Summary
Antibodies raised against the calcium-binding protein centrin, were used to identify and localise centrin containing structures in the flagellar apparatus of zoospores and cysts of the oomycetePhytophthora cinnamomi. Immunoblotting of extracts from zoospores indicates that theP. cinnamomi centrin homologue is a 20 kDa protein. Immunofluorescence microscopy with anti-centrin antibodies reveals labelling in the flagella, the basal body connector and co-localisation along the microtubular R1 root (formerly called AR3) that runs from the right side of the basal body of the anterior flagellum into the anterior of the zoospore close to the ventral surface. The centrin (R1cen) and tubulin components of the R1 root split into four loops on the right hand side of the ventral groove and rejoin along the left hand side of the groove. The R1 root continues down the left hand side of the zoospore past the basal bodies and parallel to the R4 root. We propose that at least inP. cinnamomi there is no R2 root. Immunogold labelling confirms that centrin is a component of the basal body connector complex. When the zoospores become spherical during encystment, the R1cen pivots by approximately 90 ° with respect to the nucleus.
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References
Andersen RA (1991) The cytoskeleton of chromophyte algae. Protoplasma 164: 143–159
—, Barr DJS, Lynn DH, Melkonian M, Moestrup O, Sleigh MA (1991) Terminology and nomenclature of the cytoskeletal elements associated wih the flagellar/ciliary apparatus in protists. Protoplasma 164: 1–8
Baron AT, Salisbury JL (1992) Role of centrin in spindle pole dynamics. In: Kalnins VI (ed) The centrosome. Academic Press, San Diego, pp 167–195
—, Greenwood T, Bazinet C, Salisbury JL (1993) Centrin is a component of the pericentriolar lattice. Biol Cell 76: 383–388
Barr DJS, Allan PME (1985) A comparison of the flagellar apparatus inPhytophthora, Saprolegnia, Thraustochytrium andRhizidiomyces. Can J Bot 63: 138–154
—, Désaulniers NL (1987) Ultrastructure of theLagena radicola zoospore, including a comparison with the primary and secondarySaprolegnia zoospores. Can J Bot 65: 2161–2176
— — (1989) The flagellar apparatus of the Oomycetes and Hyphochytridiomycetes. In: Green JC, Leadbetter BSC, Diver WL (eds) The chromophyte algae: problems and perspectives. Clarendon, Oxford, pp 343–355
— — (1992) The flagellar apparatus in zoospores ofPhytophthora, Pythium, andHalophytophthora. Can J Bot 70: 2163–2169
Beakes GW (1989) Oomycete fungi: their phylogeny and relationship to chromophyte algae. In: Green JC, Leadbeater BSC, Diver WL (eds) The chromophyte algae: problems and perspectives. Clarendon, Oxford, pp 325–342
Carlile MJ (1983) Motility, taxis, and tropism inPhytophthora. In: Erwin DC, Bartnicki-Garcia S, Tsao P (eds)Phytophthora. Its biology, taxonomy, ecology, and pathology. American Phytopathology Society, St Paul, pp 95–107
Davis FM, Tsao TY, Fowler SK, Rao PN (1983) Monoclonal antibodies to mitotic cells. Proc Natl Acad Sci USA 80: 2926–2930
Deacon JW, Donaldson SP (1993) Molecular recognition in the homing response of zoosporic fungi, which special reference toPythium andPhytophthora. Mycol Res 97: 1153–1171
Errabolu R, Sanders MA, Salisbury JL (1994) Cloning of a cDNA encoding human centrin, an EF-hand protein of centrosomes and mitotic spindle poles. J Cell Sci 107: 9–16
Gubler F, Jablonsky PP, Duniec J, Hardham AR (1990) Localization of calmodulin in flagella of zoospores ofPhytophthora cinnamomi. Protoplasma 155: 233–238
Gundersen JH, Elwood H, Ingold A, Kindle K, Sogin ML (1987) Phylogenetic relationships between chlorophytes, chrysophytes and oomycetes. Proc Natl Acad Sci USA 84: 5823–5827
Hardham AR (1987) Microtubules and the flagellar apparatus in zoospores and cysts of the fungusPhytophthora cinnamoml. Protoplasma 137: 109–124
— (1992) Cell biology and pathogenesis. Annu Rev Plant Physiol Plant Mol Biol 43: 491–526
—, Gubler F, Duniec J, Elliott J (1991) A review of methods for the production and use of monoclonal antibodies to study zoosporic plant pathogens. J Microsc 162: 305–318
Harper JDI, Sanders MA, Salisbury JL (1993) Phosphorylation of nuclear and flagellar basal apparatus proteins during flagellar regeneration inChlamydomonas reinhardtii. J Cell Biol 122: 877–886
Hemmes DE (1983) Cytology ofPhytophthora. In: Erwin DC, Bartnicki-Garcia S, Tsao PH (eds)Phytophthora. Its biology, taxonomy, ecology, and pathology. American Phytopathological Society, St Paul, pp 9–40
