Project Number: 6435-53000-001-00
Project Type: Appropriated

Start Date: Mar 20, 2006
End Date: Apr 30, 2009

Objective:
1. Induction, purification, and characterization of isoflavonoids in soybean seed, root, plant and cell culture incubated with A. sojae and A. sojae cell wall extracts. Induced isoflavonoids will be produced in soybean using several different biotic elicitors developed from A. sojae. Elicitor experiments will initially focus on soybean seeds, roots and cell cultures. Once an effective elicitor system is developed in the laboratory, elicitor experiments to induce isoflavonoids will be conducted on soybean seeds within the pod of a developing plant. 2. Determine hormonal and antioxidant activities of individual and combinations of induced isoflavonoids with in vitro bioassays. Determine hormonal and antioxidant activities of induced isoflavones purified under Objective 1 using in vitro bioassays. Three established assays for the determination of estrogenic and antiestrogenic activity will be utilized. Additionally, the antioxidant activities of induced isoflavones will be determined using three assays. 3. Determine hormonal and anticancer effects of individual and combinations of induced isoflavonoids with in vivo animal model systems. Determine estrogenic, antiestrogenic and anticancer activities of induced isoflavones (discovered in Objective 1) in two in vivo mouse models. Induced isoflavones that display in vitro activity in Objective 2 will be lead candidates for in vivo testing. The glyceollins represent potential beneficial antiestrogens that need to be tested in animal models to confirm activity observed in vitro. Two different mouse model systems will be utilized to confirm estrogenic, antiestrogenic and anticancer activity (breast and ovarian cancer). 4. Develop methods for the organic syntheses of the glyceollins I, II and III. Also, precursors and synthetic analogues of the glyceollins will be produced and characterized for beneficial hormonal activities. Computer modeling methods will be developed to assist in the development of structure-activity relationships that point toward the most active forms of the molecule. Accomplishment of objectives 2-4 will require seeking out additional scientific cooperators with the particular expertise needed, either within ARS or in other organizations.

Approach:
Induced isoflavonoids will be produced in soybean using several different biotic elicitors developed from A. sojae. Elicitor experiments will initially focus on soybean seeds, roots and cell cultures. Once an effective elicitor system is developed in the laboratory, elicitor experiments to induce isoflavonoids will be conducted on soybean seeds within the pod of a developing plant. Isolated induced isoflavones will be tested for hormonal and antioxidant activities using several in vitro bioassays. Three established assays for the determination of estrogenic and antiestrogenic activity will be utilized. Additionally, the antioxidant activities of induced isoflavones will be determined using three assays. Induced isoflavones that display in vitro activity will be lead candidates for in vivo testing. The glyceollins represent potential beneficial antiestrogens that need to be tested in animal models to confirm activity observed in vitro. Two different mouse model systems will be utilized to confirm estrogenic, antiestrogenic and anticancer activity (breast and ovarian cancer). Also, organic syntheses methods for the glyceollins I, II and III will be developed. Precursors and synthetic analogues of the glyceollins will be produced and characterized for beneficial hormonal activities. Computer modeling methods will be developed to assist in the development of structure-activity relationships that point toward the most active forms of the molecule.