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Regulation of attachment, germination, and appressorium formation by zoospores ofLagenidium giganteum and related oomycetes by chitin, chitosan, and catecholamines

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Summary

Lagenidium giganteum (Oomycetes: Lagenidiales), a facultative parasite of mosquito larvae, infects the larval stage of most species of mosquitoes and a very limited number of alternate hosts. Host infection by this and other members of Oomycetes is initiated by motile, laterally biflagellate zoospores. Chemical bases for the various degrees of host specificity exhibited by these parasites is not known, but presumably involves receptors on the zoospore surface recognizing compounds either secreted by or on the surface of their hosts. Surface topography had no detectable effect onL. giganteum encystment or appressorium formation. Scanning electron microscopy documented the detachment of flagella during zoospore encystment. Bulbous knobs at the basal end of the detached flagellum were interpreted as encysting zoospores dropping the axoneme and/or the basal body and associated structures to which flagella are attached. Multiple signals appear to be involved in the initial steps ofL. giganteum host invasion. Zoospores of this parasite did not encyst on powdered preparations of chitin or chitosan (deacetylated chitin). Upon dissolution of chitosan in dilute acid followed by drying these solutions to form thin, transparent films, zoospores readily encysted. The degree of reacetylation of these films and the spacing of acetylated and deacetylated residues had no significant effect on zoospore encystment. Zoospores of a strain ofLagenidium myophilum isolated from marine shrimp, that also infects mosquito larvae, encysted on chitosan films. No encystment of spores of the plant parasitePhytophthora capsici was observed on chitin or chitosan films. Simulation of cuticle sclerotization by incubating chitosan films with different catecholamines and tyrosinase significantly reduced zoospore encystment. Zoospores that encysted on chitosan films did not germinate in distilled water. Germination could be induced by adding microgram quantities of bovine serum albumin or proteins secreted by motile zoospores into the water, and to a lesser degree by some amino acids, but not by various cations. Zoospores encysted and germinated on the pupal stage of some mosquito species. Appressoria were occasionally formed, but most subsequently sent out another mycelial branch, apparently without attempting to pierce the pupal cuticle. Methylation of pupal exuviae with ethereal diazomethane or methanol/HCl significantly increased zoospore encystment. Modification of chitin by catecholamines, lipids and protein on the epicuticular larval surface all affected host invasion.

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Abbreviations

BSA:

bovine serum albumin

CID:

collision-induced dissociation

DOPA:

3,4-dihydroxyphenylalanine

ESI-MS:

electrospray mass spectrometry

ESI-MS/MS:

tandem electrospray mass spectrometry

SDS-PAGE:

sodium dodecyl sulphate polyacrylamide gel electrophoresis

WGA:

wheat germ agglutinin

ZAP:

zoospore aggregation pheromone

References

  • Aiba S (1989) Studies on chitosan: 2. Solution stability and reactivity of partially N-acetylated chitosan derivatives in aqueous media. Int J Biol Macromol 11: 249–252

    PubMed  Google Scholar 

  • — (1991) Studies on chitosan: 3. Evidence for the presence of random and block copolymer structures in partially N-acetylated chitosans. Int J Biol Macromol 13: 40–44

    PubMed  Google Scholar 

  • Alien EA, Hazen BE, Hoch HC, Leinhos GME, Staples RC, Stumpf MA, Terhune BT (1991) Appressorium formation in response to topographical signals by 27 rust species. Phytopathology 81: 323–331

    Google Scholar 

  • Andersen SO (1985) Sclerotization and tanning of the cuticle. In: Kerkut GA, Gilbert LI (eds) Comprehensive insect physiology, biochemistry and pharmacology, vol 3: integument, respiration and circulation. Pergamon, Oxford, pp 59–74

    Google Scholar 

  • —, Jacobsen JP, Roepstorff (1992) Coupling reactions between amino compounds and N-acetyldopamine catalyzed by cuticular enzymes. Insect Biochem Mol Biol 22: 517–527

