Skip to main content Accessibility help
×
Home

Expression in Plants of a Bacterial Gene Coding for Glyphosate Resistance

  • Gregory A. Thompson (a1), William R. Hiatt (a1), Daniel Facciotti (a1), David M. Stalker (a1) and Luca Comai (a1)...

Abstract

The target site of glyphosate [N-(phosphonomethyl)glycine] inhibition in plants and bacteria is 5-enolpyruvylshikimate 3-phosphate (EPSP) synthase. Our strategy for developing glyphosate-resistant crops has been to genetically engineer plants with a gene that codes for EPSP synthase with low sensitivity in glyphosate. We cloned such a gene from the aroA locus of a glyphosate-resistant mutagenized strain of Salmonella typhimurium. The enzyme encoded by this gene has a single amino acid change resulting in lower affinity for glyphosate and higher affinity for substrates than either plant or wild-type bacterial counterpart. A chimaeric gene containing the mutant aroA gene behind the octopine synthase promoter was constructed and integrated into Agrobacterium T-DNA vectors. Analysis of gall tissue from Brassica campestris L. (turnip rape) infected with A. tumefaciens K12 containing this chimaera showed mRNA and protein expressed from the bacterial gene; 50% of the total EPSP synthase activity present had kinetic properties of the mutant bacterial enzyme. Tobacco (Nicotiana tabacum L. ‘Xanthi′) plants have been regenerated from cocultivation with A. rhizogenes containing the same construct; analysis indicates expression of the gene and enhanced tolerance to glyphosate.

Copyright

References

Hide All
1. Amrhein, N., Johanning, D., Schab, J., and Schulz, A. 1983. Biochemical basis for glyphosate tolerance in a bacterium and a plant tissue culture. FEBS Lett. 157:191196.
2. Bickel, H., Palme, L., and Schultz, G. 1978. Incorporation of shikimate and other precursors into aromatic amino acids and phenylquinones of isolated spinach chloroplasts. Phytochemistry 17:119124.
3. Boocock, M. R. and Coggins, J. R. 1983. Kinetics of 5-enolpyruvylshikimate 3-phosphate synthase inhibition by glyphosate. FEBS Lett. 154:127133.
4. Comai, L., Schilling-Cordaro, C., Mergia, A., and Houck, C. M. 1983. A new technique for genetic engineering of Agrobacterium Ti plasmid. Plasmid 10:2130.
5. Comai, L., Sen, L. C., and Stalker, D. M. 1983. An altered aroA gene product confers resistance to the herbicide glyphosate. Science 221:370371.
6. Comai, L., Facciotti, D., Hiatt, W. R., Thompson, G., Rose, R. E., and Stalker, D. M. 1985. Expression in plants of a mutant aroA gene from Salmonella typhimurium confers tolerance to glyphosate. Nature 317:741744.
7. Gollub, E., Zalkin, H., and Sprinson, D. B. 1967. Correlation of genes and enzymes, and studies on relation of the aromatic pathway in Salmonella . J. Biol. Chem. 242:53235328.
8. Gresshoff, P. M. 1979. Growth inhibition of glyphosate and reversal of its action by phenylalanine and tyrosine. Aust. J. Plant Physiol. 6:177185.
9. Hiatt, W. R., Comai, L., Huang, L. -J., Rose, R., Thompson, G., and Stalker, D. 1985. Introduction and expression in plants of a glyphosate resistant aroA gene isolated from Salmonella typhimurium . Pages 479488 in Van Vloten-Doting, L., Groot, G.S.P., and Hall, T. C., eds. Molecular Form and Function of the Plant Genome. Plenum Publishing, New York.
10. Mousdale, D. M. and Coggins, J. R. 1984. Purification and properties of 5-enolpyruvyl-shikimate 3-phosphate synthase from seedlings of Pisum sativum L. Planta 160:7883.
11. Nafziger, E. D., Widholm, J. M., Steinrücken, H. C., and Killmer, J. L. 1984. Selection and characterization of a carrot cell line tolerant to glyphosate. Plant Physiol. 76:571574.
12. Rubin, J. L., Gaines, C. G., and Jensen, R. 1984. Glyphosate inhibition of 5-enolpyruvyl-shikimate 3-phosphate synthase from suspension-cultured cells of Nicotiana silvestris . Plant Physiol. 75:839845.
13. Rogers, S. G., Brand, L. A., Holder, S. B., Sharp, E. S., and Brackin, M. M. 1983. Amplification of the aroA gene from E. coli results in tolerance to the herbicide glyphosate. Appl. Environ. Microbiol. 46:3743.
14. Stalker, D. M., Hiatt, W. R., and Comai, L. 1985. A single amino acid substitution in the enzyme 5-enolpyruvylshikimate 3-phosphate synthase confers resistance to the herbicide glyphosate. J. Biol. Chem. 260:47244728.
15. Steinrücken, H. C. and Amrhein, N. 1983. 5-Enolpyruvyl-shikimate 3-phosphate synthase of Klebsiella pneumoniae. 2. Inhibition by glyphosate [N-(phosphonomethyl)glycine). Eur. J. Biochem. 143:351357.

Keywords

Metrics

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed