Skip to main content
Log in

A comparison of individual and combined l-phenylalanine ammonia lyase and cationic peroxidase transgenes for engineering resistance in tobacco to necrotrophic pathogens

  • Original Article
  • Published:
Plant Biotechnology Reports Aims and scope Submit manuscript

Abstract

This study tested the relative and combined efficacy of ShPx2 and ShPAL transgenes by comparing Nicotiana tabacum hybrids with enhanced levels of l-phenylalanine ammonia lyase (PAL) activity and cationic peroxidase (Prx) activity with transgenic parental lines that overexpress either transgene. The PAL/Prx hybrids expressed both transgenes driven by the 35S CaMV promoter, and leaf PAL and Prx enzyme activities were similar to those of the relevant transgenic parent and seven- to tenfold higher than nontransgenic controls. Lignin levels in the PAL/Prx hybrids were higher than the PAL parent and nontransgenic controls, but not significantly higher than the Prx parent. All transgenic plants showed increased resistance to the necrotrophs Phytophthora parasitica pv. nicotianae and Cercospora nicotianae compared to nontransgenic controls, with a preponderance of smaller lesion categories produced in Prx-expressing lines. However, the PAL/Prx hybrids showed no significant increase in resistance to either pathogen relative to the Prx parental line. These data indicate that, in tobacco, the PAL and Prx transgenes do not act additively in disease resistance. Stacking with Prx did not prevent a visible growth inhibition from PAL overexpression. Practical use of ShPAL will likely require more sophisticated developmental control, and we conclude that ShPx2 is a preferred candidate for development as a resistance transgene.

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.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Bate NJ, Orr J, Ni W, Meromi A, Nadler-Hassar T, Doerner PW, Dixon RA, Lamb CJ, Elkind Y (1994) Quantitative relationship between phenylalanine ammonia-lyase levels and phenylpropanoid accumulation in transgenic tobacco identifies a rate-determining step in natural product synthesis. Proc Natl Acad Sci USA 91:7608–7612

    Article  PubMed  CAS  Google Scholar 

  • Bauer N, Fulgosi H, Jelaska S (2011) Overexpression of phenylalanine ammonia-lyase in transgenic roots of Coleus blumei alters growth and rosmarinic acid synthesis. Food Technol Biotechnol 49:24–31

    CAS  Google Scholar 

  • Bindschedler LV, Dewdney J, Blee KA, Stone JM, Asai T, Plotnikov J, Denoux C, Hayes T, Gerrish C, Davies DR, Ausebel FM, Bolwell GP (2006) Peroxidase-dependent apoplastic oxidative burst in Arabidopsis required for pathogen resistance. Plant J 47:851–863

    Article  PubMed  CAS  Google Scholar 

  • Bonawitz ND, Chapple C (2010) The genetics of lignin biosynthesis: connecting genotype to phenotype. Ann Rev Genet 44:337–363

    Article  PubMed  CAS  Google Scholar 

  • Dellaporta SL (1983) A plant minipreparation: version II. Plant Mol Biol Rep 1:19–21

    Article  CAS  Google Scholar 

  • Dixon RA (2001) Natural products and disease resistance. Nature 411:843–847

    Article  PubMed  CAS  Google Scholar 

  • Dowd PF, Lagrimini LM (1997) The role of peroxidase in host insect defenses. In: Carrozi N, Koziel M (eds) Advances in insect control: the role of transgenic plants. Taylor and Francis, London, pp 195–223

    Google Scholar 

  • Dowd PF, Lagrimini LM (2006) Examination of the biological effects of high anionic peroxidase production in tobacco plants grown under field conditions I. Insect damage. Transgenic Res 15:197–204

    Article  PubMed  CAS  Google Scholar 

  • Dowd PF, Holmes RA, Pinkerton TS, Johnson ET, Lagrimini LM, Boston RS (2006) Relative activity of a tobacco hybrid expressing high levels of a tobacco anionic peroxidase and maize ribosome-inactivating protein against Helicoverpa zea and Lasioderma serricorne. J Agric Food Chem 54:2629–2634

    Article  PubMed  CAS  Google Scholar 

  • Duroux L, Welinder KG (2003) The peroxidase gene family in plants: a phylogenetic overview. J Mol Evol 57:397–407

    Article  PubMed  CAS  Google Scholar 

  • Elfstrand M, Sitbon F, Lapierre C, Bottin A, von Arnold S (2002) Altered lignin structure and resistance to pathogens in spi-2-expressing tobacco plants. Planta 214:708–716

