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Cloning, characterization and localization of three novel class III peroxidases in lignifying xylem of Norway spruce (Picea abies)

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Abstract

Plant class III peroxidases (POXs) take part in the formation of lignin and maturation of plant cell walls. However, only a few examples of such peroxidases from gymnosperm tree species with highly lignified xylem tracheids have been implicated so far. We report here cDNA cloning of three xylem-expressed class III peroxidase encoding genes from Norway spruce (Picea abies). The translated proteins, PX1, PX2 and PX3, contain the conserved amino acids required for heme-binding and peroxidase catalysis. They all begin with putative secretion signal propeptide sequences but diverge substantially at phylogenetic level, grouping to two subclusters when aligned with other class III plant peroxidases. In situ hybridization analysis on expression of the three POXs in Norway spruce seedlings showed that mRNA coding for PX1 and PX2 accumulated in the cytoplasm of young, developing tracheids within the current growth ring where lignification is occurring. Function of the putative N-terminal secretion signal peptides for PX1, PX2 and PX3 was confirmed by constructing chimeric fusions with EGFP (enhanced green fluorescent protein) and expressing them in tobacco protoplasts. Full-length coding region of px1 was also heterologously expressed in Catharanthus roseus hairy root cultures. Thus, at least the spruce PX1 peroxidase is processed via the endoplasmic reticulum (ER) most likely for secretion to the cell wall. Thereby, PX1 displays correct spatiotemporal localization for participation in the maturation of the spruce tracheid secondary cell wall.

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Abbreviations

AP:

alkaline phosphatase

DEPC:

diethylpyrocarbonate

DIG:

digoxigenin

EGFP:

enhanced green fluorescent protein

ER:

endoplasmic reticulum

FBPA:

fructose-1,6-bisphosphate aldolase

IEF:

isoelectric focusing

ORF:

open reading frame

PBS:

phosphate-buffered saline

POX:

class III plant peroxidase

RT:

room temperature

SS:

signal sequence

UPM:

Universal Primer mix

References

  • KA Blee JW Choi AP O’Connell W Schung NG Lewis GP Bolwell (2003) ArticleTitleA lignin specific peroxidase in tobacco whose antisense suppression leads to vascular tissue modification Phytochemistry 64 163–176 Occurrence Handle12946415 Occurrence Handle1:CAS:528:DC%2BD3sXms1OkurY%3D Occurrence Handle10.1016/S0031-9422(03)00212-7

    Article  PubMed  CAS  Google Scholar 

  • W Boerjan J Ralph M Baucher (2003) ArticleTitleLignin biosynthesis Annu Rev Plant Biol 54 519–546 Occurrence Handle14503002 Occurrence Handle1:CAS:528:DC%2BD3sXntFSnsrg%3D Occurrence Handle10.1146/annurev.arplant.54.031902.134938

    Article  PubMed  CAS  Google Scholar 

  • JH Christensen G Bauw KG Welinder M Montagu ParticleVan W Boerjan (1998) ArticleTitlePurification and characterization of peroxidases correlated with lignification in poplar xylem Plant Physiol 118 125–135 Occurrence Handle9733532 Occurrence Handle1:CAS:528:DyaK1cXmtV2mtLY%3D Occurrence Handle10.1104/pp.118.1.125

    Article  PubMed  CAS  Google Scholar 

  • JH Christensen S Overney A Rohde WA Diaz G Bauw P Simon M Montagu ParticleVan W Boerjan (2001) ArticleTitleThe syringaldazine-oxidizing peroxidase PXP 3–4 from poplar xylem: cDNA isolation, characterization and expression Plant Mol Biol 47 581–593 Occurrence Handle11725944 Occurrence Handle1:CAS:528:DC%2BD3MXovVylu74%3D Occurrence Handle10.1023/A:1012271729285

    Article  PubMed  CAS  Google Scholar 

  • TE Creighton (1993) Proteins, structures and molecular properties EditionNumber2 W.H. Freeman New York

