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Agrobacterium rhizogenes Versus Agrobacterium tumefaciens T-DNA Transformation in Brachypodium distachyon

CSEF · 2023 Plant Biology First Award

Hsu Evelyn · 11

Overview

Brachypodium distachyon is a monocotyledonous grass species that has emerged as a valuable model organism due to its close relationship to cereal and bioenergy crops, small genome, rapid generation time, and ease of transformation. This study aims to enhance the efficiency of Agrobacterium-mediated transformation in the inbred B. distachyon line Bd21-3 using two bacterial strains with similar virulence factors: A. tumefaciens strain AGL1 and A. rhizogenes strain 18r12v. Conflicting reports on the efficiency of these strains have been published. Plasmid constructs containing GUS (β-glucuronidase) and hygromycin resistance were introduced into the bacteria via electrical transformation and used to transform Brachypodium embryogenic calli. This was achieved by inserting DNA encoding the tumor-inducing (Ti) plasmid in A. tumefaciens or the root-inducing (Ri) plasmid in A. rhizogenes into the plant cells. The calli were co-cultivated with bacteria and subjected to selection media containing Hygromycin B. Plantlets were generated from embryogenic callus after 15 weeks, and T1 seeds were harvested 6-12 weeks later. The transformation efficiency, defined as the percentage of co-cultivated calli that generated plants, was higher in A. tumefaciens strain AGL1 (39%) than in A. rhizogenes strain 18r12v, which peaked at 96% before declining sharply after selection medium. Although further analysis is required to explain this observation, the results suggest that A. tumefaciens is a more reliable option for Brachypodium transformation, and that using full-concentration hygromycin medium can result in higher viability of embryogenic calli. These results could accelerate research in Brachypodium and have implications for the genetic improvement of biofuel crops.

Source coverage

This record comes from a published award list, not a complete project archive. Its abstract comes from CSEF's public project showcase as archived by the Internet Archive before judging (https://web.archive.org/web/20230401224130/https://ca-csef.zfairs.com/showcase/ShowcaseInfo?f=838e60b7-ea75-46e8-865c-fde4864244b3); the version presented may differ.

Awards (1)

Competition history

  • CSEF 2023 Plant Biology · Entry S1803

Resources

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