caa a22 4500 467894051 CHVBK 20180406152757.0 cr unu---uuuuu 170328e20060701xx s 000 0 eng 10.1007/s11103-006-0043-6 doi (NATIONALLICENCE)springer-10.1007/s11103-006-0043-6 Cloning, characterization and localization of three novel class III peroxidases in lignifying xylem of Norway spruce ( Picea abies ) [Elektronische Daten] [Kaisa Marjamaa, Kristiina Hildén, Eija Kukkola, Mikko Lehtonen, Heidi Holkeri, Pekka Haapaniemi, Sanna Koutaniemi, Teemu Teeri, Kurt Fagerstedt, Taina Lundell] 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. Springer Science+Business Media B.V., 2006 Lignin biosynthesis nationallicence Class III plant peroxidases nationallicence Picea abies nationallicence Gymnosperm nationallicence In situ hybridization nationallicence Secretion signal peptide nationallicence GFP nationallicence Endoplasmic reticulum nationallicence AP : alkaline phosphatase nationallicence DEPC : diethylpyrocarbonate nationallicence DIG : digoxigenin nationallicence EGFP : enhanced green fluorescent protein nationallicence ER : endoplasmic reticulum nationallicence FBPA : fructose-1,6-bisphosphate aldolase nationallicence IEF : isoelectric focusing nationallicence ORF : open reading frame nationallicence PBS : phosphate-buffered saline nationallicence POX : class III plant peroxidase nationallicence RT : room temperature nationallicence SS : signal sequence nationallicence UPM : Universal Primer mix nationallicence Marjamaa Kaisa Department of Biological and Environmental Sciences, University of Helsinki, P.O. Box 65, 00014, Helsinki, Finland aut Hildén Kristiina Department of Applied Chemistry and Microbiology, University of Helsinki, P.O. Box 56, 00014UH, Helsinki, Finland aut Kukkola Eija Department of Biological and Environmental Sciences, University of Helsinki, P.O. Box 65, 00014, Helsinki, Finland aut Lehtonen Mikko Department of Applied Biology, University of Helsinki, P.O. Box 27, 00014UH, Helsinki, Finland aut Holkeri Heidi VTT Technical Research Centre of Finland, P.O. Box 1000, 02044 VTT, Helsinki, Finland aut Haapaniemi Pekka Department of Biological and Environmental Sciences, University of Helsinki, P.O. Box 65, 00014, Helsinki, Finland aut Koutaniemi Sanna Department of Applied Biology, University of Helsinki, P.O. Box 27, 00014UH, Helsinki, Finland aut Teeri Teemu Department of Applied Biology, University of Helsinki, P.O. Box 27, 00014UH, Helsinki, Finland aut Fagerstedt Kurt Department of Biological and Environmental Sciences, University of Helsinki, P.O. Box 65, 00014, Helsinki, Finland aut Lundell Taina Department of Applied Chemistry and Microbiology, University of Helsinki, P.O. Box 56, 00014UH, Helsinki, Finland aut Plant Molecular Biology Kluwer Academic Publishers 61/4-5(2006-07-01), 719-732 0167-4412 61:4-5<719 2006 61 11103 https://doi.org/10.1007/s11103-006-0043-6 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11103-006-0043-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Marjamaa Kaisa Department of Biological and Environmental Sciences, University of Helsinki, P.O. Box 65, 00014, Helsinki, Finland aut NATIONALLICENCE 700 1- Hildén Kristiina Department of Applied Chemistry and Microbiology, University of Helsinki, P.O. Box 56, 00014UH, Helsinki, Finland aut NATIONALLICENCE 700 1- Kukkola Eija Department of Biological and Environmental Sciences, University of Helsinki, P.O. Box 65, 00014, Helsinki, Finland aut NATIONALLICENCE 700 1- Lehtonen Mikko Department of Applied Biology, University of Helsinki, P.O. Box 27, 00014UH, Helsinki, Finland aut NATIONALLICENCE 700 1- Holkeri Heidi VTT Technical Research Centre of Finland, P.O. Box 1000, 02044 VTT, Helsinki, Finland aut NATIONALLICENCE 700 1- Haapaniemi Pekka Department of Biological and Environmental Sciences, University of Helsinki, P.O. Box 65, 00014, Helsinki, Finland aut NATIONALLICENCE 700 1- Koutaniemi Sanna Department of Applied Biology, University of Helsinki, P.O. Box 27, 00014UH, Helsinki, Finland aut NATIONALLICENCE 700 1- Teeri Teemu Department of Applied Biology, University of Helsinki, P.O. Box 27, 00014UH, Helsinki, Finland aut NATIONALLICENCE 700 1- Fagerstedt Kurt Department of Biological and Environmental Sciences, University of Helsinki, P.O. Box 65, 00014, Helsinki, Finland aut NATIONALLICENCE 700 1- Lundell Taina Department of Applied Chemistry and Microbiology, University of Helsinki, P.O. Box 56, 00014UH, Helsinki, Finland aut NATIONALLICENCE 773 0- Plant Molecular Biology Kluwer Academic Publishers 61/4-5(2006-07-01), 719-732 0167-4412 61:4-5<719 2006 61 11103 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer