Maize Transformation Using the Morphogenic Genes Baby Boom and Wuschel2

Abstract

Despite the fact that maize transformation has been available for over 25 years, the technology has remained too specialized, labor-intensive, and inefficient to be useful for the majority of academic labs. Compounding this problem, future demands in maize genome engineering will likely require a step change beyond what researchers view as "traditional" maize transformation methods. Recently, we published on our use of constitutively expressed morphogenic transcription factors Baby Boom (Bbm) and Wuschel2 (Wus2) to improve maize transformation, which requires CRE-mediated excision before regeneration of healthy, fertile T0 plants. Moving beyond this first-generation system, we have developed a new expression system for Bbm and Wus2, using a non-constitutive maize phospholipid transferase protein promoter (Pltp _pro) driving Bbm expression and a maize auxin-inducible promoter (Axig1 _pro) for WUS2 expression. Using this combination of expression cassettes, abundant somatic embryos rapidly form on the scutella of Agrobacterium-transformed zygotic immature embryos. These somatic embryos are uniformly transformed and can be directly germinated into plants without a callus phase. Transformed plants are sent to the greenhouse in as little as 1 month, and these T0 plants match the seed-derived phenotype for the inbred and are fertile. T1 seeds germinate normally and have a uniformly wild-type inbred phenotype. This new system represents a rapid, user-friendly transformation process that can potentially facilitate high-throughput production of transgenic T0 plants in B73, Mo17, and the recently developed Fast-Flowering Mini-Maize.

Keywords: Axig1 pro; Bbm; Maize; Morphogenic genes; PLTP pro; Transformation; Wus2.