Skip to main content

Advertisement

Springer Nature Link
Log in

Antifungal mechanism of an anti-Pythium protein (SAP) from the marine bacterium Streptomyces sp. strain AP77 is specific for Pythium porphyrae, a causative agent of red rot disease in Porphyra spp.

  • Original Paper
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

Previously we reported an antifungal protein specific to Pythium porphyrae, a causative agent of red rot disease afflicting seaweed Porphyra spp. This study was carried out to identify the antifungal mechanism of the antifungal protein to P. porphyrae. When we first examined the effect of an anti-Pythium protein (SAP) on the P. porphyrae cell walls, SAP did not decompose the six structural polysaccharides in Pythium cell walls. However, hyphal growth was significantly inhibited in Pythium cells treated with 50 μg/ml of SAP by MTT assay. Protoplasmic leakage was observed in P. porphyrae hyphae treated with SAP for 1 h, followed by hyphal swelling and disintegration, using SYTOX Green, and SAP permeabilized the membrane of P. porphyrae in a dose-dependent manner. Treating P. porphyrae cells with SAP in the presence of carbonyl cyanide m-chlorophenylhydrazone (CCCP), a membrane-depolarizing agent, significantly reduced the membrane permeability to SYTOX Green. Moreover, a similar effect was observed when the P. porphyrae cells were treated with SAP in the presence of MgCl2. In contrast, identical treatment in the presence of KCl significantly increased the membrane permeability to SYTOX Green. These results suggested that anti-Pythium mechanism of SAP was related to alteration of the membrane permeability in P. porphyrae.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1A–D.
Fig. 2.
Fig. 3A, B.
Fig. 4.
Fig. 5A, B.
Fig. 6.
Fig. 7A, B.

Similar content being viewed by others

Explore related subjects

Discover the latest articles and news from researchers in related subjects, suggested using machine learning.

References

  • Arasaki S, Akino K, Tomiyama T (1968) A comparison of some physiological aspects in a marine Pythium on the host and on the artificial medium. Bull Misaki Marine Biol Inst Tokyo Univ 12:203–206

    Google Scholar 

  • Aronson JM, Cooper BA, Fuller MS (1967) Glucans of oomycete cell walls. Science 155:332–335

    CAS  Google Scholar 

  • Bartnicki-Garcia S (1968) Cell wall chemistry, morphogenesis, and taxonomy of fungi. Annu Rev Microbiol 22:87–108

    CAS  PubMed  Google Scholar 

  • Broekaert WF, Terras FRG, Cammue BPA, Osborn RW (1995) Plant defensins: novel antimicrobial peptides as components of the host defense system. Plant Physiol 108:1353–1358

    CAS  PubMed  Google Scholar 

  • Chet I, Henis Y, Mitchell R (1967) Chemical composition of hyphal and sclerotial walls of Sclerotium rolfsii SACC. Can J Microbiol 13:137–141

    CAS  PubMed  Google Scholar 

  • Cooper BA, Aronson JM (1967) Cell wall structure of Pythium debaryanum. Mycologia 59:658–670

    CAS  PubMed  Google Scholar 

  • Falla TJ, Karunaratne DN, Hancock REW (1996) Mode of action of the antimicrobial peptide indolicidin. J Biol Chem 271:19298–19303

    Article  CAS  PubMed  Google Scholar 

  • Fujita Y (1978) Studies on pathogenic Pythium of laver red rot in Ariake Sea farm-V: germination of Pythium porphyrae oospores. Bull Japan Soc Sci Fish 44:15–19

    Google Scholar 

  • Fuller MS, Lewis B, Cook P (1966) Occurrence of Pythium sp. on the marine alga Porphyra. Mycologia 58:313–318

    Google Scholar 

  • Hagihara B (1953) Proteolytic enzyme. In: Egami F, Ishimoto M, Maruo F, Miura Y, Sekine T, Suda M, Tamiya N (eds) Standard biochemistry experiment, (in Japanese). Bunkoudou, Tokyo, pp 208–225

  • Kitamura E, Myouga H, Kamei Y (2002) Polysaccharolytic activities of bacterial enzymes which degrade the cell walls of Pythium porphyrae, a causative fungus of red rot disease in Porphyra yezoensis. Fisheries Sci 68:436–445

    Article  CAS  Google Scholar 

  • Lebaron P, Catala P, Parthuisot N (1998) Effectiveness of SYTOX Green stain for bacterial viability assessment. Appl Environ Microbiol 64:2697–2700

    CAS  PubMed  Google Scholar 

  • Love J, Percival E (1964) The polysaccharides of the green seaweed codium fragile. III. A β-1, 4-linked mannan. J Chem Soc 3345–3350

