Skip to main content
SpringerLink
Log in

Strategies for metabolic pathway engineering with multiple transgenes

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

The engineering of metabolic pathways in plants often requires the concerted expression of more than one gene. While with traditional transgenic approaches, the expression of multiple transgenes has been challenging, recent progress has greatly expanded our repertoire of powerful techniques making this possible. New technological options include large-scale co-transformation of the nuclear genome, also referred to as combinatorial transformation, and transformation of the chloroplast genome with synthetic operon constructs. This review describes the state of the art in multigene genetic engineering of plants. It focuses on the methods currently available for the introduction of multiple transgenes into plants and the molecular mechanisms underlying successful transgene expression. Selected examples of metabolic pathway engineering are used to illustrate the attractions and limitations of each method and to highlight key factors that influence the experimenter’s choice of the best strategy for multigene engineering.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1

Similar content being viewed by others

References

  • Altpeter F, Baisakh N, Beachy R, Bock R, Capell T, Christou P, Daniell H, Datta K, Datta S, Dix PJ, Fauquet C, Huang N, Kohli A, Mooibroek H, Nicholson L, Nguyen TT, Nugent G, Raemakers K, Romano A, Somers DA, Stoger E, Taylor N, Visser R (2005) Particle bombardment and the genetic enhancement of crops: myths and realities. Mol Breeding 15:305–327

    Article  Google Scholar 

  • Andrianov V, Borisjuk N, Pogrebnyak N, Brinker A, Dixon J, Spitsin S, Flynn J, Matyszczuk P, Andryszak K, Laurelli M, Golovkin M, Koprowski H (2010) Tobacco as a production platform for biofuel: overexpression of Arabidopsis DGAT and LEC2 genes increases accumulation and shifts the composition of lipids in green biomass. Plant Biotechnol J 8:277–287

    Article  PubMed  CAS  Google Scholar 

  • Apel W, Bock R (2009) Enhancement of carotenoid biosynthesis in transplastomic tomatoes by induced lycopene-to-provitamin A conversion. Plant Physiol 151:59–66

    Article  PubMed  CAS  Google Scholar 

  • Apel W, Schulze WX, Bock R (2010) Identification of protein stability determinants in chloroplasts. Plant J 63:636–650

    Article  PubMed  CAS  Google Scholar 

  • Barkan A (2011) Expression of plastid genes: organelle-specific elaborations on a prokaryotic scaffold. Plant Physiol 155:1520–1532

    Article  PubMed  CAS  Google Scholar 

  • Barkan A, Walker M, Nolasco M, Johnson D (1994) A nuclear mutation in maize blocks the processing and translation of several chloroplast mRNAs and provides evidence for the differential translation of alternative mRNA forms. EMBO J 13:3170–3181

    PubMed  CAS  Google Scholar 

  • Beauregard M, Hefford MA (2006) Enhancement of essential amino acid contents in crops by genetic engineering and protein design. Plant Biotechnol J 4:561–574

    PubMed  CAS  Google Scholar 

  • Birch-Machin I, Newell CA, Hibberd JM, Gray JC (2004) Accumulation of rotavirus VP6 protein in chloroplasts of transplastomic tobacco is limited by protein stability. Plant Biotechnol J 2:261–270

    Article  PubMed  CAS  Google Scholar 

  • Bock R (2007) Plastid biotechnology: prospects for herbicide and insect resistance, metabolic engineering and molecular farming. Curr Opin Biotechnol 18:100–106

    Article  PubMed  CAS  Google Scholar 

  • Bock R, Warzecha H (2010) Solar-powered factories for new vaccines and antibiotics. Trends Biotechnol 28:246–252

    Article  PubMed  CAS  Google Scholar 

  • Bohmert K, Balbo I, Kopka J, Mittendorf V, Nawrath C, Poirier Y, Tischendorf G, Trethewey RN, Willmitzer L (2000) Transgenic Arabidopsis plants can accumulate polyhydroxybutyrate to up to 4% of their fresh weight. Planta 211:841–845

