Plant Physiol. Drug Metab Dispos
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Plant Physiology 84:1385-1390 (1987)
© 1987 American Society of Plant Biologists

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Membranes and Bioenergetics

Pontentiometric Cyanine Dyes Are Sensitive Probes for Mitochondria in Intact Plant Cells 1

Kinetin Enhances Mitochondrial Fluorescence

Zhanjiang Liu, W. R. Bushnell and Robert Brambl

Department of Plant Pathology, University of Minnesota, St. Paul, Minnesota 55108, Cereal Rust Laboratory, Agricultural Research Service, United States Department of Agriculture, University of Minnesota, St. Paul, Minnesota 55108, Plant Molecular Genetics Institute and Department of Plant Pathology, University of Minnesota, St. Paul, Minnesota 55108

Selected fluorescent dyes were tested for uptake by mitochrondria in intact cells of barley, maize, and onion. The cationic cyanine dye 3,3'-diheptyloxacarbocyanine iodide [DiOC7(3)] accumulated in mitochondria within 15 to 30 minutes without appreciable staining of other protoplasmic constituents. The number, shape, and movement of the fluorescent mitochondria could be seen readily, and the fluorescence intensity of the mitochondria could be monitored with a microscope photometer. Fluorescence was eliminated in 1 to 5 minutes by the protonophore carbonyl cyanide m-chlorophenylhydrazone (CCCP) indicating that maintenance of dye concentration was dependent on the inside-negative transmembrane potential maintained by functional mitochondria. Fluorescence of prestained mitochondria was enhanced within 5 to 10 minutes after addition of 0.1 millimolar kinetin to cells. The fluorescence in kinetintreated cells was dissipated by CCCP. These results suggest that kinetin interacted with respiratory processes resulting in higher potential across the mitochondrial membrane.

1 This research was supported in part by grants from the McKnight Foundation, The National Institute of General Medical Sciences (GM-19398), and the United States Department of Agriculture (85-CRCR-1-1596). Cooperative investigation of Agricultural Research Service, United States Department of Agriculture and the Minnesota Agricultural Experiment Station. Paper No. 15,060, Scientific Journal Series, Minnesota Agricultural Experiment Station. Mention of a trademark name or proprietary product does not constitute a guarantee or warranty by the United States Department of Agriculture or imply its approval to the exclusion of other products that may also be suitable.




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Copyright © 1987 by the American Society of Plant Biologists