Skip to main content
SpringerLink
Log in

Phosphite accelerates programmed cell death in phosphate-starved oilseed rape (Brassica napus) suspension cell cultures

  • Original Article
  • Published:
Planta Aims and scope Submit manuscript

Abstract

Phosphite (H2PO3 −, Phi) prevents the acclimation of plants and yeast to orthophosphate (Pi, HPO4 2−) deprivation by specifically obstructing the derepression of genes encoding proteins characteristic of their Pi-starvation response. In this study, we report that prolonged (i.e., 3–4 weeks) culture of Brassica napus L. suspension cells in Pi-deficient (−Pi) media leads to programmed cell death (PCD). However, when the B. napus cells were subcultured into −Pi media containing 2 mM Phi, they initiated PCD within 5 days, with 95% cell death observed by day 9. Dying cells exhibited several morphological and biochemical features characteristic of PCD, including protoplast shrinkage, chromatin condensation, and fragmentation of nuclear DNA. Immunoblotting indicated that B. napus cells undergoing PCD upregulated a 30-kDa cysteine endoprotease that is induced during PCD in the inner integument cells of developing B. napus seeds. It is concluded that PCD in B. napus suspension cells is triggered by extended Pi starvation, and that Phi treatment greatly accelerates this process. Our results also infer that the adaptive value of acclimating at the molecular level to Pi-stress is to extend the viability of −Pi B. napus cell cultures by about 3 weeks.

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. 1a, b.
Fig. 2.
Fig. 3a–c.
Fig. 4a–e.
Fig. 5.
Fig. 6a, b.

Similar content being viewed by others

Abbreviations

APase:

acid phosphatase (EC 3.1.3.2)

BnCysP:

B. napus cysteine proteinase

DAPI:

4′,6′-diamidino-2-phenylindole

FDA:

fluorescein diacetate

PCD:

programmed cell death

Phi:

phosphite

+Pi and −Pi:

Pi-sufficient and -deficient, respectively

PI:

propidium iodide

PSI:

Pi-starvation inducible

References

  • Abel S, Ticconi CA, Delatorre CA (2002) Phosphate sensing in higher plants. Physiol Plant 115:1–8

    CAS  PubMed  Google Scholar 

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

    CAS  PubMed  Google Scholar 

  • Bollag DM, Rozycki MD, Edelstein SJ (1996) Protein methods. Wiley–Liss, New York, pp 62–70

  • Carswell MC, Grant BR, Niere J, Theodorou ME, Plaxton WC (1996) The fungicide phosphonate disrupts the phosphate starvation response of Brassica nigra seedlings. Plant Physiol 110:105–115

    CAS  PubMed  Google Scholar 

  • Carswell MC, Grant BR, Plaxton WC (1997) Disruption of the phosphate-starvation response of oilseed rape suspension cells by the fungicide phosphonate. Planta 203:67–74

    Article  CAS  PubMed  Google Scholar 

  • De Jong AJ, Hoeberichts FA, Yakimova ET, Maximova E, Woltering EJ (2000) Chemical-induced apoptotic cell death in tomato cells: involvement of caspase-like proteases. Planta 211:656–662

    Article  PubMed  Google Scholar 

  • Devaux C, Baldet P, Joubès J, Dieuaide-Noubhani M, Just D, Chevalier C, Raymond P (2003) Physiological, biochemical and molecular analysis of sugar-starvation responses in tomato roots. J Exp Bot (in press)

  • Foerster H, Adaskaveg JE, Kim DH, Stanghellini ME (1998) Effect of phosphite on tomato and pepper plants and on susceptibility of pepper to Phytophthora root and crown rot in hydroponic culture. Plant Dis 82:1165–1170

    Google Scholar 

  • Guest D, Grant BR (1991) The complex action of phosphonates as antifungal agents. Biol Rev 66:159–158

    Google Scholar 

  • Hoeberichts FA, Orzaez D, van der Plas LHW, Woltering EJ (2001) Changes in gene expression during programmed cell death in tomato cell suspensions. Plant Mol Biol 45:641–654

    Article  CAS  PubMed  Google Scholar 

  • Jones KH, Senft JA (1985) An improved method to determine cell viability by simultaneous staining with fluorescein diacetate–propidium iodide. J Histochem Cytochem 33:77–79

    CAS  PubMed  Google Scholar 

  • Krishnamurthy KV, Krishnaraj R, Chozhavendan R, Christopher FS (2000) The programme of cell death in plants and animal — a comparison. Curr Sci 79:1169–1181

    CAS  Google Scholar 

  • McCabe PF, Leaver CJ (2000) Programmed cell death in cell cultures. Plant Mol Biol 44:350–368