    Article  PubMed  CAS  Google Scholar 

  • Bohmert-Tatarev K, McAvoy S, Daughtry S, Peoples OP, Snell KD (2011) High levels of bioplastic are produced in fertile transplastomic tobacco plants engineered with a synthetic operon for the production of polyhydroxybutyrate. Plant Physiol 155:1690–1708

    Article  PubMed  CAS  Google Scholar 

  • Capell T, Bassie L, Christou P (2004) Modulation of the polyamine biosynthetic pathway in transgenic rice confers tolerance to drought stress. Proc Natl Acad Sci USA 101:9909–9914

    Article  PubMed  CAS  Google Scholar 

  • Carrer H, Maliga P (1995) Targeted insertion of foreign genes into the tobacco plastid genome without physical linkage to the selectable marker gene. Bio/Technology 13:791–794

    Article  CAS  Google Scholar 

  • Craig W, Lenzi P, Scotti N, De Palma M, Saggese P, Carbone V, McGrath Curran N, Magee AM, Medgyesy P, Kavanagh TA, Dix PJ, Grillo S, Cardi T (2008) Transplastomic tobacco plants expressing a fatty acid desaturase gene exhibit altered fatty acid profiles and improved cold tolerance. Transgenic Res 17:769–782

    Article  PubMed  CAS  Google Scholar 

  • Daniell H (2006) Production of biopharmaceuticals and vaccines in plants via the chloroplast genome. Biotechnol J 1:1071–1079

    Article  PubMed  CAS  Google Scholar 

  • Day A, Goldschmidt-Clermont M (2011) The chloroplast transformation toolbox: selectable markers and marker removal. Plant Biotechnol J 9:540–553

    Article  PubMed  CAS  Google Scholar 

  • Dever TE (1999) Translation initiation: adept at adapting. Trends Biochem Sci 24:398–403

    Article  PubMed  CAS  Google Scholar 

  • Domergue F, Abbadi A, Heinz E (2005) Relief for fish stocks: oceanic fatty acids in transgenic oilseeds. Trends Plant Sci 10:112–116

    Article  PubMed  CAS  Google Scholar 

  • Drechsel O, Bock R (2010) Selection of Shine-Dalgarno sequences in plastids. Nucleic Acids Res 39:1427–1438

    Article  PubMed  CAS  Google Scholar 

  • Dufourmantel N, Pelissier B, Garcon F, Peltier G, Ferullo J-M, Tissot G (2004) Generation of fertile transplastomic soybean. Plant Mol Biol 55:479–489

    Article  PubMed  CAS  Google Scholar 

  • Ebinuma H, Sugita K, Matasunaga E, Endo S, Yamada K, Komamine A (2001) Systems for the removal of a selection marker and their combination with a positive marker. Plant Cell Rep 20:383–392

    Article  CAS  Google Scholar 

  • Elghabi Z, Karcher D, Zhou F, Ruf S, Bock R (2011a) Optimization of the expression of the HIV fusion inhibitor cyanovirin-N from the tobacco plastid genome. Plant Biotechnol J 9:599–608

    Article  PubMed  CAS  Google Scholar 

  • Elghabi Z, Ruf S, Bock R (2011b) Biolistic co-transformation of the nuclear and plastid genomes. Plant J 67:941–948

    Article  PubMed  CAS  Google Scholar 

  • Farré G, Bai C, Twyman RM, Capell T, Christou P, Zhu C (2011) Nutritious crops producing multiple carotenoids—a metabolic balancing act. Trends Plant Sci 16:532–540

    Article  PubMed  CAS  Google Scholar 

  • Felder S, Meierhoff K, Sane AP, Meurer J, Driemel C, Plücken H, Klaff P, Stein B, Bechtold N, Westhoff P (2001) The nucleus-encoded HCF107 gene of Arabidopsis provides a link between intercistronic RNA processing and the accumulation of translation-competent psbH transcripts in chloroplasts. Plant Cell 13:2127–2141