    Article  PubMed  CAS  Google Scholar 

  • Felton GW, Korth KL, Bi JL, Wesley SV, Huhman DV, Mathews MC, Murphy JB, Lamb C, Dixon RA (1999) Inverse relationship between systemic resistance of plants to micro-organisms and to insect herbivory. Curr Biol 9:317–320

    Article  PubMed  CAS  Google Scholar 

  • Harrison SJ, Curtis MD, McIntyre CL, Maclean DJ, Manners JM (1995) Differential expression of peroxidase isogenes during the early stages of infection of the tropical forage legume Stylosanthes humilis by Colletotrichum gloeosporioides. Mol Plant–Microbe Interact 8:398–406

    Article  PubMed  CAS  Google Scholar 

  • He C, Nourse JP, Kelemu S, Irwin JAG, Manners JM (1996) CgT1: a non-LTR retrotransposon with restricted distribution in the fungal phytopathogen Colletotrichum gloeosporoides. Mol Gen Genet 252:320–331

    PubMed  CAS  Google Scholar 

  • Higgins C, Manners JM, Scott KJ (1985) Decrease in three messenger RNA species coding for chloroplast proteins in leaves of barley infected with Erysiphe graminis f.sp. hordei. Plant Physiol 78:891–894

    Article  PubMed  CAS  Google Scholar 

  • Howles PA, Sewalt VJH, Paiva NL, Elkind Y, Bate NJ, Lamb C, Dixon RA (1996) Over-expression of l-phenylalanine ammonia lyase in transgenic tobacco plants reveals control points for flux into phenylpropanoid biosynthesis. Plant Physiol 112:1617–1624

    PubMed  CAS  Google Scholar 

  • Jung HG, Mertens DR, Payne AJ (1997) Correlation of acid detergent lignin and klason lignin with digestibility of forage dry matter and neutral detergent fiber. J Dairy Sci 80:1622–1628

    Article  PubMed  CAS  Google Scholar 

  • Kazan K, Goulter KC, Way HM, Manners JM (1998a) Expression of a pathogenesis-related peroxidase of Stylosanthes humilis in transgenic tobacco and canola and its effect on disease development. Plant Sci 136:207–217

    Article  CAS  Google Scholar 

  • Kazan K, Murray F, Goulter KC, Llewellyn D, Manners JM (1998b) Induction of cell death in transgenic plants expressing a fungal glucose oxidase. Mol Plant–Microbe Interact 11:555–562

    Article  CAS  Google Scholar 

  • Kobayashi A, Koguchi Y, Kanzaki H, Kajiyama S, Kawazu K (1994) Production of a new type of bioactive phenolic compound. Biosci Biotech Biochem 58:133–134

    Article  CAS  Google Scholar 

  • Maher EA, Bate NJ, Ni W, Elkind Y, Dixon RA, Lamb CJ (1994) Increased disease susceptibility of transgenic tobacco plants with suppressed levels of preformed phenylpropanoid products. Proc Natl Acad Sci USA 91:7802–7806

    Article  PubMed  CAS  Google Scholar 

  • Manners JM, McIntyre CL, Nourse JP (1995) Cloning and sequence of a cDNA encoding phenylalanine ammonia-lyase from the tropical forage legume Stylosanthes humilis. Plant Physiol 108:1301–1302

    Article  PubMed  CAS  Google Scholar 

  • Pallas JA, Paiva NL, Lamb C, Dixon RA (1996) Tobacco plants epigenetically suppressed in phenylalanine ammonia-lyase expression do not develop systemic acquired resistance in response to infection by tobacco. Plant J 10:281–293

    Article  CAS  Google Scholar 

  • Passardi F, Longet D, Penel C, Dunand C (2004a) The class III peroxidase multigenic family in rice and its evolution in land plants. Phytochemistry 65:1879–1893

    Article  PubMed  CAS  Google Scholar 

  • Passardi F, Penel C, Dunand C (2004b) Performing the paradoxical: how plant peroxidases modify the cell wall. Trends Plant Sci 9:534–540

    Article  PubMed  CAS  Google Scholar 

  • Passardi F, Theiler G, Zamocky M, Cosio C, Rouhier N, Teixera F, Margis-Pinheiro M, Ionnidis V, Penel C, Falquet L, Dunand C (2007) Peroxibase: the peroxidase database. Phytochemistry 68:1605–1611

    Article  PubMed  CAS  Google Scholar 

  • Robin C, Guest D (1994) Characterisation of pathogenicity of Phytophthora parasitica isolates by stem and detached leaf inoculations in four tobacco cultivars. NZ J Crop Hortic Sci 22:159–166

