Using Iron-rich Fertilizer to increase Hemoglobin in

Rhizobium Nodules Danny Muslin

Need

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

                                                                                    

                                                         

Nitrogen Cycle

Rhizobium

• Rod shaped Gram -ve bacteria

• Fixes nitrogen after infecting host plants

• Has a mutually beneficial relationship with the host

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 )

Hemoglobin

• An iron-containing oxygen-transport metalloprotein

Depiction of hemoglobin in a human adult

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

• Exogenous iron increases hemoglobin production in beta-thalassemic mice

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

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

FeFeFe

Methionine Implication In Rhizobial Iron Acquisition from Hemoglobin

(Battistoni, 2006)

• Rhizboia can utilize hemi, hemoglobin, mioglobin, leghemoglobin, and siderophores as iron sources and transporters

Chelated Iron

• Iron that has undergone chelation

• Easier for plants to absorb

Method of Testing for Hemoglobin in Rhizobium Nodules

• Method used will be:

Cyanmethemoglobin Method for Determining Hemoglobin Concentration  

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

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

Methodology FlowchartIron-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

Budget