In vitro selection of transgenic sugarcane callus utilizing a plant gene encoding a mutant form of acetolactate synthase

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dc.contributor.authorVan Der Vyver, Christellen_ZA
dc.contributor.authorConradie, Tobieen_ZA
dc.contributor.authorKossmann, Jensen_ZA
dc.contributor.authorLloyd, Jamesen_ZA
dc.date.accessioned2013-02-18T08:17:57Z
dc.date.available2013-02-18T08:17:57Z
dc.date.issued2013-02-05
dc.descriptionCITATION: Van Der Vyver, C., et al. 2013. In vitro selection of transgenic sugarcane callus utilizing a plant gene encoding a mutant form of acetolactate synthase. In Vitro Cellular and Developmental Biology - Plant, 49(2):198–206, doi:10.1007/s11627-013-9493-0.en_ZA
dc.descriptionThe original publication is available at http://www.springer.com/life+sciences/plant+sciences/journal/11627en_ZA
dc.description.abstractSelection genes are routinely used in plant genetic transformation protocols to ensure the survival of transformed cells by limiting the regeneration of non-transgenic cells. In order to find alternatives to the use of antibiotics as selection agents, we followed a targeted approach utilizing a plant gene, encoding a mutant form of the enzyme acetolactate synthase, to convey resistance to herbicides. The sensitivity of sugarcane callus (Saccharum spp. hybrids, cv. NCo310) to a number of herbicides from the sulfonylurea and imidazolinone classes was tested. Callus growth was most affected by sulfonylurea herbicides, particularly 3.6 μg/l chlorsulfuron. Herbicide-resistant transgenic sugarcane plants containing mutant forms of a tobacco acetolactate synthase (als) gene were obtained following biolistic transformation. Post-bombardment, putative transgenic callus was selectively proliferated on MS medium containing 3 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D), 20 g/l sucrose, 0.5 g/l casein, and 3.6 μg/l chlorsulfuron. Plant regeneration and rooting was done on MS medium lacking 2,4-D under similar selection conditions. Thirty vigorously growing putative transgenic plants were successfully ex vitro-acclimatized and established under glasshouse conditions. Glasshouse spraying of putative transgenic plants with 100 mg/l chlorsulfuron dramatically decreased the amount of non-transgenic plants that had escaped the in vitro selection regime. PCR analysis showed that six surviving plants were als-positive and that five of these expressed the mutant als gene. This report is the first to describe a selection system for sugarcane transformation that uses a selectable marker gene of plant origin targeted by a sulfonylurea herbicide.en_ZA
dc.description.versionPublisher's versionen_ZA
dc.format.extent11 pages
dc.identifier.citationVan Der Vyver, C., et al. 2013. In vitro selection of transgenic sugarcane callus utilizing a plant gene encoding a mutant form of acetolactate synthase. In Vitro Cellular and Developmental Biology - Plant, 49(2):198–206, doi:10.1007/s11627-013-9493-0en_ZA
dc.identifier.issn1475-2689 (online)
dc.identifier.issn1054-5476 (print)
dc.identifier.otherdoi:10.1007/s11627-013-9493-0
dc.identifier.urihttp://hdl.handle.net/10019.1/79439
dc.identifier.urihttp://link.springer.com/article/10.1007/s11627-013-9493-0
dc.language.isoen_ZAen_ZA
dc.publisherSpringeren_ZA
dc.rights.holderAuthors retain copyrighten_ZA
dc.subjectAcetolactate synthaseen_ZA
dc.subjectAcetolactate synthase mutationen_ZA
dc.subjectSugarcane transformationen_ZA
dc.subjectSulfonylurea herbicideen_ZA
dc.titleIn vitro selection of transgenic sugarcane callus utilizing a plant gene encoding a mutant form of acetolactate synthaseen_ZA
dc.typeArticleen_ZA
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