    Google Scholar 

  • Anonymous (1992)Lagenidium giganteum: exemptions from the requirement of a tolerance. Fed Regist 57: 53570

    Google Scholar 

  • Bartnicki-Garcia S (1970) Cell wall composition and other biochemical markers in fungal phytogeny. In: Harborne JB (ed) Phytochemical phylogeny. Academic Press, New York, pp 81–103

    Google Scholar 

  • Beakes G (1983) A comparative account of cyst coat ontogeny in saprophytic and fish-lesion (pathogenic) isolates of theSaprolegnia diclina-parasitica complex. Can J Bot 61: 603–625

    Google Scholar 

  • Berbee ML, Kerwin JL (1993) Ultrastructural and light microscopic localization of carbohydrates and peroxidase/catalases inLagenidium giganteum zoospores. Mycologia 85: 734–743

    Google Scholar 

  • Bertke CC, Aronson JM (1992) Hyphal wall composition ofLagenidium callinectes andLagenidium chthamalophilium. Bot Mar 35: 147–152

    Google Scholar 

  • Bidochka MJ, Khachatourians GG (1994) Basic proteases of entomopathogenic fungi differ in their adsorption properties to insect cuticle. J Invertebr Pathol 64: 26–32

    Google Scholar 

  • Blomquist GJ, Nelson DR, de Renobales M (1987) Chemistry, biochemistry and physiology of insect cuticular lipids. Arch Insect Biochem Physiol 6: 227–265

    Google Scholar 

  • Borders CL Jr, Zurcher JA (1979) Rabbit muscle enolase also has essential arrginyl residues. FEBS Lett 108: 415–418

    PubMed  Google Scholar 

  • Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254

    PubMed  Google Scholar 

  • Brey PT, Leburn RA (1987) External morphology ofLagenidium giganteum zoospores. Mycopathology 97: 45–49

    Google Scholar 

  • — —, Papierok B, Ohayon H, Vennavalli S, Hafez J (1988) Defense reactions by larvae ofAedes aegypti during infection by the aquatic fungusLagenidium giganteum (Oomycete). Cell Tissue Res 253: 245–250

    PubMed  Google Scholar 

  • Burr AW, Beakes GW (1994) Characterization of zoospore and cyst surface structure in saprophytic and fish pathogenicSaprolegnia species (oomycete fungal protists). Protoplasma 181: 142–163

    Google Scholar 

  • Byrt PN, Irving HR, Grant BR (1982) The effect of cations on zoospores of the fungusPhytophthora cinnamomi. J Gen Microbiol 128: 1189–1198

    Google Scholar 

  • Cho CW, Fuller MS (1989) Ultrastructural organization of freeze-substituted zoospores ofPhytophthora palmivora. Can J Bot 67: 1493–1499

    Google Scholar 

  • Deacon JW, Donaldson SP (1993) Molecular recognition in the homing responses of zoosporic fungi, with special reference toPythium andPhytophthora. Mycol Res 97: 1153–1171

    Google Scholar 

  • Domard A, Rinaudo M (1983) Preparation and characterization of fully deacetylated chitosan. Int J Biol Macromol 5: 49–52

    Google Scholar 

  • Domnas AJ, Srebro JP, Hicks BF (1977) Sterol requirement for zoospore formation in the mosquito-parasitizing fungusLagenidium giganteum. Mycologia 69: 875–886

    Google Scholar 

  • Domzsy JG, Roberts GAF (1985) Evaluation of infrared spectroscopic techniques for analysing chitosan. Makromol Chem 186: 1671–1677

    Google Scholar 

  • Donaldson SP, Deacon JW (1992) Role of calcium in adhesion and germination of zoospore cysts ofPythium: a model to explain infection of host plants. J Gen Microbiol 138: 2051–2059

