The combination of morphogenic regulators BABY BOOM and GRF-GIF improves maize transformation efficiency

Transformation is an indispensable tool for plant genetics and functional genomic studies. Although stable transformation no longer represents a major technology bottleneck in maize, there is still need for easily accessible and efficient transformation methods in most academic labs. Here we present the GGB transformation system, a rapid and highly efficient transformation system optimized for the immature embryo transformation of two maize genetic backgrounds, including the inbred line B104. The combination of distinct morphogenetic factors, the maize BABY BOOM transcriptional regulator (ZmBBM/EREB53) and the wheat GRF4-GIF1 (GROWTH REGULATING FACTOR4 - GRF-INTERACTING FACTOR1) chimera, together with a modified QuickCorn protocol, regenerated transformed maize seedlings in approximately two months with an efficiency of 26 to 37%; notably, the efficiency was 7-fold higher than with using either component in isolation. Additionally, ectopic expression of both morphogenetic factors did not show obvious effects on B104 development, and in particular fertility was not affected, obviating the need to remove the morphogenetic regulators post Agrobacterium infections. The GGB transformation system is designed for CRISPR-Cas9 editing but can be adapted for other purposes and should be easy to implement in most academic labs with little transformation experience. ### Competing Interest Statement J.M.D. is co-inventor in patent US2017/0362601A1 that describes the use of chimeric GRF GIF proteins with enhanced effects on plant growth (Universidad Nacional de Rosario Consejo Nacional de Investigaciones Cientificas y Tecnicas). J.D. and J.M.D. are co-inventors in UC Davis patent application WO2021007284A2 that describes the use of GRF GIF chimeras to enhance regeneration efficiency in plants.