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Archives of Microbiology

, Volume 155, Issue 2, pp 191–198 | Cite as

Differentiation-related proteins of the broad bean rust fungus Uromyces viciae-fabae, as revealed by high resolution two-dimensional polyacrylamide gel electrophoresis

  • H. Deising
  • P. R. Jungblut
  • K. Mendgen
Original Papers

Abstract

On artificial polyethylene membranes providing a thigmotropic signal, uredospores of the broad bean rust fungus Uromyces viciae-fabae differentiated a series of infection structures which in nature are necessary to invade the host tissue through the stomata. Within 24 h germ tubes, appressoria, substomatal vesicles, infection hyphae and haustorial mother cells were developed successively. Alterations in protein metabolism during infection structure differentiation of this obligate plant pathogen were analyzed in the absence of the host plant by high resolution two-dimensional polyacrylamide gel electrophoresis (2-DE) and silver staining. The norm pattern representing the 2-DE protein patterns of the whole developmental sequence of infection structures of U. viciae-fabae showed 733 spots. During infection structure differentiation 55 proteins were newly formed, altered in quantity, or disappeared. Major alterations in the protein pattern occurred during uredospore germination and when infection hyphae were formed. Uredospore germination was characterized by a decrease of acidic proteins and an increase mainly of proteins with isoelectric points ranging from weakly acidic to basic.

Key words

Infection structure differentiation Protein metabolism Rust fungi Thigmo-differentiation Two-dimensional polyacrylamide gel electrophoresis Uromyces vicae-fabae 

