Cloning and Characterization of Responsible Gene for Erucic Acid Biosynthesis in Brassica napus

The Brassica genus classified into two basic groups: High erucic acid rapeseed (HEAR) and Low erucic acid rapeseed (LEAR). Oils having high levels of erucic acid have found widespread applications for non-edible purposes and oils with low levels of erucic acid have used for edible purposes. In this study, the gene and its promoter of β -ketoacyl-CoA synthase (KCS) enzyme that responsible for erucic acid biosynthesis were isolated and their effect was investigated on rate of erucic acid content of transgenic canola. Specific primers were used to amplify fae gene and its specific promoter from genomic DNA by PCR technique. The putative gene and promoter from B. napus were cloned into pSK+ vector and sequenced in both directions. Sense construct from fae gene and its promoter cloned into a plant expression vector, pBI121. Recombinant vector transferred to rapeseed plant via Agrobacterium-mediated transformation method. Presence of gene construct in transgenic plants was confirmed with PCR, dot blot and Southern blotting analyses. Finally the transgene function and its effect on erucic acid content were measured with gas chromatography in transgenic and control plants. Four putative transgenic plants were shown an increase in erucic acid level. Maximum of increasing was observed in one sample (No. 4) with 13% C22:1 in comparison with 1.1% in control plant.