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Sequencing and functional validation of the JGI Brachypodium distachyon T‐DNA collection
Authors:Mon Mandy Hsia  Ronan O'Malley  Amy Cartwright  Rita Nieu  Sean P. Gordon  Sandra Kelly  Tina G. Williams  Delilah F. Wood  Yunjun Zhao  Jennifer Bragg  Mark Jordan  Markus Pauly  Joseph R. Ecker  Yong Gu  John P. Vogel
Affiliation:1. USDA ARS Western Regional Research Center, Albany, CA, USA;2. Genomic Analysis Laboratory, The Salk Institute for Biological Studies and the Howard Hughes Medical Institute, La Jolla, CA, USA;3. DOE Joint Genome Institute, Walnut Creek, CA, USA;4. Cereal Research Centre, Agriculture and Agri‐Food Canada, Morden, MB, Canada;5. Department of Plant and Microbial Biology, University of California, Berkeley, CA, USA
Abstract:Due to a large and growing collection of genomic and experimental resources, Brachypodium distachyon has emerged as a powerful experimental model for the grasses. To add to these resources we sequenced 21 165 T‐DNA lines, 15 569 of which were produced in this study. This increased the number of unique insertion sites in the T‐DNA collection by 21 078, bringing the overall total to 26 112. Thirty‐seven per cent (9754) of these insertion sites are within genes (including untranslated regions and introns) and 28% (7217) are within 500 bp of a gene. Approximately 31% of the genes in the v.2.1 annotation have been tagged in this population. To demonstrate the utility of this collection, we phenotypically characterized six T‐DNA lines with insertions in genes previously shown in other systems to be involved in cellulose biosynthesis, hemicellulose biosynthesis, secondary cell wall development, DNA damage repair, wax biosynthesis and chloroplast synthesis. In all cases, the phenotypes observed supported previous studies, demonstrating the utility of this collection for plant functional genomics. The Brachypodium T‐DNA collection can be accessed at http://jgi.doe.gov/our-science/science-programs/plant-genomics/brachypodium/brachypodium-t-dna-collection/ .
Keywords:insertional mutagenesis     Brachypodium distachyon     functional genomics  T‐DNA  mutant sequencing  model grass
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