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Role of hydroxyproline-rich glycoproteins in resistance of pearl millet against downy mildew pathogen Sclerospora graminicola

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Abstract

Hydroxyproline-rich glycoproteins (HRGPs) are important plant cell wall components involved in plant defense response to pathogen attack. In the present study, a resistant pearl millet (Pennisetum glaucum) cultivar, IP18292, was compared with a susceptible cultivar, 7042S, to investigate the contribution of HRGPs in the successful defense against the phytopathogenic oomycete S. graminicola. Northern hybridization using MeHRGP cDNA, a heterologous probe from cassava, indicated steady accumulation of HRGP transcripts, from 2 h.p.i. onwards with a maximum at 6 h.p.i., in the resistant cultivar. This is followed by HRGPs accumulation at about 8 h.p.i. as revealed by Western-blot analysis. Immunocytochemical localization by tissue printing and confocal immunofluorescence microscopy indicated cell walls of parenchymatic cells and the vascular tissue of coleoptile as sites of HRGP deposition. In vitro studies in the presence of horseradish peroxidase and H2O2 showed cross-linking of pearl millet HRGPs, which occurred parallel to isodityrosine accumulation. Inducible high isodityrosine content was also observed in vivo in the resistant cultivar. Here, H2O2 was found to accumulate as twin burst at 1 and 6 h.p.i., whereas in the susceptible cultivar only an early single peak was detectable. Moreover, the amount of hydroxyproline in HRGPs was about twice as high in the resistant as in the susceptible cultivar. These results suggest that cell wall strengthening in S. graminicola-infected resistant pearl millet is brought about by a combination of polypeptide cross-linking of isodityrosine as well as by the high content of hydroxyproline in HRGPs, and H2O2, in contrast to the susceptible plant.

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Abbreviations

DTT:

Dithiothreitol

h.p.i.:

Hours post infection

HRGP:

Hydroxyproline-rich glycoproteins

Hyp:

Hydroxyproline

IDT:

Isodityrosine

PAGE:

Polyacrylamide gel electrophoresis

References

  • Arrigoni Q, Arrigoni-Liso R, Calabrese G (1977) Ascorbic acid requirement for biosynthesis of hydroxyproline containing proteins in plants. FEBS Lett 82:135–138

    Article  PubMed  CAS  Google Scholar 

  • Benhamou N (1996) Elicitor-induced plant defense pathways. Trends Plant Sci 1:233–240

    Google Scholar 

  • Bestwick CS, Bennett MH, Mansfield JW (1995) Hrp mutant of Pseudomonas syringae pv phaseolicola induces cell wall alterations but not membrane damage leading to the hypersensitive reaction in lettuce. Plant Physiol 108:503–516

    PubMed  CAS  Google Scholar 

  • Bindshedler LV, Whitelegge JP, Millar DJ, Bolwell GP (2006) A two component chitin binding protein from French bean association of proline rich protein with a cysteine rich polypeptide. FEBS Lett 580:1541–1546

    Article  CAS  Google Scholar 

  • Bradley DJ, Kjellbom P, Lamb CJ (1992) Elicitor and wound induced oxidative cross of a proline rich plant cell wall protein: a novel rapid defense response. Cell 70:21–30

    Article  PubMed  CAS  Google Scholar 

  • Brady JD, Sadler IH, Fry SC (1996) Dityrosine a novel tetrameric derivative of tyrosine derivative of tyrosine in plant cell wall proteins: a new potential cross-link. Biochem J 315:323–327

    PubMed  CAS  Google Scholar 

  • Brown I, Trethowan J, Kerry M, Mansfield J, Bolwell GP (1998) Localization of components of the oxidative cross-linking of glycoproteins and of callose synthesis in papillae formed during the interaction between non-pathogenic strains of Xanthomonas campestris and French bean mesophyll cells. Plant J 15:333–343

    Article  CAS  Google Scholar 

  • Cassab GI, Varner JE (1987) Immunocytolocalization of extensin in developing soybean seed coats by immunogold-sliver staining and by tissue printing on nitrocellulose paper. J Cell Biol 105:2581–2588

