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The use of  Baby boom  ( Bbm ) and  Wuschel2  ( Wus2 ) has made maize transformation more efficient across an increasingly wide range of inbreds. However, the benefits have come with the requirement of excising these transformation helper components to enable plant regeneration, which adds size to the T-DNA, and complexity to the transformation system. A new system with the advantages of smaller size and simplicity for the selectable marker gene-containing T-DNA is described. First, expression of  Zm-Wus2  alone driven by the maize  Pltp  promoter ( Zm - Pltp   pro  ), was determined to be sufficient to induce rapid somatic embryo formation from the scutella of maize immature embryos. It was also demonstrated that co-infecting with two strains of  Agrobacterium , one with a  Wus2  expression cassette, and the other with a combination of both selectable and visual marker cassettes, produced transformed T0 plants that contained only a single copy of the selectable marker T-DNA, without the integration of  Wus2 . Furthermore, the process was optimized by varying the ratio of the two  Agrobacterium  strains, and by modulating  Wus2  expression to enable high-frequency recovery of selectable marker-containing T0 plants that did not contain  Wus2 . Several factors may have contributed to this outcome.  Wus2  expression in localized cell(s) appeared to stimulate somatic embryogenesis in neighboring cells, including those that had integrated the selectable marker. In addition, in cells in which the  Wus2  T-DNA did not integrate but the selectable marker T-DNA did, transient  Wus2  expression stimulated somatic embryo formation and regeneration of stable T0 plants that contained the selectable marker. In addition, augmenting the  Pltp  promoter with three viral enhancer elements to increase  Wus2  expression stimulated embryogenesis while precluding their regeneration. The phenomenon has now been designated as “altruistic transformation.”

Use of non-integrating Zm-Wus2 vectors to enhance maize transformation