Developmental regulation of two tomato lipoxygenase promoters in transgenic tobacco and tomato.
Two lipoxygenase (LOX) genes (tomloxA and tomloxB) are expressed in ripening tomato fruit, and tomloxA is also expressed in germinating seedlings. The 5'-upstream regions of these genes were isolated to study the regulatory elements involved in coordinating tomlox gene expression. Sequence analysis of the promoters did not reveal any previously characterized regulatory elements except for TATA and CAAT boxes. However, the sequence motif GATAcAnnAAtnTGATG was found in both promoters. Chimeric gene fusions of each tomlox promoter with the beta-glucuronidase reporter gene (gus) were introduced into tobacco and tomato plants via Agrobacterium-mediated transformation. GUS activity in tomloxA-gus plants during seed germination peaked at day 5 and was enhanced by methyl jasmonate (MeJa) treatment. No GUS activity was detected in tomloxB-gus seedlings. Neither wounding nor abscisic acid (ABA) treatment of transgenic seedlings modified the activity of either promoter. During fruit development, GUS expression in tomloxA-gus tobacco fruit increased 5 days after anthesis (DAA) and peaked at 20 DAA. In tomloxB-gus tobacco fruit, GUS activity increased at 10 DAA and peaked at 20 DAA. In transgenic tomato fruit, tomloxA-gus expression was localized to the outer pericarp during fruit ripening, while tomloxB-gus expression was localized in the outer pericarp and columella. These data demonstrate that the promoter regions used in these experiments contain cis-acting regulatory elements required for proper regulation of tomlox expression during development and for MeJa-responsiveness.
Beaudoin
N
N
Department of Molecular Biology and Genetics, University of Guelph, Ontario, Canada.
Rothstein
S J
SJ
D000040:Abscisic Acid / Q000494:pharmacology; D000085:Acetates / Q000494:pharmacology; D003517:Cyclopentanes / Q000494:pharmacology; D018507:Gene Expression Regulation, Developmental; D017343:Genes, Plant; D017930:Genes, Reporter; D006651:Histocytochemistry; D008084:Lipoxygenase / Q000235:genetics*; D018551:Solanum lycopersicum / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D054883:Oxylipins; D010937:Plant Growth Regulators / Q000494:pharmacology; D018520:Plant Shoots / Q000254:growth & development; D030821:Plants, Genetically Modified; D010947:Plants, Toxic; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D017421:Sequence Analysis; D015398:Signal Transduction; D014018:Tissue Distribution; D014026:Nicotiana / Q000235:genetics
D000085:Acetates; D003517:Cyclopentanes; D054883:Oxylipins; D010937:Plant Growth Regulators; D011993:Recombinant Fusion Proteins; D000040:Abscisic Acid; C072239:methyl jasmonate; D008084:Lipoxygenase