    PubMed  CAS  Google Scholar 

  • Fitzpatrick TB, Basset GJC, Borel P, Carrari F, DellaPenna D, Fraser PD, Hellmann H, Osorio S, Rothan C, Valpuesta V, Caris-Veyrat C, Fernie AR (2012) Vitamin deficiencies in humans: can plant science help? Plant Cell 24:395–414

    Article  PubMed  CAS  Google Scholar 

  • Fütterer J, Hohn T (1991) Translation of a polycistronic mRNA in the presence of the cauliflower mosaic virus transactivator protein. EMBO J 10:3887–3896

    PubMed  Google Scholar 

  • Garcia-Rios M, Fujita T, LaRosa PC, Locy RD, Clithero JM, Bressan RA, Csonka LN (1997) Cloning of a polycistronic cDNA from tomato encoding γ-glutamyl kinase and γ-glutamyl phosphate reductase. Proc Natl Acad Sci USA 94:8249–8254

    Article  PubMed  CAS  Google Scholar 

  • Hamilton AJ, Baulcombe DC (1999) A species of small antisense RNA in posttranscriptional gene silencing in plants. Science 286:950–952

    Article  PubMed  CAS  Google Scholar 

  • Hare PD, Chua N-H (2002) Excision of selectable marker genes from transgenic plants. Nat Biotechnol 20:575–580

    Article  PubMed  CAS  Google Scholar 

  • Harjes CE, Rocheford TR, Bai L, Brutnell TP, Kandianis CB, Sowinski SG, Stapleton AE, Vallabhaneni R, Williams M, Wurtzel ET, Yan J, Buckler ES (2008) Natural genetic variation in lycopene epsilon cyclase tapped for maize biofortification. Science 319:330–333

    Article  PubMed  CAS  Google Scholar 

  • Hashimoto M, Endo T, Peltier G, Tasaka M, Shikanai T (2003) A nucleus-encoded factor, CRR2, is essential for the expression of chloroplast ndhB in Arabidopsis. Plant J 36:541–549

    Article  PubMed  CAS  Google Scholar 

  • Hasunuma T, Miyazawa S-I, Yoshimura S, Shinzaki Y, Tomizawa K-I, Shindo K, Choi S-K, Misawa N, Miyake C (2008) Biosynthesis of astaxanthin in tobacco leaves by transplastomic engineering. Plant J 55:857–868

    Article  PubMed  CAS  Google Scholar 

  • Hirose T, Sugiura M (1997) Both RNA editing and RNA cleavage are required for translation of tobacco chloroplast ndhD mRNA: a possible regulatory mechanism for the expression of a chloroplast operon consisting of functionally unrelated genes. EMBO J 16:6804–6811

    Article  PubMed  CAS  Google Scholar 

  • Hirose T, Suguira M (2004) Functional Shine-Dalgarno-like sequences for translational initiation of chloroplast mRNAs. Plant Cell Physiol 45:114–117

    Article  PubMed  CAS  Google Scholar 

  • Hirschberg J (1999) Production of high-value compounds: carotenoids and vitamin E. Curr Opin Biotechnol 10:186–191

    Article  PubMed  CAS  Google Scholar 

  • Holmström K-O, Mäntylä E, Welin B, Mandal A, Palva ET, Tunnela OE, Londesborough J (1996) Drought tolerance in tobacco. Nature 379:683–684

    Article  Google Scholar 

  • Igloi GL, Kössel H (1992) The transcriptional apparatus of chloroplasts. Crit Rev Plant Sci 10:525–558

    Article  CAS  Google Scholar 

  • Jetter R, Kunst L (2008) Plant surface lipid biosynthetic pathways and their utility for metabolic engineering of waxes and hydrocarbon biofuels. Plant J 54:670–683

