Abstract
We are studying the interaction between the roots of Norway spruce seedlings (Picea abies L. Karst) and a highly pathogenic isolate ofPythium dimorphum. Here, we report the isolation of a cDNA from spruce roots encoding a protein with high sequence similarity to plant defensins, designated as SPI1 (Spruce Pathogen Induced No. 1). The transcript hybridizing to theSPI1 cDNA probe is highly induced in uninfected roots when the seedlings are transferred from solid to liquid incubating malt medium (hypoxic conditions). However, when the seedlings are transferred from solid to liquid malt media containing a saturating amount ofP. dimorphum hyphae, the amount of transcript is unchaged the first day after infection, but then decreases on day 1, and is no longer detectable 2 days after infection. Since plant defensins may play a role in plant defence, their negative regulation upon infection might reflect a strategy employed by this pathogenic fungus to evade the effect of toxic gene products.
References
Andresen I, Becker W, Schlüter K, Burges J, Parthier B, Apel K: The identification of leaf thionin as one of the main jasmonate-induced proteins of barley (Hordeum vulgare). Plant Mol Biol 19: 193–204 (1992).
Apel K, Bohlmann H, Reimann-Philip U: Leaf thionins: a novel class of putative defence factors. Physiol Plant 80: 315–321 (1990).
Bloch C, Richardson M: A new family of small (5 kDa) protein inhibitors of insect α-amylases from seeds or sorghum (Sorghum bicolor (L.) Moench) have sequence homologies with wheat γ-purothionins. FEBS Lett 279: 101–104 (1991).
Bohlmann H, Apel K: Thionins. Annu Rev Plant Physiol Plant Mol Biol 42: 227–240 (1991).
Bohlmann H, Clausen S, Behnke S, Giese H, Hiller C, Reimann-Phillip U, Schrader G, Barkholt V, Apel K: Leaf-specific thionins of barley: a novel class of cell wall proteins toxic to plant-pathogenic fungi and possibly involved in the defence mechanism of plant. EMBO J 7: 1559–1565 (1988).
Børja I, Sharma P, Krekling T, Lönneborg A: Cytopathological response in roots ofPicea abies seedlings infected withPythium dimorphum. Phytopathology 85: 495–501 (1995).
Bowles DJ: Defense-related proteins in higher plants. Annu Rev Biochem 59: 873–907 (1990).
Bruix M, Jimenéz MA, Santoro J, Gonzaléz C, Colilla FJ, Mendez E, Rico M: Solution structure of γ1-H and γ1-P thionins from barley and wheat endosperm determined by1H-NMR: a structural motif common to toxic arthropod proteins. Biochemistry 32: 715–724 (1993).
Carmona MJ, Molina A, Fernandez JA, Lopez-Fando JJ, Garcia-Olmedo F: Expression of the α-thionin gene from barley in tobacco confers enhanced resistance to bacterial pathogens. Plant J 3: 457–462 (1993).
Carrasco L, Vásquez D, Hernández-Lucas C, Carbonero P, Garcia-Olmedo F: Thionins, plant peptides that modify membrane permeability in cultured mammalian cells. Eur J Biochem 116: 185–189 (1981).
Colilla FJ, Rocher A, Mendez E. γ-purothionins: amino acid sequences of two polypeptides of a new family of thionins from wheat endosperm. FEBS Lett 270: 191–194 (1990).
Collinge DB, Slusarenko AJ: Plant gene expression in response to pathogens. Plant Mol Biol 9: 389–410 (1987).
Cramer CL, Ryder TB, Bell JN, Lamb CJ: Rapid switching of plant gene expression induced by fungal elicitor. Science 227: 1240–1243 (1985).
Dalkin K, Jorrin J, Dixon RA: Stress responses in alfalfa (Medicago sativa L.). VII. Induction of defense related mRNAs in elicitor-treated cell suspension cultures. Physiol Mol Plant Path 37: 293–307 (1990).
Dixon RA, Lamb CJ: Molecular communication in interactions between plants and microbial pathogens. Annu Rev Plant Physiol Plant Mol Biol 41: 339–367 (1990).
Ebrahim-Nesbat F, Behnke S, Kleinhofs A, Apel K: Cultivarrelated differences in the distribution of cell-wall-bound thionins in compatible and incompatible interactions between barley and powdery mildew. Planta 179: 203–210 (1989).
Florack DEA, Stiekema WJ: Thionins: properties, possible biological roles and mechanisms of action. Plant Mol Biol 26: 25–37 (1994).
Fernandezde Caleya R, Gonzalez-Pascual B, Garcia-Olmedo F, Carbonero P: Susceptibility of phytopathogenic bacteria to wheat purothioninsin vitro. Appl Microbiol 23: 998–1000 (1972).
Garcia-Olmedo F, Carbonero P, Hernández-Lucas C, Paz-Ares J, Ponz F, Vicento O, Sierra JM: Inhibition of eukaryotic cell-free protein synthesis by thionins from wheat endosperms. Biochim Biophys Acta 740: 52–56 (1983).