    Article  PubMed  CAS  Google Scholar 

  • Cassab GI, Varner JE (1988) Cell wall Proteins. Annu Rev Plant Physiol 39:321–353

    Article  CAS  Google Scholar 

  • Cooper JB, Varner JE (1984) Cross-linking of soluble extensin in isolated cell walls. Plant Physiol 76:414–417

    PubMed  CAS  Google Scholar 

  • Davis HA, Daniels MJ, Dow JW (1997) Induction of extracellular matrix glycoproteins in Brassica petioles by wounding and in response to Xanthomonas campestris. Mol Plant Microbe Interact 10:812–820

    Article  Google Scholar 

  • De Tullio MC, Paciolla C, Vecchia FD, Rascin N, D’Emerico S, De Gara L, Liso R, Arrigoud O (1999) Changes in onion root development induced by the inhibition of peptidylproly hydroxylase and influence of the ascorbate system on cell division and elongation. Planta 209:424–434

    Article  PubMed  Google Scholar 

  • Deepak S, Shailasree S, Sujeeth N, Kini RK, Shetty HS, Mithöfer A (2007) Purification and characterization of proline/hydroxyproline-rich glycoprotein from pearl millet coleoptiles infected with downy mildew pathogen Sclerospora graminicola. Phytochemistry 68:298–305

    Article  PubMed  CAS  Google Scholar 

  • Epstein L, Lamport DTA (1984) An intramolecular linkage involving isodityrosine in extensin. Phytochemistry 23:1241–1246

    Article  CAS  Google Scholar 

  • Frew JE, Jones P, Scholes G (1983) Spectrophotometric determination of hydrogen peroxide and organic hydroperoxides at low concentrations in aqueous solution. Anal Chim Acta 1555:139–150

    Article  Google Scholar 

  • Fritz SE, Hood KR, Hood EE (1991) Localization of soluble and insoluble fractions of hydroxyproline rich glycoproteins during maize kernel development. J Cell Sci 98:545–550

    CAS  Google Scholar 

  • Fry SC (1982) Isodityrosine a new cross linking amino acid from plant cell wall glycoproteins. Biochem J 204:449–455

    PubMed  CAS  Google Scholar 

  • Fry SC (1984) Isodityrosine a biphenyl ether cross link in plant cell wall glycoprotein: Identification assay and chemical synthesis. Methods Enzymol 107:388–397

    CAS  Google Scholar 

  • Fry SC (1986) Cross linking of matrix polymers in the growing cell walls of angiosperms. Annu Rev Plant Physiol 37:165–186

    Article  CAS  Google Scholar 

  • Garcia Muniz N, Martinez-Isquierdo JAM, Puigodomenech P (1998) Induction of mRNA accumulation corresponding to gene encoding a cell wall hydroxyproline rich glycoprotein by fungal elicitors. Plant Mol Biol 38:623–632

    Article  PubMed  CAS  Google Scholar 

  • Geetha HM, Shetty HS (2002) Expression of oxidative burst in cultured cells of pearl millet cultivars against Sclerospora graminicola inoculation and elicitor treatment. Plant Sci 163:653–660

    Article  CAS  Google Scholar 

  • Han YH, Gomez-Vasquez R, Reilly K, Li HY, Tohme J, Cooper RM, Beeching JR (2001) Hydroxyproline-rich glycoproteins expressed during stress responses in cassava. Euphyta 120:59–70

    Article  CAS  Google Scholar 

  • Hause B, Demus U, Teichmann C, Parthier B, Wasternack C (1996) Developmental and tissue-specific expression of JIP−23, a jasmonate-inducible protein of barley. Plant Cell Physiol 37:641–649

    PubMed  CAS  Google Scholar 

  • Hood KR, Baasiri RA, Fritz SE, Hood EE (1991) Biochemical and tissue print analyses of hydroxyproline rich glycoproteins in cell walls of sporophytic maize tissue. Plant Physiol 96:1214–1219

    PubMed  CAS  Google Scholar 

  • Isayenkov S, Mrosk C, Stenzel I, Strack D, Hause B (2005) Suppression of allene oxide cyclase in hairy roots of Medicago truncatula reduces jasmonate levels and the degree of mycorrhization with Glomus intraradices. Plant Physiol 139:1401–1410

