P309
A Genome Engineering Toolbox for Legume Functional Genomics

Date: Monday, January 13, 2014
Room: Grand Exhibit Hall
Shaun J Curtin , Department of Plant Pathology, University of Minnesota, St. Paul, MN
Roxanne Denny , Department of Plant Pathology, University of Minnesota, St. Paul, MN
Joseph Guhlin , Department of Plant Pathology, University of Minnesota, St. Paul, MN
Jean-Michel Michno , Department of Agronomy and Plant Genetics, University of Minnesota, St. Paul, MN
Benjamin W. Campbell , Department of Agronomy and Plant Genetics, University of Minnesota, St. Paul, MN
Ryan C. Donohue , University of Minnesota
Tomas Cermak , University of Minnesota, Saint Paul, MN
Paul Atkins , University of Minnesota
Nicholas Baltes , University of Minnesota
Colby Starker , University of Minnesota, Saint Paul, MN
Yong Zhang , University of Electronic Science and Technology of China
Yiping Qi , East Carolina University
Peter L. Tiffin , Department of Plant Biology, University of Minnesota, Saint Paul, MN
Dan Voytas , University of Minnesota, Saint Paul, MN
Robert M. Stupar , Department of Agronomy and Plant Genetics, University of Minnesota, St. Paul, MN
Nevin D. Young , Department of Plant Pathology, University of Minnesota, St. Paul, MN
Advances in legume research have been frustrated by a lack of mutants and mutagenesis methodologies. This is especially relevant as scientists seek to explore the biological functions of gene candidates discovered through methods such as RNA-seq or genome-wide association studies (GWAS).  Here we demonstrate a suite of mutagenesis platforms including; Tnt1 retro-transposons, hairpin constructs, zinc-finger (ZFNs), TAL-effector (TALENs) and CRISPRs (RNA-guided) nucleases to engineer Dicer-like and nodulation mutants in soybean and Medicago truncatula respectively.  We have used the genetic transformation of Tnt1, hairpin, ZFNs, TALENs and CRISPR to knock-out/down single copy and duplicated legume genes, both in hairy root and whole plant systems. Some of the target genes were chosen in order to explore and optimize the features of knock-out/down systems (e.g., phytoene desaturase), while others were identified by soybean homology to Arabidopsis Dicer-like genes or by GWAS of nodulation in Medicago. These developments demonstrate the utility of these mutagenesis platforms for functional genomic studies and for potential trait improvement strategies in legumes.
Back to: Genome Mapping, Tagging & Characterization: Legumes, Soybeans, Common Beans
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