Alteration of fatty acid biosynthesis pathway to reduce erucic acid in mustard (Brassica rapa L.) by genome editing approach

Oilseed Brassicas are the 3^rd largest edible oil source in the world. Despite being so, their fatty acid profiles have shown high levels of erucic acid content which is detrimental for human health. Erucic acid can cause myocardial damage and muscle lesions in the heart if taken frequently. This issue prevents the worldwide use of mustard oil as an ideal edible oil and its global commercial export. Genome editing technology have been a pioneer in producing smart varieties of crop which are either adaptable to the current changing climate or capable of ensuring public health. Previous reports show that, the Fatty Acyl CoA Elongase 1 (FAE1) gene is the key gene for the synthesis of erucic acid. The product of which is 3-ketoacyl-CoA synthase, one of the four enzyme complexes involved in the synthesis of erucic acid from oleic acid. Brassica rapa has two functional homologues of this gene on its A08 and C03 chromosomes. By using CRISPR (Clustered Regularly Interspaced Palindromic Repeats) associated protein Cas9 genome editing technology it is possible to knockout the expression of this gene and to produce low erucic acid producing novel B. rapa plants. To do so, analysis of fatty acid profile of the local Tori-7 variety of B. rapa has been done along with in-silico analysis and molecular characterization of the said genotype. SgRNAs have been designed to target our two homologues of the FAE1 gene and integrated into the pAGM8031_fae1 vector via modular cloning method. Through Agrobacterium-mediated genetic transformation method, novel mutant lines are being developed with the goal to conduct molecular analysis on the transgenic lines for our desired mutation and establishing a homozygous line of genome edited, low erucic acid producing B. rapa plants which can ensure safety of public health if released commercially.