Direct modification of multiple gene homoeologs in Brassica oleracea and Brassica napus using doubled haploid inducer-mediated genome-editing system.

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Thediploid Brassica oleracea and allopolyploidy Brassica napus are predominant members of commonly consumed vegetables and plant oil, respectively. B. oleracea vegetables mainly include Broccoli, Cauliflower, Cabbage, Brussels sprouts and Kohlrabi. The complex genome structure and gene function redundancy are the main obstacles for gene stacking through traditional cross breeding approach. To solve this problem, high-efficiency CRISPR/Cas9 genome editing technologies have been established (Lawrenson et al., 2015; Li et al., 2018). However, an open question is that most of these established approaches employed Agrobacterium-mediated T-DNA transformation to deliver CRISRP/Cas9 components into plant cells, which would unavoidably introduce exogenous large DNA fragments. Moreover, Agrobacterium-based strategy deeply relies on transformation efficiency of the recipient genotype, which extremely restricts rapid application of CRISRP/Cas9 in the majority of elite commercial verities.