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Plant Physiol, August 2000, Vol. 123, pp. 1583-1592
Detection of Expansin Proteins and Activity during Tomato Fruit
Ontogeny1
Jocelyn K.C.
Rose,
Daniel J.
Cosgrove,
Peter
Albersheim,
Alan G.
Darvill, and
Alan B.
Bennett*
Complex Carbohydrate Research Center and Department of Biochemistry
and Molecular Biology, University of Georgia, 220 Riverbend Road,
Athens, Georgia 30602-4712 (J.K.C.R., P.A., A.G.D.); Department of
Biology, 208 Mueller Laboratory, The Pennsylvania State University,
University Park, Pennsylvania 16802 (D.J.C.); and Mann Laboratory,
Department of Vegetable Crops, University of California, Davis,
California 95616 (A.B.B.)
Expansins are plant proteins that have the capacity to induce
extension in isolated cell walls and are thought to mediate pH-dependent cell expansion. J.K.C. Rose, H.H. Lee, and A.B. Bennett ([1997] Proc Natl Acad Sci USA 94: 5955-5960) reported the
identification of an expansin gene (LeExp1) that is
specifically expressed in ripening tomato (Lycopersicon
esculentum) fruit where cell wall disassembly, but not cell
expansion, is prominent. Expansin expression during fruit ontogeny was
examined using antibodies raised to recombinant LeExp1 or a cell
elongation-related expansin from cucumber (CsExp1). The LeExp1
antiserum detected expansins in extracts from ripe, but not preripe
tomato fruit, in agreement with the pattern of LeExp1
mRNA accumulation. In contrast, antibodies to CsExp1 cross-reacted with
expansins in early fruit development and the onset of ripening, but not
at a later ripening stage. These data suggest that ripening-related and
expansion-related expansin proteins have distinct antigenic epitopes
despite overall high sequence identity. Expansin proteins were detected
in a range of fruit species and showed considerable variation in
abundance; however, appreciable levels of expansin were not present in
fruit of the rin or Nr tomato mutants
that exhibit delayed and reduced softening. LeExp1 protein accumulation
was ethylene-regulated and matched the previously described expression
of mRNA, suggesting that expression is not regulated at the level of
translation. We report the first detection of expansin activity in
several stages of fruit development and while characteristic creep
activity was detected in young and developing tomato fruit and in ripe pear, avocado, and pepper, creep activity in ripe tomato showed qualitative differences, suggesting both hydrolytic and expansin activities.
1
This work was supported by the U.S. Department
of Energy (grant nos. DE-FG02-96ER20220 to J.K.C.R., P.A., and
A.G.D. and DE-FG02-84ER13179 to D.J.C.).
*
Corresponding author; e-mail abbennett{at}ucdavis.edu; fax
530-752-4554.
© 2000 American Society of Plant Physiologists
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