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Comparison of selective agents for use with the selectable marker gene bar in maize transformation
Authors:Briana K Dennehey  William L Retersen  Colleen Ford-Santino  Mark Pajeau  Charles L Armstrong
Institution:(1) Monsanto Company, 700 Chesterfield Parkway North, 63198 St. Louis, MO;(2) Present address: Department of Biology, Indiana University, 47405 Bloomington, IN
Abstract:The effectiveness of four phosphinothricin (PPT)-based selective agents were evaluated for use in maize transformation: glufosinate, bialaphos, Basta® and Herbiace®. Glufosinate and its commercial formulation, Basta®, were less effective in controlling growth of non-transgenic corn callus than the tripeptide, bialaphos, or its commercial formulation, Herbiace®. Addition of 25 mM l-proline had no significant effect on selection when using bialaphos. However, when l-proline was included with the selective agent glufosinate, selection was inhibited and callus growth was enhanced. At four weeks, callus growth on 0.3, 1.0 and 3.0 mg l-1 glufosinate in the presence of proline was 76, 43, and 21% of control growth, respectively, and in the absence of proline was only 32, 9, and 6% of control growth. Optimized selection protocols for Basta® and bialaphos yielded comparable numbers of transformants. Using these protocols, fertile transgenic plants were regenerated from transformed callus cultures.Abbreviations AA amino acid - GS glutamine synthetase - PAT phosphinothricin acetyl transferase - PPT 2-amino-(methylphosphinyl)-butanoic acid (phosphinothricin) - 2,4-d 2,4-dichlorophenoxyacetic acid
Keywords:bialaphos  glufosinate  phosphinothricin  l-proline" target="_blank">l-proline  Zea mays
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