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Auxin Deprivation Induces Synchronous Golgi Differentiation in Suspension-Cultured Tobacco BY-2 Cells1

Zev M. Winicur2, *, Guo Feng Zhang3, and L. Andrew Staehelin

Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, Colorado 80309-0347

To date, the lack of a method for inducing plant cells and their Golgi stacks to differentiate in a synchronous manner has made it difficult to characterize the nature and extent of Golgi retailoring in biochemical terms. Here we report that auxin deprivation can be used to induce a uniform population of suspension-cultured tobacco (Nicotiana tabacum cv BY-2) cells to differentiate synchronously during a 4-d period. Upon removal of auxin, the cells stop dividing, undergo elongation, and differentiate in a manner that mimics the formation of slime-secreting epidermal and peripheral root-cap cells. The morphological changes to the Golgi apparatus include a proportional increase in the number of trans-Golgi cisternae, a switch to larger-sized secretory vesicles that bud from the trans-Golgi cisternae, and an increase in osmium staining of the secretory products. Biochemical alterations include an increase in large, fucosylated, mucin-type glycoproteins, changes in the types of secreted arabinogalactan proteins, and an increase in the amounts and types of molecules containing the peripheral root-cap-cell-specific epitope JIM 13. Taken together, these findings support the hypothesis that auxin deprivation can be used to induce tobacco BY-2 cells to differentiate synchronously into mucilage-secreting cells.

1   This work was supported by a National Institutes of Health grant (no. 18639 to L.A.S.).
2   Present address: Plant Biology Department, University of Minnesota, 220 Biological Sciences Center, 1445 Gortner Avenue, St. Paul, MN 55108.
3   Present address: Department of Biochemistry and Biophysics, University of California, San Francisco, CA 94143-0554.
*   Corresponding author; e-mail zwinicur{at}biosci.cbs.umn.edu; fax 1-612-625-1738.

Plant Physiol. (1998) 117: 501-513
Copyright Clearance Center:   0032-0889/98/117/0501/13
© 1998 American Society of Plant Physiologists




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