Genotype-Independent Transformation and Genome Editing of Brassica napus Using a Novel Explant Material

Abstract

Agrobacterium -mediated transformation of canola ( Brassica napus ) via hypocotyl segments has been a commonly used method for the past 30 years. While the hypocotyl-based method is well-established, it is not readily adapted to elite germplasm and the prolonged process is not ideal for a production transformation setting. We developed an Agrobacterium -mediated transformation method based on epicotyl and higher stem (internodal) segments that is efficient, rapid and amenable for high-throughput transformation and genome editing. The method has been successfully implemented in multiple canola genotypes. The method appears to be genotype-independent, with varying transformation efficiencies. Internodal segment transformation was used to generate transgenic events as well as CRISPR-Cas9-mediated frameshift gene knockouts.

Editorial: Genome editing technology in polyploid crops

“…). Similarly, to achieve genotype-independent transformation, a comparable strategy has been implemented in Brassica napus , utilizing epicotyl and higher stem (internodal) segments as recipient material ( Chu et al., 2020 ). Furthermore, the utilization of regeneration-related genes, such as WUSCHEL ( WUS2 ), BABY BOOM ( BBM ), and SHOOT MERISTEMLESS ( STM ), holds great promise for substantially improving the efficiency of plant transformation ( Lowe et al., 2016 ).…”

“…One article featured in this Research Topic presents an improved CRISPR/Cas9-based transformation protocol in highbush blueberry, a polyploid woody species, through the utilization of de novo shoot organogenesis (Vaia et al). Similarly, to achieve genotype-independent transformation, a comparable strategy has been implemented in Brassica napus, utilizing epicotyl and higher stem (internodal) segments as recipient material (Chu et al, 2020). Furthermore, the utilization of regeneration-related genes, such as WUSCHEL (WUS2), BABY BOOM (BBM), and SHOOT MERISTEMLESS Frontiers in Plant Science frontiersin.org 01…”