Advertisement

Protoplasma

, Volume 181, Issue 1–4, pp 213–232 | Cite as

Cell surface antigens ofPhytophthora spores: biological and taxonomic characterization

  • A. R. Hardham
  • D. M. Cahill
  • M. Cope
  • B. K. Gabor
  • F. Gubler
  • G. J. Hyde
Article

Summary

The oomycetes are a class of protists that produce biflagellate asexual zoospores. Members of the oomycetes have close phylogenetic affinities with the chromophyte algae and are widely divergent from the higher fungi. This review focuses on two genera,Phytophthora andPythium, which belong to the family Pythiaceae, and the order Peronosporales. These two genera contain many species that cause serious diseases in plants. Molecules on the surface of zoospores and cysts of these organisms are likely to play crucial roles in the infection of host plants. Knowledge of the properties of the surface of these cells should thus help increase our understanding of the infection process. Recent studies ofPhytophthora cinnamomi andPythium aphanidermatum have used lectins to analyse surface carbohydrates and have generated monoclonal antibodies (MAbs) directed towards a variety of zoospore and cysts surface components. Labelling studies with these probes have detected molecular differences between the surface of the cell body and of the flagella of the zoospores. They have been used to follow changes in surface components during encystment, including the secretion of an adhesive that bonds the spores to the host surface. Binding of lectin and antibody probes to the surface of living zoospores can induce encystment, giving evidence of cell receptors involved in this process. Freeze-substitution and immunolabelling studies have greatly augmented our understanding of the synthesis and assembly of the zoospore surface during zoosporogenesis. Synthesis of a variety of zoospore components begins when sporulation is induced. Cleavage of the multinucleate sporangium is achieved through the progressive extension of partitioning membranes, and a number of surface antigens are assembled onto the zoospore surface during cleavage. Comparisons of antibody binding to many isolates and species ofPhytophthora andPythium have revealed that surface components on zoospores and cysts exhibit a range of taxonomic specificities. Surface antigens or epitopes may occur on only a few isolates of a species; they may be species-specific, genus-specific or occur on the spores of both genera. Spore surface antigens thus promise to be of significant value for studies of the taxonomy and phylogeny of these protists, as well as for disease diagnosis.

Keywords

Oomycetes Pythium Phytophthora Monoclonal antibodies Surface antigens Immunocytochemistry 