    Article  PubMed  CAS  Google Scholar 

  • Kang Z, Buchenauer H (2003) Immunocytochemical localization of cell wall-bound thionins and hydroxyproline-rich glycoproteins in Fusarium culmorum-infected wheat spikes. J Phytopathol 151:120–129

    Article  CAS  Google Scholar 

  • Keller B, Tepleton MD, Lamb CJ (1988) Glycine rich cell wall protein in bean; gene structure and association of the protein with the vascular system. EMBO J 3:3625–3633

    Google Scholar 

  • Kumudini BS, Vasanthi NS, Shetty HS (2001) Hypersensitive response, cell death and histochemical localisation of hydrogen peroxide in the host and nonhost seedlings infected with downy mildew pathogen Sclerospora graminicola. Ann Appl Biol 139:217–225

    Article  Google Scholar 

  • Leach JE, Cantrell MA, Sequiera L (1982) Hydroxyproline rich bacterial agglutinin from potato. Plant Physiol 70:1353–1358

    PubMed  CAS  Google Scholar 

  • Mazau D, Rumeau D, Esquerre-Tugaye MT (1987) Molecular approaches to understanding cell surface interactions between plants and fungal pathogens. Plant Physiol Biochem 25:337–343

    CAS  Google Scholar 

  • Millar DJ, Slabas AR, Sidebottom C, Smith CG, Allen AK, Bolwell GP (1992) A major stress inducible Mr- 42kDa wall glycoprotein of French bean (Phaseolus vulgaris L.). Planta 187:176–184

    Article  CAS  Google Scholar 

  • Narvez-Vásquez J, Pearce G, Ryan CA (2005) The plant cell wall matrix harbors a precursor of defense signaling peptides. Proc Natl Acad Sci USA 102:12974–12977

    Article  CAS  Google Scholar 

  • Otte O, Barz W (2000) Characterization and oxidative in vitro cross linking of an extensin like protein and a proline-rich protein purified from chick pea cell walls. Phytochemistry 53:1–5

    Article  PubMed  CAS  Google Scholar 

  • Prockop DJ, Udenfriend SA (1960) Specific method for the analysis of hydroxyproline in tissues and urine. Anal Biochem 1:228–239

    Article  PubMed  CAS  Google Scholar 

  • Raggi V (2000) Hydroxyproline rich glycoprotein accumulation in tobacco leaves protected against Erysiphe cichotacearum by potato virus Y infection. Plant Pathol 49:179–186

    Article  CAS  Google Scholar 

  • Rumeau D, Mazau D, Panabieres F, Delseny M, Esquerre-Tugaye M (1988) Accumulation of hydroxyproline rich glycoprotein mRNAs in infected or ethylene treated melon plants. Physiol Mol Plant Pathol 33:419–428

    Article  CAS  Google Scholar 

  • Safeeulla KM (1976) Biology and control of the downy mildew of pearl millet, sorghum and finger millet. Wesley Press, Mysore

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor

    Google Scholar 

  • Shailasree S, Kini KR, Deepak S, Kumudini BS, Shetty HS (2004) Accumulation of hydroxyproline-rich glycoproteins in pearl millet seedlings in response to Sclerospora graminicola infection. Plant Sci 167:1227–1234

    Article  CAS  Google Scholar 

  • Shivakumar PD, Geetha HM, Shetty HS (2003) Peroxidase activity and isozyme analysis of pearl millet seedlings and their implications in downy mildew disease resistance. Plant Sci 164:85–93

    Article  CAS  Google Scholar 

  • Smallhood M, Martin H, Knox JP (1995) An epitope of rice threonine and HRGP is common to cell wall and hydrophobic plasma membrane glycoproteins. Planta 196:510–522

    Google Scholar 

  • Sommer-Knudsen J, Bacic A, Clarke AE (1998) Hydroxyproline-rich plant glycoproteins. Phytochemistry 47:483–497

    Article  CAS  Google Scholar 

  • Stafstrom JP, Staehelin LA (1986) Cross linking patterns on salt extractable extensin from carrot cell walls. Plant Physiol 81:234–241

