Skip to main content
SpringerLink
Log in

Expression of a metacaspase gene of Nicotiana benthamiana after inoculation with Colletotrichum destructivum or Pseudomonas syringae pv. tomato, and the effect of silencing the gene on the host response

  • Biotic and Abiotic Stress
  • Published:
Plant Cell Reports Aims and scope Submit manuscript

Abstract

Metacaspases are cysteine proteinases that have homology to caspases, which play a central role in signaling and executing programmed cell death in animals. A type II metacaspase cDNA, NbMCA1, was amplified from Nicotiana benthamiana infected with Colletotrichum destructivum. It showed a peak in expression at 72 h post-inoculation corresponding with the switch to necrotrophy by C. destructivum. Inoculation of N. benthamiana with an incompatible bacterium, Pseudomonas syringae pv. tomato, which should induce a non-host hypersensitive response (HR), did not result in an increase in NbMCA1 expression at the time of necrosis development at 20–24 h postinoculation. Virus-induced silencing of NbMCA1 resulted in three to four times more lesions due to C. destructivum compared with leaves inoculated with the PVX vector without the cloned metacaspase gene or inoculated with water only. However, virus-induced silencing of NbMCA1 did not affect the HR necrosis or population levels of P. syringae pv. tomato. Although this metacaspase gene does not appear to be involved in the programmed cell death of non-host HR resistance to P. syringae, it does affect the susceptibility of N. benthamiana to C. destructivum indicating a function in a basal defense response. Possible roles of NbMCA1could be in degrading virulence factors of the pathogen, processing pro-proteins involved in stress responses, eliminating damaged proteins created during stress, and/or degrading proteins to remobilize amino acids to fuel de novo synthesis of proteins involved in stress adaptations.

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
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3789–3402

    Article  Google Scholar 

  • Armando B, Julio M, Mario RS, Helena P (2003) Metacaspase related to programmed cell death in Arabidopsis thaliana. In: Proceedings of the 7th International Conference on Plant Molecular Biology, pp S21–72

  • Baulcombe D, Hamilton A, Voinnet O, Lu R, Peart JR, Malcuit I, Moffett P (2002) Sense and susceptibility: dissecting disease resistance using viruses and silencing. In: Leong SA, Allen C, Triplett EW (eds) Biology of plant–microbe interactions. International Society for Molecular Plant–Microbe Interactions, St. Paul, pp 1–10

    Google Scholar 

  • Beers EP, Woffenden BJ, Zhao C (2000) Plant proteolytic enzymes: possible roles during programmed cell death. Plant Mol Biol 44:399–415

    Article  PubMed  CAS  Google Scholar 

  • Bozhkov PV, Suarez MF, Filonova LH, Daniel G, Zamyatnin AA, Rodriguez-Nieto S, Zhivotovsky B, Smertenko A (2005) Cysteine protease mcll-Pa executes programmed cell death during plant embryogenesis. Proc Natl Acad Sci USA 102:14463–14468

    Article  PubMed  CAS  Google Scholar 

  • Chen GYJ, Jin S, Goodwin PH (2000) An improved method for the isolation of total RNA from Malva pusilla tissues infected with Colletotrichum gloeosporioides. J Phytopathol 148:57–60

    Article  CAS  Google Scholar 

  • Chen N, Hsiang T, Goodwin PH (2002) Use of green fluorescent protein to quantify the growth of Colletotrichum during infection of tobacco. J Microbiol Methods 53:113–122

    Article  CAS  Google Scholar 

  • Cohen GM (1997) Caspases: the executioners of apoptosis. Biochem J 326:1–16

    PubMed  CAS  Google Scholar 

  • Cryns V, Yuan J (1998) Proteases to die for. Genes Dev 12:1551–1570

    PubMed  CAS  Google Scholar 

  • Dean JD, Goodwin PH, Hsiang T (2002) Comparison of relative RT-PCR and northern blot analyses to measure expression of β-1,3-glucanase in Nicotiana benthamiana infected with Colletotrichum destructivum. Plant Mol Biol Rep 20:347–356

    Article  CAS  Google Scholar 

  • del Pozo O, Lam E (1998) Caspases and programmed cell death in the hypersensitive response of plants to pathogens. Curr Biol 8:1129–1132

    Article  PubMed  Google Scholar 

  • Dickman MB, Park YK, Oltersdorf T, Li W, Clemente T, French R (2001) Abrogation of disease development in plants expressing animal antiapoptotic genes. Proc Natl Acad Sci USA 98:6957–62

