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Hauptmann RM, Vasil V, Ozias-Akins P, et al. Evaluation of selectable markers for obtaining stable transformants in the gramineae. Plant Physiol. 1988;86(2):602-6doi: 10.1104/pp.86.2.602.
Hauptmann, R. M., Vasil, V., Ozias-Akins, P., Tabaeizadeh, Z., Rogers, S. G., Fraley, R. T., Horsch, R. B., & Vasil, I. K. (1988). Evaluation of selectable markers for obtaining stable transformants in the gramineae. Plant physiology, 86(2), 602-6. https://doi.org/10.1104/pp.86.2.602
Hauptmann, R M, et al. "Evaluation of selectable markers for obtaining stable transformants in the gramineae." Plant physiology vol. 86,2 (1988): 602-6. doi: https://doi.org/10.1104/pp.86.2.602
Hauptmann RM, Vasil V, Ozias-Akins P, Tabaeizadeh Z, Rogers SG, Fraley RT, Horsch RB, Vasil IK. Evaluation of selectable markers for obtaining stable transformants in the gramineae. Plant Physiol. 1988 Feb;86(2):602-6. doi: 10.1104/pp.86.2.602. PMID: 16665953; PMCID: PMC1054530.
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Plant Physiol. 1988 Feb;86(2):602-6. doi: 10.1104/pp.86.2.602.

Evaluation of selectable markers for obtaining stable transformants in the gramineae.

Plant physiology

R M Hauptmann, V Vasil, P Ozias-Akins, Z Tabaeizadeh, S G Rogers, R T Fraley, R B Horsch, I K Vasil

Affiliations

  1. Department of Vegetable Crops, 511 Bartram Hall West, University of Florida, Gainesville, Florida 32611.

PMID: 16665953 PMCID: PMC1054530 DOI: 10.1104/pp.86.2.602

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.

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