Enhanced production of single copy backbone-free transgenic plants in multiple crop species using binary vectors with a pRi replication origin in <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis> |
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Authors: | Xudong Ye Edward J Williams Junjiang Shen Susan Johnson Brenda Lowe Sharon Radke Steve Strickland James A Esser Michael W Petersen Larry A Gilbertson |
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Institution: | (1) Agracetus Campus, Monsanto Company, 8520 University Green, P. O. Box 620999, Middleton, WI 53562, USA;(2) Mystic Research, Monsanto Company, 62 Maritime Drive, Mystic, CT 06355, USA;(3) Calgene Campus, Monsanto Company, 1920 Fifth St, Davis, CA 95616, USA;(4) Monsanto Company, 700 Chesterfield Pkwy, St. Louis, MO 63017, USA |
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Abstract: | Single transgene copy, vector backbone-free transgenic crop plants are highly desired for functional genomics and many biotechnological
applications. We demonstrate that binary vectors that use a replication origin derived from the Ri plasmid of Agrobacterium rhizogenes (oriRi) increase the frequency of single copy, backbone-free transgenic plants in Agrobacterium tumefaciens mediated transformation of soybean, canola, and corn, compared to RK2-derived binary vectors (RK2 oriV). In large scale soybean
transformation experiments, the frequency of single copy, backbone-free transgenic plants was nearly doubled in two versions
of the oriRi vectors compared to the RK2 oriV control vector. In canola transformation experiments, the oriRi vector produced
more single copy, backbone-free transgenic plants than did the RK2 oriV vector. In corn transformation experiments, the frequency
of single copy backbone-free transgenic plants was also significantly increased when using the oriRi vector, although the
transformation frequency dropped. These results, derived from transformation experiments using three crops, indicate the advantage
of oriRi vectors over RK2 oriV binary vectors for the production of single copy, backbone-free transgenic plants using Agrobacterium-mediated transformation. |
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