Summary
Major crop losses occur annually as a result of biotic and abiotic stresses. The ability to hyperexpress foreign proteins, single-step multigene engineering, lack of positive effect and gene silencing, vector sequences and pleiotropic effects have resulted in several hundred-fold more tolerance to the environmental stresses via chloroplast genetic engineering than nuclear genetic engineering. Maternal inheritance of chloroplast expressed transgenes renders the technology environmentally safe and promotes public acceptance. This review provides protocols for engineering agronomic traits like insect, herbicide and disease resistance; salt and drought tolerance; and phytoremediation via chloroplast genome.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Daniell, H. (2002) Molecular strategies for gene containment in transgenic crops. Nat. Biotechnol. 20, 581–586.
Daniell, H. and Parkinson, C. L. (2003) Jumping genes and containment. Nat Biotechnol. 21, 374–375.
Daniell, H. and Dhingra, A. (2002) Multigene engineering: dawn of an exciting new era in biotechnology. Curr. Opin. Biotechnol. 13, 136–141.
Daniell, H., Khan, M. S, and Alison, L. (2002) Milestones in chloroplast genetic engineering: an environmentally friendly era in biotechnology. Trends Plant Sci. 7, 84–91.
Palmer, J. D. (1985) Comparative organization of chloroplast genomes. Annu. Rev. Genet. 19, 325–354.
Lilly, J. W., Havey, M. J., Jackson, S. A., and Jiang, J. M. (2001) Cytogenomic analyses reveal the structural plasticity of the chloroplast genome in higher plants. Plant Cell 13, 245–254.
Daniell, H., Vivekananda, J., Nielsen, B. L., Ye, G. N., Tewari, K. K., and Sanford, J. C. (1990) Transient foreign gene expression in chloroplasts of cultured tobacco cells after biolistic delivery of chloroplast vectors. Proc. Natl. Acad. Sci. USA 87, 88–92.
Daniell, H., Datta, R., Varma, S., Gray, S., and Lee, S. B. (1998) Containment of herbicide resistance through genetic engineering of the chloroplast genome. Nat. Biotechnol. 16, 345–348.
Guda, C., Lee, S. B., and Daniell, H. (2000) Stable expression of biodegradable protein based polymer in tobacco chloroplasts. Plant Cell Rep. 19, 257–262.
Kota, M., Daniell, H., Varma, S., Garczynski, S. F., Gould, F., and William, M. J. (1999) Overexpression of the Bacillus thuringiensis (Bt) Cry2Aa2 protein in chloroplasts confers resistance to plants against susceptible and Bt-resistant insects. Proc. Natl. Acad. Sci. USA 96, 1840–1845.
DeCosa, B., Moar, W., Lee, S. B., Miller, M., and Daniell, H. (2001) Overexpression of the Bt Cry2Aa2 operon in chloroplasts leads to formation of insecticidal crystals. Nat. Biotechnol. 19, 71–74.
Ruiz, O. N., Hussein, H., Terry, N., and Daniell, H. (2003) Phytoremediation of organomercurial compounds via chloroplast genetic engineering. Plant Physiol. 132, 1344–1352.
Bizily, S. P., Rugh, C.L., and Meagher, R. B. (2000) Phytodetoxification of hazardous organomercurials by genetically engineered plants. Nat. Biotechnol. 18, 213–217.
DeGray, G., Kanniah, R., Franzine, S., John, S., and Daniell, H. (2001) Expression of an antimicrobial peptide via the chloroplast genome to control phytopathogenic bacteria and fungi. Plant Physiol. 127, 852–862.
Iamtham, S. and Day, A. (2000) Removal of antibiotic resistance genes from transgenic tobacco plastids. Nat. Biotechnol. 18, 1172–1176.
Lee, S. B., Kwon, H. B., Kwon, S. J., et al. (2003) Accumulation of trehalose within transgenic chloroplasts confers drought tolerance. Mol. Breed. 11, 1–13.
McBride, K. E., Svab, Z., Schaaf, D. J., Hogan, P. S., Stalker, D. M., and Maliga, P. (1995) Amplification of a chimeric Bacillus gene in chloroplasts leads to an extraordinary level of an insecticidal protein in tobacco. Biotechnology 13, 362–365.
Staub, J. M., Garcia, B., Graves, J., etal. (2000) High-yield production of a human therapeutic protein in tobacco chloroplasts. Nat. Biotechnol. 18, 333–338.
Daniell, H., Lee, S. B., Panchal, T., and Wiebe, P. O. (2001) Expression of the native cholera toxin B subunit gene and assembly as functional oligomers in transgenic tobacco chloroplasts. J. Mol. Biol. 311, 1001–1009.
Ruiz, G. (2002) Optimization of codon composition and regulatory elements for expression of the human IGF-1 in transgenic chloroplasts. MS thesis, University of Central Florida, Orlando, FL.
Torres, M. (2002) Expression of interferon α5 in transgenic chloroplasts of tobacco. MS thesis, University of Central Florida, Orlando, FL.
