Skip to main content
Log in

Transformation of poplar (Populus alba) plastids and expression of foreign proteins in tree chloroplasts

  • Original paper
  • Published:
Transgenic Research Aims and scope Submit manuscript

Abstract

Plastid transformation offers several unique advantages compared with nuclear genome transformation, such as high level of transgene expression within plastids, expressing multiple transgenes as operons, lack of position effect due to site-specific transgene integration, and reducing risks of gene flow via pollen due to maternal inheritance of the plastid genome. Plastid transformation has been applied to several herbal species, but as yet there are no applications to tree species. We report here the first successful plastid transformation in a tree species, Populus alba. A vector for plastid transformation of poplar (Populus alba) was constructed, which carried the spectinomycin resistance gene and the green fluorescence protein gene as marker genes. In the regenerated shoots, the site-specific integration of foreign genes and the establishment of a high homoplastomic state were confirmed. Immunoblot analysis and histological observations corroborated the accumulation of green fluorescence protein in chloroplasts. The establishment of a plastid transformation system in poplar provides a novel tool for tree biotechnology.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Boerjan W (2005) Biotechnology and the domestication of forest trees. Curr Opin Biotech 16:159–166

    Article  PubMed  CAS  Google Scholar 

  • Bogorad L (2000) Engineering chloroplasts: an alternative site for foreign genes, proteins, reactions and products. Trends Biotechnol 18:257–263

    Article  PubMed  CAS  Google Scholar 

  • Brink MF, Brink G, Verbeet MP, de Boer HA (1994) Spectinomycin interacts specifically with the residues G1064 and 01192 in 16S rRNA thereby potentially freezing this molecule into an inactive conformation. Nucleic Acids Res 22:325–331

    Article  PubMed  CAS  Google Scholar 

  • Brunner AM, Mohamed R, Meilen R, Sheppard LA, Rottman WH, Strauss SH (1998) Genetic engineering of sexual sterility in shade trees. J Arboriculture 25:263–272

    Google Scholar 

  • Daniell H, Datta R, Varma S, Gray S, Lee SB (1998) Containment of herbicide resistance through genetic engineering of the chloroplast genome. Nat Biotechnol 16:345–348

    Article  PubMed  CAS  Google Scholar 

  • Daniell H, Kumara S, Dufourmantel N (2005) Breakthrough in chloroplast genetic engineering of agronomically important crops. Trends Biotechnol 23: 238–245

    Article  PubMed  CAS  Google Scholar 

  • De Cosa B, Moar W, Lee S-B, Miller M, Daniell H (2001) Overexpression of the Bt cry2Aa2 operon in chloroplasts leads to formation of insecticidal crystals. Nat Biotechnol 19:71–74

    Article  PubMed  Google Scholar 

  • DeGray G, Rajasekaran K, Smith F, Sanford J, Daniell H (2001) Expression of an antimicrobial peptide via the chloroplast genome to control phytopathogenic bacteria and fungi. Plant Physiol 127:852–862

    Article  PubMed  CAS  Google Scholar 

  • Donahue RA, Davis TD, Michler CH, Riemenschneider TE, Carter DR, Marquardt PE, Sankhla N, Sankhla D, Haissig BE, Isebrands JG (1994) Growth, photosynthesis and herbicide tolerance of genetically modified hybrid poplar. Can J For Res 24:2377–2383

    Article  Google Scholar 

  • Dufourmantel N, Pelissier B, Garçon F, Peltier G, Ferullo JM, Tissot G (2004) Generation of fertile transplastomic soybean. Plant Mol Biol 55:479–89

    Article  PubMed  CAS  Google Scholar 

  • Eckenwalder JE (1996) Systematics and evolution of Populus. In: Stettler RF, Bradshaw HD Jr Heilman PE, Hinckley TM (eds) Biology of Populus and its implications for management and conservation. NRC Research Press, Ottawa, Ontario Canada, pp 7–32

    Google Scholar 

  • Eriksson ME, Israelsson M, Olsson O, Moritz T (2000) Increased gibberellin biosynthesis in transgenic trees promotes growth biomass production and xylem fiber length. Nat Biotechnol 18:784–788

    Article  PubMed  CAS  Google Scholar 

  • Fromm H, Edelman M, Aviv D, Galun E (1987) The molecular basis for rRNA-dependent spectinomycin resistance in Nicotiana chloroplasts. EMBO J 6:3233–3237

