A proteinase inhibitor from Nicotiana alata inhibits the normal development of light-brown apple moth, Epiphyas postvittana in transgenic apple plants |
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Authors: | Gowri Maheswaran Lucinda Pridmore Peter Franz Marilyn A Anderson |
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Institution: | (1) Primary Industries Research Victoria, Victorian AgriBiosciences Centre, 1 Park Drive, Bundoora, Victoria, 3083, Australia;(2) School of Biochemistry, La Trobe University, Bundoora, Victoria, 3086, Australia |
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Abstract: | Insecticidal proteins are a potential resource to enhance resistance to insect pests in transgenic plants. Here, we describe
the generation and analysis of the apple cultivar ‘Royal Gala’ transgenic for Nicotiana alata (N. alata) proteinase inhibitor (PI) and the impact of this PI on the growth and development of the Epiphyas postvittiana (light-brown apple moth). A cDNA clone encoding a proteinase inhibitor precursor from N. alata (Na-PI) under the control of either a double 35S promoter or a promoter from a ribulose-1,5-bisphosphate carboxylase small
sub-unit gene (rbcS-E9 promoter) was stably incorporated into ‘Royal Gala’ apple using Agrobacterium-mediated transformation. A 40.3 kDa Na-PI precursor protein was expressed and correctly processed into 6-kDa proteinase inhibitors
in the leaves of transgenic apple lines. The 6-kDa polypeptides accumulated to levels of 0.05 and 0.1% of the total soluble
protein under the control of the rbc-E9 promoter and the double 35S promoter, respectively. Light-brown apple moth larvae fed with apple leaves expressing Na-PI
had significantly reduced body weight after 7 days of feeding and female pupae were 19–28% smaller than controls. In addition,
morphological changes such as pupal cases attached to the wing, deformed wings, deformed body shape, and pupal cases and curled
wings attached to a deformed body were observed in adults that developed from larvae fed with apple leaves expressing Na-PI,
when compared to larvae fed with the non-transformed apple leaves. |
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Keywords: | Apple Epiphyas postvittiana Insect pest resistance Nicotiana alata Proteinase inhibitor Transgenic Malus × domestica Borkh |
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