1. Using Iron-rich Fertilizer to increase Hemoglobin in Rhizobium Nodules Danny Muslin

2. Need World fertilizer prices surged by more than 200 percent in 2007 (international Center for Soil Fertility and Agricultural Development)

3. Nitrogen Cycle

4. Rhizobium Rod shaped Gram -ve bacteria Fixes nitrogen after infecting host plants Has a mutually beneficial relationship with the host

5. Rhizobium Nodules Inside the nodules is where nitrogen fixation occurs Nodules have a finite lifespan. (in the case of soybean, estimated at 50-60 days )

6. Hemoglobin An iron-containing oxygen-transport metalloprotein Depiction of hemoglobin in a human adult

7. Exogenous iron increases hemoglobin in beta-thalassemic mice. (Ginzburg et al., 2008) Exogenous iron increases hemoglobin production in beta-thalassemic mice

8. THE INFLUENCE OF EXOGENOUS IRON ON FORMATION OF HEMOGLOBIN IN THE PREMATURE INFANT (Hammond, 1960) Found that extra iron supplements can be utilized by infants to increase hemoglobin concentration

9. Iron Nutrition and Interactions in Plants By Y. Chen, Y. Hadar, Seagram Center for Soil and Water Sciences (1991) State that nodulation and nitrogen fixation occur at a more efficient rate when a steady supply of iron is present Fe

10. Methionine Implication In Rhizobial Iron Acquisition from Hemoglobin (Battistoni, 2006) Rhizboia can utilize hemi, hemoglobin, mioglobin, leghemoglobin, and siderophores as iron sources and transporters

11. Chelated Iron Iron that has undergone chelation Easier for plants to absorb

12. Method of Testing for Hemoglobin in Rhizobium Nodules Method used will be: Cyanmethemoglobin Method for Determining Hemoglobin Concentration

13. Purpose The purpose of this experiment is to increase the rate of nitrogen fixation in rhizobium nodules by using chelated iron- rich fertilizer to increase hemoglobin production in the nodules. The amount of hemoglobin in a nodule is correlated to the amount of nitrogen it is fixing

14. Hypothesis Alternate: The group that is exposed to chelated iron will have a higher concentration of hemoglobin and a higher rate of nitrogen fixation Null: The groups will not be different with respect to hemoglobin levels and amount of nitrogen fixed

15. Methodology Flowchart Iron-rich fertilizer to increase hemoglobin in rhizobium nodules Soybean Plants ( Glycine max) Control (200) Soybean plants without R. Leguminosarum or chelated iron fertilizer Rhizobium (200) Soybean plants with R. Leguminosarum, without chelated iron fertilizer Rhizobium /w Iron (200) Soybean plants with R. Leguminosarum and chelated iron fertilizer The two groups that have nodulation will be tested for nodule hemoglobin using the cyanomethemeglobin method once a week (this allows for nodule regrowth). After the first week (allowing nitrogen to be fixed) all groups will then also be tested once a week, (on the same day as the hemoglobin test), for amount of nitrogen fixed using a LaMotte nitrogen test kit The amount of nitrogen fixed by each group will be measured in pounds, and the amount of hemoglobin in rhizobium nodules will be measured in Hgb Values of g/100ml. The results for nitrogen fixation and hemoglobin levels, will both separately be analyzed with a one-way ANOVA followed by a scheffe post-hoc test

16. Budget