For the purpose of reducing lipoxygenase 3 synthesis, a construct was developed in which two complementary, internal fragments of the SnLOX3 gene were arranged in sense and in antisense orientation, separated by a spacer and transformed back into S. nigrum plants. These fragments induce RNA-silencing, which reduces the steady-state levels of SnLOX3 mRNA in the transgenic plants, thus reducing the production of the SnLOX3 protein. Silencing has been engineered and is effected by constitutive expression (enabled by the CaMV 35S promoter) of an antisense-intron-sense gene cassette. Briefly, the mechanism for gene silencing in this GMO include: (1) splicing of the intron 3 from the pyruvate-orthophosphate-dikinase gene (pdk i3) from Flaveria trinervia; (2) hybridization of the RNA of the two SnLOX3 fragments (sense and anti-sense) forming double-stranded RNA molecules (which are unstable in the cells); and (3) degradation of the double-stranded RNA molecules leading to the so-called post-transcriptional gene silencing which is expected to limit the formation of endogenous lipoxygenase 3 and, therewith, the formation of jasmonate.

Agrobacterium tumefaciens was used to insert T-DNA into plant nuclei. A hygromycin resistance gene aph4 from Escherichia coli under the control of the Pnos promoter was utilized to select for transgenic plants. This transgenic genotype bears one copy of the T-DNA and does not contain complete sequences of the nptIII gene.

There were no previous modifications of these plants.