within in the anther.

• Although EMS1 expression results in sterility, the EMS1 plants still exhibit normal floral and vegetative properties (other then within the anther).

The EMS1 GeneEXCESS MICROSPOROCYTES1

Low molecular weight ladder by New England Biolabs.

“Mother Plant”, Expressing EMS1 Gene

e e50% of the offspring are male sterile.

“Father Plant”, EMS1 Carrier

E

e

EeEe

ee ee

Genetically modified crops have been pollinating wild plants in the environment as well as nearby commercial farms for years. This cross-pollination reduces bio-diversity and may also infringe upon patent laws. Male Sterility would eliminate this problem because the pollen would no longer reach unwanted areas. A sterile EMS1 plant would not be able to cross-pollinate wild plants in the environment, thus preserving biodiversity. By first expressing the gene in the Petunia x hybrida, it may later prove possible to express the gene other commercial plants. If successful the EMS1 gene could become a valuable tool for commercial farmers across the nation.

Purpose

Methods

• Extract RNA from the petunia plant.

• Extract DNA from the Petunia plant (to be used as a control)

• Conduct Reverse Transcription to form cDNA from the extracted RNA

• Perform a Polymerase Chain Reaction

• Run the results of the PCR reaction on an agarose gel.

First Polymerase Chain Reaction: Reagents10x Buffer- 2ul DNTp- .5ulTaq- .1ulTemplate- 1ulForward Primer- .5ul(From 12.5uM dilution)Reverse Primer- .5ul(From 12.5uM dilution)Water-14.8ulTotal= 20ul

Program94o - 5min94o - 30s Repeat 55o - 30s 3972o - 30s Times72o - 5min10o - Pause

EMS1 degenerate primers:Forward:EMETLGKGGN ATG GA(A/G) ACN (T/C)TN GGN AA(A/G) (21mer. degeneracy 2-11,Tm=63+7)

GGN ATG GA(A/G) ACN (T/C)TN GGN (18mer, degeneracy 2-10, Tm=56+6)

Reverse:GGNLVGWGGN GGN AA(C/T) (T/C)TN GTN GGN TGGCCA NCC NAC NA(G/A) (G/A)TT NCC NCC (21mer, degeneracy 2-11, Tm=67+7)

CCA NCC NAC NA(G/A) (G/A)TT NCC (degeneracy 2-10, Tm 56=6)Assume Tm for G/C is 4, A/T is 2 and N is 3.

Data Test 1The far left column is the Petunia cDNA, the next is the DNA from Columbia Arabidopsis and the

third is the control cDNA. The fourth column is the marker.

Data Test 2The ladder is to the far left—starting from the next

column, Petunia cDNA, Columbia DNA, and Control cDNA with a 50 degree annealing

temperature. Next, the same templates but with 56 degree annealing temperature and finally the

last three are the touchdown.

7/14/08

Data Test 4The second, third and fourth column after the ladder revealed nothing; therefore the

PCR was a unsuccessful.

8/6/08

Data Test 3After the marker, a column is skipped but

then there is Petunia cDNA, Petunia DNA, and Control cDNA with 200nM primer concentration. The next three

have 1uM primer concentration. The last three have 4uM primer concentration.

Data Test 6The first two columns are 4uM cDNA and DNA at 45 degree annealing temperature. The next two are 8uM primer concentrations, with a 45 degree annealing temperature. The next two are extracted DNA with 4uM and 8uM primer concentration at a 45 degree annealing. The final two are ten fold dilutions of cDNA. This new reaction had a 45 degree annealing temperature and used both 4uM and 8uM primer concentrations. Petunia cDNA, Petunia DNA and some DNA from the last gel extraction were used as templates. In two more tubes, Petunia cDNA was diluted ten fold.

Data Test 5After the ladder, the first two columns are Petunia cDNA and Petunia DNA with 45 degree annealing and 4uM primer concentration. The next two are 8uM concentration with a 45 degree annealing temperature. Fifth and sixth columns are 4uM

primers with 50 degree annealing temperature and the last column is a 50 degree annealing

temperature with 8uM primer concentration.

