ABSTRACT
The cloning of promoter sequences of two invertase genes from potato (Solanum tuberosum L.) is described. Histochemical analysis of series of reporter transgenic lines reveals phloem-specific expression from both promoters, with one expressed preferentially in internal phloem and the other in external phloem of stem vascular bundles.
Subject(s)
Gene Expression Regulation, Plant , Glycoside Hydrolases/genetics , Solanum tuberosum/genetics , Artificial Gene Fusion , Cloning, Molecular , Genes, Reporter , Glycoside Hydrolases/metabolism , Histocytochemistry , Plant Structures/anatomy & histology , Plant Structures/enzymology , Plant Structures/genetics , Plants, Genetically Modified , Promoter Regions, Genetic , Solanum tuberosum/anatomy & histology , Solanum tuberosum/enzymology , beta-FructofuranosidaseABSTRACT
The organisation of two invertase genes (invGE and invGF) linked in direct tandem repeat within the potato genome is detailed. The genes exhibit a similar intron/exon structure which differs from previously described plant invertase genes; while intron locations are conserved between the genes, minor differences in exon length are seen. Both genes encode enzymes with putative extracellular location. Biochemical analysis of gene expression showed expression in floral tissues for both genes, with expression of the upstream gene (invGE) also detected in leaf tissue. Promoter sequences from both genes have been fused to the beta-glucuronidase (GUS) reporter gene (uidA) and transformed into potato. One promoter-GUS reporter construct was also transformed into tobacco. Histochemical analysis of transgenic lines defined specific expression from the downstream (invGF) promoter in potato and tobacco pollen, with expression first detected in the late uninucleate stage of tobacco microspore development. The invGE promoter determined expression in pollen and other floral tissues, but also at lateral nodes in stem, root and tuber. An association of invertase expression with generative tissue, both in vegetative and sexual modes of growth, is indicated.