Construction of Plant Expression Vectors Harboring WRI1 and LPAAT Genes and Its Transformation in Tobacco Plants

In oilseed crops, a number of genes involved in the production of triacylglycerol have been identified that changes in their expression have increase the seed oil content. WRI1 and LPAAT are key genes in this synthetic pathway that their overexpression can increase the oil content. In this study, the expression vectors carrying WRI1 and LPAAT genes were designed and constructed for genetic transformation of tobacco (Nicotiana tabaccum) plants. The synthetic WRI1 and LPAAT genes were isolated from the PGH.WRI1 and PGH.LPAAT cloning vector using specific restriction enzymes and then cloned in the PGH.O3.2.2 intermediate vector under the control of SBP and Napin promoters and E9 terminator. Finally, the genetic cassettes were transferred to the plant transformation pBin19 binary vector. The resulting constructs were transferred to Agrobacterium tumefacience strain EHA105 and were used for genetic transformation of tobacco plants. Molecular analysis of transgenic plants confirmed the presence and activity of WRI1 and LPAAT genes. Seeds from transgenic plants were selected on the medium containing kanamycin and developed strong and healthy seedlings.