Glyphosate is a potent herbicide. It works by competitive inhibition of the enzyme 5-enol-pyruvyl shikimate-3-phosphate synthase (EPSPS), which catalyzes an essential step in the aromatic amino acid biosynthetic pathway. We report the genetic engineering of herbicide resistance by stable integration of the petunia EPSPS gene into the tobacco chloroplast genome using the tobacco or universal vector. Southern blot analysis confirms stable integration of the EPSPS gene into all of the chloroplast genomes (5000–10,000 copies per cell) of transgenic plants. Seeds obtained after the first self-cross of transgenic plants germinated and grew normally in the presence of the selectable marker, whereas the control seedlings were bleached. While control plants were extremely sensitive to glyphosate, transgenic plants survived sprays of high concentrations of glyphosate. Chloroplast transformation provides containment of foreign genes because plastid transgenes are not transmitted by pollen. The escape of foreign genes via pollen is a serious environmental concern in nuclear transgenic plants because of the high rates of gene flow from crops to wild weedy relatives.
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Shah, D.M., Horoh, R.B.,, Klee, H.J., Kishore, G.M., Winter, J.A., E.Tumer, N. et al. 1986. Engineering herbicides tolerance in transgenic plants. Science 233: 478–481.
Cioppa, G.D., Baner, S.C.,, Tayler, M.L., Roshester, D.E., Klein, B.K., Shah, D.M. et al. 1987. Targeting a herbicide resistant enzyme from Escherichia coli to chloroplasts of higher plants. Bio/Technology 5: 579–584.
Llewellyn, D. and Fitt, G. 1996. Pollen dispersal from two field trials of trans-genic cotton in the Namoi valley, Australia. Molecular Breeding 2: 157–166.
Umbeck, P.F., Barton, K.A.,, Nordheim, E.V., McCarty, J.C., Parrot, W.L., and Jenkins, J.N. 1991. Degree of pollen dispersal by insects from a field test of genetically engineered cotton. J. Econ. Entomol. 84: 1943–1950.
King, J. 1996. Could transgenic supercrops one day breed superweeds? Science 274: 180–181.
Mikkelsen, T.R., Anderson, B., and, Jörgensen, R.B. 1996. The risk of crop transgene spread. Nature 380: 31.
Daniell, H. and Rebeiz, C.A. 1982. Chloroplast culture IX: chlorphyll(ide) A biosynthesis in vitro at rates higher than in vivo. Biochem. Biophys. Res. ComMun. 106: 466–471.
Daniell, H., Ramanujan, P.,, Krishnan, M., Gnanam, A., and Rebeiz, C.A. 1983. In vitro synthesis of photosynthetic membranes: I. Development of photosystem I activity and cyclic phosphorylation. Biochem. Biophys. Res. ComMun. 111: 740–749.
Daniell, H., Krishnan, M.,, Umabai, U., and Gnanam, A. 1986. An efficient and prolonged in vitro translational system from cucumber etioplasts. Biochem. Biophys. Res. ComMun. 135: 248–255.
Daniell, H. and McFadden, B.A. 1987. Uptake and expression of bacterial and cyanobacterial genes by isolated cucumber etioplasts. Proc. Natl. Acad. Sci. USA 84: 6349–6353.
Carlson, P.S. 1973. The use of protoplasts for genetic research. Proc. Natl. Acad. Sci. USA 70: 598–602.
Daniell, H. 1993. Foreign gene expression in chloroplasts of higher plants mediated by tungsten particle bombardment. Methods Enzymol. 217: 536–556.
Daniell, H., Vivekananda, J.,, Nielsen, B.L., Ye, G.N., Tewari, K.K., and Sanford, J.C. 1990. Transient foreign gene expression in chloroplast of cultured tobacco cells following biolistic delivery of chloroplast vectors. Proc. Natl. Acad. Sci. USA 87: 88–92.
Ye, G.N., Daniell, H., and, Sanford, J.C. 1990. Optimization of delivery of foreign DNA into higher plant chloroplasts. Plant Mol. Biol. 15: 809–819.
Daniell, H., Krishnan, M., and, McFadden, B.A. 1991. Expression of β-glucuronidase gene in different cellular compartments following biolistic delivery of foreign DNA into wheat leaves and calli. Plant Cell Reports 9: 615–619.
Svab, Z. and Maliga, P. 1993. High frequency plastid transformation in tobacco by selection for a chimeric aadA gene. Proc. Natl. Acad. Sci. USA 90: 913–917.
McBride, K.E., Svab, Z.,, Schaaf, D.J., Hogen, P.S., Stalker, D.M., and Maliga, P. 1995. Amplification of a chimeric Bacillus gene in chloroplasts leads to extraordinary level of an insecticidal protein in tobacco. Bio/Technology 13: 362–365.
Maier, R.M., Neckermann, K.,, Igloi, G.L., and Kössel, H. 1995. Complete sequence of the maize chloroplast genome: gene content, hotspots of divergence and fine tuning of genetic information by transcript editing. J. Mol. Biol. 251: 614–628.
Brixey, P.J., Guda, C. and Daniell, H. 1997. The chloroplast psbA promoter is more efficient in E. coli than the T7 promoter for hyper expression of a foreign protein. Biotechnology Letters 19: 395–399.
Svab, Z., Hajdukiewicz, P. and Maliga, P. 1990. Stable transformation of plastids in higher plants. Proc. Natl. Acad. Sci. USA 87: 8526–8530.
Della-Cioppa, G., Bauer, S.C., Klein, B.K., Shaw, D.M., Fraley, R.T., and Kishore, G.M. 1986. Translocation of the precursor of 5-enolpyruvylshikimate-3-phosphate synthase into chloroplasts of higher plants in vitro. Proc. Natl. Acad. Sci. USA 83: 6873–6877.
Rogers, S.G., Brand, L.A., Holder, S.B., Sharps, E.S., and Brackin, M.J. 1983. Amplification of the aro A gene from Escherichia coli results in tolerance to the herbicide glyphosate. Appl. Environ. Microbiol. 46: 37–43.
Daniell, H. 1997. Transformation and foreign gene expression in plants mediated by microprojectile bombardment. Methods Mol. Biol. 62: 463–489.
Edwards, K., Johnstone, C., and Thompson, C. 1991. A simple and rapid method for preparation of plant genomic DNA for PCR analysis. Nucl. Acids. Res. 19: 1349.
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Daniell, H., Datta, R., Varma, S. et al. Containment of herbicide resistance through genetic engineering of the chloroplast genome. Nat Biotechnol 16, 345–348 (1998). https://doi.org/10.1038/nbt0498-345
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