    Google Scholar 

  • G-P Di Sansebastiano N Paris S Marc-Martin J-M Neuhaus (1998) ArticleTitleSpecific accumulation of GFP in a non-acidic compartment via C-terminal propeptide-mediated sorting pathway Plant J 15 449–457 Occurrence Handle9753772 Occurrence Handle1:CAS:528:DyaK1cXmtFOitrk%3D Occurrence Handle10.1046/j.1365-313X.1998.00210.x

    Article  PubMed  CAS  Google Scholar 

  • M Elfstrand CG Fossdal F Sitbon O Olsson A Lönneborg S Arnold Particlevon (2001) ArticleTitleOverexpression of endogenous peroxidase-like spi 2 in transgenic Norway spruce plants results in increased total peroxidase activity and reduced growth Plant Cell Rep 20 596–603 Occurrence Handle1:CAS:528:DC%2BD3MXnvVCrsrk%3D Occurrence Handle10.1007/s002990100360

    Article  CAS  Google Scholar 

  • M Elfstrand F Sitbon C Lapierre A Bottin S Arnold Particlevon (2002) ArticleTitleAltered lignin structure and resistance to pathogens in spi 2-expressing tobacco plants Planta 214 708–716 Occurrence Handle11882939 Occurrence Handle1:CAS:528:DC%2BD38Xhs1yqs7c%3D Occurrence Handle10.1007/s00425-001-0681-5

    Article  PubMed  CAS  Google Scholar 

  • I El Mansouri JA Mercado N Santiago-Dómenech F Pliego-Alfaro V Valpuestra MA Quesada (1999) ArticleTitleBiochemical and phenotypical characterization of transgenic tomato plants overexpressing a basic peroxidase Physiol Plant 106 355–362 Occurrence Handle1:CAS:528:DyaK1MXmvFejsrk%3D Occurrence Handle10.1034/j.1399-3054.1999.106401.x

    Article  CAS  Google Scholar 

  • O Emanuelsson H Nielsen S Brunak G Heijne Particlevon (2000) ArticleTitlePredicting subcellular localization of proteins based on their N-terminal amino acid sequence J Mol Biol 300 1005–1016 Occurrence Handle10891285 Occurrence Handle1:CAS:528:DC%2BD3cXks1OntrY%3D Occurrence Handle10.1006/jmbi.2000.3903

    Article  PubMed  CAS  Google Scholar 

  • F Diaz-DeLeon KL Klotz LM Lagrimini (1993) ArticleTitleNucleotide sequence of the tobacco (Nicotiana tabacum) anionic peroxidase gene Plant Physiol 101 1117–1118 Occurrence Handle1:CAS:528:DyaK3sXit1Ohs7w%3D Occurrence Handle10.1104/pp.101.3.1117

    Article  CAS  Google Scholar 

  • CG Fossdal P Sharma A Lönneborg (2001) ArticleTitleIsolation of the first putative peroxidase cDNA from a conifer and the local and systemic accumulation of related proteins upon pathogen infection Plant Mol Biol 47 423–435 Occurrence Handle11587513 Occurrence Handle1:CAS:528:DC%2BD3MXntlGju7s%3D Occurrence Handle10.1023/A:1011615328684

    Article  PubMed  CAS  Google Scholar 

  • C Gabaldón M López-Serrano M Pedreño A Ros Barceló (2005) ArticleTitleCloning and molecular characterization of the basic peroxidase isoenzyme from Zinnia elegans, an enzyme involved in lignin biosynthesis Plant Physiol 139 1138–1154 Occurrence Handle16258008 Occurrence Handle10.1104/pp.105.069674

    Article  PubMed  Google Scholar 

  • M Gajhede DJ Schuller A Henriksen AT Smith TL Poulos (1997) ArticleTitleCrystal structure of horseradish peroxidase C at 2.15 A resolution Nat Struct Biol 4 1032–1038 Occurrence Handle9406554 Occurrence Handle1:CAS:528:DyaK2sXnvV2nsrs%3D Occurrence Handle10.1038/nsb1297-1032

