Skip to main content
Log in

Programmed cell death of plant tracheary elements differentiating in vitro

  • Published:
Protoplasma Aims and scope Submit manuscript

Summary

We used various microscopic and labeling techniques to examine events occurring during the programmed cell death (PCD) of plant tracheary elements (TEs) developing in vitro. TEs differentiating in vitro synthesize a secondary cell wall which is complex in composition and pattern at approximately 72 h after hormone manipulation. The timing of PCD events was established relative to this developmental marker. Cytoplasmic streaming continues throughout secondary wall synthesis, which takes 6 h to complete in a typical cell. Vital dye staining and ultrastructural analysis show that the vacuole and plasma membrane are intact during secondary cell wall synthesis, but the cytoplasm becomes less dense in appearance, most likely through the action of confined hydrolysis by small vacuoles which are seen throughout the cell at this time. The final, preeminent step of TE PCD is a rapid collapse of the vacuole occurring after completion of secondary cell wall synthesis. Vacuole collapse is an irreversible commitment to death which results in the immediate cessation of cytoplasmic streaming and leads to the complete degradation of cellular contents, which is probably accomplished by release of hydrolytic enzymes sequestered in the vacuole. This event represents a novel form of PCD. The degradation of nuclear DNA is detectable by TUNEL, an in situ labeling method, and appears to occur near or after vacuole collapse. Our observations indicate that the process of cellular degradation that produces the hollow TE cell corpse is an active and cell-autonomous process which is distinguishable morphologically and kinetically from necrosis. Although TE PCD does not resemble apoptosis morphologically, we describe the production of spherical protoplast fragments by cultured cells that resemble apoptotic bodies but which are not involved in TE PCD. We also present evidence that, unlike the hypersensitive response (HR), TE PCD does not involve an oxidative burst. While this evidence does not exclude a role for reactive oxygen intermediates in TE PCD, it does suggest TE PCD is mechanistically distinct from cell death during the HR.

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.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

BA:

6-benzylamino-purine

DAPI:

4′,6-diamidino-2-phenylindole diacetate

DCF:

2,7-dichlorofluorescein diacetate

DPI:

diphenyleneiodonium

FDA:

fluorescein diacetate

HR:

hypersensitive response

NAA:

α-naphthalene-acetic acid

PCD:

programmed cell death

ROI:

reactive oxygen intermediate

TE:

tracheary element

TUNEL:

TdT-mediated dUTP nick end labeling

References

  • Apostol I, Heinstein P, Low P (1989) Rapid stimulation of an oxidative burst during elicitation of cultured plant cells: role in defense and signal transduction. Plant Physiol 90: 109–116

    Google Scholar 

  • Auh C, Murphy T (1995) Plasma membrane redox enzyme is involved in the synthesis of Superoxide and hydrogen peroxide byPhytophthora elicitor-stimulated rose cells. Plant Physiol 107: 1241–1247

    Google Scholar 

  • Bestwick C, Bennett M, Mansfield J (1995)Hrp mutant ofPseudomonas syringae pv.phaseolicola induces cell wall alterations but not membrane damage leading to the hypersensitive reaction in lettuce. Plant Physiol 108: 503–516

    Google Scholar 

  • Bornman C, Spurr A, Addicott F (1967) Abscisin, auxin, and gibberellin effects on the developmental aspects of abscission in cotton. Am J Bot 54: 125–135

    Google Scholar 

  • Bortner C, Oldenburg N, Cidlowski J (1995) The role of DNA fragmentation in apoptosis. Trends Cell Biol 5: 21–26

    Google Scholar 

  • Burgess J, Linstead P (1984a) Comparison of tracheary element differentiation in intact leaves and isolated mesophyll cells ofZinnia elegans. Micron Microsc Acta 15: 153–160

    Google Scholar 

  • — — (1984b) In vitro tracheary element formation: structural studies and the effect of tri-iodobenzoic acid. Planta 160: 481–489

    Google Scholar 

  • Clarke P (1990) Developmental cell death: morphological diversity and multiple mechanisms. Anat Embryol 181: 195–213

