1. The effect of Paecilomyces lilacinus, Pasteuria Penetrans, chitin and clove oil on Resistant soy, inoculated with Rhizobium Leguminosarum, and C.elegans. Demitri Dedousis Jonathan Labib Joe Toole

2. Need http://www.greenfacts.org/en/biodi versity/images/bdv-3-17-n- fertilizers.jpg http://i23.ebayimg.com/06/i/000/ae/2f/b02d_12.JPG

3. SCN /C.elegans www.epa,gov

4. Resistant Soy http://www.biochem.arizona.edu http://www.itmonline.org

5. Rhizobium http://cache.viewimages.com/xc/3353096.jpg?v=1&c=ViewImages&k=2 &d=0629904139C22E587A5A3CC511DB2D78A55A1E4F32AD3138

6. Paecilomyces lilacinus http://www.im.ac.

7. Fluid Mosaic Model http://www.wmc.ac.uk/biology/Image2.jpg

8. Pasteuria Penetrans www1.montpellier.inra.fr

9. Chitin Stimulates growth of bacterial microbiota and actinomycetes Limits number of fungal species naturaly produced non-chemical substance http://www.wmc.ac.uk/biology/Image2.jpg

10. Clove Oil a natural substance Levels of clove oil up to.4% is needed to decrease egg count.12% kills half of all eggs and hatched nematodes www.koihealth.org

11. Inheritance of resistance to soybean cyst nematode in soybean RIL and F Tilka http://www.nature.com Resistant Strain V. Susceptible Strain Tilka confirms resistant soy are unaffected by SCN SCN can circumvent soy resistance in under 3 years (36 generations Creation of new resistant strains is more difficult

12. In-vitro Assays of Meloidogyne incognita and Heterodera glycines for Detection of Nematode-antagonistic Fungal Compounds By Nital Nital Concludes that parasitic nematodes were reduced significantly by fungus Nital discuses how fungi effects hatching of nematodes http://www.ars.usdahttp://www.ars.usda.

13. Pathogenicity of Fungi to Eggs of Heterodera Glycine Dickson Dickson discusses parasitism of nematodes in cyst/egg level Dickson suggests that egg parasitism greatly reduces levels of adult population http://fulltextt10.fcla.edu/DLData/SN/SN0022300X/0028_002/96_1 9.pdf

14. Detection of the nematophagous fungus Paecilomyces lilacinus in soil by real-time PCR and parasitism bioassay By Senyu Chen Chen suggests temperatures and acidity for optimal growth of fungi Chen also suggests soil composition for growth of fungi

15. Characteristics and Efficacy of a Sterile Hyphomycete (ARF18), a New Biocontrol Agent for Heterodera glycine and Other Nematodes By Kim Kim developed a method for counting nematode eggs and adults www.fao.org

16. Biocontrol of Nematodes by Pasteuria Penetrans by Hewlett, Dickson, Serracin www1.montpellier.inra.fr Pasteuria endospores are best cultured by inoculating infected nematodes into host plant ( soy) Endospores can be stored in solution 104-105 endospores per gram of soil needed to reduce nematode population

17. Life Cycle Ultrastructure and Host Specifity of Pasteuria that Parasitizes the Soybean Cyst Nematode, Heterodera Glycines by G. R. Noel endospores only attach themselves and develop in female juvenile nematodes 100,000 endospores/g reduced nematode population by 95% Pod yields increased 94% with 100,000 endospores/g trc.ucdavis.edu

18. Purpose The purpose of the experiment would be to test how biological controls to parasitic nematodes (i.e. SCN) would work with a SCN resistant variety of soy to limit both damage of SCN to soy and the development of resistance among parasitic nematodes, the experiment would also test how fungi effects the local micro biota.

19. Hypothesis Hypothesis: H(o)- Resistant soy inoculated with biological controls will have no significant difference in C.elegan count or Rhizobium population from that in plain soy. H(a)- Resistant soy inoculated with biological controls will have significantly lower C.elegan count than that in plain soy and similar to that in soy exposed to Copper Sulfate. Resistant soy inoculated with biological controls will have statistically similar Rhizobium populations to plain soy and statistically higher Rhizobium populations than the soy with resistant Rhizobium and copper sulfate.