Falconer, R. (2002) Expression of interferon α2b in transgenic chloroplasts of a low-nicotine tobacco. MS thesis, University of Central Florida, Orlando, FL.
Fernandez-San Millan, A., Mingo-Castel, A., and Daniell, H. (2003) A chloroplast transgenic approach to hyper-express and purify human serum albumin, a protein highly susceptible to proteolytic degradation. Plant Biotechnol. J. 1, 71–79.
Leelavathi S. and Reddy V. S. (2003) Chloroplast expression of His-tagged GUS-fusions: a general strategy to overproduce and purify foreign proteins using transplastomic plants as bioreactors. Mol. Breed. 11, 49–58.
Daniell, H., Dhingra, A., and San-Milan, A. F. (2001) 12th International Congress on Photosynthesis, Vol. S40-04 1–6, CSIRO, Brisbane, Australia.
Daniell, H., Watson, J., Koya, V., and Leppla, S. (2004) Expression of Bacillus anthracis protective antigen in transgenic chloroplasts of tobacco, a non-food/feed crop. Vaccine, in press.
Singleton, M. L. (2003) Expression of CaF1 and LcrV as a fusion protein for a vaccine against Yersinia pestis via chloroplast genetic engineering. MS thesis, University of Central Florida, Orlando, FL.
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.
Daniell, H., Krishnan, M., and McFadden, B. F. (1991) Transient expression of beta-glucuronidase in different cellular compartments following biolistic delivery of foreign DNA into wheat leaves and calli. Plant Cell Rep. 9, 615–619.
Goldschmidt Clermont, M. (1991) Transgenic expression of aminoglycoside adenine transferase in the chloroplast—a selectable marker for site-directed transformation of Chlamydomonas. Nucleic Acids Res. 19, 4083–4089.
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.
Carrer, H., Hockenberry, T. N., Svab, Z., and Maliga, P. (1993) Kanamycin resistance as a selectable marker for plastid transformation in tobacco. Mol. Gen. Genet. 241, 49–56.
Bateman, J. M. and Purton, S. (2000) Tools for chloroplast transformation in Chlamydomonas: expression vectors and a new dominant selectable marker. Mol. Gen. Genet. 263, 404–410.
Huang, F. C., Klaus, S. M. J., Herz, S., Zou, Z., Koop, H. U., and Golds, T. J. (2002) Efficient plastid transformation in tobacco using the aphA-6 gene and kanamycin selection. Mol. Genet. Genom. 268, 19–27.
Hibberd, J. M., Linley, P. J., Khan, M. S., and Gray, J. C. (1998) Transient expression of green fluorescent protein in various plastid types following microprojectile bombardment. Plant J. 16, 627–632.
Sidorov, V. A., Kasten, D., Pang, S. Z., Hajdukiewicz, P. T. J., Staub, J. M., and Nehra, N. S. (1999) Stable chloroplast transformation in potato: use of green fluorescent protein as a plastid marker. Plant J. 19, 209–216.
Khan, M. S. and Maliga, P. (1999) Fluorescent antibiotic resistance marker for tracking plastid transformation in higher plants. Nat. Biotechnol. 17, 910–915.
Daniell, H., Muthukumar, B., and Lee, S. B. (2001) Marker free transgenic plants: engineering the chloroplast genome without the use of antibiotic selection. Curr. Genet. 39, 109–116.
Devine, A. L. and Daniell, H. (2004) Plastids, in Annual Plant Reviews, Vol. 13, (Moller, E., ed.) Blackwell, Oxford, UK, Chapter 10, pp. 283–323.
Dhingra, A. and Daniell, H. (2004) Chloroplast genetic engineering via organogenesis or somatic embryogenesis, in Arabidopsis Protocols, 2nd Ed., in press.
Molina, A., Hervas-Stubbs, S., Daniell, H., Mingo-Castel, A. M., and Veramendi, J., (2004) High-yield expression of a viral peptide animal vaccine in transgenic tobacco chloroplasts. Plant Biotechnol. J. 2, 141–153.
Dhingra, A. (2000) Analysis of components involved in regulation of plastid gene expression, plastome organization and its manipulation with emphasis on rice (Oryza sativa L.). PhD thesis, University of Delhi South Campus, New Dehli, India.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2005 Humana Press Inc.
About this protocol
Cite this protocol
Daniell, H., Ruiz, O.N., Dhingra, A. (2005). Chloroplast Genetic Engineering to Improve Agronomic Traits. In: Peña, L. (eds) Transgenic Plants: Methods and Protocols. Methods in Molecular Biology™, vol 286. Humana Press. https://doi.org/10.1385/1-59259-827-7:111
Download citation
DOI: https://doi.org/10.1385/1-59259-827-7:111
Publisher Name: Humana Press
Print ISBN: 978-1-58829-263-6
Online ISBN: 978-1-59259-827-4
eBook Packages: Springer Protocols