Regulation of expression of a wound-inducible tomato inhibitor I gene in transgenic nightshade plants

Plant Molecular Biology
R JohnsonC A Ryan

Abstract

A wound-inducible proteinase Inhibitor I gene from tomato containing 725 bp of the 5' region and 2.5 kbp of the 3' region was stably incorporated into the genome of black nightshade plants (Solanum nigrum) using an Agrobacterium Ti plasmid-derived vector. Transgenic nightshade plants were selected that expressed the tomato Inhibitor I protein in leaf tissue. The leaves of the plants contained constitutive levels of the inhibitor protein of up to 60 micrograms/g tissue. These levels increased by a factor of about two in response to severe wounding. Only leaves and petioles exhibited the presence of the inhibitor, indicating that the gene exhibited the same tissue specificity of expression found in situ in wounded tomato leaves. Inhibitor I was extracted from leaves of wounded transformed nightshade plants and was partially purified by affinity chromatography on a chymotrypsin-Sepharose column. The affinity-purified protein was identical to the native tomato Inhibitor I in its immunological reactivity and in its inhibitory activity against chymotrypsin. The protein exhibited the same Mr of 8 kDa as the native tomato Inhibitor I and its N-terminal amino acid sequence was identical to that of the native tomato inhibitor I, indicati...Continue Reading

References

Apr 1, 1990·Plant Molecular Biology·R Johnson, C A Ryan
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Oct 1, 1971·Immunochemistry·R TrautmanG G Wagner
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Mar 1, 1988·Plant Molecular Biology·J C Rogers

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Citations

Sep 10, 2014·PloS One·Shu-Ye JiangSrinivasan Ramachandran

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