    Google Scholar 

  • McDonald AE, Grant BR, Plaxton WC (2001a) Phosphite (phosphorous acid): its relevance in the environment and agriculture, and influence on the plant phosphate starvation response. J Plant Nutr 24:1505–1519

    Article  CAS  Google Scholar 

  • McDonald AE, Niere JO, Plaxton WC (2001b) Phosphite disrupts the acclimation of Saccharomyces cerevisiae to phosphate starvation. Can J Microbiol 47:969–978

    Article  CAS  PubMed  Google Scholar 

  • Mlejnek P, Procházka S (2002) Activation of caspase-like proteases and induction of apoptosis by isopentenyladenosine in tobacco BY-2 cells. Planta 215:158–166

    Article  CAS  PubMed  Google Scholar 

  • Moraes TF, Plaxton WC (2000) Purification and characterization of phosphoenolpyruvate carboxylase from Brassica napus (rapeseed) suspension cell cultures. Implications for phosphoenolpyruvate carboxylase regulation during phosphate starvation and the integration of glycolysis with nitrogen assimilation. Eur J Biochem 267:4465–4476

    Article  CAS  PubMed  Google Scholar 

  • Pennel RI, Lamb C (1997) Programmed cell death in plants. Plant Cell 9:1157–1168

    CAS  Google Scholar 

  • Plaxton WC (1999) Metabolic aspects of phosphate starvation in plants. In: Deikman J, Lynch J (eds) Phosphorus in plant biology: regulatory roles in molecular, cellular, organismic, and ecosystem processes. ASPP, Rockville, MD, pp 164–176

  • Rickard DA (2000) Review of phosphorous acid and its salts as fertilizer materials. J Plant Nutr 23:161–180

    CAS  Google Scholar 

  • Smith CR, Knowles VL, Plaxton WC (2000) Purification and characterization of cytosolic pyruvate kinase from Brassica napus (rapeseed) suspension cell cultures. Implications for the integration of glycolysis with nitrogen assimilation. Eur J. Biochem 267:4477–4485

    Article  CAS  Google Scholar 

  • Swidzinski JA, Sweetlove LJ, Leaver CJ (2002) A custom microarray analysis of gene expression during programmed cell death in Arabidopsis thaliana. Plant J 30:431–446

    Article  CAS  PubMed  Google Scholar 

  • Ticconi CA, Delatorre CA, Abel S (2001) Attenuation of phosphate starvation responses by phosphite in Arabidopsis. Plant Physiol 127:963–972

    Article  CAS  PubMed  Google Scholar 

  • Varadarajan DK, Karthikeyan AS, Matilda PD, Raghothama KG (2002) Phosphite, an analog of phosphate, suppresses the coordinated expression of genes under phosphate starvation. Plant Physiol 129:1–9

    Article  Google Scholar 

  • Wan L, Xia Q, Selvaraj G (2002) Early stages of seed development in Brassica napus: a seed coat-specific cysteine proteinase associated with programmed cell death of the inner integument. Plant J 30:1–10

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We are very grateful to Drs. Steven Huber (USDA Plant Science Research, North Carolina State Univ.), Paul McCabe (Dept. of Botany, Univ. College Dublin), and Phillipe Raymond (Unité Mixte Rech. Physiol. Biotech. Vég., Centre de Recherché de Bordeaux) for helpful discussions, to Drs. William Newcomb and Paul Young (Dept. of Biology, Queens Univ.) for providing microscopy facilities, and to Dr. Gopalan Selvaraj (Plant Biotech. Inst, National Research Council of Canada) for the gift of rabbit anti-(BnCysP) immune serum. Financial support was generously provided by research and equipment grants from the Natural Sciences and Engineering Research Council of Canada (NSERC).

Author information

Authors and Affiliations

  1. Departments of Biology and Biochemistry, Queen's University, Kingston, Ontario, K7L 3N6, Canada

    William C. Plaxton

  2. Department of Biology, Queen's University, Kingston, Ontario, K7L 3N6, Canada

    Vinay K. Singh, Susan M. Wood & Vicki L. Knowles

Authors
  1. Vinay K. Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Susan M. Wood
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Vicki L. Knowles
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. William C. Plaxton
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to William C. Plaxton.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Singh, V.K., Wood, S.M., Knowles, V.L. et al. Phosphite accelerates programmed cell death in phosphate-starved oilseed rape (Brassica napus) suspension cell cultures. Planta 218, 233–239 (2003). https://doi.org/10.1007/s00425-003-1088-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00425-003-1088-2

Keywords

Search

Navigation