    PubMed  CAS  Google Scholar 

  • Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158

    Article  PubMed  CAS  Google Scholar 

  • Gordon MP, Choe N, Duffy J, Ekuan G, Heilman P, Muiznieks I, Ruszaj M, Shurtleff BB, Strand S, Wilmoth J, Newman LA (1998) Phytoremediation of trichloroethylene with hybrid poplars. Environ Health Perspect 106:1001–1012

    Article  PubMed  CAS  Google Scholar 

  • Heuchelin SA, McNabb HS, Klopfenstenin NB (1997) Agrobacterium mediated transformation of Populus × euramericana “Ogy” using the chimeric CaMV 35S–Pin2 gene fusion. Can J For Res 27:1041–1048

    Article  CAS  Google Scholar 

  • Hu W-J, Harding SA, Lung J, Popko JL, Ralph J, Stokke DD, Tsai C-J, Vincent L, Chiang VL (1999) Repression of lignin biosynthesis promotes cellulose accumulation and growth in transgenic trees. Nat Biotechnol 17:808–812

    Article  PubMed  CAS  Google Scholar 

  • Johanson U, Hughes D (1995) A new mutation in 16S rRNA of Escherichia coli conferring spectinomycin resistance. Nucleic Acids Res 23:464–466

    Article  PubMed  CAS  Google Scholar 

  • Kanamoto H, Yamashita A, Asao H, Okumura S, Takase H, Hattori M, Yokota A, Tomizawa K-I (2006) Efficient and stable transformation of Lactuca sativa L. cv Cisco (lettuce) plastids. Transgenic Res 15:205–217

    Google Scholar 

  • Khan MS, Maliga P (1999) Fluorescent antibiotic resistance marker for tracking plastid transformation in higher plants. Nat Biotechnol 17:910–915

    Article  PubMed  CAS  Google Scholar 

  • Köhler RH, Cao J, Zipfel WR, Webb WW, Hanson MR (1997) Exchange of protein molecules through connections between higher plant plastids. Science 276:2039–2042

    Article  PubMed  Google Scholar 

  • Kumar S, Dhingra A, Daniell H (2004a) Plastid-expressed betaine aldehyde dehydrogenase gene in carrot cultured cells, roots, and leaves confers enhanced salt tolerance. Plant Physiol 136:2843–2854

    Article  CAS  Google Scholar 

  • Kumar S, Dhingra A, Daniell H (2004b) Stable transformation of the cotton plastid genome and maternal inheritance of transgenes. Plant Mol Biol 56:203–216

    Article  CAS  Google Scholar 

  • Lutz KA, Knapp JE, Maliga P (2001) Expression of bar in the plastid genome confers herbicide resistance. Plant Physiol 125:1585–1590

    Article  PubMed  CAS  Google Scholar 

  • Madoka Y, Tomizawa K, Mizoi J, Nishida I, Nagano Y, Sasaki Y (2002) Chloroplast transformation with modified accD operon increases acetyl-CoA carboxylase and causes extension of leaf longevity and increase in seed yield in tobacco. Plant Cell Physiol 43:1518–1525

    Article  PubMed  CAS  Google Scholar 

  • Maliga P (2004) Plastid transformation in higher plants. Annu Rev Plant Biol 55:289–313

    Article  PubMed  CAS  Google Scholar 

  • McCown BH, McCabe DE, Russell DR, Robison DJ, Barton KA, Raffa KF (1991) Stable transformation of Populus and incorporation of pest resistance by electric discharge particle acceleration. Plant Cell Rep 9:590–594

    Article  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco cultures. Physiol Plant 15:473–479

    Article  CAS  Google Scholar 

  • Newman LA, Strand SE, Choe N, Duffy J, Ekuan G, Ruszaj M, Shurtleff BB, Wilmoth J, Heilman P, Gordon MP (1997) Uptake and biotransformation of trichloroethylene by hybrid poplars. Environ Sci Technol 31:1062–1067

    Article  CAS  Google Scholar 

  • Peña L, Séguin A (2001) Recent advances in the genetictransformation of trees. Trensds Biotechnology 19:500–506

    Article  Google Scholar 

  • Petit RJ, Duminil J, Fineschi S, Hampe A, Salvini D, Vendramin GG (2005) Comparative organization of chloroplast mitochondrial and nuclear diversity in plant populations. Mol Ecol 14:689–701

    Article  PubMed  CAS  Google Scholar 

  • Pyke KA, Howells CA (2002) Plastid and stromule morphogenesis in tomato. Ann Bot 90:559–566

