Agrobacterium ‐Mediated Transformation of Brachypodium distachyon

Journal Article · Tue Mar 12 00:00:00 EDT 2019 · Current Protocols in Plant Biology

DOI:https://doi.org/10.1002/cppb.20088· OSTI ID:1499072

Chen, Fengjuan ^[1]; Liu, Qi ^[1]; P. Vogel, John ^[2]; Wu, Jiajie ^[1]

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1. State Key Laboratory of Crop Biology, Shandong Agricultural University Tai'an Shandong China, College of Agronomy, Shandong Agricultural University Tai'an Shandong China
2. DOE Joint Genome Institute Walnut Creek California, University of California Berkeley Berkeley California

Abstract Brachypodium distachyon is an excellent model system for the grasses and has been adopted as a research organism by many laboratories around the world. It has all of the biological traits required for a model system, including small stature, short life cycle, small genome, simple growth requirements, and a close relationship to major crop plants (cereals). In addition, numerous resources have been developed for working with this species, including genome sequences for many lines, sequenced mutant collections, and a large, freely available germplasm collection. Fortunately, among grasses B. distachyon is one of the most easily transformed species, an absolute necessity for a model system. Agrobacterium ‐mediated transformation is the preferred method to transform plants because it usually results in simple insertions of target DNA. In this article, we describe a method for Agrobacterium ‐mediated transformation of the inbred B. distachyon lines Bd21 and Bd21‐3. Embryogenic callus induced from immature embryos is co‐cultivated with Agrobacterium tumefaciens strain AGL1 or Agrobacterium rhizogenes strain 18r12v. Hygromycin and paromomycin are used as selective agents, with comparable transformation efficiencies (defined as the percentage of co‐cultivated callus that produce transgenic plants) of 40% to 70%. It takes 20 to 30 weeks to obtain T ₁ seeds starting from the initial step of dissecting out immature embryos. This protocol has been shown to be efficient and facile in several studies that resulted in the creation of over 22,000 T‐DNA mutants. © 2019 by John Wiley & Sons, Inc.

• Sponsoring Organization: USDOE
• Grant/Contract Number: DE‐AC02‐05CH1123
• OSTI ID: 1499072
• Journal Information: Current Protocols in Plant Biology, Journal Name: Current Protocols in Plant Biology Vol. 4 Journal Issue: 2; ISSN 2379-8068
• Publisher: Wiley Blackwell (John Wiley & Sons)
• Country of Publication: United States
• Language: English