    Article  PubMed  CAS  Google Scholar 

  • Kebeish R, Niessen M, Thiruveedhi K, Bari R, Hirsch H-J, Rosenkranz R, Stäbler N, Schönfeld B, Kreuzaler F, Peterhänsel C (2007) Chloroplastic photorespiratory bypass increases photosynthesis and biomass production in Arabidopsis thaliana. Nat Biotechnol 25:593–599

    Article  PubMed  CAS  Google Scholar 

  • Kohli A, Leech M, Vain P, Laurie DA, Christou P (1998) Transgene organization in rice engineered through direct DNA transfer supports a two-phase integration mechanism mediated by the establishment of integration hot spots. Proc Natl Acad Sci USA 95:7203–7208

    Article  PubMed  CAS  Google Scholar 

  • Kooter JM, Matzke MA, Meyer P (1999) Listening to the silent genes: transgene silencing, gene regulation and pathogen control. Trends Plant Sci 4:340–347

    Article  PubMed  Google Scholar 

  • Kozak M (1994) Features in the 5′ non-coding sequences of rabbit α and β-globin mRNAs that affect translational efficiency. J Mol Biol 235:95–110

    Article  PubMed  CAS  Google Scholar 

  • Kozak M (1999) Initiation of translation in prokaryotes and eukaryotes. Gene 234:187–208

    Article  PubMed  CAS  Google Scholar 

  • Kozak M (2005a) A second look at cellular mRNA sequences said to function as internal ribosome entry sites. Nucleic Acids Res 33:6593–6602

    Article  PubMed  CAS  Google Scholar 

  • Kozak M (2005b) Regulation of translation via mRNA structure in prokaryotes and eukaryotes. Gene 361:13–37

    Article  PubMed  CAS  Google Scholar 

  • Krichevsky A, Meyers B, Vainstein A, Maliga P, Citovsky V (2010) Autoluminescent plants. PLoS ONE 5:e15461

    Article  PubMed  CAS  Google Scholar 

  • Lee S-B, Kwon H-B, Kwon S-J, Park S-C, Jeong M-J, Han S-E, Byun M-O, Daniell H (2003) Accumulation of trehalose within transgenic chloroplasts confers drought tolerance. Mol Breeding 11:1–13

    Article  CAS  Google Scholar 

  • Lelivelt CLC, McCabe MS, Newell CA, de Snoo CB, van Dun KMP, Birch-Machin I, Gray JC, Mills KHG, Nugent JM (2005) Stable plastid transformation in lettuce (Lactuca sativa L.). Plant Mol Biol 58:763–774

    Article  PubMed  CAS  Google Scholar 

  • Liere K, Börner T (2007) Transcription and transcriptional regulation in plastids. Top Curr Genet 19:121–174

    Article  CAS  Google Scholar 

  • Lössl A, Eibl C, Harloff H-J, Jung C, Koop H-U (2003) Polyester synthesis in transplastomic tobacco (Nicotiana tabacum L.): significant contents of polyhydroxybutyrate are associated with growth reduction. Plant Cell Rep 21:891–899

    PubMed  Google Scholar 

  • Lössl A, Bohmert K, Harloff H, Eibl C, Mühlbauer S, Koop H-U (2005) Inducible trans-activation of plastid transgenes: expression of the R. eutropha phb operon in transplastomic tobacco. Plant Cell Physiol 46:1462–1471

    Article  PubMed  CAS  Google Scholar 

  • Lu Y, Rijzaani H, Karcher D, Ruf S, Bock R (2013) Efficient metabolic pathway engineering in transgenic tobacco and tomato plastids with synthetic multigene operons. Proc Natl Acad Sci USA 110:E623–E632

    Google Scholar 

  • Lutz KA, Maliga P (2007) Construction of marker-free transplastomic plants. Curr Opin Biotechnol 18:107–114

    Article  PubMed  CAS  Google Scholar 

  • Ma S, Tang N, Tian J (2012) DNA synthesis, assembly and applications in synthetic biology. Curr Opin Chem Biol 16:260–267