    PubMed  CAS  Google Scholar 

  • Sutherland MW (1991) The generation of oxygen radicals during host plant responses to infection. Physiol Mol Plant Pathol 39:79–93

    Article  CAS  Google Scholar 

  • Templeton MD, Dixon RA, Lamb CJ, Lawton MA (1990) Hydroxyproline rich glycoprotein transcripts exhibit different spatial patterns of accumulation in compatible and incompatible interactions between Phaseolus vulgaris and Colletotrichum lindimuthianum. Plant Physiol 94:1265–1269

    Article  PubMed  CAS  Google Scholar 

  • Tenhaken R, Levine A, Brisson LF, Dixon R, Lamb C (1995) Function of the oxidative burst in disease resistance. Proc Natl Acad Sci USA 92:4158–4163

    Article  PubMed  CAS  Google Scholar 

  • Tschank G, Sanders J, Baringhaus KH, Dallacker F, Kivirikko KI (1994) Structural requirements for the utilization of ascorbate analogues in the prolyl 4-hydroxylase reaction. Biochem J 300:75–79

    PubMed  CAS  Google Scholar 

  • Vandenbosch KA, Bradley DJ, Knox JP, Perotto S, Butcher GW, Brewin NJ (1989) Common components of the infection thread matrix and the intercellular space identified by immunocytochemical analysis of pea nodules and uninfected roots. EMBO J 8:335–341

    PubMed  CAS  Google Scholar 

  • Winston S, Fuller S, Huller J (1987) Western blotting. In: Ausubel EM (ed) Current protocols in molecular biology. John Wiley, New York, pp 10.8.1–10.8.6

    Google Scholar 

  • Wojtaszek P, Trethowan J, Bolwell GP (1995) Specificity in the immobilization of cell wall proteins in response to different elicitor molecules in suspension-cultured cells of French bean (Phaseolus vulgaris L.). Plant Mol Biol 28:1075–1087

    Article  PubMed  CAS  Google Scholar 

  • Wojtaszek P, Trethowan J, Bolwell GP (1997) Reconstitution in vitro of the components and conditions required for the oxidative cross linking of extracellular proteins in French bean (Phaseolus vulgaris L.). FEBS Lett 405:95–98

    Article  PubMed  CAS  Google Scholar 

  • York WS, Darvill AG, McNeil M, Stevensen TT, Albersheim P (1986) Isolation and characterization of plant cell walls and cell wall components. Methods Enzymol 118:3–41

    Article  CAS  Google Scholar 

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Acknowledgments

The authors are grateful to the DANIDA-ENRECA for the facilities made available for the research program. We also thank the Department of Science and Technology, Government of India and Indian Council of Agricultural Research, New Delhi and the Max-Planck-Society for financial support. The authors thank E.A. Rathbun for providing MAC 265 monoclonal antibody and K. Reilly for cMeHRGP1 cDNA probe. DS thanks the German Academic Exchange Service (DAAD) for financial support to carry out research work at the Max-Planck-Institute for Chemical Ecology, Jena, Germany. SS acknowledges the research fellowship received from Council of Scientific and Industrial Research, New Delhi, India.

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Authors and Affiliations

  1. Department of Studies in Applied Botany and Biotechnology, University of Mysore, Manasagangotri, Mysore, 570006, Karnataka, India

    Shantharaj Deepak, Sekhar Shailasree, Ramachandra K. Kini & Shekar H. Shetty

  2. Department of Bioorganic Chemistry, Max-Planck-Institute for Chemical Ecology, Hans-Knöll-Straße 8, 07745, Jena, Germany

    Shantharaj Deepak & Axel Mithöfer

  3. Department of Secondary Metabolism, Leibniz Institute of Plant Biochemistry, 06120, Halle/Saale, Germany

    Bettina Hause

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  1. Shantharaj Deepak
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  2. Sekhar Shailasree
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Correspondence to Axel Mithöfer.

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Deepak, S., Shailasree, S., Kini, R.K. et al. Role of hydroxyproline-rich glycoproteins in resistance of pearl millet against downy mildew pathogen Sclerospora graminicola . Planta 226, 323–333 (2007). https://doi.org/10.1007/s00425-007-0484-4

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