    Article  Google Scholar 

  • Sarowar S, Kim YJ, Kim EN, Kim KD, Choi JY, Hyung NI, Shin JS (2006) Constitutive expression of two pathogenesis-related genes in tomato plants enhanced resistance to oomycete pathogen Phytophthora capsici. Plant Cell Tissue Organ Culture 86:7–14

    Article  CAS  Google Scholar 

  • Sasaki K, Iwai T, Hiraga S, Kuroda K, Seo S, Mitsuhara I, Miyasaka A, Iwano M, Ito H, Matsui H, Ohashi Y (2004) Ten rice peroxidases redundantly respond to multiple stresses including infection with rice blast fungus. Plant Cell Physiol 45:1442–1452

    Article  PubMed  CAS  Google Scholar 

  • Schweizer P (2008) Tissue-specific expression of a defence-related peroxidase in transgenic wheat potentiates cell death in pathogen-attacked leaf epidermis. Mol Plant Pathol 9:45–57

    Article  PubMed  CAS  Google Scholar 

  • Sewalt VJH, Ni W, Blount JW, Jung HG, Masoud SA, Howles PA, Lamb C, Dixon RA (1997) Reduced lignin content and altered lignin composition in transgenic tobacco down-regulated in expression of l-phenylalanine ammonia lyase or cinnamate 4-hydroxylase. Plant Physiol 115:41–50

    PubMed  CAS  Google Scholar 

  • Shadle GL, Wesley SV, Korth KL, Chen F, Lamb C, Dixon RA (2003) Phenylpropanoid compounds and disease resistance in transgenic tobacco with altered expression of l-phenylalanine ammonia lyase. Phytochemistry 64:153–161

    Article  PubMed  CAS  Google Scholar 

  • Stephenson SA, Green JR, Manners JM, Maclean DJ (1997) Cloning and characterisation of glutamine synthetase from Colletotrichum gloeosporioides and demonstration of elevated expression during pathogenesis on Stylosanthes guianensis. Curr Genet 31:447–454

    Article  PubMed  CAS  Google Scholar 

  • Tucker CC, Chakraborty S (1997) Quantitative assessment of lesion characteristics and disease severity using digital image processing. J Phytopathol 145:273–278

    Article  Google Scholar 

  • Van Soest PJ (1963) Use of detergents in analysis of fibrous feeds II. A rapid method for determination of fiber and lignin. J Assoc Off Agric Chem 46:829–835

    Google Scholar 

  • Vogt T (2010) Phenylpropanoid biosynthesis. Mol Plant 3:2–20

    Article  PubMed  CAS  Google Scholar 

  • Wally O, Punja ZK (2010) Enhanced disease resistance in transgenic carrot (Daucus carota L.) plants over-expressing a rice cationic peroxidase. Planta 232:1229–1239

    Article  PubMed  CAS  Google Scholar 

  • Way HM, Kazan K, Goulter KG, Birch RG, Manners JM (2000) Expression of the Shpx2 peroxidase gene of Stylosanthes humilis in transgenic tobacco leads to enhanced resistance to Phytophthora parasitica pv. nicotianae and Cercospora nicotianae. Mol Plant Pathol 4:223–232

    Article  Google Scholar 

  • Way HM, Kazan K, Mitter N, Goulter KG, Birch RG, Manners JM (2002) Constitutive expression of a phenylalanine ammonia-lyase gene from Stylosanthes humilis in transgenic tobacco leads to enhanced disease resistance but impaired plant growth. Physiol Mol Plant Pathol 60:275–282

    CAS  Google Scholar 

  • Weng J-K, Li X, Bonawitz ND, Chapple C (2008) Emerging strategies of lignin engineering and degradation for cellulosic biofuel production. Curr Opin Biotechnol 19:166–172

    Article  PubMed  CAS  Google Scholar 

  • Wernsman EA, Matzinger DF (1980) Tobacco. In: Fehr WR, Hadley HH (eds) Hybridisation of crop plants. American Society of Agronomy and Crop Science of America, Madison, pp 657–668

    Google Scholar 

Download references

Acknowledgments

HW acknowledges support from the Grains and Development Corporation and the Cooperative Research Centre for Tropical Plant Pathology. The expert advice and assistance of Drs. Kemal Kazan and Ken Goulter are gratefully acknowledged. Dr. J. Hendrickz of the University of Queensland provided essential advice on statistical analysis.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to John M. Manners.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Way, H.M., Birch, R.G. & Manners, J.M. A comparison of individual and combined l-phenylalanine ammonia lyase and cationic peroxidase transgenes for engineering resistance in tobacco to necrotrophic pathogens. Plant Biotechnol Rep 5, 301–308 (2011). https://doi.org/10.1007/s11816-011-0183-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11816-011-0183-2

Keywords

Navigation