Abbreviations

MAbs

monoclonal antibodies

ConA

Concanavalin A

SBA

soybean agglutinin

WGA

wheat germ agglutinin

gps

glycoproteins

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Adler J (1976) The sensing of chemicals by bacteria. Scient Am 234: 40–47Google Scholar
  2. Ali-Shtayeh MS, MacDonald JD, Kabashima J (1991) A method for using commercial ELISA tests to detect zoospores ofPhytophthora andPythium species in irrigation water. Plant Dis 75: 305–311Google Scholar
  3. Anderson MA, Sandrin MS, Clarke AE (1984) A high proportion of hybridomas raised to a plant extract secrete antibody to arabinose or galactose. Plant Physiol 75: 1013–1016Google Scholar
  4. Arms K, Camp PS (1979) Biology. Holt, Rinehart and Winston, WB Saunders, New YorkGoogle Scholar
  5. Bacic A, Williams ML, Clarke AE (1985) Studies on the cell surface of zoospores and cysts of the fungusPhytophthora cinnamomi: nature of the surface saccharides as determined by quantitative lectin binding studies. J Histochem Cytochem 33: 384–388Google Scholar
  6. Bailey AM, Mena GL, Herrera-Estrella L (1991) Genetic transformation of the plant pathogensPhytophthora capsici andPhytophthora parasitica. Nucleic Acids Res 19: 4273–4278Google Scholar
  7. — — — (1993) Transformation of four pathogenicPhytophthora spp. by microprojectile bombardment on intact mycelia. Curr Genet 23: 42–46Google Scholar
  8. Barr DJS (1981) The phylogenetic and taxonomic implications of flagellar rootlet morphology among zoosporic fungi. BioSystems 14: 359–370Google Scholar
  9. — (1983) The zoosporic grouping of plant pathogens. Entity or non-entity? In: Buczacki ST (ed) Zoosporic plant pathogens. A modern perspective. Academic Press, London, pp 43–83Google Scholar
  10. Bartnicki-Garcia S (1970) Cell wall composition and other biochemical markers in fungal phytogeny. In: Harborne JB (ed) Phytochemical phytogeny, Academic Press, London, pp 81–103Google Scholar
  11. — (1987) The cell wall: a crucial structure in fungal evolution. In: Rayner ADM, Brasier CM, Moore D (eds) Evolutionary biology of the fungi. Cambridge University Press, Cambridge, pp 389–403Google Scholar
  12. —, Wang MC (1983) Biochemical aspects of morphogenesis inPhytophthora. In: Erwin DC, Bartnicki-Garcia S, Tsao P (eds)Phytophthora. Its biology, taxonomy, ecology, and pathology. American Phytopathological Society, St. Paul, MN, pp 121–137Google Scholar
  13. Bouck GB (1971) The structure, origin, isolation, and composition of the tubular mastigonemes of theOchromonas flagellum. J Cell Biol 50: 362–384Google Scholar
  14. Bu'Lock JD, Osagie AU (1976) Sterol biosynthesis via cycloartenol inSaprolegnia. Phytochemistry 15: 1249–1251Google Scholar
  15. Burr AW, Beakes GW (1994) A comparative study of zoospore and cysts surface structure in saprophytic and fish pathogenicSaprolegnia species (oomycetes fungi) using lectins and monoclonal antibodies. Protoplasma 181: 142–163Google Scholar
  16. Burrell RG, Clayton CW, Gallegly ME, Lilly VG (1966) Factors affecting the antigenicity of the mycelium of three species ofPhytophthora. Phytopathology 56: 422–426Google Scholar
  17. Cahill DM, Hardham AR (1994) Exploitation of zoospore taxis in the development of a novel dipstick immunoassay for the specific detection ofPhytophthora cinnamomi. Phytopathology 84: 193–200Google Scholar
  18. Carlile MJ (1966) The orientation of zoospores and germ-tubes. In: Madelin MF (ed) The fungal spore, Butterworths, London, pp 175–186Google Scholar
  19. — (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, MN, pp 95–107Google Scholar
  20. Cavalier-Smith T (1981) Eukaryote kingdoms: seven or nine? BioSystems 14: 461–481Google Scholar
  21. — (1986) The kingdom Chromista: origin and systematics. Prog Phycol Res 4: 309–347Google Scholar
