Characterization of ppEG1, a member of a multigene family which encodes endo-beta-1,4-glucanase in peach.
Three cDNA clones (pCel 10, pCel 20 and pCel 30), each encoding different endo-beta-1,4-glucanases in peach, were obtained by RT-PCR and their expression investigated by northern analysis during leaf and fruit abscission and during fruit development. This analysis allowed the detection of only the pCel 10-related mRNA. A 2.2 kb transcript accumulated in ethylene activated abscission zones of leaves and fruits, and ppEG1 (Prunus persica endoglucanase 1) the gene coding for pCel 10, was isolated and characterized. A cDNA (termed pCel 1), containing the entire open reading frame of ppEGC1, was obtained and its sequence used to define the structure of the gene and the exon/intron boundaries. ppEG1 consists of 7 exons and encodes a 497 amino acid polypeptide including a putative signal peptide at the N-terminus. The similarity of this peach endo-beta-1,4-glucanase (EGase, EC 3.2.1.4) is high (76.3%) with the ripening avocado and low (47.3%) with the bean abscission EGase. A 1639 bp region at the 5' of the transcription start site shows regulatory functions in transgenic tobacco plants, as judged by its ability to drive GUS expression in cell separation-related events.
Trainotti
L
L
Department of Biology, University of Padua, Italy.
Spolaore
S
S
Ferrarese
L
L
Casadoro
G
G
D000595:Amino Acid Sequence; D001483:Base Sequence; D002480:Cellulase / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics*; D018076:DNA, Complementary / Q000096:biosynthesis / Q000302:isolation & purification; D005638:Fruit; D017343:Genes, Plant; D017930:Genes, Reporter; D005966:Glucuronidase / Q000235:genetics; D008969:Molecular Sequence Data; D005810:Multigene Family; D016366:Open Reading Frames; D018515:Plant Leaves; D030821:Plants, Genetically Modified; D010947:Plants, Toxic; D011401:Promoter Regions, Genetic; D014026:Nicotiana / Q000235:genetics
D018076:DNA, Complementary; D005966:Glucuronidase; D002480:Cellulase