Brefeldin A Affects Growth, Endoplasmic Reticulum, Golgi Bodies, Tubular Vacuole System, and Secretory Pathway in Pisolithus tinctorius

Abstract

Cole, L., Davies, D., Hyde, G. J., and Ashford, A. E. 2000. Brefeldin A affects growth, endoplasmic reticulum, golgi bodies, tubular vacuole system, and secretory pathway in Pisolithus tinctorius. Brefeldin A (BFA) reduced radial growth in Pisolithus tinctorius at a concentration as low as 2 μM. Use of endoplasmic reticulum (ER)-Tracker dye, unconjugated BFA, and fluorescent BFA (BODIPY-BFA) allowed comparison of the effects of BFA on the endomembrane system of P. tinctorius at the light and electron microscope levels. Both ER-Tracker dye and BODIPY-BFA have been shown previously to label the ER. Unconjugated BFA and BODIPY-BFA modified the ER network and disrupted the tubular vacuole system in the tip region. The ultrastructure in freeze-substituted hyphae showed that BFA treatment resulted in (i) disruption of the Spitzenkörper, (ii) reduction in number of apical vesicles, (iii) redistribution and mild dilation of ER, and (iv) persistence and increased size and complexity of Golgi bodies. The effects of BFA on the ER were only partially reversible in the time period examined. We conclude that in P. tinctorius, BFA as the free metabolite or BODIPY-BFA affects the tubular vacuole system as well as anterograde membrane flow between the ER and the Golgi bodies and post-Golgi transport.