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THE PLANT CELL, Vol 2, Issue 2 95-106, Copyright © 1990 by American Society of Plant Biologists
Analysis of Stress-Induced or Salicylic Acid-Induced Expression of the Pathogenesis-Related 1a Protein Gene in Transgenic Tobacco
M. Ohshima, H. Itoh, M. Matsuoka, T. Murakami and Y. Ohashi
National Institute of Agrobiological Resources, Tsukuba Science City, Tsukuba, Ibaraki 305, Japan
The cis-acting elements for regulating gene expression of the tobacco
pathogenesis-related 1a protein gene were analyzed in transgenic plants.
The 5[prime]-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[prime]-deleted
chimeric genes was constructed and transformed into tobacco plants.
Transgenic plants with a 0.3-kilobase fragment of the 5[prime]-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.
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