Cryo-FE-SEM & TEM immuno-techniques reveal new details for understanding white-rot decay of lignocellulose

Geoffrey Daniel (Corresponding Author), Jindrich Volc, Marja-Leena Niku-Paavola

Research output: Contribution to journalArticleScientificpeer-review

29 Citations (Scopus)

Abstract

High-resolution Cryo-Field Emission Scanning Electron Microscopy (HR-Cryo-FE-SEM) and immuno-cytochemistry were used to reveal novel details on the morphological events and spatial distribution of oxidoreductive enzymes during the degradation of birch wood by the white-rot fungi Phlebia radiata and mutant strain P. radiata Cel 26. Cryo-observations of fractured fibres showed degradation across the cell wall by P. radiata (wild) to progress by delamination and removal of concentric orientated aggregates from the secondary S2 cell wall. Decay by P. radiata Cel 26 progressed by removal of materials (lignin and hemicelluloses) between the aggregates (primarily cellulose) that remained even after advanced decay. With both decay patterns, extracellular slime materials were present uniting lumina hyphae with the attacked fibre wall. The extracellular slime material had two morphological forms: viz a fibrillar (often tripartite) and a ‘gel-form’, the former found in discrete bands progressing across the lumen onto the fibre wall. Using TEM immunocytochemistry, laccase, manganese peroxidase (MnP) and diarylpropane enzymes were localized in the periplasmic space of luminal hyphae, in association with the cell membrane, periplasmic vesicles and fungal cell wall. Extracellularly, the three enzymes were found associated with the slime and tripartite membranes and with the birch cell walls at all stages of attack through to middle lamella corner decay. Enzyme distribution was correlated with morphological changes in cell wall structure. The association of extracellular slime with these enzymes and sites of decay strongly suggests a major role for this matrix in fibre cell wall decomposition. To cite this article: G. Daniel et al., C. R. Biologies 327 (2004).

Original languageEnglish
Pages (from-to)861 - 871
Number of pages11
JournalComptes Rendus: Biologies
Volume327
Issue number9-10
DOIs
Publication statusPublished - 2004
MoE publication typeA1 Journal article-refereed

Fingerprint

lignocellulose
Cell Wall
deterioration
Cells
cell walls
Transmission electron microscopy
Scanning electron microscopy
Enzymes
enzymes
Betula
dietary fiber
Hyphae
Fibers
manganese peroxidase
immunocytochemistry
cell aggregates
hyphae
degradation
Phlebia
methodology

Keywords

  • white rot
  • fungi
  • white-rot fungi
  • Phlebia radiata
  • HR-cryo FE-SEM
  • TEM immunocytochemistry
  • laccase
  • manganese peroxidase
  • diarylpropane peroxidase
  • extracellular slime
  • biodegradation
  • birch

Cite this

Daniel, Geoffrey ; Volc, Jindrich ; Niku-Paavola, Marja-Leena. / Cryo-FE-SEM & TEM immuno-techniques reveal new details for understanding white-rot decay of lignocellulose. In: Comptes Rendus: Biologies. 2004 ; Vol. 327, No. 9-10. pp. 861 - 871.
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abstract = "High-resolution Cryo-Field Emission Scanning Electron Microscopy (HR-Cryo-FE-SEM) and immuno-cytochemistry were used to reveal novel details on the morphological events and spatial distribution of oxidoreductive enzymes during the degradation of birch wood by the white-rot fungi Phlebia radiata and mutant strain P. radiata Cel 26. Cryo-observations of fractured fibres showed degradation across the cell wall by P. radiata (wild) to progress by delamination and removal of concentric orientated aggregates from the secondary S2 cell wall. Decay by P. radiata Cel 26 progressed by removal of materials (lignin and hemicelluloses) between the aggregates (primarily cellulose) that remained even after advanced decay. With both decay patterns, extracellular slime materials were present uniting lumina hyphae with the attacked fibre wall. The extracellular slime material had two morphological forms: viz a fibrillar (often tripartite) and a ‘gel-form’, the former found in discrete bands progressing across the lumen onto the fibre wall. Using TEM immunocytochemistry, laccase, manganese peroxidase (MnP) and diarylpropane enzymes were localized in the periplasmic space of luminal hyphae, in association with the cell membrane, periplasmic vesicles and fungal cell wall. Extracellularly, the three enzymes were found associated with the slime and tripartite membranes and with the birch cell walls at all stages of attack through to middle lamella corner decay. Enzyme distribution was correlated with morphological changes in cell wall structure. The association of extracellular slime with these enzymes and sites of decay strongly suggests a major role for this matrix in fibre cell wall decomposition. To cite this article: G. Daniel et al., C. R. Biologies 327 (2004).",
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Cryo-FE-SEM & TEM immuno-techniques reveal new details for understanding white-rot decay of lignocellulose. / Daniel, Geoffrey (Corresponding Author); Volc, Jindrich; Niku-Paavola, Marja-Leena.