  22. Dewey FM (1990) The use of monoclonal antibodies to detect plant invading fungi. In: Schots A (eds) Monoclonal antibodies in agriculture. Pudoc, Wageningen, pp 21–25Google Scholar
  23. Dick MW (1989) Phylum Oomycota. In: Margulis L, Corliss JO, Melkonian M, Chapman DJ (eds) Handbook of Protoctista. Jones and Bartlett, Boston, pp 661–685Google Scholar
  24. Dudler R (1990) The single-copy actin gene ofPhytophthora megasperma encodes a protein considerably diverged from any other known actin. Plant Mol Biol 14: 415–422Google Scholar
  25. Erwin DC, Bartnicki-Garcia S, Tsao PH (eds) (1983)Phytophthora. Its biology, taxonomy, ecology, and pathology. American Phytopathological Society, St. Paul, MNGoogle Scholar
  26. Estrada Garcia MT, Green JR, Booth JM, White JG, Callow JA (1989) Monoclonal antibodies to cell surface components of zoospores and cysts of the fungusPythium aphanidermatum reveal species-specific antigens. Exp Mycol 13: 348–355Google Scholar
  27. —, Callow JA, Green JR (1990 a) Monoclonal antibodies to the adhesive cell coat secreted byPythium aphanidermatum zoospores recognise 200×103 Mr glycoproteins stored within large peripheral vesicles. J Cell Sci 95: 199–206Google Scholar
  28. —, Ray TC, Green JR, Callow JA, Kennedy JF (1990 b) Encystment ofPythium aphanidermatum zoospores is induced by root mucilage polysaccharides, pectin and a monoclonal antibody to a surface antigen. J Exp Biol 41: 693–699Google Scholar
  29. Evans PT, Holaway BL, Malmberg RL (1988) Biochemical differentiation in the tobacco flower probed with monoclonal antibodies. Planta 175: 259–269Google Scholar
  30. Foster H, Coffey MD, Elwood H, Sogin ML (1990) Sequence analysis of the small subunit ribosomal RNAs of three zoosporic fungi and implications for fungal evolution. Mycologia 82: 306–312Google Scholar
  31. Gabor BK, O'Gara ET, Philip BA, Horan DP, Hardham AR (1993) Specificities of monoclonal antibodies toPhytophthora cinnamomi in two rapid diagnostic assays. Plant Dis 77: 1189–1197Google Scholar
  32. Gallegly ME (1983) New criteria for classifyingPhytophthora and critique of existing approaches. In: Erwin DC, Bartnicki-Garcia S, Tsao P (eds)Phytophthora. Its biology, taxonomy, ecology, and pathology. American Phytopathological Society, St. Paul, MN, pp 167–172Google Scholar
  33. Gubler F, Hardham AR (1988) Secretion of adhesive material during encystment ofPhytophthora cinnamomi zoospores, characterized by immunogold labelling with monoclonal antibodies to components of peripheral vesicles. J Cell Sci 90: 225–235Google Scholar
  34. — — (1990) Protein storage in large peripheral vesicles inPhytophthora zoospores and its breakdown after cyst germination. Exp Mycol 14: 393–404Google Scholar
  35. — — (1991) The fate of peripheral vesicles in zoospores ofPhytophthora cinnamomi during infection of plants. In: Mendgen K, Lesemann D-E (eds) Electron microscopy of plant pathogenesis. Springer, Berlin Heidelberg New York Tokyo, pp 197–210Google Scholar
  36. — —, Duniec J (1989) Characterising adhesiveness ofPhytophthora cinnamomi zoospores during encystment. Protoplasma 149: 24–30Google Scholar
  37. Gunderson JH, Elwood H, Ingold A, Kindle K, Sogin ML (1987) Phylogenetic relationships between chlorophytes, chrysophytes, and oomycetes. Proc Natl Acad Sci USA 84: 5823–5827Google Scholar
  38. Halsall DM (1976) Specificity of cytoplasmic and cell-wall antigens from four species ofPhytophthora. J Gen Microbiol 94: 149–158Google Scholar
  39. Hardham AR (1985) Studies on the cell surface of zoospores and cysts of the fungusPhytophthora cinnamomi: the influence of fixation on patterns of lectin binding. J Histochem Cytochem 33: 110–118Google Scholar
  40. — (1987 a) Ultrastructure and serial section reconstruction of zoospores of the fungusPhytophthora cinnamomi. Exp Mycol 11: 297–306Google Scholar
  41. — (1987 b) Microtubules and the flagellar apparatus in zoospores and cysts of the fungusPhytophthora cinnamomi. Protoplasma 137: 109–124Google Scholar
