Transgenic tobacco cell cultures expressing a Trichoderma harzianum endochitinase gene release the enzyme into the medium |
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Authors: | A Brants E D Earle |
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Institution: | (1) Cornell University, Department of Plant Breeding, Ithaca, NY 14853-1901, USA e-mail: ede3@cornell.edu Fax: +1-607-2556683, US |
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Abstract: | Calli and cell suspensions were obtained from tobacco plants transformed with an endochitinase-encoding cDNA from the biocontrol
fungus Trichoderma harzianum. Calli from four primary transformants had high levels of endochitinase activity, like the plants from which they were derived.
Endochitinase activity was also detected in the medium surrounding the calli and in the medium from transgenic cell suspensions.
Western blots demonstrated the presence of the expected 40-kDa T. harzianum protein in transgenic samples but not in controls. These results indicate that the fungal enzyme is secreted and that the
fungal signal peptide in the cDNA construct functions in plant cells. A cell suspension medium in which the protein concentration
was increased up to 34-fold by ammonium sulfate precipitation inhibited germination of Penicillium digitatum spores. Some inhibition of spore germination was also observed in concentrated medium from control suspensions, probably
due to the secretion and concentration of endogenous enzymes.
Received: 6 May 2000 / Revision received: 6 September 2000 · Accepted: 14 September 2000 |
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Keywords: | Callus Cell suspension Endochitinase Transgenic tobacco Trichoderma harzianum |
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