Establishing And Optimization Of Genetic Transformation System In Orychophragmus Violaceus

Orychophragmus violaceus is a crucifer with53.17%linoleic acid in its seed oil. Linoleic acid is the precursor for some unusual high-value fatty acids such as epoxy and acetylenic fatty acids. These fatty acids have unique potential performance and are precious resources of the renewable raw materials in the chemical industry. However, plant species with rich unusual fatty acids usually have very poor agronomic performance.In this study, we chose hypocotyls of Orychophragmus violaceus as explants and regenerated shoots on MS basic medium added with certain concentrations of NAA,6-BA, TDZ, ZT and AgNO3, and established an efficient regeneration system for Orychophragmus violaceus. Genes for enzymes of Epoxygenase and Acetylase that regulate the synthesis of epoxy and acetylenic fatty acids were introduced into Orychophragmus violaceus by using Agrobacterium-mediated transformation method. We also focused on some of the important factors such as types of explants, the concentration of kanamycin, the selection of infection time and concentration of Agrobacteria, the time length of pre-cultivation and co-cultivation and the selection of antibacterial agents that influence the efficiency of genetic transformation. In this study we established an efficient genetic transformation system by using hypocotyls of Orychophragmus violaceus for as explants for transformation. Finally, the genomic DNA of regenerated plantlets resistant to kanamycin were extracted and NPT Ⅱ fragment were amplified by PCR, suggesting that we might have obtained transgenic Orychophragmus violaceus.