Abbreviations

2-DE

two-dimensional polyacrylamide gel electrophoresis

DAPI

4,6-diamino-phenylindol

kDa

kilo Dalton

pl

isoelectric point

PMSF

phenylmethylsulfonyl fluoride

SDS-PAGE

sodium dodecyl sulfate polyacrylamide gel electrophoresis

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References

  1. BhairiSM, StaplesRC, FreveP, YoderOC (1989) Characterization of an infection structure-specific gene from the rust fungus Uromyces appendiculatus. Gene 81: 237–243CrossRefGoogle Scholar
  2. BhairiSM, LaccettiL, StaplesRC (1990) Effect of heat shock on expression of thigmo-specific genes from a rust fungus. Exp Mycol 14: 94–98CrossRefGoogle Scholar
  3. BradfordMM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72: 248–254CrossRefGoogle Scholar
  4. CarlileMJ (1975) Taxes and tropisms: Diversity, biological significance and evolution. In: CarlileMJ (ed) Primitive sensory and communication systems. The taxes and tropisms of microorganisms and cells. Academic Press, London New York, pp 1–28Google Scholar
  5. DickinsonS (1949) Studies in the physiology of obligate parasitism. II. The behaviour of the germ tubes of certain rusts in contact with various membranes. Ann Bot 13: 219–236CrossRefGoogle Scholar
  6. DunnGA, HealthJP (1976) A new hypothesis of contact guidance in tissue cells. Exp Cell Res 101: 1–14CrossRefGoogle Scholar
  7. EckerskornC, JungblutP, MewesW, KloseJ, LottspeichF (1988) Identification of mouse brain proteins after two-dimensional electrophoresis and electroblotting by microsequence analysis and amino acid composition analysis. Electrophoresis 9: 830–838CrossRefGoogle Scholar
  8. FreytagS, BruscaglioniL, GoldRE, MendgenK (1988) Basidiospores of rust fungi (Uromyces species) differentiate infection structure in vitro. Exp Mycol 12: 275–283CrossRefGoogle Scholar
  9. HochHC, StaplesRC (1987) Structural and chemical ranges among the rust fungi during appressorium development. Annu Rev Phytopathol 25: 231–247CrossRefGoogle Scholar
  10. HochHC, StaplesRC, WhiteheadB, ComeauJ, WolfED (1987) Signaling for growth orientation and cell differentiation by surface topography in Uromyces. Science 235: 1659–1662CrossRefGoogle Scholar
  11. HuangB-F, StaplesRC (1982) Synthesis of proteins during differentiation of the bean rust fungus. Exp Mycol 6: 7–14CrossRefGoogle Scholar
  12. JugblutPR, KloseJ (1986) Composition and genetic variability of Heparin-Sepharose CL-6B protein fractions obtained from the solubilized proteins of mouse organs. Biochem Gene 24: 925–939CrossRefGoogle Scholar
  13. JungblutPR, SeifertR (1990) Analysis by high resolution two-dimensional electrophoresis of differentiation-dependent alternations in cytosolic protein pattern of HL-60 leucemic cells. J Biochem Biophys Methods 21: 47–58CrossRefGoogle Scholar
  14. JungblutPR, SchneiderW, KloseJ (1985) Quantitative analysis of two-dimensional electrophoresis protein patterns: Comparison of visual evaluation with computer-assisted evaluation. In: NeuhoffV (ed) Electrophoresis '84. Verlag Chemie, Weinheim, pp 301–303Google Scholar
  15. KimWK, HowesNK, RohringerR (1982) Detergent-soluble polypeptides in germinated uredospores and differentiated uredosporelings of wheat stem rust. Can J Plant Pathol 4: 328–333CrossRefGoogle Scholar
  16. KloseJ (1975) Protein mapping by combined isoelectric focusing and electrophoresis of mouse tissue. A novel approach to testing for induced point mutations in mammals. Humangenetik 26: 231–243PubMedGoogle Scholar
  17. KunohH, NicholsonRL, KobayashiI (1991) Extracellular materials of fungal structures: Their significance at prepenetration stages of infection. In: MendgenK, LesemannDE (eds) Electronmicroscopy of plant pathogens. Springer, Berlin Heidelberg New York, pp 223–234CrossRefGoogle Scholar
  18. MendgenK, SchneiderA, SterkM, FinkW (1988) The differentiation of infection structures as a result of recognition events between some biotrophic parasites and their hosts. J Phytopathol 123: 259–272CrossRefGoogle Scholar
  19. ShawM, BoassonR, ScrubbL (1985) Effect of heat shock on protein synthesis in flax rust uredosporelings. Can J Bot 63: 2069–2076CrossRefGoogle Scholar
  20. StaplesRC, HuangB-F (1981) Gene activation during differentiation of the rusts and anthracnose fungi. In: TurianG, HohlHR (eds) The fungal spore: Morphogenetic controls. Academic Press, London New York, pp 335–353Google Scholar
  21. StaplesRC, GrambowHJ, HochHC, WynnWK (1983) Contact with membrane grooves induces wheat stem rust uredospore germlings to differentiate appressoria but not vesicles. Phytopathology 73: 1436–1439CrossRefGoogle Scholar
  22. StaplesRC, YoderOC, HochHC, EpsteinL, BhairiS (1986) Gene expression during infection structure development by germlings of the rust fungi. In: BaileyJA (ed) Biology and molecular biology of plant-pathogen interactions. NATO ASI Series, vol H1. Springer, Berlin Heidelberg New York, pp 331–341CrossRefGoogle Scholar
  23. StaplesRC, HochHC, FreveP, BourettTM (1989) Heat shockinduced development of infection structures by bean rust uredospore germlings. Exp Mycol 13: 149–157CrossRefGoogle Scholar
  24. WannerR, FörsterH, MendgenK, StaplesR (1985) Synthesis of differentiation-specific proteins in germlings of the wheat stem rust fungus after heat shock. Exp Mycoll 9: 279–283Google Scholar
  25. WynnWK (1976) Appressrium formation over stomates by the bean rust fungus: Response to a surface contact stimulus. Phytopathology 66: 136–146CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 1991

Authors and Affiliations

  • H. Deising
    • 1
  • P. R. Jungblut
    • 2
  • K. Mendgen
    • 1
  1. 1.Fakultät für Biologie, PhytopathologieUniversität KonstanzKonstanzGermany
  2. 2.Institut für Toxikologie und EmbryonalpharmakologieFreie Universität BerlinBerlin 33Germany

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