    Article  PubMed  CAS  Google Scholar 

  • Maass D, Arango J, Wüst F, Beyer P, Welsch R (2009) Carotenoid crystal formation in Arabidopsis and carrot roots caused by increased phytoene synthase protein levels. PLoS ONE 4:e6373

    Article  PubMed  CAS  Google Scholar 

  • Madoka Y, Tomizawa K-I, 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 

  • Magee AM, Horvath EM, Kavanagh TA (2004) Pre-screening plastid transgene expression cassettes in Escherichia coli may be unreliable as a predictor of expression levels in chloroplast-transformed plants. Plant Sci 166:1605–1611

    Article  CAS  Google Scholar 

  • Maliga P, Bock R (2011) Plastid biotechnology: food, fuel, and medicine for the 21st century. Plant Physiol 155:1501–1510

    Article  PubMed  CAS  Google Scholar 

  • Manichaikul A, Ghamsari L, Hom EFY, Lin C, Murray RR, Chang RL, Balaji S, Hao T, Shen Y, Chavali AK, Thiele I, Yang X, Fan C, Mello E, Hill DE, Vidal M, Salehi-Ashtiani K, Papin JA (2009) Metabolic network analysis integrated with transcript verification for sequenced genomes. Nat Methods 6:589–592

    Article  PubMed  CAS  Google Scholar 

  • Mayer JE, Pfeiffer WH, Beyer P (2008) Biofortified crops to alleviate micronutrient malnutrition. Curr Opin Plant Biol 11:166–170

    Article  PubMed  CAS  Google Scholar 

  • Mette MF, van der Winden J, Matzke MA, Matzke AJM (1999) Production of aberrant promoter transcripts contributes to methylation and silencing of unlinked homologous promoters in trans. EMBO J 18:241–248

    Article  PubMed  CAS  Google Scholar 

  • Mette MF, Aufsatz W, van der Winden J, Matzke MA, Matzke AJM (2000) Transcriptional silencing and promoter methylation triggered by double-stranded RNA. EMBO J 19:5194–5201

    Article  PubMed  CAS  Google Scholar 

  • Misawa N (2011) Pathway engineering for functional isoprenoids. Curr Opin Biotechnol 22:627–633

    Article  PubMed  CAS  Google Scholar 

  • Mozes-Koch R, Gover O, Tanne E, Peretz Y, Maori E, Chernin L, Sela I (2012) Expression of an entire bacterial operon in plants. Plant Physiol 158:1883–1892

    Article  PubMed  CAS  Google Scholar 

  • Munns R (2005) Genes and salt tolerance: bringing them together. New Phytol 167:645–663

    Article  PubMed  CAS  Google Scholar 

  • Nakashita H, Arai Y, Shikanai T, Doi Y, Yamaguchi I (2001) Introduction of bacterial metabolism into higher plants by polycistronic transgene expression. Biosci Biotechnol Biochem 65:1688–1691

    Article  PubMed  CAS  Google Scholar 

  • Nanjo T, Kobayashi M, Yoshiba Y, Kakubari Y, Yamaguchi-Shinozaki K, Shinozaki K (1999) Antisense suppression of proline degradation improves tolerance to freezing and salinity in Arabidopsis thaliana. FEBS Lett 461:205–210

    Article  PubMed  CAS  Google Scholar 

  • Naqvi S, Zhu C, Farre G, Ramessar K, Bassie L, Breitenbach J, Conesa DP, Ros G, Sandmann G, Capell T, Christou P (2009a) Transgenic multivitamin corn through biofortification of endosperm with three vitamins representing three distinct metabolic pathways. Proc Natl Acad Sci USA 106:7762–7767

    Article  PubMed  CAS  Google Scholar 

  • Naqvi S, Farré G, Sanahuja G, Capell T, Zhu C, Christou P (2009b) When more is better: multigene engineering in plants. Trends Plant Sci 15:48–56

