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l.ve-Cel. Imaging of Dual.l.bel.d Gol.i Stacks in Tobacco BY-2 Cel.s Reveal. Simil.r Behaviors for Different Cisternae during Movement and Brefel.in A Treatment

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Mol.cul.r Pl.nt 2011年第5期4卷 896-908页

作者： stephanie l. madison andreas nebenfuhr Department of Biochemistry and Cel.ul.r and Mol.cul.r Biol.gy University of Tennessee Knoxville TN 37996-0840 USA

In pl.nt cel.s, the Gol.i apparatus consists of numerous stacks that, in turn, are composed of several.fl.ttened cisternae with a cl.ar cis-to-trans pol.rity. During normal.functioning within l.ving cel.s, this unusual.organel.e displ.ys a wide range of dynamic behaviors such as whol. stack motil.ty, constant membrane fl.x through the cisternae, and Gol.i enzyme recycl.ng through the ER. In order to further investigate various aspects of Gol.i stack dynamics and integrity, we co-expressed pairs of establ.shed Gol.i markers in tobacco BY-2 cel.s to distinguish sub-compartments of the Gol.i during monensin treatments, movement, and brefel.in A （BFA）-induced disassembl.. A combination of cis and trans markers reveal.d that Gol.i stacks remain intact as they move through the cytopl.sm. The Gol.i stack orientation during these movements showed a sl.ght preference for the cis side moving ahead, but trans cisternae were al.o found at the l.ading edge. During BFA treatments, the different sub-compartments of about hal. of the observed stacks fused with the ER sequential.y; however, no consistent order coul. be detected. In contrast, the ionophore monensin resul.ed in swel.ing of trans cisternae whil. medial.and particul.rl. cis cisternae were mostl. unaffected. Our resul.s thus demonstrate a re- markabl. equival.nce of the different cisternae with respect to movement and BFA-induced fusion with the ER. In addi- tion, we propose that a combination of dual.l.bel.fl.orescence microscopy and drug treatments can provide a simpl. al.ernative approach to the determination of protein l.cal.zation to specific Gol.i sub-compartments.

关键词： Gol.i apparatus Gol.i stack integrity brefel.in A monensin tobacco BY-2 cel.s l.ve-cel. imaging.

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