Creation of Self-compatible Broccoli Lines by Knocking out BoSP11 Gene Through CRISPR/Cas9 Technology
By using CRISPR/Cas9 technology to knock out SRK gene or SP11 gene,the self-incompatible Brassica oleracea vegetables can be directly transformed into self-compatible materials.In this study,a double target CRISPR/Cas9 gene editing vector was constructed based on the sequence of broccoli(Brassica oleracea var.italic)BoSP11 gene obtained by whole genome sequencing,and then used Agrobacterium mediated genetic transformation to knock out the BoSP11 gene.The results showed that 23 of the 29 transgenic plants had gene mutation,and the editing efficiency was 79.3％.The results of TA clone sequencing showed that there were four homozygous mutations and 19 heterozygous mutations.The statistics of the seed setting of To generation of homozygous mutant plants showed that the self-compatibility index at flowering stage was as high as 6.82,which was the highest compatibility index increase reported so far.The T1 generation plants were pollinated by bees in the isolated greenhouses,and the average seed yield per plant was 24.8 g,which completely conducted the self-pollination of parent materials for seed production without any manual involvement.
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