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ISHS Acta Horticulturae 929: XXVIII International Horticultural Congress on Science and Horticulture for People (IHC2010): International Symposium on Genomics and Genetic Transformation of Horticultural Crops

EFFICIENCY OF PMI/MANNOSE AS A SELECTABLE MARKER FOR TRANSFORMATION OF KALANCHOE

Authors:   A. Ilczuk, H. Mibus, M. Schmidt , M. Serek
Keywords:   Agrobacterium tumefaciens, phosphomannose isomerase, selection agent
Abstract:
Mannose is an unsuitable carbon source for many plants. Various factors were studied to evaluate the efficacy of mannose in the selection process for production of transgenic plants using Agrobacterium tumefaciens-mediated transformation of leaf explants of Kalanchoe blossfeldiana. To establish a non-antibiotic selection system that utilizes the phosphomannose-isomerase (PMI) gene from E. coli, we determined the optimum mannose concentration for selecting transformed cells. Shoot organogenesis in K. blossfeldiana leaf explants was suppressed effectively on medium supplemented with mannose, or mannose with glucose, which indicated that Kalanchoe cells had little or no PMI activity and could not utilize mannose as a carbon source. The optimum concentration of mannose was found to eliminate non-transgenic shoots. Transformed plants were confirmed by GUS assay. Transformation efficiency ranged up to ca. 40%. The PMI/mannose system provides an efficient method for selecting transgenic plants without antibiotics or herbicides.
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