AbstractBecause of its high-quality seed protein and ability to thrive in marginal habitats, Chenopodium quinoa has been identified as an important emerging grain crop for global food security. However, the lack of an efficient and robust transformation system has been a barrier for conducting the advanced genetic studies needed to better understand and improve the species for agronomic traits. Here we present a novel transformation system based on Agrobacterium-mediated transformation of in vitro floral culture. Quinoa floral cultures were established from inflorescences that naturally formed on plants grown in vitro. When placed on a cytokinin-containing medium, chopped inflorescences rapidly generated highly meristematic floral cultures, primarily composed of floral buds, flowers at various developmental stages, inflorescence shoots, and leafy structures. Transformation of these cultures with Agrobacterium carrying selectable and visual markers (NPTII and GUS) produced independent, stably transformed meristematic cultures resistant to paromomycin after an extended selection period (about 3 mo with sub-culture occurring every 15 d). Transformation frequency was about 20% and was calculated as the number of independent transformed events per initial number of floral culture explants used for transformation. In vitro flowers and inflorescences from putative transgenic events self-pollinated naturally and produced viable seeds that germinated without dormancy. We also demonstrated that flowering shoots could be successfully grafted onto wild rootstock to increase the number of seeds generated from T0 floral shoots. Molecular and phenotypic analysis of the progeny confirmed that the transgenes were stably integrated and inherited according to expected Mendelian ratios.
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