    Article  PubMed  CAS  Google Scholar 

  • Newell CA, Birch-Machin I, Hibberd JM, Gray JC (2003) Expression of green fluorescent protein from bacterial and plastid promoters in tobacco chloroplasts. Transgenic Res 12:631–634

    Article  PubMed  CAS  Google Scholar 

  • Nikolau BJ, Perera MADN, Brachova L, Shanks B (2008) Platform biochemicals for a biorenewable chemical industry. Plant J 54:536–545

    Article  PubMed  CAS  Google Scholar 

  • Oey M, Lohse M, Kreikemeyer B, Bock R (2009a) Exhaustion of the chloroplast protein synthesis capacity by massive expression of a highly stable protein antibiotic. Plant J 57:436–445

    Article  PubMed  CAS  Google Scholar 

  • Oey M, Lohse M, Scharff LB, Kreikemeyer B, Bock R (2009b) Plastid production of protein antibiotics against pneumonia via a new strategy for high-level expression of antimicrobial proteins. Proc Natl Acad Sci USA 106:6579–6584

    Article  PubMed  CAS  Google Scholar 

  • Paine JA, Shipton CA, Chaggar S, Howells RM, Kennedy MJ, Vernon G, Writht SY, Hinchliffe E, Adams JL, Silverstone AL, Drake R (2005) Improving the nutritional value of Golden Rice through increased pro-vitamin A content. Nat Biotechnol 23:482–487

    Article  PubMed  CAS  Google Scholar 

  • Park H-S, Himmelbach A, Browning KS, Hohn T, Ryabova LA (2001) A plant viral “reinitiation” factor interacts with the host translational machinery. Cell 106:723–733

    Article  PubMed  CAS  Google Scholar 

  • Peremarti A, Twyman RM, Gómez-Galera S, Naqvi S, Farré G, Sabalza M, Miralpeix B, Dashevskaya S, Yuan D, Ramessar K, Christou P, Zhu C, Bassie L, Capell T (2010) Promoter diversity in multigene transformation. Plant Mol Biol 73:363–378

    Article  PubMed  CAS  Google Scholar 

  • Peretz Y, Mozes-Koch R, Akad F, Tanne E, Czosnek H, Sela I (2007) A universal expression/silencing vector in plants. Plant Physiol 145:1251–1263

    Article  PubMed  CAS  Google Scholar 

  • Potrykus I (1991) Gene transfer to plants: assessment of published approaches and results. Annu Rev Plant Physiol Plant Mol Biol 42:205–225

    Article  CAS  Google Scholar 

  • Powell ALT, Nguyen CV, Hill T, Cheng KLL, Figueroa-Balderas R, Aktas H, Ashrafi H, Pons C, Fernández-Munoz R, Vicente A, Lopez-Baltazar J, Barry CS, Liu Y, Chetelat R, Granell A, Van Deynze A, Giovannoni JJ, Bennett AB (2012) Uniform ripening encodes a Golden 2-like transcription factor regulating tomato fruit chloroplast development. Science 336:1711–1715

    Article  PubMed  CAS  Google Scholar 

  • Quesada-Vargas T, Ruiz ON, 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 

  • Radakovits R, Jinkerson RE, Darzins A, Posewitz MC (2010) Genetic engineering of algae for enhanced biofuel production. Eukaryot Cell 9:486–501

    Article  PubMed  CAS  Google Scholar 

  • Rhodes MJC (1994) Physiological roles for secondary metabolites in plants: some progress, many outstanding problems. Plant Mol Biol 24:1–20

    Article  PubMed  CAS  Google Scholar 

  • Ruf M, Kössel H (1988) Occurrence and spacing of ribosome recognition sites in mRNAs of chloroplasts from higher plants. FEBS Lett 240:41–44

    Article  CAS  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 

  • Ruf S, Karcher D, Bock R (2007) Determining the transgene containment level provided by chloroplast transformation. Proc Natl Acad Sci USA 104:6998–7002