  42. — (1989) Lectin and antibody labelling of surface components of spores ofPhytophthora cinnamomi. Aust J Plant Physiol 16: 19–32Google Scholar
  43. —, Gubler F (1990) Polarity of attachment of zoospores of a root pathogen and pre-alignment of the emerging germ tube. Cell Biol Int Rep 14: 947–956Google Scholar
  44. —, Suzaki E (1986) Encystment of zoospores of the fungus,Phytophthora cinnamomi, is induced by specific lectin and monoclonal antibody binding to the cell surface. Protoplasma 133: 165–173Google Scholar
  45. — — (1990) Glycoconjugates on the surface of the pathogenic fungusPhytophthora cinnamomi studied using fluorescence and electron microscopy and flow cytometry. Can J Microbiol 36: 183–192Google Scholar
  46. — —, Perkin JL (1985) The detection of monoclonal antibodies specific for surface components on zoospores and cysts ofPhytophthora cinnamomi. Exp Mycol 9: 264–268Google Scholar
  47. — — — (1986) Monoclonal antibodies to isolate-, species-and genus-specific components on the surface of zoospores and cysts of the fungusPhytophthora cinnamomi. Can J Bot 64: 311–321Google Scholar
  48. —, Gubler F, Duniec J (1991 a) Ultrastructural and immunological studies of zoospores ofPhytophthora. In: Lucas JA, Shattock RC, Shaw DS, Cooke LR (eds)Phytophthora. Cambridge University Press, Cambridge, pp 50–69Google Scholar
  49. — — —, Elliott J (1991 b) A review of methods for the production and use of monoclonal antibodies to study zoosporic plant pathogens. J Microsc 162: 305–318Google Scholar
  50. Heath IB (1980) Variant mitoses in lower eukaryotes: indicators of the evolution of mitosis? Int Rev Cytol 64: 1–80Google Scholar
  51. —, Greenwood AD, Griffiths HB (1970) The origin of flimmer inSaprolegnia, Dictyuchus, Synura andCryptomonas. J Cell Sci 7: 445–461Google Scholar
  52. 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, MN, pp 9–40Google Scholar
  53. Hill FG, Outka DE (1974) The structure and origin of mastigonemes inOchromonas minute andMonas sp. J Protozool 21: 299–312Google Scholar
  54. Hohl HR, Hamamoto ST (1967) Ultrastructural changes during zoospore formation inPhytophthora parasitica. Amer J Bot 54: 1131–1139Google Scholar
  55. Holwill MEJ (1982) Dynamics of eukaryotic flagellar movement. In: Amos WB, Duckett JG (eds) Prokaryotic and eukaryotic flagella. Cambridge University Press, Cambridge, pp 289–312Google Scholar
  56. Hutter R, DeMoss J (1967) Organization of the tryptophan pathway: a phylogenetic study of the fungi. J Bacteriol 94: 1896–1907Google Scholar
  57. Hyde GJ, Hardham AR (1993) Microtubules regulate the generation of polarity in zoospores ofPhytophthora cinnamomi. Eur J Cell Biol 62: 75–85Google Scholar
  58. —, Gubler F, Hardham AR (1991 a) Ultrastructure of zoosporogenesis inPhytophthora cinnamomi. Mycol Res 95: 577–591Google Scholar
  59. —, Lancelle S, Hepler PK, Hardham AR (1991 b) Freeze substitution reveals a new model for sporangial cleavage inPhytophthora, a result with implications for cytokinesis in other eukaryotes. J Cell Sci 100: 735–748Google Scholar
  60. Jahn TL, Landman MD, Fonseca JR (1964) The mechanism of locomotion of flagellates. II. Function of the mastigonemes ofOchromonas. J Protozool 11: 291–296Google Scholar
  61. Judelson HS, Michelmore RV (1989) Structure and expression of a gene encoding heat-shock protein Hsp 70 from the oomycete fungusBremia lactucae. Gene 79: 207–217Google Scholar
  62. — — (1991) Transient expression of genes in the oomycetePhytophthora infestans usingBremia lactucae regulatory sequences. Curr Genet 19: 453–459Google Scholar
  63. —, Tyler BM, Michelmore RW (1991) Transformation of the oomycete pathogen,Phytophthora infestans. Mol Plant Microbe Interact 4: 602–607Google Scholar
  64. — — — (1992) Regulatory sequences for expressing genes in oomycete fungi. Mol Gen Genet 234: 138–146Google Scholar
