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<OL> <LI> 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. <LI>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. <LI> 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). <LI> 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. </ol> 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.