Berg, Howard , Schmidt, Monica , Herman, Eliot .
Nitrogen/Sulfur mapping in genetically modified soybean.
Soybean as potential feed is limited by a relative amino acid deficiency in its protein content. Altering soybean so that it possesses enhanced nutritional content has been a goal of academic and industry research. We have been refining various methods to be used in assaying recombinant soybean seeds. Transgenic soybean plants with RNAi-suppressed intrinsic seed proteins were produced and analyzed for protein distribution and protein amino acids. In order to map the intracellular distribution of nitrogen/sulfur in control and transgenic cotyledons, they were prepared for electron microscopy by high pressure freezing and freeze substitution. Electron Energy Loss Spectroscopy (EELS) was used to map elemental sulfur, phosphorus and nitrogen. The resulting maps can be used to quantify the nitrogen/sulfur distribution and spatial nitrogen/sulfur ratios. Phosphorus distribution identifies structural elements including ribosomes and nuclear material as well as phosphorus sequestered in the protein storage vacuole primarily as phytin. The EELS assay provides a high resolution assay capable of identifying elemental distribution within membrane compartments in a digitized dataset that can be analyzed for spatial distribution and concentration. The mapping of nitrogen/sulfur distribution is useful in biotechnology projects designed to enhance the nutritional quality of plants such as soybean.
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1 - Danforth Plant Science Center, 975 North Warson Road, St Louis, MO, 63132, USA
2 - Danforth Plant Science Center
3 - Plant Genetics Research Unit, USDA/ARS
Presentation Type: Plant Biology Abstract
Location: Exhibit Hall (Northeast, Southwest & Southeast)/Hilton
Date: Sunday, July 8th, 2007
Time: 8:00 AM