  65. —, Coffey MD, Arredondo FR, Tyler BM (1993) Transformation of the oomycete pathogenPhytophthora megasperma f.sp.glycinea occurs by DNA integration into single or multiple chromosomes. Curr Genet 23: 211–218Google Scholar
  66. Karlovsky P, Prell HH (1991) The TRP1 gene ofPhytophthora parasitica encoding indole-3-glycerolphosphate synthase-N-(5′-phosphoribosyl)anthranilate isomerase: structure and evolutionary distance from homologous fungal genes. Gene 109: 161–165Google Scholar
  67. Kinghorn JR, Moon RP, Unkles SE, Duncan JM (1991) Gene structure and expression inPhytophthora infestans and the development of gene-mediated transformation. In: Lucas JA, Shattock RC, Shaw DS, Cooke LR (eds)Phytophthora. Cambridge University Press, Cambridge, pp 295–311Google Scholar
  68. Knox JP, Roberts K (1989) Carbohydrate antigens and lectin receptors of the plasma membrane of carrot cells. Protoplasma 152: 123–129Google Scholar
  69. Krywiencyzk J, Dorworth CE (1980) Serological relationships of some fungi of the genusPythium. Can J Bot 58: 1412–1417Google Scholar
  70. Leedale GF, Leadbetter BSC, Massalski A (1970) The intracellular origin of flagellar hairs in the Chrysophyceae and Xanthophyceae. J Cell Sci 6: 710–719Google Scholar
  71. Lehnen LR Jr, Powell MJ (1988) Cytochemical localization of carbohydrates in zoospores ofSaprolegnia ferax. Mycologia 80: 423–432Google Scholar
  72. — — (1989) The role of kinetosome-associated organelles in the attachment of encysting secondary zoospores ofSaprolegnia ferax to substrates. Protoplasma 149: 163–174Google Scholar
  73. — — (1993) Characterization of cell surface carbohydrates on asexual spores of the water moldSaprolegnia ferax. Protoplasma 175: 161–172Google Scholar
  74. Loiseaux S (1973) Ultrastructure of zoidogenesis in unilocular zoidocysts of several brown algae. J Phycol 9: 277–289Google Scholar
  75. MacDonald JD, Duniway JM (1979) Use of fluorescent antibodies to study the survival ofPhytophthora megasperma andP. cinnamomi zoospores in soil. Phytopathology 69: 436–441Google Scholar
  76. —, Stites J, Kabashima J (1990) Comparison of serological and culture plate methods for detecting species ofPhytophthora, Pythium, andRhizoctonia in ornamental plants. Plant Dis 74: 665–659Google Scholar
  77. Malajczuk N, McComb AJ, Parker CA (1975) An immunofluorescence technique for detectingPhytophthora cinnamomi Rands. Aust J Bot 23: 289–309Google Scholar
  78. Manavathu EK, Suryanarayana K, Hasnain SE, Leung W (1988) DNA-mediated transformation in the aquatic filamentous fungusAchlya ambisexualis. J Gen Bicrobiol 134: 2019–2028Google Scholar
  79. Margulis L, Schwartz KV (1988) Five kingdoms. An illustrated guide to the phyla of life on earth. WH Freeman, New YorkGoogle Scholar
  80. Merz WG, Burrell RG, Gallegly ME (1969) A serological comparison of six homothallic species ofPhytophthora. Phytopathology 59: 367–370Google Scholar
  81. Moestrup Ø (1982) Flagellar structure in algae: a review, with new observations particularly on the Chrysophyceae, Phaeophyceae (Fucophyceae), Euglenophyceae andReckertia. Phycologia 21: 427–528Google Scholar
  82. Mohan SB (1988) Evaluation of antisera raised againstPhytophthora fragariae for detecting the red core disease of strawberries by enzyme-linked immunosorbent assay (ELISA). Plant Pathol 37: 206–216Google Scholar
  83. Morris PF, Ward EWB (1992) Chemoattraction of zoospores of the soybean pathogen,Phytophthora sojae, by isoflavones. Physiol Mol Plant Pathol 40: 17–22Google Scholar
  84. Petersen FP, Maybroda AM, Grothaus GD, Miller SA (1989) Monoclonal antibodies and methods for fungal pathogen detection. United States Patent no 4,845,197Google Scholar
  85. Prell HH, Karlovsky P, Bahnweg G (1991) Towards transformation inPhytophthora nicotianae. In: Lucas JA, Shattock RC, Shaw DS, Cooke LR (eds)Phytophthora. Cambridge University Press, Cambridge, pp 312–325Google Scholar
