Cloning and Functional Analysis of Lateral Branching Regulatory Gene BoBRC1 in Broccoli
In this study,the BoBRC1 gene was cloned from the elite broccoli inbred line L461.Expression pattern analysis showed that BoBRC1 had the highest expression level in axillary bud tissues,and changed dynamically with axillary bud growth.According to the sequence of BoBRC1,two targets were designed on the first exon,and a CRISPR/Cas9 gene editing vector was constructed.The construct was introduced into L461 by Agrobacterium-mediated genetic transformation,and a total of 18 plants were confirmed as positive transgenic.The results showed that three plants harbored mutations in at least one of the targeted regions corresponding to an editing efficiency of 16％.Among them L461-3 was edited in both Target 1 and Target 2 while L461-1 and L461-11 were edited in Target 2.Compared with the wild type,the homologous editing mutants showed a significant increase in the number and length of lateral branches.In the bobrc1 mutant,qRT-PCR analysis of key genes downstream of lateral branch gene BoBRC1 showed a significant decrease in the expression of the BoHB21,BoHB53 and BoNCED3 genes while the expression of the BoHB40 significantly increased.This study verified the function of BoBRC1 gene in the regulation of lateral branch development in broccoli,and knocking out can promote the growth of broccoli lateral branches.
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