    Article  PubMed  CAS  Google Scholar 

  • Ruwe H, Schmitz-Linneweber C (2012) Short non-coding RNA fragments accumulating in chloroplasts: footprints of RNA binding proteins? Nucleic Acids Res 40:3106–3116

    Article  PubMed  CAS  Google Scholar 

  • Scharff LB, Childs L, Walther D, Bock R (2011) Local absence of secondary structure permits translation of mRNAs that lack ribosome-binding sites. PLoS Genet 7:e1002155

    Article  PubMed  CAS  Google Scholar 

  • Schmitz-Linneweber C, Small I (2008) Pentatricopeptide repeat proteins: a socket set for organelle gene expression. Trends Plant Sci 13:663–670

    Article  PubMed  CAS  Google Scholar 

  • Shiina T, Tsunoyama Y, Nakahira Y, Khan MS (2005) Plastid RNA polymerases, promoters, and transcription regulators in higher plants. Int Rev Cytol 244:1–68

    Article  PubMed  CAS  Google Scholar 

  • Staub JM, Maliga P (1995) Expression of a chimeric uidA gene indicates that polycistronic mRNAs are efficiently translated in tobacco plastids. Plant J 7:845–848

    Article  PubMed  CAS  Google Scholar 

  • Steen EJ, Kang Y, Bokinsky G, Hu Z, Schirmer A, McClure A, del Cardayre SB, Keasling JD (2010) Microbial production of fatty-acid-derived fuels and chemicals from plant biomass. Nature 463:559–562

    Article  PubMed  CAS  Google Scholar 

  • Stern DB, Goldschmidt-Clermont M, Hanson MR (2010) Chloroplast RNA metabolism. Annu Rev Plant Biol 61:125–155

    Article  PubMed  CAS  Google Scholar 

  • Sugita M, Sugiura M (1996) Regulation of gene expression in chloroplasts of higher plants. Plant Mol Biol 32:315–326

    Article  PubMed  CAS  Google Scholar 

  • Sweetlove LJ, Fernie AR (2005) Regulation of metabolic networks: understanding metabolic complexity in the systems biology era. New Phytol 168:9–24

    Article  PubMed  CAS  Google Scholar 

  • Tarczynski MC, Jensen RG, Bohnert HJ (1993) Stress protection of transgenic tobacco by production of the osmolyte mannitol. Science 259:508–510

    Article  PubMed  CAS  Google Scholar 

  • Thyssen G, Svab Z, Maliga P (2012) Exceptional inheritance of plastids via pollen in Nicotiana sylvestris with no detectable paternal mitochondrial DNA in the progeny. Plant J 72:84–88

    Article  PubMed  CAS  Google Scholar 

  • Tieman D, Bliss P, McIntyre LM, Blandon-Ubeda A, Bies D, Odabasi AZ, Rodríguez GR, van der Knaap E, Taylor MG, Goulet C, Mageroy MH, Snyder DJ, Colquhoun T, Moskowitz H, Clark DG, Sims C, Bartoshuk L, Klee HJ (2012) The chemical interactions underlying tomato flavor preferences. Curr Biol 22:1035–1039

    Article  PubMed  CAS  Google Scholar 

  • Toepfer R, Martini N, Schell J (1995) Modification of plant lipid synthesis. Science 268:681–686

    Article  CAS  Google Scholar 

  • Verhounig A, Karcher D, Bock R (2010) Inducible gene expression from the plastid genome by a synthetic riboswitch. Proc Natl Acad Sci USA 107:6204–6209

    Article  PubMed  CAS  Google Scholar 

  • Viitanen PV, Devine AL, Khan MS, Deuel DL, van Dyk DE, Daniell H (2004) Metabolic engineering of the chloroplast genome using the Escherichia 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 

