Location: Molecular Plant Pathology LaboratoryTitle: Pyramiding rice cystatin genes (OCI and OCII) in potato (Solanum tuberosum L cv. Jelica)) Author
Submitted to: Euphytica
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/14/2014
Publication Date: 4/26/2014
Citation: Cingel, A., Savic, J., Cosic, T., Momcilovic, I., Smigocki, A.C., Ninkovic, S. 2014. Pyramiding rice cystatin genes (OCI and OCII) in potato (Solanum tuberosum L cv. Jelica). Euphytica. DOI 10.1007/s10681-014-1119-z. Interpretive Summary: One of the major advances being used in current biotechnology to improve disease and pest control is the introduction of more than one trait (gene) into the plant genome, termed stacking or pyramiding. However, the manipulation of multiple genes in plants is still difficult to achieve and, therefore, more reliable gene pyramiding protocols for plants need to be developed. In the present study, we report on an adapted method for generating genetically modified potato that carries two beneficial genes. The method is based on using potato genetically modified with a single gene (OCI) which is then subjected to sequential modification with a second gene (OCII). Resulting potato shoots are rooted under highly controlled conditions that preferentially select for plants that carry both genes. Scientists will use this information to develop genetically modified plants that carry multiple traits for insect control that will increase yields and reduce usage of chemical pesticides.
Technical Abstract: One of the major advances being used in current biotechnology to improve disease and pest control is the introduction of more than one beneficial gene into transgenic plants. Proteinase inhibitors oryzacystatins I and II (OCI and OCII) show potential in controlling pests that utilize cysteine proteinases for protein digestion. To enhance resistance to pests in potato (Solanum tuberosum L cv. Jelica), OCI-expressing transgenic potato lines were re-transformed with Agrobacterium tumefaciens carrying the OCII gene. Regenerated shoots were rooted on high levels of kanaymcin (100 µg/ml). PCR analysis of 22 rooted re-transformed seedlings showed that 91% of them carried the OCII gene and had no disruption integrity of the previously introduced OCI gene. Induction of OCI and OCII gene transcripts and accumulation of biologically active OCI and OCII recombinant proteins was confirmed in all analyzed transgenic lines. This study demonstrated an efficient introduction of two different genes (OCI and OCII) by sequential re-transformation of an OCI-expressing transgenic potato line.