Evaluation of Selectable Markers for Obtaining Stable Transformants in the Gramineae
Citations Over TimeTop 10% of 1988 papers
Abstract
Cell suspension cultures of Triticum monococcum, Panicum maximum, Saccharum officinarum, Pennisetum americanum, and a double cross trispecific hybrid between Pennisetum americanum, P. purpureum, and P. squamulatum were tested for resistance to kanamycin, hygromycin, and methotrexate for use in transformation studies. All cultures showed high natural levels of resistance to kanamycin, in excess of 800 milligrams per liter, and variable levels of resistance to hygromycin. Methotrexate was a potent growth inhibitor at low concentrations with all species. Kanamycin and hygromycin were growth inhibitory only if added early (within 5 days after protoplast isolation and culture). Protoplasts of T. monococcum, P. maximum, S. officinarum, and the tri-specific hybrid were electroporated with plasmid DNA containing hygromycin (pMON410), kanamycin (pMON273), or methotrexate (pMON806) resistance genes. Resistant colonies were obtained at low frequencies (1 x 10(-5) to 2 x 10(-6)) when selected under conditions which were growth inhibitory to protoplasts electroporated without DNA. Southern blot hybridization confirmed stable integration of plasmid DNA into T. monococcum using hygromycin vectors and P. maximum using the methotrexate vector with 1 to 10 copies integrated per haploid genome.
Related Papers
- → The isopentenyl transferase gene is effective as a selectable marker gene for plant transformation in tobacco ( Nicotiana tabacum cv. Petite Havana SRI)(2001)51 cited
- → Hygromycin resistance as an efficient selectable marker for wheat stable transformation(1996)82 cited
- → Kanamycin in Cereal Biotechnology: A Screening or a Selectable Marker?(2021)1 cited
- HIBRIDISASI SOMATIK INTERGENERIK ANGGREK Dendrobium striaenopsis DAN Phalaenopsis amboinensis MELALUI FUSI PROTOPLAS(2019)
- → Kanamycin in Cereal Biotechnology: A Screening or a Selectable Marker?(2021)