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item Smigocki, Anna

Submitted to: In Vitro Cellular And Developmental Biology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/26/2003
Publication Date: 11/1/2003
Citation: Ivic, S.D., Smigocki, A.C. 2003. Transformation of sugar beet cell suspension cultures. In Vitro Cellular And Developmental Biology. 39(6):1-6.

Interpretive Summary: Disease and pest problems are responsible for decreases in production of sugar from sugar beet, an economically important crop that provides more than 35% of the world's sugar supply. Conventional breeding has had little success for improving disease resistance, therefore, we are developing genetic engineering approaches for transferring known disease resistance genes from other living organisms to sugar beet. In this report, we summarize the progress we made in the development of improved technology for transfer of genes to commercially important sugar beet varieties. We prepared suspensions of cells derived from sugar beet leaves and showed that they are more susceptible to the uptake of genes (DNA). We were able to generate a large number of these cells in a relatively short period of time and showed that foreign genes were taken up and incorporated into the sugar beet chromosomes. The progress described in this report will be of benefit to scientists interested in employing genetic engineering approaches for increasing sugar yields from sugar beet.

Technical Abstract: A sugar beet transformation method was developed using particle bombardment of short-term suspension cultures of a breeding line FC607. Highly embryogenic suspension cultures derived from leaf callus were bombarded with the uidA (GUS) reporter gene under the control of either the osmotin or proteinase inhibitor II gene promoter, and the npt II selectable marker gene. Transient GUS expression was 500 to 4000 blue units per 0.2 g of bombarded cells two days after bombardment. Stably transformed calli were recovered on both kanamycin and paromomycin media. The greatest number of GUS (+) calli was obtained when high levels of Km (50-100 mg/l) were applied two days after transformation for three to five weeks, followed by either no selection or reduced levels of the antibiotic. Several transformed embryos were detected by histochemical GUS staining. Stable incorporation of the GUS gene into the genome was confirmed by Southern blot analyses.