  • Matsuzaki T, Suzuki T, Fujikura K, Takata K (1997) Nuclear staining for laser confocal microscopy. Acta Histochem Cytochem 30:309–314

    Google Scholar 

  • Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol 65:55–63

    CAS  PubMed  Google Scholar 

  • Oppenheimer CH, ZoBell CE (1952) The growth and viability of sixty-three species of marine bacteria as influenced by hydrostatic pressure. J Mar Res 11:10-18

    Google Scholar 

  • Osborn RW, De Samblanx G.W, Thevissen K, Goderis I, Torrekens S, Van Lenven F, Attenborough S, Rees SB, Broekaert WF (1995) Isolation and characterization of plant defensins from seeds of Asteraceae, Fabaceae, Hippocastanaceae and Saxifragaceae. FEBS Lett 368:257–262

    CAS  PubMed  Google Scholar 

  • Redenbach M, Kieser HM, Denapaite D, Eichner A, Cullum J, Kinashi H, Hopwood DA (1996) A set of ordered cosmids and a detailed genetic and physical map for the 8 Mb Streptomyces coelicolor A3(2) chromosome. Mol Microbiol 21:77–96

    CAS  PubMed  Google Scholar 

  • Roth B, Poot M, Yue S, Millard P (1997) Bacterial viability and antibiotic susceptibility testing with SYTOX Green nucleic acid stain. Appl Environ Microbiol 63:2421–2431

    CAS  PubMed  Google Scholar 

  • Somogyi M (1952) Notes on sugar determination. J Biol Chem 195:19–23

    CAS  Google Scholar 

  • Tercero JA, Lacalle RA, Jimenez A (1993) The pur8 gene from the pur cluster of Streptomyces alboniger encodes a highly hydrophobic polypeptide which confers resistance to puromycin. Eur J Biochem 218:963–971

    CAS  PubMed  Google Scholar 

  • Terras FR, Schoofs HM, De Bolle MF, Van Leuven F, Rees SB, Vanderleyden J, Cammue BP, Broekaert WF (1992) Analysis of two novel classes of plant antifungal proteins from radish (Raphanus sativus L.) seeds. J Biol Chem 267:15301-15309

    CAS  PubMed  Google Scholar 

  • Thevissen K, Ghazi A, De Samblanx GW, Brownlee C, Osborn RW, Broekaert WF (1996) Fungal membrane responses induced by plant defensins and thionins. J Biol Chem 271:15018–15025

    CAS  PubMed  Google Scholar 

  • Thevissen K, Osborn RW, Acland DP, Broekaert WF (1997) Specific, high affinity binding sites for an antifungal plant defensin on Neurospora crassa hyphae and microsomal membranes. J Biol Chem 272:32176–32181

    CAS  PubMed  Google Scholar 

  • Thevissen K, Terras FRG, Broekaert WF (1999) Permeabilization of fungal membranes by plant defensins inhibits fungal growth. Appl Environ Microbiol 65:5451–5458

    CAS  PubMed  Google Scholar 

  • Uppalapati SR, Fujita Y (1999) Carbohydrate regulation of attachment, encystment, and appressorium formation by Pythium porphyrae (oomycota) zoospores on Porphyra yezoensis (rhodophyta). J Phycol 36:359–366

    Article  Google Scholar 

  • Veldhuis MJW, Cucci TL, Sieracki ME (1997) Cellular DNA content of marine phytoplankton using two new fluorochromes: taxonomic and ecological implications. J Phycol 33:527–541

    CAS  Google Scholar 

  • White JG, Lyons NF, Wakeham AJ, Mead A, Green JR (1994) Serological profiling of the fungal genus Pythium. Physiol Mol Plant Pathol 44:349–361

    Google Scholar 

  • Woo JH, Kitamura E, Myouga H, Kamei Y (2002) An antifungal protein from the marine bacterium, Streptomyces sp. strain AP77 is specific for Pythium porphyrae, a causative agent of red rot disease in Porphyra spp. Appl Environ Microbiol 68:2666–2675

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Marine and Highland Bioscience Center, Saga University, Saga 847–0021, Karatsu, Japan

    J.-H. Woo & Y. Kamei

Authors
  1. J.-H. Woo
    View author publications

    Search author on:PubMed Google Scholar

  2. Y. Kamei
    View author publications

    Search author on:PubMed Google Scholar

Corresponding author

Correspondence to Y. Kamei.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Woo, JH., Kamei, Y. Antifungal mechanism of an anti-Pythium protein (SAP) from the marine bacterium Streptomyces sp. strain AP77 is specific for Pythium porphyrae, a causative agent of red rot disease in Porphyra spp.. Appl Microbiol Biotechnol 62, 407–413 (2003). https://doi.org/10.1007/s00253-003-1235-5

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-003-1235-5

Keywords