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Protoplasma

, Volume 196, Issue 3–4, pp 197–211 | Cite as

Programmed cell death of plant tracheary elements differentiating in vitro

  • A. Groover
  • N. DeWitt
  • A. Heidel
  • A. Jones
Article

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.

Keywords

Apoptosis Programmed cell death Tracheary element Xylogenesis Zinnia elegans 

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

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Copyright information

© Springer-Verlag 1997

Authors and Affiliations

  • A. Groover
    • 1
  • N. DeWitt
    • 1
  • A. Heidel
    • 1
  • A. Jones
    • 1
  1. 1.Department of BiologyUniversity of North Carolina at Chapel HillNCUSA

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