    Google Scholar 

  • Dietrich R, Delaney T, Uknes S, Ward E, Ryals J, Dangl J (1994) Arabidopsis mutants simulating disease resistance response. Cell 77: 565–577

    Google Scholar 

  • Doyle J, Doyle J (1990) Isolation of DNA from small amounts of plant tissue. BRL Focus 12: 13–15

    Google Scholar 

  • Ellis R, Yuan J, Horvitz R (1991) Mechanisms and functions of cell death. Annu Rev Cell Biol 7: 663–698

    Google Scholar 

  • Esau K, Cheadle V, Risley E (1963) A view of the ultrastructure ofCucurbita xylem. Bot Gaz 124: 311–316

    Google Scholar 

  • Fett W, Jones S (1995) Microscopy of the interaction ofhrp mutants ofPseudomonas syringae pv.phaseolicola with a nonhost plant. Plant Sci 107: 27–39

    Google Scholar 

  • Fukuda H, Komamine A (1980a) Establishment of an experimental system for the study of tracheary element differentiation from single cells isolated from the mesophyll ofZinnia elegans. Plant Physiol 65: 57–60

    Google Scholar 

  • — — (1980b) Direct evidence for cytodifferentiation to tracheary elements without intervening mitosis in a culture of single cells isolated from the mesophyll ofZinnia elegans. Plant Physiol 65: 61–64

    Google Scholar 

  • Gahan P, Maple A (1965) The behaviour of lysosome-like particles during cell differentiation. J Exp Bot 17: 151–155

    Google Scholar 

  • Gavrieli Y, Sherman Y, Ben-Sasson S (1992) Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol 119: 493–501

    Google Scholar 

  • Glazener J, Orlandi E, Baker J (1996) The active oxygen response of cell suspensions to incompatible bacteria is not sufficient to cause hypersensitive cell death. Plant Physiol 110: 759–763

    Google Scholar 

  • Greenberg J, Guo A, Klessig D, Ausubel M (1994) Programmed cell death in plants: a pathogen-triggered response activated coordinately with multiple defense functions. Cell 77: 551–563

    Google Scholar 

  • Jabs T, Dietrich R, Dangl J (1996) Extracellular Superoxide initiates runaway cell death in an Arabidopsis mutant. Science 273: 1853–1856

    Google Scholar 

  • Jacobs W (1952) The role of auxin in differentiation of xylem around a wound. Am J Bot 39: 301–309

    Google Scholar 

  • Jones A, Dangl J (1996) Logjam at the Styx: the multiplicity of programmed cell death pathways in plants. Trends Plant Sci 1: 114–119

    Google Scholar 

  • Kane D, Sarafian T, Anton R, Hahn H, Gralla E, Valentine J, Ord T, Bredesen D (1993) Bcl-2 inhibition of neural death: decreased generation of reaction oxygen species. Science 262: 1274–1277

    Google Scholar 

  • Kerr J, Wyllie A, Currie A (1972) Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 26: 239–257

    Google Scholar 

  • Lai V, Srivastava L (1976) Nuclear changes during the differentiation of xylem vessel elements. Cytobiology 12: 220–243

    Google Scholar 

  • Levine A, Tenhaken R, Dixon R, Lamb C (1994) Hydrogen peroxide from the oxidative burst orchestrates the plant hypersensitive disease resistance response. Cell 79: 583–593

    Google Scholar 

  • —, Pennell R, Alvarez M, Palmer R, Lamb C (1996) Calcium-mediated apoptosis in a plant hypersensitive disease resistance response. Curr Biol 6: 427–437

    Google Scholar 

  • Locht U, Meier I, Hahlbrock K, Somssich I (1990) A 125 bp promoter fragment is sufficient for strong elicitor-mediated gene activation in parsley. EMBO J 9: 2945–2950

    Google Scholar 

  • Martin S, Green D, Cotter T (1994) Dicing with death: dissecting the components of the apoptosis machinery. Trends Biochem Sci 19: 26–30

    Google Scholar 

  • Masters J, Finch C, Sapolsky R (1989) Glucocorticoid endangerment of hippocampal neurons does not involve deoxyribonucleic acid cleavage. Endocrinology 124: 3083–3088

