Phosphomannose Isomerase: An Efficient Selectable Marker for Plant Transformation

• Published in: In vitro cellular & developmental biology. Plant 2001-03, Vol.37 (2), p.127-132
• Main Authors: Janet Reed, Laura Privalle, M. Luann Powell, Moez Meghji, Dawson, John, Erik Dunder, Janet Suttie, Allan Wenck, Karen Launis, Catherine Kramer, Yin-Fu Chang, Hansen, Genevieve, Martha Wright
• Format: Article
• Language: English
• Subjects: Animal digestion; Barley; Corn; Enzymes; Food allergies; Glycoproteins; Plant cells; Plants; Special: Workshop: Alternative Markers for Plant Transformation: Workshop Presentations from the 2000 World Congress on In vitro Biology; Sugar beets; Transgenic plants
• ISSN: 1054-5476; 1475-2689
• DOI: 10.1007/s11627-001-0024-z

Phosphomannose isomerase (PMI) catalyzes the reversible interconversion of mannose 6-phosphate and fructose 6-phosphate. Plant cells lacking this enzyme are incapable of surviving on synthetic medium containing mannose as a carbon source. Maize, wheat and barley plants, genetically modified to express the Escherichia coli manA gene (pmi) under the control of a plant promoter, were able to survive selection on mannose-containing medium. Transformation frequencies averaged 45% for maize transformation via Biolistics™, 35% for maize Agrobacterium-mediated transformation, 20% for wheat, 3% for barley, and 2% for watermelon transformation. Moreover, the frequencies exceeded those obtained for maize and wheat using the pat or bar gene with Basta® selection. A preliminary safety assessment has been conducted for PMI. Purified PMI protein demonstrates no adverse effects in an acute mouse toxicity test. Purified PMI protein was readily digested in simulated mammalian gastric and intestinal fluids. Plants derived from sugar beet and corn cells that had been genetically modified to express the E. coli manA gene were evaluated for biochemical changes in mannose-associated pathways. No detectable changes in glycoprotein profiles were detected in PMI-transformed plants as compared to non-transgenic controls. The yield and nutritional composition of grain from PMI-transformed corn plants compared to their non-transformed isogenic counterparts were also determined and no statistically significant differences were found. The inherent safety of a system based on simple sugar metabolism coupled with high transformation frequencies for monocots make pmi an ideal selectable marker for plant transformation.