Abstract
Cultural investigations revealed that Naemacyclus minor, Lophodermium seditiosum and Cenangium ferruginosum were the most frequent colonizers of asymptomatic and symptomatic Pinus sylvestris needles. Since ultrastructural observations showed that morphological features were not suitable to differentiate hyphae of N. minor from hyphae of other isolates, the on-section immunogold labelling technique was applied in combination with an anti-N. minor specific immunoserum. The specificity of this serum was tested against culture hyphae of all isolates. Anti-N. minor specific immunoserum was then used to identify N. minor hyphae in thin sections of green P. sylvestris needles. The infection loci identified were restricted to small tissue areas located in the vicinity of stomata. In the hypodermis, hyphae and endocell-containing hyphae were located within the lumina of host cells but outside the protoplast. The growth of hyphae from cell to cell occurred through pits. The hyphae spreat into the mesophyll intercellularly and continued with the intracellular colonization of moribund and dead mesophyll cells in a later stage of infection. The observed host-parasite interactions at cellular and ultrastructural level are discussed in connection with the still controversial interpretation of the pathogenicity of N. minor.
Similar content being viewed by others
References
Acker G (1988) Immunoelectron microscopy of surface antigens (polysaccharides) of gram-negative bacteria using pre- and post-embedding techniques. Methods Microbiol 20: 147–174
Acker G, Knapp W, Wartenberg K, Mayer H (1981) Localization of enterobacterial common antigen in Yersinia enterocolitica by the immunoferritin technique. J Bacteriol 147: 602–611
Arx JA (1981) The genera of fungi: sporulating in pure culture. AR Genter, Vaduz
Benhamou N, Ouellette GB (1987) Ultrastructural study and cytochemical investigation, by means of enzyme-gold complexes, of the fungus Ascocalyc abietina. Can J Bot 65: 168–181
Benhamou N, Ouellette GB, Lafontaine JG, Joly JR (1985) Use of monoclonal antibodies to detect a phytotoxic glycolipide produced by Ophiostoma ulmi, the Dutch elm disease pathogen. Can J Bot 63: 1177–1184
Bernstein ME, Carroll GC (1977) Internal fungi in old-growth Douglas fir foliage. Can J Bot 55: 644–653
Blanchette RA, Abad AR (1992) Immunocytochemistry of fungal infection processes in trees. In: Blanchette RA, Biggs AR (eds) Defense mechanisms of woody plants against fungi. Springer, Berlin Heidelberg New York Tokyo, pp 424–444
Bracker CE, Littlefield LJ (1973) Structural of hostpathogen interfaces. In: Byrde RJW, Cutting CV (eds) Fungal pathogenicity and the plant response. Academic Press, London New York, pp 159–317
Butin H (1989) Krankheiten der Wald- und Parkbäume, 2. Aufl. Thieme, Stuttgart New York
Darker GD (1932) The Hypodermataceae of conifers. Contrib Arnold Arbor Harv Univ 1, 1–131
Darker GD (1967) A revision of the genera of the Hypodermataceae. Can J Bot 45: 1399–1444
Ellis MB (1971) Dematiaceous Hyphomycetes. Commonwealth Mycological Institute, Kew Surrey, UK
Ellis MB (1976) More Dematiaceous Hyphomycetes. Commonwealth Mycological Institute, Kew Surrey, UK
Ellis MB, Ellis JP (1985) Microfungi on land plants. An identification handbook. Croom Helm, London Sydney
Gadgil PD (1977) How important is Naemacyclus? What's New in Forest Research 56. New Zealand Forest Research
Gadgil PD (1984) Cyclaneusma (Naemacyclus) needle cast of Pinus radiata in New Zealand. 1. Biology of Cyclaneusma minus. NZ J For Sci 14: 179–196
Gianinazzi S, Gianinazzi-Pearson V (1992) Cytology, histochemistry and immunocytochemistry as tools for studying structure and function in endomycorrhiza. Methods Microbiol 24: 109–139