  • Waterhouse PM, Graham MW, Wang M-B (1998) Virus resistance and gene silencing in plants can be induced by simultaneous expression of sense and antisense RNA. Proc Natl Acad Sci USA 95:13959–13964

    Article  PubMed  CAS  Google Scholar 

  • Westhoff P, Herrmann RG (1988) Complex RNA maturation in chloroplasts. Eur J Biochem 171:551–564

    Article  PubMed  CAS  Google Scholar 

  • Willey DL, Gray JC (1989) Two small open reading frames are co-transcribed with the pea chloroplast genes for the polypeptides of cytochrome b-559. Curr Genet 15:213–220

    Article  PubMed  CAS  Google Scholar 

  • Wurbs D, Ruf S, Bock R (2007) Contained metabolic engineering in tomatoes by expression of carotenoid biosynthesis genes from the plastid genome. Plant J 49:276–288

    Article  PubMed  CAS  Google Scholar 

  • Yan J, Kandianis CB, Harjes CE, Bai L, Kim E-H, Yang X, Skinner DJ, Fu Z, Mitchell S, Li Q, Fernandez MGS, Zaharieva M, Babu R, Fu Y, Palacios N, Li J, DellaPenna D, Brutnell T, Buckler ES, Warburton ML, Rocheford T (2010) Rare genetic variation at Zea mays crtRB1 increases β-carotene in maize grain. Nat Genet 42:322–327

    Article  PubMed  CAS  Google Scholar 

  • Ye X, Al-Babili S, Klöti A, Zhang J, Lucca P, Beyer P, Potrykus I (2000) Engineering the provitamin A (β-carotene) biosynthetic pathway into (carotenoid-free) rice endosperm. Science 287:303–305

    Article  PubMed  CAS  Google Scholar 

  • Yueh A, Schneider RJ (2000) Translation by ribosome shunting on adenovirus and hsp70 mRNAs facilitated by complementarity to 18S rRNA. Genes Dev 14:414–421

    PubMed  CAS  Google Scholar 

  • Zhelyazkova P, Hammani K, Rojas M, Voelker R, Vargas-Suárez M, Börner T, Barkan A (2012) Protein-mediated protection as the predominant mechanism for defining processed mRNA termini in land plant chloroplasts. Nucleic Acids Res 40:3092–3105

    Article  PubMed  CAS  Google Scholar 

  • Zhou F, Karcher D, Bock R (2007) Identification of a plastid intercistronic expression element (IEE) facilitating the expression of stable translatable monocistronic mRNAs from operons. Plant J 52:961–972

    Article  PubMed  CAS  Google Scholar 

  • Zhu C, Naqvi S, Gomez-Galera S, Pelacho AM, Capell T, Christou P (2007) Transgenic strategies for the nutritional enhancement of plants. Trends Plant Sci 12:548–555

    Article  PubMed  CAS  Google Scholar 

  • Zhu C, Naqvi S, Breitenbach J, Sandmann G, Christou P, Capell T (2008) Combinatorial genetic transformation generates a library of metabolic phenotypes for the carotenoid pathway in maize. Proc Natl Acad Sci USA 105:18232–18237

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

Work on plastid transformation, combinatorial transformation and metabolic pathway engineering in the author’s laboratory is supported by grants from the Bundesministerium für Bildung und Forschung (BMBF), the European Union (EU-FP7 METAPRO 244348, COST Actions FA0804 and FA1006), the Deutsche Forschungsgemeinschaft (DFG) and by the Max Planck Society.

Author information

Authors and Affiliations

  1. Max-Planck-Institut für Molekulare Pflanzenphysiologie, Am Mühlenberg 1, 14476, Potsdam-Golm, Germany

    Ralph Bock

Authors
  1. Ralph Bock
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ralph Bock.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bock, R. Strategies for metabolic pathway engineering with multiple transgenes. Plant Mol Biol 83, 21–31 (2013). https://doi.org/10.1007/s11103-013-0045-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11103-013-0045-0

Keywords

Search

Navigation