    Article  PubMed  CAS  Google Scholar 

  • Espinosa A, Guo M, Tam VC, Fu ZQ, Alfano JR (2003) The Pseudomonas syringae type III-secreted protein HopPtoD2 possesses protein tyrosine phosphatase activity and suppresses programmed cell death in plants. Mol Microbiol 49:377–387

    Article  PubMed  CAS  Google Scholar 

  • Forsthoefel NR, Cushman MAF, Ostrem JA, Cushman JC (1998) Induction of a cysteine protease cDNA from Mesembryanthemum crystallinum leaves by environmental stress and plant growth regulators. Plant Sci 136:195–206

    Article  CAS  Google Scholar 

  • Hao L, Hsiang T, Goodwin PH (2006) Role of two cysteine proteinases in the susceptible response of Nicotiana benthamiana to infection by Colletotrichum destructivum and the hypersensitive response to Pseudomonas syringae pv. tomato. Plant Sci 170:1001–1009

    Article  CAS  Google Scholar 

  • Hatsugai N, Kuroyanagi M, Yamada K, Meshi T, Tsuda S, Kondo M, Nishimura M, Hara-Nishimura I (2004) A plant vacuolar protease, VPE, mediates virus-induced hypersensitive cell death. Science 305:855–858

    Article  PubMed  CAS  Google Scholar 

  • Heath MC (2000) Hypersensitive response-related death. Plant Mol Biol 44:321–334

    Article  PubMed  CAS  Google Scholar 

  • Hedges SB (2002) The origin and evolution of model organisms. Nat Rev Genet 3:838–849

    Article  PubMed  CAS  Google Scholar 

  • Hoeberichts FA, ten Have A, Woltering EJ (2003) A tomato metacaspase gene is upregulated during programmed cell death in Botrytis cinerea-infected leaves. Planta 217:517–522

    Article  PubMed  CAS  Google Scholar 

  • Jones L, Hamilton AJ, Voinnet O, Thomas CL, Maule AJ, Baulcombe D (1999) RNA-DNA interactions and DNA methylation in post-transcriptional gene silencing. Plant Cell 11:2291–2301

    Article  PubMed  CAS  Google Scholar 

  • King EO, Ward MK, Raney DE (1954) Two simple media for the demonstration of pyocyanin and fluorescin. J Lab Clin Med 44:301–307

    PubMed  CAS  Google Scholar 

  • Klement Z (1963) Rapid detection of the pathogenicity of phytopathogenic Pseudomonads. Nature 199:299–300

    Article  PubMed  CAS  Google Scholar 

  • Lam E, del Pozo DO (2000) Caspase-like protease involvement in the control of plant cell death. Plant Mol Biol 44:417–428

    Article  PubMed  CAS  Google Scholar 

  • Lu R, Martin-Hernandez AM, Peart JR, Malcuit I, Baulcombe DC (2003) Virus-induced gene silencing in plants. Methods 30:296–303

    Article  PubMed  CAS  Google Scholar 

  • Mandanhar J, Hartman G, Sinclair J (1986) Colletotrichum destructivum, the anamorph of Glomerella glycines. Phytopathology 76:282–285

    Article  Google Scholar 

  • Palma MJ, Sandalio ML, Javier CF, Romero-Puertas CM, McCathy I, del Rio AL (2002) Plant proteases, protein degradation, and oxidative stress: role of peroxisomes. Plant Physiol Biochem 40:521–530

    Article  CAS  Google Scholar 

  • Preston GM (2000) Pseudomonas syringae pv. tomato the right pathogen, of the right plant, at the right time. Mol Plant Pathol 1:263–275

    Article  Google Scholar 

  • Reed JC (2000) Mechanisms of apoptosis. Am J Pathol 157:1415–1430

    PubMed  CAS  Google Scholar 

  • Richael C, Lincoln JE, Bostock RM, Gilchrist DG (2001) Caspase inhibitors reduce symptom development and limit bacterial proliferation in susceptible plant tissues. Physiol Mol Plant Pathol 59:213–221

    Article  CAS  Google Scholar 

  • Robertson D (2004) VIGS vectors for gene silencing: many targets, many tools. Annu Rev Plant Biol 55:495–519

    Article  PubMed  CAS  Google Scholar 

  • Rommens CMT, Salmeron JM, Oldroyd GED, Staskawicz BJ (1995) Intergeneric transfer and functional expression of the tomato disease resistance gene Pto. Plant Cell 7:1537–1544