Garcia-Olmedo F, Rodriguez-Palenzuela P, Hernández-Lucas C, Ponz F, Mariana C, Carmona MJ, Lopez-Fando J, Fernandez JA, Carbonero P: The thionins: a protein family that includes purothionins, viscotoxins and crambins. Oxford Surv Plant Mol Cell Biol 6: 31–60 (1989).
Genetics Computer Group: Program Manual for the Wisconsin package, version 8. GCG, Madison, WI (1994).
Good AG, Crosby WL: Induction of alcohol dehydrogenase and lactate dehydrogenase in hypoxically induced barley. Plant Physiol 90: 860–866 (1989).
Hake S, Kelley PM, Taylor WC, Freeling M: Coordinate induction of alcohol dehydrogenase 1, aldolase and other anaerobic RNAs in maize. J Biol Chem 260: 5050–5054 (1985).
Ishibashi N, Yamauchi D, Minamikawa T: Stored mRNA in cotyledons ofVigna unguiculata seeds: nucleotide sequence of cloned cDNA for a stored mRNA and induction of its synthesis by precocious germination. Plant Mol Biol 15: 59–64 (1990).
Karunanandaa B, Singh A, Kao T-H: Characterization of a predominantly pistil expressed gene encoding a plant γ-thionin protein ofPetunia inflata. Plant Mol Biol 26: 459–464 (1994).
Kelley PM: Maize pyruvate decarboxylase mRNA is induced anaerobically. Plant Mol Biol 13: 213–222 (1989).
Logemann J, Mayer JE, Schell J, Willmitzer L: Differential expression of genes in potato tubers after wounding. Proc Natl Acad Sci USA 85: 1136–1140 (1988).
Lönneborg A, Sharma P, Stougaard P: Construction of subtractive cDNA library using magnetic beads and PCR. PCR Meth Appl 4: S168-S176 (1995).
Lutchke HA, Chow KC, Mickel FS, Moss KA, Kern HF, Scheele GA: Selection of AUG initiation codons differs in plants and animals. EMBO J 6: 43–48 (1987).
Mendez E, Moreno A, Colilla F, Pelaez F, Limas GG, Mendez R, Sorano F, Salina M, de Haro C: Primary structure and inhibition of protein synthesis in eukaryotic cell-free system of a novel thionin, γ-hordothionin, from barley endosperm. Eur J Biochem 194: 553–539 (1990).
Rice P: Program Manual for the EGCG Package. The Sanger Centre, Hixton Hall, Cambridge (1994).
Sauer N, Corbin DR, Keller B, Lamb CJ: Cloning and characterization of a wound-specific hydroxyproline-rich glycoprotein inPhaseolus vulgaris. Plant Cell Environ 13: 257–266 (1990).
Schultz DJ, Craig R, Cox-Foster DL, Mumma RO, Medford JI: RNA isolation from recalcitrant plant tissue. Plant Mol Biol 12: 310–316 (1994).
Sharma P, Børja D, Stougaard P, Lönneborg A: PR-proteins accumulating in spruce roots infected with a pathogenicPythium sp. isolate include chitinases, chitosanases and β-1,3-glucanases. Physiol Mol Plant Path 43: 57–67 (1993).
Sharma P, Lönneborg A, Stougaard P: PCR based construction of subtractive cDNA library using magnetic beads. Biotechniques 15: 610–611 (1993).
Staub U, Polivka H, Gross HJ: Two rapid microscale procedures for isolation of total RNA from leaves rich in polyphenols and polysaccharides: application for sensitive detection of grapevine viroids. J Virol Meth 52: 209–218 (1995).
Stiekema WJ, Heidekamp F, Dirkse WG, van Beckum J, de Haan P, ten Bosch C, Louwerse JD: Molecular cloning and analysis of four potato tuber mRNAs. Plant Mol Biol 11: 255–269 (1989).
Terras FRG, Eggermont K, Kovaleva V, Raikhel NV, Osborn RW, Kester A, Rees SB, Torrekens S, Leuven FV, Vanderleyden J, Cammue BPA, Broekaert WF: Small cysteine-rich antifungal proteins from radish: their role in host defense. Plant Cell 7: 573–588 (1995).
Titarenko E, Hargreaves J, Keon J, Gurr SJ: Defense-related gene expression in barley coleoptile cells following infection bySeptoria nodorum. In: Fritig B, Legrand M (eds) Mechanisms of Plant Defense Responses. pp. 308–311. Kluwer Academic Publishers. Dordrecht (1993).
von Heijne G: A new method for predicting signal sequence cleavage sites. Nucl Acids Res 14: 4683–4690 (1986).
Xie Y, Wu R: Rice alcohol dehydrogenase genes: anaerobic induction, organ specific expression and characterization of cDNA clones. Plant Mol Biol 13: 53–68 (1989).
Zhang S, Sheng J, Liu Y, Mehdy MC: Fungal elicitor-induced bean proline-rich protein mRNA downregulation is due to destabilization that is transcription and translation dependent. Plant Cell 5: 1089–1099 (1993).
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Sharma, P., Lönneborg, A. Isolation and characterization of a cDNA encoding a plant defensin-like protein from roots of Norway spruce. Plant Mol Biol 31, 707–712 (1996). https://doi.org/10.1007/BF00042244
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DOI: https://doi.org/10.1007/BF00042244