Deliverable 1: Soybean producers will eventually possess improved soybean cultivars based on the novel traits developed by this project. We will generate new mutations for seven soybean genes known to be involved in seed protein, amino acid, oil and/or fatty acid composition. These will add new germplasm for breeders interested in incorporating new or different seed composition traits in their breeding programs.
Deliverable 2: The training workshop will expand the use of precision genomics throughout the soybean community, resulting in a greater number of targeted mutations that will be available for soybean breeders’ efforts at developing and adapting to new markets for producers. U.S. scientists trained in the precision genomics technologies will enable these approaches to be used for a wide range of traits and applications in the future

Achieving objectives:
• Targeted mutations in whole soybean plants were obtained for three genes; two were generated by ZFNs and the third gene was mutated with a TALEN.
• CRISPR/Cas9 system has shown promise in model organisms and was optimized for use in soybean
• CRISPR/Cas9 constructs for 10 genes, including FAD2-1 and FatB, were completed. Mutagenesis screens for these genes were initiated in soybean hairy roots.
• Soybean Precision Genomics workshops were held in 2012, 2013 and 2014 and were attended by over 160 scientists. Attendance has increased each year, suggesting increasing interest in these technologies.
• Cellectis Plant Sciences has reported the recovery of targeted mutations for the soybean FAD-2 seed-expressed genes using TALENs. The lead scientists at Cellectis on this project received training in soybean genetic transformation and genome engineering technologies while members of Stupar’s lab, in conjunction with USB support.

Not achieving objectives:
• The complicated design and limited throughput of ZFNs and TALENs hindered success in this project. The researchers successfully targeted genes with both the ZFN and TALEN methods, but were not able to obtain mutations in seed composition genes. In the course of the project when the difficulties became evident and the newest method, CRISPR/Cas9, became known they rapidly shifted focus.