Gibson IAS (1979) Diseases of forest trees widely planted as Exotics in the Tropics and Southern Hemisphere, part II. The genus Pinus. Commonwealth Mycological Institute, Kew Surrey, UK
Hinton DM, Bacon CW (1985) The distribution and ultrastructure of the endophyte of toxic tall fescue. Can J Bot 63: 36–42
Huber SJ, Hock B (1985) A solid-phase enzyme immunoassay for quantitative determination of the herbicide terbutryn. J Plant Dis Prot 92: 147–156
Karadzic' D (1981) Infection of Pinus sylvestris by Naemacyclus minor. In: Millar CS (ed) Current research on conifer needle diseases. Aberdeen University, Aberdeen, pp 99–101
Kistler BR, Merrill W (1978) Etiology, symptomology, epidemiology and control of Naemacyclus needlecast of Scots pine. Phytopathology 68: 267–271
Kowalski T (1982) Fungi infecting Pinus sylvestris needles of various ages. Eur J For Pathol 12: 182–190
Kowalski T (1988) Cyclaneusma (Naemacyclus) minus an Pinus sylvestris in Polen. Eur J For Pathol 18: 176–183
Kowalski T, Lang KJ (1983) Über die Mycoflora in den Nadeln unterschiedlich alter Kiefern (Pinus sylvestris). Phytopathol Z 107: 9–21
Kunoh H, Nicholson RL, Kobayashi I (1991) Extracellular materials of fungal structures: their significance at penetration stages of infection. In: Mengden K, Lesemann D-E (eds) Electron microscopy of plant pahtogens. Springer, Berlin Heidelberg New York Tokyo, pp 223–234
Lim LL, Fineran BA, Cole ALJ (1983) Ultrastructure of intrahyphal hyphae of Glomus fasciculatum (Thaxter) Gerdamann and Trappe in roots of while clover (Trifolium repens L.). New Phytol 95: 231–239
Millar CS, Minter DW, (1980) CMI-Description of pathogenic fungi and bacteria, set 66, no. 659, Naemacyclus minor. Kew Surrey, UK
Pendland JC (1981) Resistant structures in the entomogenous hyphomycete, Nomuraea rileyi: an ultrastructural study. Can J Bot 60: 1569–1576
Perotto S, Malavasi F, Butcher GW (1992) Use of monoclonal antibodies to study mycorrhiza: present applications and perspectives. Methods Microbiol 24: 221–248
Rack K, Butin H (1984) Experimenteller Nachweis nadelbewohnender Pilze bei Koniferen. Eur J For Pathol 14: 302–310
Rack K, Scheidemann I (1987) Über Sukzession und pathogene Eigenschaften Kiefernnadeln bewohnender Pilze. Eur J For Pathol 17: 102–109
Reynolds DS (1963) The use of lead citrate at high pH as an electron opaque stain in electron microscopy. J Cell Biol 17: 208–212
Roth J (1982) The protein A-gold (pAg) technique — a qualitative and quantitative approach for antigen localization on thin sections. In: Bullock GR, Petrusz P (eds) Technique in immunocytochemistry vol 1. Academic Press, London, pp 107–133
Spurr AR (1969) A low-viscosy epoxy resin embedding medium for electron microscopy. J Ultrastruct Res 26: 31–43
Suske J, Acker G (1989a) Identification of endophytic hyphae of Lophodermium piceae in tissues of green, symptomless Norway spruce needles by immunoelectron microscopy. Can J Bot 67: 1768–1774
Suske J, Acker G (1989b) Endophytic needle fungi: culture, ultrastructural and immunocytochemical studies. In: Schulze E-D, Lange OE, Orene R (eds) Ecological studies. vol 77. Springer, Berlin Heidelberg New York, pp 121–136
Suske J, Acker G (1990) Host-endophyte interaction between Lophodermium piceae and Picea abies: cultural, ultrastructural and immunocytochemical studies. Sydowia Ann Mycol 42: 211–217
Walles B, Nyman B, Alden T (1973) On the ultrastructure of Pinus sylvestris L. Stud For Suec 106: 1–26
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Franz, F., Grotjahn, R. & Acker, G. Identification of Naemacyclus minor hyphae within needle tissues of Pinus sylvestris by immunoelectron microscopy. Arch. Microbiol. 160, 265–272 (1993). https://doi.org/10.1007/BF00292075
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00292075