Data Test 8The second column after the marker

contained 2 uM MG+2, the next 2.5 uM MG+2, 3 uM MG+2, 3.5 uM MG+2, 4 uM

MG+2, 5 uM MG+2, 6 uM MG+2, and in the final column tube 1.5 uM MG+2 was added

with 5% DMSO.

Data Test 7The first two columns after the ladder are

cDNA and DNA with 8uM primer concentration and 45 degree annealing

temperature. The next two are the same except for a 50 degree annealing

temperature.

Data Test 10The first two columns after the ladder are cDNA and Petunia DNA. Dilutions of two, four and ten fold cDNA are the next three columns and DNA dilutions are the next

three after that. The final two columns are extracted DNA templates.

Data Test 9The first column after the marker contained

2 uM MG+2, the next 2.5 uM MG+2, 3 uM MG+2, 3.5 uM MG+2, 4 uM MG+2, 5 uM

MG+2, 6 uM MG+2, and in the final column tube 1.5 uM MG+2 was added with 5%

DMSO.

Future Research • Select Transformed bacteria on LB agar media + antibiotic. • Grow up several colonies and plasmid DNA by minipreps.• Use restriction digestion on plasmids to check for expected product

size.• Sequence several gene products to confirm if one is Petunia EMS1.• Use additional primers to amplify gene product in plasmid and

incorporate new sequences at end to allow ligation into RNAi vector.• Test on agarose gel for amplification. • Ligate into RNAi vector.• Transform ligation into E.Coli bacteria.• Select transformants on appropriate antibiotic.• Grow up several colonies & isolate plasmid DNA by minipreps. • Use restriction digestion on plasmids to check for expected product

size.• Sequence several gene products to confirm amplification and

orientation is correct. • Transform plant into Agrobacterium.• Use Agrobacterium to transform Petunia.

Lacroix, Benoit et al. (2006).Will You Let Me Use Your Nucleus? How Agrobacterium Gets its T-DNA Expressed in the Host Plant Cell. Pharmacol. 84, 333-345.

Tzfira, Tzvi et al. (2006).Agrobacterium-mediated genetic transformation of plants: biology and biotechnology. Current opinion in Biotechnology. 17, 147-154.

Zhang, Wei et al. (2006).Regulation of arabidopsis tapetum development and function by DYSFUNCTIONAL TAPETUM1 (DYT1) encoding a putative bHLH transcription factor. Development. 133, 3085-3095.

Tzfira, Tzvi et al. (2004).Agrobacterium T-DNA Integration: molecules and models. TRENDS in Genetics . 20, 375-383 .

Zhao, Dazhong et al. (2003).Members of the arabidopsis- skip1-like gene family exhibit a variety of expression patterns and may play diverse roles in arabidopsis. Plant Physiology. 133, 203-217.

Zhao, Da-Zhong et al. (2002).The EXCESS MICROSPOROCYTES1 gene encodes a putative leucine-rich repeat receptor protein kinase that controls somatic and reproductive cell fates in the Arabidopsis anther . Genes and Development. 16, 2021-2031.

Bibliography

Tzfira, Tzvi et al. (2002).Partners-in-infection: host proteins involved in the transformation of plant cells by Agrobacterium . TRENDS in Cell Biology. 12, 121-129.

Azumi, Yoshitaka et al. (2002).Homolog interaction during meiotic prophase I in Arabidopsis requires the SOLO DANCERS gene encoding a novel cyclin-like protein . EMBO . 21, 3081-3095.

Zhao, Dazhong et al. (2001).The ASK1 gene regulates development and interacts with the UFO gene to control floral organ identity in Arabidopsis.. Development. 128, 2735-2746.

Yang, Ming et al. (1999).The Arabidopsis SKP1-LIKE1 gene is essential for male meiosis and may control homologue separation . Proc. Natl. Acad. Sci. USA. 96, 11416-11421.

Bibliography

A Special Thanks To…

Dr.Hong MaProfessor of BiologyDepartment of BiologyPennsylvania State University

Dr.Lawrance Hobbie Science ChairmanDepartment of Biology Adelphi University

The Biological Sterilization of Petunia xhybrida Using the

Excessmicrosporcytes1 Gene

By Michael Fahner

The 6The 6thth Annual AnnualIslip High SchoolIslip High School

ScienceScience

SymposiumSymposium