    Google Scholar 

  • Durso L, Lehnen LP Jr, Powell MJ (1993) Characteristics of extracellular adhesions produced duringSaprolegnia ferax secondary zoospore encystment and cystospore germination. Mycologia 85: 744–755

    Google Scholar 

  • Golkar L, LeBrun RA, Ohayon H, Gounon P, Papierok B, Brey PT (1993) Variation of larval susceptibility toLagenidium giganteum in three mosquito species. J Invertebr Pathol 62: 1–8

    PubMed  Google Scholar 

  • Grove SN, Bracker CE (1978) Protoplasmic changes during zoospore encystment and cyst germination inPythium aphanidermatum. Exp Mycol 2: 51–98

    Google Scholar 

  • 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: 225–235

    Google Scholar 

  • Hadley NF (1994) Water relations of terrestrial arthropods. Academic Press, San Diego

    Google Scholar 

  • Hall DA, Czerkawski JW (1961) The reaction between elastase and elastic tissue. 5. Groupings essential for activity. Biochem J 80: 128–136

    PubMed  Google Scholar 

  • 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: 110–118

    PubMed  Google Scholar 

  • —, Suzaki E (1986) Encystment of zoospores of the fungusPhytophthora cinnamomi is induced by specific lectin and monoclonal antibody binding to the cell surface. Protoplasma 133: 165–173

    Google Scholar 

  • Hatai K, Lawhavinit O (1988)Lagenidium myophilum sp. nov., a new parasite on adult northern shrimp (Pandalus borealis Krøyer). Trans Mycol Soc Japan 29: 175–184

    Google Scholar 

  • Heath IB, Kaminskyj SG, Bauchop T (1986) Basal body loss during fungal zoospore encystment: evidence against centriole autonomy. J Cell Sci 83: 135–140

    PubMed  Google Scholar 

  • Hepburn HR (1985) Structure of the integument. In: Kerkut GA, Gilbert LI (eds) Comprehensive insect physiology, biochemistry and pharmacology, vol. 3: integument, respiration and circulation. Pergamon, Oxford, pp 1–58

    Google Scholar 

  • Hirano S, Tsuneyasu S, Kondo Y (1981) Heterogeneous distribution of amino groups in partially N-acetylated derivatives of chitosan. Agric Biol Chem 45: 1335–1339

    Google Scholar 

  • Ho HH, Zachariah K, Hickman CJ (1967) Anatomy of beads on flagella of zoospores ofPhytophthora megasperma var.sojae. Can J Bot 45: 1765–1768

    Google Scholar 

  • Hoch HC, Mitchell JE (1972) The ultrastructure of zoospores ofAphanomyces euteiches and of their encystment and subsequent germination. Protoplasma 75: 113–138

    Google Scholar 

  • —, Staples RC, Whitehead B, Comeau J, Wolf ED (1987) Signaling for growth orientation and cell differentiation by surface topography inUromyces. Science 235: 1659–1661

    Google Scholar 

  • Hopps HB (1970) Preparation and reactions of diazomethane. Aldrichim Acta 3: 9–12

    Google Scholar 

  • Irving HR, Grant BR (1984) The effect of calcium on zoospore differentiation inPhytophthora cinnamomi. J Gen Microbiol 130: 1569–1576

    Google Scholar 

  • Kates M (1986) Identification of individual lipids and lipid moieties. In: Burdon RH, van Knippenberg PH (eds) Techniques of lipidology, isolation, analysis and identification of lipids, Vol 3, part 2. Elsevier, Amsterdam, pp 352–353

    Google Scholar 

  • Kelleher M, Hinch JM, Clarke AE, Bacic A (1990) Direct detection of cell surface receptors for glycosyl residues onPhytophthora cinnamomi using polysaccharide-coated fluorescent latex beads. Physiol Mol Plant Pathol 37: 377–387

