Transformation of maize cells and regeneration of fertile transgenic plants

• Published in: The Plant cell Vol. 2; no. 7; pp. 603 - 618
• Main Authors: Gordon-Kamm, W.J, Spencer, T.M, Mangano, M.L, Adams, T.R, Daines, R.J, Start, W.G, O'Brien, J.V, Chambers, S.A, Adams, W.R. Jr, Willetts, N.G
• Format: Journal Article
• Language: English
• Published: United States American Society of Plant Physiologists 01-07-1990
• Subjects: acyltransferases; bar gene; beta-glucuronidase; bilanafos; Callus; Cell lines; cell suspension culture; Corn; Cryopreservation; Cultured cells; DNA; embryo (plant); Embryonic cells; enzyme activity; fertility; gene transfer; genes; genetic transformation; genetically modified organisms; herbicide resistance; in vitro selection; microprojectile bombardment; nucleic acid hybridization; phosphothricin acetyltransferase; Plant cells; Plants; regenerative ability; reporter genes; Streptomyces; Streptomyces hygroscopicus; tissue culture; Transgenic plants; Zea mays
• ISSN: 1040-4651; 1532-298X
• DOI: 10.1105/tpc.2.7.603

A reproducible system for the generation of fertile, transgenic maize plants has been developed. Cells from embryogenic maize suspension cultures were transformed with the bacterial gene bar using microprojectile bombardment. Transformed calli were selected from the suspension cultures using the herbicide bialaphos. integration of bar and activity of the enzyme phosphinothricin acetyltransferase (PAT) encoded by bar were confirmed in all bialaphos-resistant callus lines. Fertile transformed maize plants (R0) were regenerated, and of 53 progeny (R1) tested, 29 had PAT activity. All PAT-positive progeny analyzed contained bar. Localized application of herbicide to leaves of bar-transformed R0 and R1 plants resulted in no necrosis, confirming functional activity of PAT in the transgenic plants. Cotransformation experiments were performed using a mixture of two plasmids, one encoding PAT and one containing the nonselected gene encoding beta-glucuronidase. R0 plants regenerated from cotransformed callus expressed both genes. These results describe and confirm the development of a system for introduction of DNA into maize.