caa a22 4500 46790149X CHVBK 20180406152819.0 cr unu---uuuuu 170328e20061101xx s 000 0 eng 10.1007/s00299-006-0179-5 doi (NATIONALLICENCE)springer-10.1007/s00299-006-0179-5 The use of the phosphomannose-isomerase/mannose selection system to recover transgenic apple plants [Elektronische Daten] [Juliana Degenhardt, Annika Poppe, Jurith Montag, Iris Szankowski] A selection system based on the phosphomannose-isomerase gene (pmi) as a selectable marker and mannose as the selective agent was evaluated for the transformation of apple (Malus domestica Borkh.). Mannose is an unusable carbon source for many plant species. After uptake, mannose is phosphorylated by endogenous hexokinases to mannose-6-phosphate. The accumulation of mannose-6-phosphate leads to a block in glycolysis by inhibition of phosphoglucose-isomerase, resulting in severe growth inhibition. The phosphomannose-isomerase is encoded by the manA gene from Escherichia coli and catalyzes the conversion of mannose-6-phosphate to fructose-6-phosphate, an intermediate of glycolysis. Transformed cells expressing the manA gene can therefore utilize mannose as a carbon and survive on media containing mannose. The manA gene along with a β-glucuronidase (GUS) gene was transferred into apple cv. ‘Holsteiner Cox' via Agrobacterium tumefaciens-mediated transformation. Leaf explants were selected on medium supplemented with different concentrations and combinations of mannose and sorbitol to establish an optimized mannose selection protocol. Transgenic lines were regenerated after an initial selection pressure of 1-2gl−1 mannose in combination with 30gl−1 sorbitol followed by a stepwise increase in the mannose concentration up to 10gl−1 and simultaneous decrease in the sorbitol concentration. Integration of transgenes in the apple genome of selected plants was confirmed by PCR and southern blot analysis. GUS histochemical and chlorophenol red (CPR) assays confirmed activity of both transgenes in regenerated plants. The pmi/mannose selection system is shown to be highly efficient for producing transgenic apple plants without using antibiotics or herbicides. Springer-Verlag, 2006 Agrobacterium tumefaciens nationallicence Chlorophenol red nationallicence GUS nationallicence Malus domestica Borkh nationallicence Positive selection nationallicence Degenhardt Juliana Institute of Biological Production Systems, Fruit Science Section, University of Hannover, Herrenhaeuser Str. 2, 30419, Hannover, Germany aut Poppe Annika Institute of Biological Production Systems, Fruit Science Section, University of Hannover, Herrenhaeuser Str. 2, 30419, Hannover, Germany aut Montag Jurith Institute of Biological Production Systems, Fruit Science Section, University of Hannover, Herrenhaeuser Str. 2, 30419, Hannover, Germany aut Szankowski Iris Institute of Biological Production Systems, Fruit Science Section, University of Hannover, Herrenhaeuser Str. 2, 30419, Hannover, Germany aut Plant Cell Reports Springer-Verlag 25/11(2006-11-01), 1149-1156 0721-7714 25:11<1149 2006 25 299 https://doi.org/10.1007/s00299-006-0179-5 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00299-006-0179-5 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Degenhardt Juliana Institute of Biological Production Systems, Fruit Science Section, University of Hannover, Herrenhaeuser Str. 2, 30419, Hannover, Germany aut NATIONALLICENCE 700 1- Poppe Annika Institute of Biological Production Systems, Fruit Science Section, University of Hannover, Herrenhaeuser Str. 2, 30419, Hannover, Germany aut NATIONALLICENCE 700 1- Montag Jurith Institute of Biological Production Systems, Fruit Science Section, University of Hannover, Herrenhaeuser Str. 2, 30419, Hannover, Germany aut NATIONALLICENCE 700 1- Szankowski Iris Institute of Biological Production Systems, Fruit Science Section, University of Hannover, Herrenhaeuser Str. 2, 30419, Hannover, Germany aut NATIONALLICENCE 773 0- Plant Cell Reports Springer-Verlag 25/11(2006-11-01), 1149-1156 0721-7714 25:11<1149 2006 25 299 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer