ER-tracker dye and BODIPY-brefeldin A differentiate the endoplasmic reticulum and Golgi bodies from the tubular-vacuole system in living hyphae of Pisolithus tinctorius

被引:64
作者
Davies, D. [1 ]
Hyde, G.J. [1 ]
Ashford, A.E. [1 ]
Cole, Louise [1 ]
机构
[1] School of Biological Sciences, University of New South Wales, Sydney
关键词
BODIPY-brefeldin A; Endoplasmic reticulum; ER-Tracker(TM) Blue-White DPX; Fluorescence microscopy; Freeze-substitution; Fungi; Golgi bodies; Pisolithus tinctorius; Ultrastructure; Vacuoles;
D O I
10.1046/j.1365-2818.2000.00664.x
中图分类号
学科分类号
摘要
Two fluorochromes, ER-Tracke(TM) Blue-White DPX dye and the fluorescent brefeldin A (BFA) derivative, BODIPY-BFA, label the endoplasmic reticulum (ER) in hyphal tips of Pisolithus tinctorius and allow its differentiation from the tubular-vacuole system at the light microscope level in living cells. The ER-Tracker dye labels a reticulate network similar in distribution to ER as seen in electron micrographs of freeze-substituted hyphae. BODIPY- BFA stains a thicker axially aligned structure with an expanded region at the apex, which is similar to that seen when hyphae are stained with ER-Tracker dye in the presence of unconjugated BFA. This structure is considered to be ER modified by BFA, a view supported by ultrastructural observations of the effect of BFA on the fungal ER. Both fluorescent probes also stain punctate structures, which are most likely to be Golgi bodies. Neither probe labels the tubular-vacuole system.
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页码:239 / 249
页数:10
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  • [1] Abad A.R., Cease K.R., Blanchette R.A., A rapid technique using epoxy resin Quetol 651 to prepare woody plant tissues for ultrastructural study, Can. J. Bot., 66, pp. 677-682, (1988)
  • [2] Betina V., Biological effects of the antibiotic Brefeldin a (decumbin, cyanein, ascotoxin, synergisidin): A retrospective, Folia Microbiol., 37, pp. 3-11, (1992)
  • [3] Boevink P., Santa Cruz S., Hawes C.H.N., Oparka K., Virus mediated delivery of the green fluorescent protein to the endoplasmic reticulum of plant cells, Plant J., 10, pp. 935-941, (1996)
  • [4] Boevink P., Oparka K., Santa Cruz S., Martin B., Betteridge A., Hawes C., Stacks on tracks: The plant Golgi apparatus traffics on actin/ER network, Plant J., 15, pp. 441-447, (1998)
  • [5] Bourett T.M., Howard R.J., Enhanced labelling of concanavalin A binding sites in fungal endomembrane using a double-sided, indirect method, Mycol. Res., 98, pp. 769-775, (1994)
  • [6] Bourett T.M., Howard R.J., Brefeldin a-induced structural changes in the endomembrane system of a filamentous fungus, Magnaporthe grisea, Protoplasma, 190, pp. 151-163, (1996)
  • [7] Butt T.M., Hoch H.C., Staples R.C., St Leger R.J., Use of fluorochromes in the study of fungal cytology and differentiation, Exp. Mycol., 13, pp. 303-320, (1989)
  • [8] Campbell I.M., Fungal secondary metabolism research: Past, present and future, J. Natural Products, 46, pp. 60-70, (1983)
  • [9] Cole L., Hyde G.J., Ashford A.E., Uptake and compartmentalisation of fluorescent probes by Pisolithus tinctorius hyphae: Evidence for an anion transport mechanism at the tonoplast but not for fluid-phase endocytosis, Protoplasma, 199, pp. 18-29, (1997)
  • [10] Cole L., Orlovich D.A., Ashford A.E., Structure, function, and motility of vacuoles in filamentous fungi, Fungal Genet. Biol., 24, pp. 86-100, (1998)