    Article  PubMed  CAS  Google Scholar 

  • Rajora OP, Dancik BP (1992) Chloroplast DNA inheritance in Populus. Theor Appl Genet 84:280–285

    Google Scholar 

  • Quesada-Vargas T, Ruiz ON, Henry Daniell H (2005) Characterization of heterologous multigene operons in transgenic chloroplasts. Transcription, processing, and translation. Plant Physiol 138:1746–1762

    Article  PubMed  CAS  Google Scholar 

  • Rishi AS, Nelson ND, Goyal (2001) A Genetic modification for improvement of Populus. Physiol Mol Biol Plants 7:7–21

    Google Scholar 

  • Ruf S, Hermann M, Berger IJ, Carrer H, Bock R (2001) Stable genetic transformation of tomato plastids and expression of a foreign protein in fruit. Nat Biotechnol 19:870–875

    Article  PubMed  CAS  Google Scholar 

  • Sikdar SR, Serino G, Chaudhuri S, Maliga P (1998) Plastid transformation in Arabidopsis thaliana. Plant Cell Rep 18:20–24

    Article  CAS  Google Scholar 

  • Slavov GT, Difazio SP, Strauss SH (2004) Gene flow in forest trees gene migration patterns and landscape modeling of transgene dispersal in hybrid poplar. In: Nijs HCM, Bartsch D, Sweet J (eds) Introgression from genetically modified plants into wild relatives. CABI Pub, Cambridge, pp 89–106

    Google Scholar 

  • Staub JM, Garcia B, Graves J, Hajdukiewicz PTJ, Hunter P, Nehra N, Paradkar V, Schlittler M, Carroll JA, Spatola L, Ward D, Ye G, Russell DA (2000) High-yield production of a human therapeutic protein in tobacco chloroplasts. Nat Biotechnol 18:333–338

    Article  PubMed  CAS  Google Scholar 

  • Stettler RF, Zsuffa L, Wu R (1996) The role of hybridization in the genetic manipulation of Populus. In: Stettler RF, Bradshaw HD Jr, Heilman PE, Hinckley TM (eds) Biology of Populus and its implications for management and conservation. NRC Research Press, Ottawa, Ontario, Canada, pp 87–112

    Google Scholar 

  • Svab Z, Hajdukiewicz P, Maliga P (1990) Stable transformation of plastids in higher plants. Proc Natl Acad Sci USA 87:8526–8530

    Article  PubMed  CAS  Google Scholar 

  • Svab Z, Maliga P (1993) High-frequency plastid transformation in tobacco by selection for a chimeric aadA gene. Proc Natl Acad Sci USA 90:913–917

    Article  PubMed  CAS  Google Scholar 

  • Tuskan GA, Walsh ME (2001) Short-rotation woody crop systems atmospheric carbon dioxide and carbon management a US case study. For Chron 77:259–264

    Google Scholar 

  • Viitanen PV, Devine AL, Khan MS, Deuel DL, Van Dyk DE, Daniell H (2004) Metabolic engineering of the chloroplast genome using the Echerichia coli ubiC gene reveals that chorismate is a readily abundant plant precursor for p-Hydroxybenzoic acid biosynthesis. Plant Physiol 136:4048–4060

    Article  PubMed  CAS  Google Scholar 

  • Wang L, Han Y, Hu J (2004) Transgenic forest trees for insectresistance. In. Kumar S, Fladung M (eds) Molecular genetics and breeding of forest trees. Food and Products Press, New York, pp 243–261

    Google Scholar 

  • Williams CG (2005) Framing the issues on transgenic forests. Nat Biotechnol 23:530–532

    Article  PubMed  CAS  Google Scholar 

  • Yin TM, Difazio SP, Gunter LE, Riemenschneider D, Tuskan GA (2004) Large-scale heterospecific segregation distortion in Populus revealed by a dense genetic map. Theor Appl Genet 109:451–463

    Article  PubMed  CAS  Google Scholar 

  • Zhang Q, Liu Y, Sodmergen (2003) Examination of the cytoplasmic DNA in male reproductive cells to determine the potential for cytoplasmic inheritance in 295 angiosperm species. Plant Cell Physiol 44:941–951

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the Ministry of Economy, Trade and Industry (METI) of Japan.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Ken-Ichi Tomizawa.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Okumura, S., Sawada, M., Park, Y.W. et al. Transformation of poplar (Populus alba) plastids and expression of foreign proteins in tree chloroplasts. Transgenic Res 15, 637–646 (2006). https://doi.org/10.1007/s11248-006-9009-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11248-006-9009-3

Keywords

Navigation