20. Methedology SCN Resistant Soy inoculated with Rhizobium, C.elegans and Paecilomyces lilacinus SCN Resistant Soy inoculated with Rhizobium and C.elegans SCN Resistant Soy inoculated with Resistant Rhizobium, C.elegans and copper sulfate The soy will be grown in individual pots with a sand soil mixture and the varidables will be measured: ｷ Nodule number ｷ C.elegan count ｷ Nitrogen Content ｷ Leaf Number ｷ Leaf Quality ｷ Plant Height X5 SCN Resistant Soy inoculated with Rhizobium, C.elegans and Chitin SCN Resistant Soy inoculated with Rhizobium, C.elegans and Clove Oil SCN Resistant Soy inoculated with Rhizobium, C.elegans and Pasteria Penetrans The mean will be calculated + standard deviation and the data will be analysed with and ANOVA and Sheffe post hoc test

21. Budget C.elgans (Carolina) 9.85 Nitrogen Test Kit (Carolina) 23.95 Rhizobium Inoculum (Carolina) 12 Rhizobium agar (Carolina) 5.25 Rhizobium Culture (Carolina)8.50 Petri Dishes (Carolina)36.10 E.Coli (Carolina) 8.20 Luria Broth Agar (Carolina) 15.50 Copper Sulfate Pentahydrate 250g 162.50 Biostat Paecilomyces lilacinus (Lymaz Corp) Arthrobotrys conoides Carnivorous Fungus Kit, Carolina 38.95 SCN Resistant Soy (Soybean Depository )

22. Do Ability Techniques for culturing soy and rhizobium have previously been mastered Senyu Chen a top expert has offered to help with protocols on culturing parasitic nematodes

23. Bibliography Abril, Adriana B. "The Importance of Phyllosphere Microbial Population in Nitrogen Cycling." Abstract. Tropical Ecology 21 (2005): 103-107. Bjedov, Ivana. "Stress-Induced Mutagenesis in Bacteria." Science 300 (2003): 1404- 1409. 13 May 2007. Biggs, Alton. Biology. Columbus: McGraw-Hill Companies, New York, 2004. 293-300. Bleifield, Maurice. Barron's Biology. New York: Barron_Educational Series. 217-220. Boven, Van. Segregation Distortion in a Deme-Structured Population: Opposing Bremer, Eric. "Nitrogen Benefits From Rhizobia." Micro Biology 14 (1996). Demands of Gene, Individual and Group Selection. Evolutionary Biology 12 (1999): 80- 93. Abstract. De Bruin, Jason. "Soil Microbiota Effects on Rye Growth Implications for Integration of a Rye Cover Crop Into Temperate Cropping Systems." Abstract. Renewable Agriculture and Food Systems 21 (2007): 245-252. Driscoll, Charles. "Nitrogen Pollution in the U.S." BioScience 53 (2003): 357-361. Keymer, Juan. "Bacterial Metapopulations in Nanofabricated Landscape." Physical Sciences 103 (2006): 172-185. 16 May 2007. Laird, Patricia. "Root Nodulation." Microbes 68 (2002): 143-146. Laguserre,G. Microbiology and Ecology 52 (2006): 3. Abstract. Response of Rhizobial Populations to Moderate Copper Stress Applied to an Agricultural Soil Epub (2006): 426-435

24. Bibliography Natsch, Andreas. "Importance of Preferential Flow and Soil Managment in Transport of a Biocontrol Strain of Pseudomonas Flurecens Instructure Field Soil." Enviromental Microbiology 62 (1995): 33-40. 12 May 2007. Nicholson, P.S. "The Effects of Pesticides on the Diversity of Culturable Soil Bacteria." 84 (1998): 551. Abstract. Journal of Applied Microbiology. Musser, James M. "Molecular Population Genetic Analysis of Emerged Bacterial Pathogens: Selected Insights." Perspective 2 (1996). 13 May 2007. Sibly, Richard M. "Efficient Experimental Designs for Studing Stress and Population Density in Animal Populations." School of Animal and Microbial Sciences. Abstract. Ecological Applications 9 (1998): 496-503. Rouse, Jeremy. "Nitrogen Pollution Assesment." Enviromental Health 107 (1999): 905- 1001. Whitford, Walter G. "The Importance of Soil Biota in Maintaining Ecosystem Integrity." Abstract. Enviromental Monitoring 23 (1994). Zaharn, Hambi. "Rhisobium-Legume Symbiosis and Nitrogen Fixation Under Severe Conditions." Department of Botany. Abstract. Microbial and Molecular Biology 63 (1999): 968-989.