  86. Pscheidt JW, Burkett JZ, Fisher SL, Hamm PB (1992) Sensitivity and clinical use ofPhytophthora-specific immunoassay kits. Plant Dis 76: 928–932Google Scholar
  87. Raven PH, Evert RF, Curtis H (1981) Biology of plants. Worth Publishers, New YorkGoogle Scholar
  88. Ray PM, Steeves TA, Fultz SA (1983) Botany. WB Saunders, Philadelphia, pp 784Google Scholar
  89. Reichle RE (1969) Fine structure ofPhytophthora parasitica zoospores. Mycologia 61: 30–51Google Scholar
  90. Sachay DJ, Hudspeth DSS, Nadler SA, Hudspeth MES (1993) Oomycete mtDNA:Phytophthora genes for cytochrome c oxidase use an unmodified genetic code and encode proteins most similar to those of plants. Exp Mycol 17: 7–23Google Scholar
  91. Sadowski LA, Powell MJ (1990) Cytochemical detection of polysaccharides in zoospores ofAphanomyces euteiches. Can J Bot 68: 1379–1388Google Scholar
  92. Sansome ER (1987) Fungal chromosomes as observed with the light microscope. In: Rayner ADM, Brasier CM, Moore D (eds) Evolutionary biology of the fungi. Cambridge University Press, Cambridge, pp 97–113Google Scholar
  93. Smith E, Roberts K, Hutchings A, Galfre G (1984) Monoclonal antibodies to the major structural glycoprotein of theChlamydomonas cell wall. Planta 161: 330–338Google Scholar
  94. Stamps DJ, Waterhouse GM, Newhook FJ, Hall GS (1990) Revised tabular key to the species ofPhytophthora. CAB International, WallingfordGoogle Scholar
  95. Unkles SE, Moon RP, Hawkins AR, Duncan JM, Kinghorn JR (1991) Actin in the oomycetous fungusPhytophthora infestons is the product of several genes. Gene 100: 105–112Google Scholar
  96. Villee CA, Solomon EP, Martin CE, Martin DW, Berg LR, Davis PW (1985) Biology. WB Saunders, Fort Worth, pp 1412Google Scholar
  97. Vogel HJ (1965) Lysine biosynthesis and evolution. In: Bryson V, Vogel HJ (eds) Evolving genes and proteins. Academic Press, New York, pp 25–40Google Scholar
  98. Vujicic R, Colhoun J, Chapman JA (1968) Some observations on the zoospores ofPhytophthora erythroseptica. Trans Br Mycol Soc 51: 125–127Google Scholar
  99. Waterhouse GM, Newhook FJ, Stamps DJ (1983) Present criteria for classification ofPhytophthora. In: Erwin DC, Bartnicki-Garcia S, Tsao P (eds)Phytophthora. Its biology, taxonomy, ecology, and pathology. American Phytopathological Society, St. Paul, MN, pp 139–147Google Scholar
  100. Werres S (1988) Enzyme-linked immunosorbent assay (ELISA) as a method for detection ofPhytophthora fragariae Hickman in strawberry roots. Nachrichtenbl Deutsch Planzenschutz 40: 146–150Google Scholar
  101. White DG (1976) The preparation and use of a fluorescent antibody reagent for the detection ofPhythium graminicola. Phytopathology 66: 523–525Google Scholar
  102. Wolters J, Erdmann VA (1988) Cladistic analysis of ribosomal RNAs —the phylogeny of eukaryotes with respect to the endosymbiotic theory. BioSystems 21: 209–214Google Scholar
  103. Wycoff KL, Ayers AR (1990) Monoclonal antibodies to surface and extracellular antigens of a fungal plant pathogen,Phytophthora megasperma f. sp.glycinea, recognize specific carbohydrate epitopes. Physiol Mol Plant Pathol 37: 55–79Google Scholar
  104. —, Jellison J, Ayers AR (1987) Monoclonal antibodies to glycoprotein antigens of a fungal plant pathogen,Phytophthora megasperma f. sp.glycinea. Plant Physiol 85: 508–515Google Scholar
  105. Zentmyer GA (1980)Phytophthora cinnamomi and the diseases it causes. American Phytopathological Society, St. Paul, MNGoogle Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • A. R. Hardham
    • 1
  • D. M. Cahill
    • 1
  • M. Cope
    • 1
  • B. K. Gabor
    • 1
  • F. Gubler
    • 1
  • G. J. Hyde
    • 1
  1. 1.Plant Cell Biology Group, Research School of Biological SciencesAustralian National UniversityCanberraAustralia

Personalised recommendations