    Article  PubMed  CAS  Google Scholar 

  • A Groover JR Fontana G Dupper C Ma R Martienssen S Strauss R Meilan (2004) ArticleTitleGene and enhancer trap tagging of vascular-expressed genes in poplar trees Plant Physiol 134 1742–1751 Occurrence Handle15051863 Occurrence Handle1:CAS:528:DC%2BD2cXjsFKmtr4%3D Occurrence Handle10.1104/pp.103.034330

    Article  PubMed  CAS  Google Scholar 

  • JM Harkin JR Obst (1973) ArticleTitleLignification in trees: indication of exclusive peroxidase participation Science 180 296–298 Occurrence Handle1:CAS:528:DyaE3sXhsFKrsL0%3D

    CAS  Google Scholar 

  • S Hawkins J Samaj V Lauvergeat A Boudet J Grima-Pettenati (1997) ArticleTitleCinnamyl alcohol dehydrogenase: identification of new sites of promoter activity in transgenic poplar Plant Physiol 113 321–325 Occurrence Handle12223610 Occurrence Handle1:CAS:528:DyaK2sXht1Cltr4%3D

    PubMed  CAS  Google Scholar 

  • S Hawkins A Boudet J Grima-Pettenati (2003) ArticleTitleCharacterisation of caffeic acid O-methyltransferase and cinnamyl alcohol dehydrogenase gene expression patterns by in situ hybridisation in Eucalyptus gunnii Hook. plantlets Plant Sci 164 165–173 Occurrence Handle1:CAS:528:DC%2BD3sXivVCmsw%3D%3D Occurrence Handle10.1016/S0168-9452(02)00361-8

    Article  CAS  Google Scholar 

  • T Higuchi (1997) Biochemistry and molecular biology of wood Springer-Verlag Berlin Heidelberg

    Google Scholar 

  • S Hiraga K Sasaki H Ito Y Ohashi H Matsui (2001) ArticleTitleA large family of plant III peroxidases Plant Cell Physiol 42 462–468 Occurrence Handle11382811 Occurrence Handle1:CAS:528:DC%2BD3MXktVWntrY%3D Occurrence Handle10.1093/pcp/pce061

    Article  PubMed  CAS  Google Scholar 

  • UC Kalluri CP Joshi (2004) ArticleTitleDifferential expression patterns of two cellulose synthase genes are associated with primary and secondary cell wall development in aspen trees Planta 220 47–55 Occurrence Handle15278458 Occurrence Handle1:CAS:528:DC%2BD2cXptlSmur8%3D Occurrence Handle10.1007/s00425-004-1329-z

    Article  PubMed  CAS  Google Scholar 

  • M Karimi D Inze A Depicker (2002) ArticleTitleGateway vectors for Agrobacterium-mediated plant transformation Trends Plant Sci 7 IssueID5 193–195 Occurrence Handle11992820 Occurrence Handle1:CAS:528:DC%2BD38Xjtl2ktrk%3D Occurrence Handle10.1016/S1360-1385(02)02251-3

    Article  PubMed  CAS  Google Scholar 

  • S Koutaniemi MM Toikka A Kärkönen M Mustonen T Lundell LK Simola IA Kilpeläinen TH Teeri (2005) ArticleTitleCharacterization of basic p-coumaryl and coniferyl alcohol oxidasing peroxidases from lignin forming Picea abies suspension culture Plant Mol Biol 58 141–157 Occurrence Handle16027971 Occurrence Handle1:CAS:528:DC%2BD2MXmt1ygtbg%3D Occurrence Handle10.1007/s11103-005-5345-6

