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Jasmonate-inducible expression of a potato cathepsin D inhibitor-GUS gene fusion in tobacco cells

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Abstract

A potato gene encoding cathepsin D inhibitor (CDI) is expressed constitutively in tubers and flower buds and it is inducible in leaves upon wounding of the tissue or by treatment with methyl jasmonate (MJA). A fusion gene (CDI:GUS) in which the 2.4 kb long promoter of the CDI gene was translationaly fused with the coding sequence for β-glucuronidase (GUS) showed MJA-inducible expression in transformed tobacco cells in suspension. The maximum level of induction by MJA was obtained in the absence of auxin and repression of MJA-inducible expression of the fusion gene by auxin was released by aphidicolin, the results suggesting that MJA-inducible expression is repressed during active cell division. JA and MJA showed similar activities in inducing the expression of the fusion gene, while other JA-related compounds such as cucurbic acid, tuberonic acid and dihydrojasmonic acid neither induced expression of the fusion gene nor inhibited the MJA-inducible expression of the fusion gene. Methyl dihydrojasmonate specifically stimulated the MJA-inducible expression of the fusion gene. The MJA-inducible expression of the fusion gene was observed even with a 100 bp long promoter of the CDI gene albeit with significantly decreased level of expression compared to the 2.4 kb long promoter. The 100 bp long CDI promoter did not contain a G-box or hexamer motif that had been implicated in the MJA-responsive expression of several other plant genes. Further mutagenesis of the 100 bp long promoter by deletion or oligonucleotide insertion suggested that although a sequence between −100 and −82 is required for the MJA-responsive expression, the presence of this sequence alone does not confer the MJA-responsive expression.

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Abbreviations

BA:

N6-benzyladenine

CA:

cucurbic acid

CaMV:

cauliflower mosaic virus

CDI:

cathepsin D inhibitor

DMSO:

dimethyl sulfoxide

GUS:

β-glucuronidase

HJA:

dihydrojasmonic acid

JA:

Jasmonic acid

MCA:

methyl cucurbate

MJA:

methyl jasmonate

MHJA:

methyl dihydrojasmonate

MTA:

methyl tuberonate

PI-II:

proteinase inhibitor II

TA:

tuberonic acid

2,4-D:

2,4-dichlorophenoxyacetic acid.

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  1. Atsushi Ishikawa

    Present address: Faculty of Biotechnology, Fuhui Prefectural University, 4-1-1 Kenjojuna, Matsuoka-cho, 910-11, Yoshida-gun, Fuhui, Japan

Authors and Affiliations

  1. Laboratory of Biochemistry, School of Agriculture, Nagoya University, Chikusa-ku, 464-01, Nagoya, Japan

    Atsushi Ishikawa & Kenzo Nakamura

  2. Department of Bioscience and Chemistry, Faculty of Agriculture, Hokkaido University, 060, Sapporo, Japan

    Teruhiko Yoshihara

Authors
  1. Atsushi Ishikawa
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  2. Teruhiko Yoshihara
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  3. Kenzo Nakamura
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Ishikawa, A., Yoshihara, T. & Nakamura, K. Jasmonate-inducible expression of a potato cathepsin D inhibitor-GUS gene fusion in tobacco cells. Plant Mol Biol 26, 403–414 (1994). https://doi.org/10.1007/BF00039549

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  • DOI: https://doi.org/10.1007/BF00039549

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