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Develop gene transfer system for cotton by improving protocols developed in laboratory for biolistic and Ti plasmid-based transformation of seedling-derived explant materials from elite cotton cultivars. Incorporate genes for known antifungal proteins/enzymes (chitinases, glucanases, antifungal peptides, etc.) into cotonseed under the control of various promoters functional in cotton. Test transformed seed tissues for expression of A. flavus resistance genes and antifungal activity.
A gene transfer system will be established based on the biolistic and Ti plasmid vector techniques for stable transformation of cotton with genes under the control of variou promoters functional in cotton. Current protocols will be used involving Ti plasmid-based vectors for transformation of cotton hypocotyl or meristematic tissues with one or more plasmids carrying a dominant selectable marker (such as resistance to an antibiotic). Following transformation,tissues will be cultured on selective media to obtain sexually functional plants with stably incorporated foreign genes expressed in seed under the control of various promoters (e.g., those yielding seed specific expression).Once a successful cottonseed transformation system is developed, cotton will be transformed with availbale genes for antifungal proteins/enzymes (e.g., chitinases, glucanases, antifungal peptides, etc.); transgenic cottonseed will be tested for expression of resistance genes and antifuntgal activity against A. flavus.