    Article  PubMed  CAS  Google Scholar 

  • E Kukkola S Koutaniemi M Gustafsson T Karhunen K Ruel T Lundell P. G Saranpää Brunow T Teeri K Fagerstedt (2003) ArticleTitleLocalization of dibenzodioxocin substructures in lignifying Norway spruce xylem by transmission electron microscopy-immunogold labeling Planta 217 229–237 Occurrence Handle12783330 Occurrence Handle1:CAS:528:DC%2BD3sXkt1ajsbY%3D

    PubMed  CAS  Google Scholar 

  • AR Leitch T Schwarzacher D Jackson IJ Leitch (1994) Microscopy handbook: in situ hybridisation Royal Microscopical Society Oxford

    Google Scholar 

  • Y Li S Kajita S Kawai Y Katayama N Morohoshi (2003) ArticleTitleDown regulation of an anionic peroxidase in transgenic aspen and its effect on lignin characteristics J Plant Res 116 175–182 Occurrence Handle12836039 Occurrence Handle1:CAS:528:DC%2BD3sXltVWgs7Y%3D Occurrence Handle10.1007/s10265-003-0087-5

    Article  PubMed  CAS  Google Scholar 

  • K Marjamaa M Lehtonen T Lundell M Toikka KV Fagerstedt (2003) ArticleTitleDevelopmental lignification and seasonal variation in β-glucosidase and peroxidase activities in xylem of Scots pine, Norway spruce and silver birch Tree Physiol 23 977–986 Occurrence Handle12952784 Occurrence Handle1:CAS:528:DC%2BD3sXptVCmu74%3D

    PubMed  CAS  Google Scholar 

  • K Marjamaa E Kukkola T Lundell P Karhunen P Saranpää KV Fagerstedt (2006) ArticleTitleMonolignol oxidation by xylem peroxidase isoforms of Norway spruce (Picea abies) and silver birch (Betula pendula) Tree Physiol 26 605–611 Occurrence Handle16452074 Occurrence Handle1:CAS:528:DC%2BD28XltFWqs7o%3D

    PubMed  CAS  Google Scholar 

  • S Martens T Teeri G Forkmann (2002) ArticleTitleHeterologous expression of dihydroflavonol 4-reductase from various plants FEBS Lett 531 453–458 Occurrence Handle12435592 Occurrence Handle1:CAS:528:DC%2BD38XosFCksLs%3D Occurrence Handle10.1016/S0014-5793(02)03583-4

    Article  PubMed  CAS  Google Scholar 

  • T Matsui H Nakayama K Yoshida A Shinmyo (2003) ArticleTitleVesicular transport route of horseradish C1a peroxidase is regulated by N- and C-terminal propeptides in tobacco cells Appl Microbiol Biotechnol 62 517–522 Occurrence Handle12879300 Occurrence Handle1:CAS:528:DC%2BD3sXos1Snu70%3D Occurrence Handle10.1007/s00253-003-1273-z

    Article  PubMed  CAS  Google Scholar 

  • GJ McDougal (2001) ArticleTitleCell-wall proteins from Sitka spruce xylem are selectively insolubilised during formation of dehydrogenation polymers of coniferyl alcohol Phytochem 57 157–163 Occurrence Handle10.1016/S0031-9422(01)00027-9

    Article  Google Scholar 

  • S Munro HR Pelham (1987) ArticleTitleA C-terminal signal prevents secretion of luminal ER proteins Cell 48 899–907 Occurrence Handle3545499 Occurrence Handle1:CAS:528:DyaL1cXht1Kqsb8%3D Occurrence Handle10.1016/0092-8674(87)90086-9

    Article  PubMed  CAS  Google Scholar 

  • U Nehls A Wiese M Guttenberger R Hampp (1998) ArticleTitleCarbon allocation in ectomycorrhizas: identification and expression analysis of an Amanita muscaria monosaccharide transporter Mol Plant Micr Inter 11 167–176 Occurrence Handle1:CAS:528:DyaK1cXht12htbs%3D