    Google Scholar 

  • Kerwin JL, Washino RK (1983) Sterol induction of sexual reproduction inLagenidium giganteum. Exp Mycol 7: 109–115

    Google Scholar 

  • —, Dritz DA, Washino RK (1988) Nonmammalian safety tests forLagenidium giganteum (Oomycetes: Lagenidiales). J Econ Entomol 81: 158–171

    PubMed  Google Scholar 

  • — (1990) Confirmation of the safety ofLagenidium giganteum (Oomycetes: Lagenidiales) to mammals. J Econ Entomol 83: 374–376

    PubMed  Google Scholar 

  • —, Grant DF, Berbee ML (1991) Specific induction of encystment ofLagenidium giganteum zoospores by concanavalin A and derivatives of chitin and chitosan. Protoplasma 161: 43–51

    Google Scholar 

  • —, Johnson LM, Whisler HC, Tuiniga AR (1992) Infection and morphogenesis ofPythium marinum in species of Porphyra and other red algae. Can J Bot 70: 1017–1024

    Google Scholar 

  • —, Dritz DA, Washino RK (1994) Pilot scale production and application in wildlife ponds ofLagenidium giganteum (Oomycetes: Lagenidiales). J Am Mosq Control Assoc 10: 451–455

    PubMed  Google Scholar 

  • Kole AP (1965) Flagella. In: Ainsworth GC, Sussman AS (eds) The fungi: an advanced treatise. Academic Press, New York, pp 77–93

    Google Scholar 

  • Kramer KJ, Hopkins TL, Schaefer J, Morgan TD, Garbow JR, Jacob GS, Stejskal EO, Speirs RD (1987) Mechanisms of insect cuticle stabilization. How do tobacco hornworms do it? In: Law JH (ed) Molecular entomology. Liss, New York, pp 331–355

    Google Scholar 

  • Kremer AB, Egan RM, Sable MZ (1980) The active site of transketolase. Two arginine residues are essential for activity. J Biol Chem 255: 2405–2411

    PubMed  Google Scholar 

  • Lewin RA, Meinhart JO (1953) Studies on the flagella of algae. III. Electron micrographs ofChlamydomonas moewusii. Can J Bot 31: 711–717

    Google Scholar 

  • Lewis SD, Shafer JA (1973) Conversion of exposed aspartyl and glutamyl residues in proteins to asparaginyl and glutaminyl residues. Biochim Biophys Acta 303: 284–291

    PubMed  Google Scholar 

  • Li J, Christensen BM (1993) Involvement of L-tyrosine and phenol oxidase in the tanning ofAedes aegypti eggs. Insect Biochem Mol Biol 23: 739–748

    Google Scholar 

  • Lockey KH (1988) Lipids of the insect cuticle: origin, composition and function. Comp Biochem Physiol 89B: 595–645

    Google Scholar 

  • Lundblad RL, Noyes CM (1984) Chemical reagents for protein modification, vol 2. CRC Press, Boca Raton

    Google Scholar 

  • Min H-K, Hatai K, Bai S (1994) Some inhibitory effects of chitosan on fish-pathogenic oomycete,Saprolegnia parasitica. Fish Pathol 29: 73–77

    Google Scholar 

  • Muzzarelli RAA, Ilari P, Xia W, Pinotti M, Tomasetti M (1994) Tyrosinase-mediated quinone tanning of chitinous materials. Carbohydr Polymers 24: 295–300

    Google Scholar 

  • Nagata Y, Burger MM (1974) Wheat germ agglutinin — molecular characteristics and specificity for sugar binding. J Biol Chem 249: 3116–3122

    PubMed  Google Scholar 

  • Nestrud LB, Anderson RL (1994) Aquatic safety ofLagenidium giganteum: effects on freshwater fish and invertebrates. J Invertebr Pathol 64: 228–233

    PubMed  Google Scholar 

  • Powell MJ, Lehnen LP, Bortnick RN (1985) Microbody-like organelles as taxonomic markers among oomycetes. BioSystems 18: 321–334

