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PLANT PHYSIOLOGY , Vol 106, Issue 1 53-60, Copyright © 1994 by American Society of Plant Biologists
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METABOLISM AND ENZYMOLOGY |
Apoplastic Peroxidases and Lignification in Needles of Norway Spruce (Picea abies L.)
A. Polle, T. Otter and F. Seifert
Institut fur Forstbotanik und Baumphysiologie, Professur fur Baumphysiologie, Albert-Ludwigs-Universitat Freiburg, Am Flughafen 17, D-79085 Freiburg, Germany (A.P., T.O.)
The objective of the present study was to investigate the correlation of
soluble apoplastic peroxidase activity with lignification in needles of
field-grown Norway spruce (Picea abies L.) trees. Apoplastic peroxidases
(EC 1.11.1.7) were obtained by vacuum infiltration of needles. The lignin
content of isolated cell walls was determined by the acetyl bromide method.
Accumulation of lignin and seasonal variations of apoplastic peroxidase
activities were studied in the first year of needle development. The major
phase of lignification started after bud break and was terminated about 4
weeks later. This phase correlated with a transient increase in apoplastic
guaiacol and coniferyl alcohol peroxidase activity. NADH oxidase activity,
which is thought to sustain peroxidase activity by production of H2O2,
peaked sharply after bud break and decreased during the lignification
period. Histochemical localization of peroxidase with guaiacol indicated
that high activities were present in lignifying cell walls. In mature
needles, lignin was localized in walls of most needle tissues including
mesophyll cells, and corresponded to 80 to 130 [mu]mol lignin monomers/g
needle dry weight. Isoelectric focusing of apoplastic washing fluids and
activity staining with guaiacol showed the presence of strongly alkaline
peroxidases (isoelectric point [greater than or equal to] 9) in all
developmental stages investigated. New isozymes with isoelectric points of
7.1 and 8.1 appeared during the major phase of lignification. These
isozymes disappeared after lignification was terminated. A strong increase
in peroxidase activity in autumn was associated with the appearance of
acidic peroxidases (isoelectric point [less than or equal to] 3). These
results suggest that soluble alkaline apoplastic peroxidases participate in
lignin formation. Soluble acidic apoplastic peroxidases were apparently
unrelated to developmentally regulated lignification in spruce needles.
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