title: OptimizedAgrobacterium-mediated sorghum transformation protocol and molecular data of transgenic sorghum plants.
abstract: Agrobacterium-mediated sorghum transformation frequency has been enhanced significantlyviamedium optimization using immature embryos from sorghum variety TX430 as the target tissue. The new transformation protocol includes the addition of elevated copper sulfate and 6-benzylaminopurine in the resting and selection media. UsingAgrobacteriumstrain LBA4404, the transformation frequency reached over 10% using either of two different selection marker genes, moPAT or PMI, and any of three different vectors in large-scale transformation experiments. WithAgrobacteriumstrain AGL1, the transformation frequencies were as high as 33%. Using quantitative PCR analyses of 1,182 T0transgenic plants representing 675 independent transgenic events, data was collected for T-DNA copy number, intact or truncated T-DNA integration, and vector backbone integration into the sorghum genome. A comparison of the transformation frequencies and molecular data characterizing T-DNA integration patterns in the transgenic plants derived from LBA4404 versus AGL1 transformation revealed that twice as many transgenic high-quality events were generated when AGL1 was used compared to LBA4404. This is the first report providing molecular data for T-DNA integration patterns in a large number of independent transgenic plants in sorghum.
authors: DuPont Agricultural Biotechnology, DuPont Pioneer, 8305 NW 62nd Avenue, P. O. Box 7060, Johnston, IA 50131 USA.
authors: Emily
authors: E
authors: Wu
authors: DuPont Agricultural Biotechnology, DuPont Pioneer, 8305 NW 62nd Avenue, P. O. Box 7060, Johnston, IA 50131 USA.
authors: Brian
authors: B
authors: Lenderts
authors: DuPont Agricultural Biotechnology, DuPont Pioneer, 8305 NW 62nd Avenue, P. O. Box 7060, Johnston, IA 50131 USA.
authors: Kimberly
authors: K
authors: Glassman
authors: 1034 Cambridge Way Drive, Chesterfield, MO 63017 USA.
authors: Maya
authors: M
authors: Berezowska-Kaniewska
authors: DuPont Agricultural Biotechnology, DuPont Pioneer, 8305 NW 62nd Avenue, P. O. Box 7060, Johnston, IA 50131 USA.
authors: Heather
authors: H
authors: Christensen
authors: DuPont Agricultural Biotechnology, DuPont Pioneer, 8305 NW 62nd Avenue, P. O. Box 7060, Johnston, IA 50131 USA.
authors: Tracy
authors: T
authors: Asmus
authors: DuPont Agricultural Biotechnology, DuPont Pioneer, 8305 NW 62nd Avenue, P. O. Box 7060, Johnston, IA 50131 USA.
authors: Shifu
authors: S
authors: Zhen
authors: DuPont Agricultural Biotechnology, DuPont Pioneer, 8305 NW 62nd Avenue, P. O. Box 7060, Johnston, IA 50131 USA.
authors: Uyen
authors: U
authors: Chu
authors: DuPont Agricultural Biotechnology, DuPont Pioneer, 4010 Point Eden Way, Hayward, CA 94545 USA.
authors: Myeong-Je
authors: MJ
authors: Cho
authors: DuPont Agricultural Biotechnology, DuPont Pioneer, 8305 NW 62nd Avenue, P. O. Box 7060, Johnston, IA 50131 USA.
authors: Zuo-Yu
authors: ZY
authors: Zhao
keywords: Agrobacterium; Sorghum; T-DNA insertion pattern; Transformation; Transgenic plants