Isolation and characterization of a gene encoding endo-beta-1,4-glucanase from pepper (Capsicum annuum L.).

Abstract

The endo-beta-1,4-glucanases, or cellulases, of higher plants are cell wall-associated enzymes believed to function in cell wall changes associated with the diverse processes of fruit ripening, organ abscission and cell elongation. We have isolated and characterized cDNA and genomic clones encoding a cellulase, PCEL1, which is abundant in ripening pepper fruit. Genomic analysis indicates that PCEL1 is encoded by a single gene, PCEL1, which belongs to a small, structurally divergent gene family. In ripening fruit, PCEL1 transcription is initiated at two distinct sites which yields overlapping mRNA species of 1.7 and 2.1 kb. High-level accumulation of both transcripts occurs in red fruit, while the 1.7 kb transcript is detected at a much lower level in stem and petiolar tissue. The increase in cellulase activity which is measured during fruit ripening is the product of PCEL1 expression and is tightly coupled to fruit reddening. High-level applications of ethylene serve to enhance the rate of ripening and the accumulation of PCEL1 mRNA. A direct role for ethylene in regulating PCEL1 expression is shown by the exclusive induction, in immature green fruit, of the 1.7 kb transcript in response to prolonged high-level exposure to ethylene--a pattern of expression not observed in fruit development on the vine.