In: Comptes Rendus: Biologies, Vol. 327, No. 9-10, 2004, p. 861 - 871.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Cryo-FE-SEM & TEM immuno-techniques reveal new details for understanding white-rot decay of lignocellulose

AU - Daniel, Geoffrey

AU - Volc, Jindrich

AU - Niku-Paavola, Marja-Leena

PY - 2004

Y1 - 2004

N2 - High-resolution Cryo-Field Emission Scanning Electron Microscopy (HR-Cryo-FE-SEM) and immuno-cytochemistry were used to reveal novel details on the morphological events and spatial distribution of oxidoreductive enzymes during the degradation of birch wood by the white-rot fungi Phlebia radiata and mutant strain P. radiata Cel 26. Cryo-observations of fractured fibres showed degradation across the cell wall by P. radiata (wild) to progress by delamination and removal of concentric orientated aggregates from the secondary S2 cell wall. Decay by P. radiata Cel 26 progressed by removal of materials (lignin and hemicelluloses) between the aggregates (primarily cellulose) that remained even after advanced decay. With both decay patterns, extracellular slime materials were present uniting lumina hyphae with the attacked fibre wall. The extracellular slime material had two morphological forms: viz a fibrillar (often tripartite) and a ‘gel-form’, the former found in discrete bands progressing across the lumen onto the fibre wall. Using TEM immunocytochemistry, laccase, manganese peroxidase (MnP) and diarylpropane enzymes were localized in the periplasmic space of luminal hyphae, in association with the cell membrane, periplasmic vesicles and fungal cell wall. Extracellularly, the three enzymes were found associated with the slime and tripartite membranes and with the birch cell walls at all stages of attack through to middle lamella corner decay. Enzyme distribution was correlated with morphological changes in cell wall structure. The association of extracellular slime with these enzymes and sites of decay strongly suggests a major role for this matrix in fibre cell wall decomposition. To cite this article: G. Daniel et al., C. R. Biologies 327 (2004).

AB - High-resolution Cryo-Field Emission Scanning Electron Microscopy (HR-Cryo-FE-SEM) and immuno-cytochemistry were used to reveal novel details on the morphological events and spatial distribution of oxidoreductive enzymes during the degradation of birch wood by the white-rot fungi Phlebia radiata and mutant strain P. radiata Cel 26. Cryo-observations of fractured fibres showed degradation across the cell wall by P. radiata (wild) to progress by delamination and removal of concentric orientated aggregates from the secondary S2 cell wall. Decay by P. radiata Cel 26 progressed by removal of materials (lignin and hemicelluloses) between the aggregates (primarily cellulose) that remained even after advanced decay. With both decay patterns, extracellular slime materials were present uniting lumina hyphae with the attacked fibre wall. The extracellular slime material had two morphological forms: viz a fibrillar (often tripartite) and a ‘gel-form’, the former found in discrete bands progressing across the lumen onto the fibre wall. Using TEM immunocytochemistry, laccase, manganese peroxidase (MnP) and diarylpropane enzymes were localized in the periplasmic space of luminal hyphae, in association with the cell membrane, periplasmic vesicles and fungal cell wall. Extracellularly, the three enzymes were found associated with the slime and tripartite membranes and with the birch cell walls at all stages of attack through to middle lamella corner decay. Enzyme distribution was correlated with morphological changes in cell wall structure. The association of extracellular slime with these enzymes and sites of decay strongly suggests a major role for this matrix in fibre cell wall decomposition. To cite this article: G. Daniel et al., C. R. Biologies 327 (2004).

KW - white rot

KW - fungi

KW - white-rot fungi

KW - Phlebia radiata

KW - HR-cryo FE-SEM

KW - TEM immunocytochemistry

KW - laccase

KW - manganese peroxidase

KW - diarylpropane peroxidase

KW - extracellular slime

KW - biodegradation

KW - birch

U2 - 10.1016/j.crvi.2004.08.003

DO - 10.1016/j.crvi.2004.08.003

M3 - Article

VL - 327

SP - 861

EP - 871

JO - Comptes Rendus: Biologies

JF - Comptes Rendus: Biologies

SN - 1631-0691

IS - 9-10

ER -