Differential ethylene-inducible expression of cellulase in pepper plants.
Ethylene promotes the abscission of leaves and the ripening of fruits in pepper plants, and in both events an increase in cellulase activity is observed. However, two enzyme isoforms (pI 7.2 and 8.5, respectively) are differentially involved in the two physiological phenomena. The pI 8.5 form has been purified from ripe fruits. It is a glycoprotein with an apparent molecular mass of 54 kDa. Two short peptides were sequenced and a very high homology to a tomato cellulase was observed. Polyclonal antibodies, raised against the purified enzyme, have allowed us to demonstrate that the observed ethylene-induced increase in cellulase activity is paralleled by de novo synthesis of protein. Three cDNAs (CX1, CX2 and CX3), encoding different cellulases, were obtained and characterized and their expression investigated. Accumulation of all three mRNAs is induced by ethylene treatment, though to different levels. CX1 is mainly expressed in ripe fruits while CX2 is especially found in abscission zones. CX3 accumulates at very low levels in activated abscission zones. Comparisons with other known cellulases demonstrate clear heterogeneity within the higher plant cellulases. Differences in ethylene inducibility and molecular structure suggest different physiological roles for cellulase in pepper plants.
Department of Biology, University of Padua, Padova, Italy.
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Ferrarese
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Trainotti
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Moretto
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Polverino de Laureto
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Rascio
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Casadoro
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D002212:Capsicum / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D002480:Cellulase / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D003001:Cloning, Molecular; D003429:Cross Reactions; D005030:Ethylenes / Q000494:pharmacology*; D018506:Gene Expression Regulation, Plant; D015151:Immunoblotting; D007527:Isoenzymes / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D008969:Molecular Sequence Data; D010937:Plant Growth Regulators / Q000494:pharmacology*; D018515:Plant Leaves / Q000201:enzymology; D018547:Plant Stems / Q000201:enzymology; D010946:Plants, Medicinal; D016133:Polymerase Chain Reaction; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid
D005030:Ethylenes; D007527:Isoenzymes; D010937:Plant Growth Regulators; C036216:ethylene; D002480:Cellulase