    Article  PubMed  CAS  Google Scholar 

  • Ruiz M, Voinnet O, Baulcombe D (1998) Initiation and maintenance of virus-induced gene silencing. Plant Cell 10: 937–946

    Article  PubMed  CAS  Google Scholar 

  • Scofield SR, Tobias CM, Rathjen JP, Chang JH, Lavelle DT, Michelmore RW, Staskawicz BJ (1996) Molecular basis of gene-for-gene specificity in bacterial speck disease of tomato. Science 274:2063–2065

    Article  PubMed  CAS  Google Scholar 

  • Sessa G, Martin GB (2000) Signal recognition and transduction mediated by the tomato Pto kinase: a paradigm of innate immunity in plants. Microbes Infect 2:1591–1597

    Article  PubMed  CAS  Google Scholar 

  • Shen S, Goodwin PH, Hsiang T (2001) Hemibiotrophic infection and identity of the fungus, Colletotrichum destructivum, causing anthracnose of tobacco. Mycol Res 105:1340–1347

    Google Scholar 

  • Shimada T, Hiraiwa N, Nishimura M, Hara-Nishimura I (1994) Vacuolar processing enzyme of soybean that converts proprotein to the corresponding mature forms. Plant Cell Physiol 35:713–718

    PubMed  CAS  Google Scholar 

  • Solomon M, Belenghi B, Delledonne M, Menachem E, Levine A (1999) The involvement of cysteine proteases and protease inhibitor genes in the regulation of programmed cell death in plants. Plant Cell 11:431–443

    Article  PubMed  CAS  Google Scholar 

  • Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 24:4876–4882

    Article  Google Scholar 

  • Takken LWF, Luderer R, Gabriels HEJS, Westerink N, Lu R, Joosten HAJM (2000) A functional cloning strategy, based on a binary PVX-expression vector, to isolate HR-inducing cDNAs of plant pathogens. Plant J 24:275–283

    Article  PubMed  CAS  Google Scholar 

  • Uren AG, O’Rourke K, Aravind LA, Pisabarro MT, Seshagiri S, Koonin EV, Dixit VM (2000) Identification of paracaspases and metacaspases: two ancient families of caspase-like proteins, one of which plays a key role in MALT lymphoma. Mol Cell 6:961–967

    PubMed  CAS  Google Scholar 

  • van Baarlen P, Woltering EJ, Staats M, van Kan JAL (2007) Histochemical and genetic analysis of host and non-host interactions of Arabidopsis with three Botrytis species: an important role for cell death control. Mol Plant Pathol 8:41–54

    Article  Google Scholar 

  • Vercammen D, van de Cotte B, De Jaeger G, Eeckhout D, Casteels P, Vandepoele K, Vandenberghe I, Van Beeumen J, Inze D, Van Breusegem F (2004) Type II metacaspases Atmc4 and Atmc9 of Arabidopsis thaliana cleave substrates after arginine and lysine. J Biol Chem 279:45329–45336

    Article  PubMed  CAS  Google Scholar 

  • Watanabe N, Lam E (2004) Recent advance in the study of caspase-like proteases and Bax inhibitor-1 in plants: their possible roles as regulator of programmed cell death. Mol Plant Pathol 5:65–70

    Article  CAS  Google Scholar 

  • Watanabe N, Lam E (2005) Two arabidopsis metacaspases AtMCP1b and AtMCP2b are arginine/lysine-specific cysteine proteases and activate apoptosis-like cell death in yeast. J Biol Chem 280:14691–14699

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

Funding for this study was provided by the Natural Science and Engineering Research Council of Canada. Pseudomonas syringae pv. tomato DC3000 was kindly provided by Dr. Dianne Cuppels, Agriculture and Agri-Food Canada, London, ON, Canada.

Author information

Authors and Affiliations

  1. Department of Environmental Biology, University of Guelph, Guelph, ON, Canada, N1G 2W1

    L. Hao, P. H. Goodwin & T. Hsiang

Authors
  1. L. Hao
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. P. H. Goodwin
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. T. Hsiang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to P. H. Goodwin.

Additional information

Communicated by H. Judelson.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hao, L., Goodwin, P.H. & Hsiang, T. Expression of a metacaspase gene of Nicotiana benthamiana after inoculation with Colletotrichum destructivum or Pseudomonas syringae pv. tomato, and the effect of silencing the gene on the host response. Plant Cell Rep 26, 1879–1888 (2007). https://doi.org/10.1007/s00299-007-0387-7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00299-007-0387-7

Keywords

Search

Navigation