Developing a rapid and highly efficient cowpea regeneration, transformation and genome editing system using embryonic axis explants

• DOI: 10.1111/tpj.15202
• Publication Date: 2021-02-18T03:38:12Z
• Authors: Ping Che; Shujun Chang; Marissa K. Simon; Zhifen Zhang; Ahmed Shaharyar; Jesse Ourada; Dennis O’Neill; Mijael Torres‐Men...; Yinping Guo; Kathleen M. Maras...; Jean‐Philippe Vie...; Peggy Ozias‐Akins; Marc C. Albertsen; Todd J. Jones
• Keywords: Gene Editing; 0301 basic medicine; 2. Zero hunger; Vigna; Gene Transfer Techniques; Agrobacterium; Original Articles; Plants, Genetically Modified; 03 medical and health sciences; Transformation, Genetic; Seeds; Cotyledon; Genome, Plant; Plant Shoots

SUMMARYCowpea (Vigna unguiculata (L.) Walp.) is one of the most important legume crops planted worldwide, but despite decades of effort, cowpea transformation is still challenging due to inefficient Agrobacterium‐mediated transfer DNA delivery, transgenic selection and in vitro shoot regeneration. Here, we report a highly efficient transformation system using embryonic axis explants isolated from imbibed mature seeds. We found that removal of the shoot apical meristem from the explants stimulated direct multiple shoot organogenesis from the cotyledonary node tissue. The application of a previously reported ternary transformation vector system provided efficient Agrobacterium‐mediated gene delivery, while the utilization of spcN as selectable marker enabled more robust transgenic selection, plant recovery and transgenic plant generation without escapes and chimera formation. Transgenic cowpea plantlets developed exclusively from the cotyledonary nodes at frequencies of 4% to 37% across a wide range of cowpea genotypes. CRISPR/Cas‐mediated gene editing was successfully demonstrated. The transformation principles established here could also be applied to other legumes to increase transformation efficiencies.