title: Rapid and efficient Agrobacterium-mediated transformation of sorghum (Sorghum bicolor) employing standard binary vectors and bar gene as a selectable marker.
abstract: A rapid and efficient Agrobacterium -mediated transformation system in sorghum has been developed employing standard binary vectors and bar gene as a selectable marker. Sorghum (Sorghum bicolor) is an important food and biofuel crop worldwide, for which improvements in genetic transformation are needed to study its biology and facilitate agronomic and commercial improvement. Here, we report optimization of regeneration and transformation of public sorghum genotype P898012 using standard binary vectors and bar gene as a selectable marker. The tissue culture regeneration time frame has been reduced to 7-12 weeks with a yield of over 18 plants per callus, and the optimized transformation system employing Agrobacterium tumefaciens strain AGL1 and the bar with a MAS promoter achieved an average frequency over 14 %. Of randomly analyzed independent transgenic events, 40-50 % carry single copy of integrated T-DNA. Some independent transgenic events were derived from the same embryogenic callus lines, but a 3:1 Mendelian segregation ratio was found in all transgenic events with single copy as estimated by Southern blots. The system described here should facilitate studies of sorghum biology and agronomic improvement.
authors.affiliation: Plant Transformation Core Facility, Division of Plant Sciences, University of Missouri, 1-33 Agriculture Building, Columbia, MO, 65211, USA.
authors.forename: Phat T
authors.initials: PT
authors.lastname: Do
authors.affiliation: Plant Transformation Core Facility, Division of Plant Sciences, University of Missouri, 1-33 Agriculture Building, Columbia, MO, 65211, USA.
authors.forename: Hyeyoung
authors.initials: H
authors.lastname: Lee
authors.affiliation: Plant Transformation Core Facility, Division of Plant Sciences, University of Missouri, 1-33 Agriculture Building, Columbia, MO, 65211, USA.
authors.forename: Muruganantham
authors.initials: M
authors.lastname: Mookkan
authors.affiliation: Department of Biochemistry, University of Missouri, Columbia, MO, 65211, USA.
authors.forename: William R
authors.initials: WR
authors.lastname: Folk
authors.affiliation: Plant Transformation Core Facility, Division of Plant Sciences, University of Missouri, 1-33 Agriculture Building, Columbia, MO, 65211, USA. zhangzh@missouri.edu.
authors.forename: Zhanyuan J
authors.initials: ZJ
authors.lastname: Zhang
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics*; D015139:Blotting, Southern; D020090:Chromosome Segregation / Q000235:genetics; D005798:Genes, Bacterial; D005819:Genetic Markers; D005822:Genetic Vectors / Q000378:metabolism*; D005838:Genotype; D005966:Glucuronidase / Q000378:metabolism; D006540:Herbicides / Q000633:toxicity; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic / Q000235:genetics; D012038:Regeneration; D045868:Sorghum / Q000235:genetics; D013194:Staining and Labeling; D046509:Tissue Culture Techniques; D014170:Transformation, Genetic
chemical_list: D005819:Genetic Markers; D006540:Herbicides; D005966:Glucuronidase
keywords: Agrobacterium; Bar; Plant transformation; Sorghum; Standard binary vectors