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Rise in chlorotetracycline fluorescence accompanies tracheary element differentiation in suspension cultures ofZinnia

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Summary

Developing tracheary elements in suspension cultures ofZinnia elegans fluoresce intensely relative to non-differentiating cells when stained with chlorotetracycline (CTC), a fluorescent chelate probe for membrane associated calcium. This suggests that a change in calcium uptake or subcellular distribution accompanies the onset of tracheary element differentiation. A few cells in early differentiating cultures were brightly fluorescent, but did not have visible cell wall thickenings, suggesting that a rise in sequestered calcium may precede visible differentiation. Diffuse CTC fluorescence in early differentiation most likely results from sequestration of calcium in the endoplasmic reticulum. Late in differentiation, CTC fluorescence becomes punctate in appearance, probably due to loss of plasma membrane integrity occurring at the onset of autolysis.Zinnia suspension culture cells were found to be very sensitive to CTC and low concentrations (10 μM) were used to assure accurate localization of membrane-associated calcium in healthy cells.

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Abbreviations

CTC:

chlorotetracycline

DIC:

differential interference contrast

DiOC6 :

3,3′-dihexyloxacarbocyanine iodide

ER:

endoplasmic reticulum

EGTA:

ethylene glycol bis-(amino-ethyl ether) N,N,N1N1-tetraacetic acid

NPN:

n-phenylnaphthylamine

OsFeCN:

osmium tetroxide and potassium ferricyanide

TE:

tracheary element

TEM:

transmission electron microscopy

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Authors and Affiliations

  1. Department of Biological Sciences, Texas Tech University, Lubbock, Texas

    Alison W. Roberts & Candace H. Haigler

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  1. Alison W. Roberts
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  2. Candace H. Haigler
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Roberts, A.W., Haigler, C.H. Rise in chlorotetracycline fluorescence accompanies tracheary element differentiation in suspension cultures ofZinnia . Protoplasma 152, 37–45 (1989). https://doi.org/10.1007/BF01354238

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  • DOI: https://doi.org/10.1007/BF01354238

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