    CAS  Google Scholar 

  • H Nielsen J Engelbrecht S Brunak G Heijne Particlevon (1997) ArticleTitleIdentification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites Prot Eng 10 1–6 Occurrence Handle1:CAS:528:DyaK2sXhsVersrs%3D Occurrence Handle10.1093/protein/10.1.1

    Article  CAS  Google Scholar 

  • L Østergaard K Teilum O Mirza O Mattsson M Petersen KG Welinder J Mundy M Gajhede A Henriksen (2000) ArticleTitleArabidopsis ATP A2 peroxidase. Expression and high-resolution structure of a plant peroxidase with implications for lignification Plant Mol Biol 44 231–243 Occurrence Handle11117266 Occurrence Handle10.1023/A:1006442618860

    Article  PubMed  Google Scholar 

  • F Passardi D Longet C Penel C Dunand (2004) ArticleTitleThe class III peroxidase multigenic family in rice and its evolution in land plants Phytochem 65 1879–1893 Occurrence Handle1:CAS:528:DC%2BD2cXmtVCjtrc%3D Occurrence Handle10.1016/j.phytochem.2004.06.023

    Article  CAS  Google Scholar 

  • HR Pelham (1988) ArticleTitleEvidence that luminal ER proteins are sorted from secreted proteins in a post-ER compartment EMBO J 7 913–918 Occurrence Handle3402439 Occurrence Handle1:CAS:528:DyaL1cXitVSgsbY%3D

    PubMed  CAS  Google Scholar 

  • M Quiroga C Guerrero MA Botella A Barceló I Amaya MI Medina FJ Alonso S Milrad Forchetti Particlede H Tigier V Valpuesta (2000) ArticleTitleA tomato peroxidase involved in the synthesis of lignin and suberin Plant Physiol 122 1119–1127 Occurrence Handle10759507 Occurrence Handle1:CAS:528:DC%2BD3cXktFSqtbc%3D Occurrence Handle10.1104/pp.122.4.1119

    Article  PubMed  CAS  Google Scholar 

  • S Regan V Bourquin H. Tuominen B Sundberg (1999) ArticleTitleAccurate and high resolution in situ hybridization analysis of gene expression in secondary stem tissues Plant J 19 363–369 Occurrence Handle10476083 Occurrence Handle1:CAS:528:DyaK1MXmt1Olt7s%3D Occurrence Handle10.1046/j.1365-313X.1999.00536.x

    Article  PubMed  CAS  Google Scholar 

  • S Regan B Sundberg (2002) High resolution in situ hybridisation in woody tissues N Chaffey (Eds) Wood formation in trees Taylor & Francis New York 297–317

    Google Scholar 

  • DJ Schuller N Ban RB Huystee A McPherson TL Poulos (1996) ArticleTitleThe crystal structure of peanut peroxidase Structure 4 311–321 Occurrence Handle8805539 Occurrence Handle1:CAS:528:DyaK28XitVensr8%3D Occurrence Handle10.1016/S0969-2126(96)00035-4

    Article  PubMed  CAS  Google Scholar 

  • L Simola J Lemmetyinen A Santanen (1992) ArticleTitleLignin release and photomixotrophism in suspension cultures of Picea abies Physiol Plantarum 84 374–379 Occurrence Handle1:CAS:528:DyaK38XisFahs78%3D Occurrence Handle10.1111/j.1399-3054.1992.tb04678.x

    Article  CAS  Google Scholar 

  • AT Smith NC Veitch (1998) ArticleTitleSubstrate binding and catalysis in heme peroxidases Curr Opin Chem Biol 2 269–278 Occurrence Handle9667928 Occurrence Handle1:CAS:528:DyaK1cXjs1OltLg%3D Occurrence Handle10.1016/S1367-5931(98)80069-0

    Article  PubMed  CAS  Google Scholar 

  • T Talas-Oğraş K Kazan N Gözükirmizi (2001) ArticleTitleDecreased peroxidase activity in transgenic tobacco and its effects on lignification Biotech Lett 23 267–273 Occurrence Handle10.1023/A:1005602213270

