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

K. Marjamaa (Corresponding Author), K. Hilden, E. Kukkola, M. Lehtonen, Heidi Holkeri, P. Haapaniemi, S. Koutaniemi, Teemu Teeri, K. Fagerstedt, T. Lundell

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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.
Original languageEnglish
Pages (from-to)719-732
Number of pages14
JournalPlant Molecular Biology
Volume61
Issue number4-5
DOIs
Publication statusPublished - 2006
MoE publication typeA1 Journal article-refereed

Fingerprint

Abies
Picea
Peroxidases
Xylem
peroxidases
Norway
Picea abies
xylem
Organism Cloning
molecular cloning
Cell Wall
Peroxidase
peroxidase
tracheids
cell walls
secretion
Protein Sorting Signals
signal peptide
Gymnosperms
Catharanthus

Keywords

  • Class III plant peroxidases
  • Endoplasmic reticulum
  • GFP
  • Gymnosperm
  • In situ hybridization
  • Lignin biosynthesis
  • Picea abies
  • Secretion signal peptide

Cite this

Marjamaa, K. ; Hilden, K. ; Kukkola, E. ; Lehtonen, M. ; Holkeri, Heidi ; Haapaniemi, P. ; Koutaniemi, S. ; Teeri, Teemu ; Fagerstedt, K. ; Lundell, T. / Cloning, characterization and localization of three novel class III peroxidases in lignifying xylem of Norway spruce (Picea abies). In: Plant Molecular Biology. 2006 ; Vol. 61, No. 4-5. pp. 719-732.
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title = "Cloning, characterization and localization of three novel class III peroxidases in lignifying xylem of Norway spruce (Picea abies)",
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.",
keywords = "Class III plant peroxidases, Endoplasmic reticulum, GFP, Gymnosperm, In situ hybridization, Lignin biosynthesis, Picea abies, Secretion signal peptide",
author = "K. Marjamaa and K. Hilden and E. Kukkola and M. Lehtonen and Heidi Holkeri and P. Haapaniemi and S. Koutaniemi and Teemu Teeri and K. Fagerstedt and T. Lundell",
year = "2006",
doi = "10.1007/s11103-006-0043-6",
language = "English",
volume = "61",
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Marjamaa, K, Hilden, K, Kukkola, E, Lehtonen, M, Holkeri, H, Haapaniemi, P, Koutaniemi, S, Teeri, T, Fagerstedt, K & Lundell, T 2006, 'Cloning, characterization and localization of three novel class III peroxidases in lignifying xylem of Norway spruce (Picea abies)', Plant Molecular Biology, vol. 61, no. 4-5, pp. 719-732. https://doi.org/10.1007/s11103-006-0043-6

Cloning, characterization and localization of three novel class III peroxidases in lignifying xylem of Norway spruce (Picea abies). / Marjamaa, K. (Corresponding Author); Hilden, K.; Kukkola, E.; Lehtonen, M.; Holkeri, Heidi; Haapaniemi, P.; Koutaniemi, S.; Teeri, Teemu; Fagerstedt, K.; Lundell, T.

In: Plant Molecular Biology, Vol. 61, No. 4-5, 2006, p. 719-732.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Cloning, characterization and localization of three novel class III peroxidases in lignifying xylem of Norway spruce (Picea abies)

AU - Marjamaa, K.

AU - Hilden, K.

AU - Kukkola, E.

AU - Lehtonen, M.

AU - Holkeri, Heidi

AU - Haapaniemi, P.

AU - Koutaniemi, S.

AU - Teeri, Teemu

AU - Fagerstedt, K.

AU - Lundell, T.

PY - 2006

Y1 - 2006

N2 - 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.

AB - 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.

KW - Class III plant peroxidases

KW - Endoplasmic reticulum

KW - GFP

KW - Gymnosperm

KW - In situ hybridization

KW - Lignin biosynthesis

KW - Picea abies

KW - Secretion signal peptide

U2 - 10.1007/s11103-006-0043-6

DO - 10.1007/s11103-006-0043-6

M3 - Article

VL - 61

SP - 719

EP - 732

JO - Plant Molecular Biology

JF - Plant Molecular Biology

SN - 0167-4412

IS - 4-5

ER -