    Google Scholar 

  • Mehdy M (1994) Active oxygen species in plant defense against pathogens. Plant Physiol 105: 467–472

    Google Scholar 

  • Mittler R, Lam E (1995) In situ detection of nDNA fragmentation during the differentiation of tracheary elements in higher plants. Plant Physiol 108: 489–493

    Google Scholar 

  • —, Schulaer V, Lam E (1995) Coordinated activation of programmed cell death and defense mechanisms in transgenic tobacco plants expressing a bacterial proton pump. Plant Cell 7: 29–42

    Google Scholar 

  • Nagl W (1977) Plastolysosomes — plastids involved in the autolysis of the embryo-suspensor inPhaseolis. Z Pflanzenphysiol 85: 45–51

    Google Scholar 

  • Roberts L (1960) Experiments on xylem regeneration in stem wound responses inColeus. Bot Gaz 121: 201–208

    Google Scholar 

  • —, Fosket D (1962) Further experiments on wound vessel formation in stem wounds inColeus. Bot Gaz 123: 247–254

    Google Scholar 

  • Rosl F (1992) A simple and rapid method for detection of apoptosis in human cells. Nucleic Acid Res 20: 5243

    Google Scholar 

  • Ryerson D, Heath M (1996) Cleavage of nuclear DNA into oligonu-cleosomal fragments during cell death induced by fungal infection or by abiotic treatment. Plant Cell 8: 393–402

    Google Scholar 

  • Schwartz L, Smith S, Jones M, Osborne B (1993) Do all programmed cell deaths occur via apoptosis? Proc Natl Acad Sci USA 90: 980–984

    Google Scholar 

  • Srivastava L, Singh A (1972) Certain aspects of xylem differentiation in corn. Can J Bot 50: 1795–1804

    Google Scholar 

  • Stewart B (1994) Mechanisms of apoptosis: integration of genetic, biochemical, and cellular indicators. J Natl Cancer Inst 86: 1286–1296

    Google Scholar 

  • Thelen M, Northcote D (1989) Identification and purification of a nuclease fromZinnia elegans: a potential molecular marker for xylogenesis. Planta 179: 181–195

    Google Scholar 

  • Tomei L, Shapiro J, Cope F (1993) Apoptosis in C3H/10T1/2 mouse embryonic cells: evidence for internucleosomal DNA modification in the absence of double-strand cleavage. Proc Natl Acad Sci USA 90: 853–857

    Google Scholar 

  • Walbot V, Hoisington D, Neuffer M (1983) Disease lesions mimic mutations. In: Kosuge T, Meredith C, Hallaender A (eds) Genetic engineering of plants. Plenum, New York, pp 431–442

    Google Scholar 

  • Wang H, Li J, Bostock R, Gilchrist D (1996) Apoptosis: a functional paradigm for programmed cell death induced by a host-selective phytotoxin and invoked during development. Plant Cell 8: 375–391

    Google Scholar 

  • Wodzicki T, Brown C (1973) Organization and breakdown of the protoplast during maturation of pine tracheids. Am J Bot 60: 632–640

    Google Scholar 

  • —, Humphreys W (1972) Cytodifferentiation of maturing pine tracheids: the final stage. Tissue Cell 4: 525–528

    Google Scholar 

  • Wyllie A, Kerr J, Currie A (1980) Cell death: the significance of apoptosis. Int Rev Cytol 68: 251–306

    Google Scholar 

  • Ye Z, Droste D (1996) Isolation and characterization of cDNAs encoding xylogenesis-associated and wounding-induced ribonucleases inZinnia elegans. Plant Mol Biol 30: 697–709

    Google Scholar 

  • —, Varner J (1993) Gene expression patterns associated with in vitro tracheary element formation in isolated single mesophyll cells ofZinnia elegans. Plant Physiol 103: 805–813

    Google Scholar 

  • — — (1996) Induction of cysteine and serine proteases during xylogenesis inZinnia elegans. Plant Mol Biol 30: 1233–1246

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Groover, A., DeWitt, N., Heidel, A. et al. Programmed cell death of plant tracheary elements differentiating in vitro. Protoplasma 196, 197–211 (1997). https://doi.org/10.1007/BF01279568

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01279568

Keywords

Navigation