    Article  Google Scholar 

  • MT Tarkka TA Nyman N Kalkkinen M Raudaskoski (2000) ArticleTitleScots pine expresses short root specific peroxidases during development Eur J Biochem 267 1–8 Occurrence Handle10.1046/j.1432-1327.2000.02000.x

    Article  Google Scholar 

  • B Theilade SK Rasmussen I Rosenkrands H Frokjaer J Hejgaard J Theilade K Pihakaski-Maunsbach AB Maunsbach (1993) Subcellular localization of barley grain peroxidase BP2 by immuno-electron microscopy KG Welinder SK Rasmussen C Penel H Greppin (Eds) Plant peroxidases Biochemistry and Physiology University of Geneva Switzerland 321–324

    Google Scholar 

  • R Töpfer V Matzcit B Gronenborn J Schell HH Steinbiss (1987) ArticleTitleA set of plant expression vectors for transcriptional and translational fusions Nucleic Acid Res 24 5890

    Google Scholar 

  • KG Welinder (1992) Plant peroxidases: Structure and function relationships C Penel T Gaspar H Greppin (Eds) Plant peroxidases 1980–1990. Topics and detailed literature on biochemical aspects University of Geneva Switzerland 1–24

    Google Scholar 

  • KG Welinder AF Justesen IVH Kjaersgård RB Jensen SK Rasmussen HM Jespersen L Duroux (2002) ArticleTitleStructural diversity and transcription of class III peroxidases from Arabidopsis thaliana Eur J Biochem 269 6063–6081 Occurrence Handle12473102 Occurrence Handle1:CAS:528:DC%2BD3sXht1GntQ%3D%3D Occurrence Handle10.1046/j.1432-1033.2002.03311.x

    Article  PubMed  CAS  Google Scholar 

  • R Whetten Y-H Sun Y Zhang R Sederoff (2001) ArticleTitleFunctional genomics and cell wall biosynthesis in loblolly pine Plant Mol Biol 47 275–291 Occurrence Handle11554476 Occurrence Handle1:CAS:528:DC%2BD3MXmslGmtr4%3D Occurrence Handle10.1023/A:1010652003395

    Article  PubMed  CAS  Google Scholar 

  • L Wu SP Joshi VL Chiang (2000) ArticleTitleA xylem-specific cellulose synthase gene from aspen (Populus tremuloides) is responsive to mechanical stress Plant J 22 495–502 Occurrence Handle10886769 Occurrence Handle1:CAS:528:DC%2BD3cXltFKksLk%3D Occurrence Handle10.1046/j.1365-313x.2000.00758.x

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Biological and Environmental Sciences, University of Helsinki, P.O. Box 65, 00014, Helsinki, Finland

    Kaisa Marjamaa, Eija Kukkola, Pekka Haapaniemi & Kurt Fagerstedt

  2. Department of Applied Chemistry and Microbiology, University of Helsinki, P.O. Box 56, 00014UH, Helsinki, Finland

    Kristiina Hildén & Taina Lundell

  3. Department of Applied Biology, University of Helsinki, P.O. Box 27, Helsinki, 00014UH, Finland

    Mikko Lehtonen, Sanna Koutaniemi & Teemu H. Teeri

  4. VTT Technical Research Centre of Finland, P.O. Box 1000, 02044 VTT, Helsinki, Finland

    Heidi Holkeri

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  1. Kaisa Marjamaa
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  2. Kristiina Hildén
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  3. Eija Kukkola
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  4. Mikko Lehtonen
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Correspondence to Kaisa Marjamaa.

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Marjamaa, K., Hildén, K., Kukkola, E. et al. Cloning, characterization and localization of three novel class III peroxidases in lignifying xylem of Norway spruce (Picea abies). Plant Mol Biol 61, 719–732 (2006). https://doi.org/10.1007/s11103-006-0043-6

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