Holloway SA, Heath IB (1977) Morphogenesis and the role of microtubules in synchronous populations ofSaprolegnia zoospores. Exp Mycol 1: 9–29
Höhfeld I, Otten J, Melkonian M (1988) Contractile eukaryotic flagella: centrin is involved. Protoplasma 147: 16–24
Hyde GJ, Hardham AR (1993) Microtubules regulate the generation of polarity in zoospores ofPhytophthora cinnamomi. Eur J Cell Biol 62: 75–85
Katsaros CI, Maier I, Melkonian M (1993) Immunolocalization of centrin in the flagellar apparatus of male gametes ofEctocarpus siliculosus (Phaeophyceae) and other brown algal motile cells. J Phycol 29: 787–797
Koutoulis A, McFadden GI, Wetherbee R (1988) Spine-scale reorientation inApedinella radians (Pedinellales, Chrysophyceae): the microarchitecture and immunocytochemistry of the associated cytoskeleton. Protoplasma 147: 25–41
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685
McFadden GI, Schulze D, Surek B, Salisbury JL, Melkonian M (1987) Basal body reorientation mediated by a Ca2+-modulated contractile protein. J Cell Biol 105: 903–912
Melkonian M, Beech PL, Katsaros C, Schulze D (1992) Centrinmediated cell motility in algae. In: Melkonian M (ed) Algal cell motility. Chapman and Hall, New York, pp 179–221
Moudjou M, Paintrand M, Vigues B, Bornens M (1991) A human centrosomal protein is immunologically related to basal body-associated proteins from lower eucaryotes and is involved in the nucleation of microtubules. J Cell Biol 115: 129–140
Piperno G, Mead K, Shestak W (1992) The inner dynein arms I2 interact with a “dynein regulatory complex” inChlamydomonas flagella. J Cell Biol 118: 1455–1463
Preisig HR (1989) The flagellar base ultrastructure and phylogeny of chromophytes. In: Green JC, Leadbetter BSC, Diver WL (eds) The chromophyte algae: problems and perspectives. Clarendon, Oxford, pp 179–221
Roberts KR, Roberts JE (1991) The flagellar apparatus and cytoskeleton of the dinoflagellates. A comparative review. Protoplasma 164: 105–122
— —, Cormier SA (1992) The dinoflagellate cytoskeleton. In: Menzel D (ed) The cytoskeleton of the algae. CRC Press, Boca Raton, pp 19–38
Salisbury JL (1989) Algal centrin: calcium-sensitive contractile organelles. In: Coleman AW, Goff LJ, Stein-Taylor JR (eds) Algae as experimental systems. AR Liss, New York, pp 19–37
— (1989) Centrin and the algal flagellar apparatus. J Phycol 25: 201–206
—, Floyd GL (1978) Calcium-induced contraction of the rhizoplast of a quadriflagellate green alga. Science 202: 975–977
—, Baron A, Surek B, Melkonian M (1984) Striated flagellar roots: isolation and partial characterization of a calcium-modulated contractile organelle. J Cell Biol 99: 962–970
—, Sanders MA, Harpst L (1987) Flagellar root contraction and nuclear movement during flagellar regeneration inChlamydomonas reinhardtii. J Cell Biol 105: 1799–1805
—, Baron AT, Sanders MA (1988) The centrin-based cytoskeleton ofChlamydomonas reinhardtii: distribution in interphase and mitotic cells. J Cell Biol 107: 635–641
Sanders MA, Salisbury JL (1994) Centrin plays an essential role in microtubule severing during flagellar excision inChlamydomonas reinhardtii. J Cell Biol 124: 795–805
Sherwin T, Gull K (1989) Visualization of detyrosination along single microtubules reveals novel mechanisms of assembly during cytoskeletal duplication inTrypanosomes. Cell 57: 211–221
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354
Vaughn KC, Sherman TD, Renzaglia KS (1993) A centrin homologue is a component of the multilayered structure in bryophytes and pteridophytes. Protoplasma 175: 58–66
Wright RL, Salisbury JL, Jarvik JW (1985) A nucleus basal body connector inChlamydomonas reinhardtii that may function in basal body localization or segregation. J Cell Biol 101: 1903–1912
—, Adler SA, Spanier JG, Jarvik JW (1989) Nucleus-basal body connector inChlamydomonas: evidence for a role in basal body segregation and against essential roles in mitosis or in determining cell polarity. Cell Motil Cytoskeleton 14: 516–526
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Harper, J.D.I., Gubler, F., Salisbury, J.L. et al. Centrin association with the flagellar apparatus in spores ofPhytophthora cinnamomi . Protoplasma 188, 225–235 (1995). https://doi.org/10.1007/BF01280374
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DOI: https://doi.org/10.1007/BF01280374