    PubMed  Google Scholar 

  • Randolph LR, Powell MJ (1992) Ultrastructure of zoospores of the oomyceteApodachlya pyrifera. Mycologia 84: 768–780

    Google Scholar 

  • Rinaudo M, Domard A (1989) Solution properties of chitosan. In: Skjak-Braek G, Anthonson T, Sanford P (eds) Chitin and chitosans: sources, chemistry, biochemistry, physical properties and applications. Elsevier, London, pp 71–86

    Google Scholar 

  • Riordan JF, Wacker WEC, Vallee BL (1965) N-acetylimidazole: a reagent for determination of “free” tyrosyl residues of proteins. Biochemistry 4: 1758–1765

    Google Scholar 

  • Royle DJ, Hickman CJ (1964) Analysis of factors governing in vitro accumulation of zoospores ofPythium aphanidermatum on roots. I. Behavior of zoospores. Can I Microbiol 10: 151–162

    Google Scholar 

  • Sadowski LA, Powell MJ (1990) Cytochemical detection of polysaccharides in zoospores ofAphanomyces euteiches. Can J Bot 68: 1379–1388

    Google Scholar 

  • Santilli V (1958) Zoospore morphology of the Californian strain ofPlasmopara viticola. Nature 181: 924

    Google Scholar 

  • Schnellman J, Zeltins A, Blaak H, Schrempf H (1994) The novel lectin-like protein CHB1 is encoded by a chitin-inducibleStreptomyces olivaceoviridis gene and binds specifically to crystalline α-chitin of fungi and other organisms. Mol Microbiol 13: 807–819

    PubMed  Google Scholar 

  • Siegel JP, Shadduck JA (1987) Safety of the entomopathogenic fungusLagenidium giganteum (Oomycetes: Lagenidiales) to mammals. J Econ Entomol 80: 994–997

    PubMed  Google Scholar 

  • Sing VO, Bartnicki-Garcia S (1975) Adhesion ofPhytophthora palmivora zoospores: detection and ultrastructural visualization of concanavalin A-receptor sites appearing during encystment. J Cell Sci 19: 11–20

    PubMed  Google Scholar 

  • Sugumaran M (1991) Molecular mechanisms for mammalian melanogenesis — comparison with insect cuticular sclerotization. FEBS Lett 293: 4–10

    Google Scholar 

  • Vujicic R, Colhoun J, Chapman JA (1968) Some observations on the zoospores ofPhytophthora erythroseptica. Trans Br Mycol Soc 51: 125–127

    Google Scholar 

  • Yamada H, Imoto T (1981) A convenient synthesis of glycolchitin, a substrate of lysozyme. Carbohydr Res 92: 160–162

    PubMed  Google Scholar 

  • Zeltins A, Schrempf H (1995) Visualization ofa-chitin with a specific chitin-binding protein (CHB1) fromStreptomyces olivaceoviridis. Anal Biochem 231: 287–294

    PubMed  Google Scholar 

  • Zomer E, Lipke H (1981) Tyrosine metabolism inAedes aegypti II. Arrest of sclerotization by MON 0585 and diflubenzuron. Pestic Biochem Physiol 16: 28–37

    Google Scholar 

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  1. Botany Department, University of Washington, Box 351330, 98195, Seattle, WA, USA

    E. E. Petersen, M. J. Semon, J. L. Kerwin & J. M. Brower

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  1. E. E. Petersen
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  2. M. J. Semon
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  3. J. L. Kerwin
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Petersen, E.E., Semon, M.J., Kerwin, J.L. et al. Regulation of attachment, germination, and appressorium formation by zoospores ofLagenidium giganteum and related oomycetes by chitin, chitosan, and catecholamines. Protoplasma 197, 96–110 (1997). https://doi.org/10.1007/BF01279888

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