Analysis of stress-induced or salicylic acid-induced expression of the pathogenesis-related 1a protein gene in transgenic tobacco.
The cis-acting elements for regulating gene expression of the tobacco pathogenesis-related 1a protein gene were analyzed in transgenic plants. The 5'-flanking 2.4-kilobase fragment from the pathogenesis-related 1a protein gene was joined to the bacterial beta-glucuronidase gene and introduced into tobacco cells by Agrobacterium-mediated gene transfer. Promoter activity was monitored by quantitative and histochemical assay of beta-glucuronidase activity in leaves of regenerated transgenic plants. The level of beta-glucuronidase activity was clearly increased by treatment with salicylic acid, by cutting stress, and by local lesion formation caused by tobacco mosaic virus infection. Cytochemical studies of the induced beta-glucuronidase activity revealed tissue-specific and developmentally regulated expression of the pathogenesis-related 1a gene after stress or chemical treatment and after pathogen attack. To identify the cis-acting element more precisely, a series of 5'-deleted chimeric genes was constructed and transformed into tobacco plants. Transgenic plants with a 0.3-kilobase fragment of the 5'-flanking region of the pathogenesis-related 1a gene had the same qualitative response as those with the 2.4-kilobase fragment upon treatment with salicylic acid or infection with TMV. Thus, the 0.3-kilobase DNA sequence fragment was sufficient to allow the regulated expression of the pathogenesis-related 1a gene.
Ohshima
M
M
National Institute of Agrobiological Resources, Tsukuba Science City, Ibaraki, Japan.
Itoh
H
H
Matsuoka
M
M
Murakami
T
T
Ohashi
Y
Y
D001483:Base Sequence; D002872:Chromosome Deletion; D004790:Enzyme Induction / Q000187:drug effects; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D005966:Glucuronidase / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D006651:Histocytochemistry; D007613:Kanamycin Resistance / Q000235:genetics; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000096:biosynthesis / Q000187:drug effects / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D010947:Plants, Toxic; D012045:Regulatory Sequences, Nucleic Acid; D012459:Salicylates / Q000494:pharmacology*; D020156:Salicylic Acid; D014026:Nicotiana / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development
D010940:Plant Proteins; D012459:Salicylates; C053376:pathogenesis-related proteins, plant; D005966:Glucuronidase; D020156:Salicylic Acid