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Transgenic sugarcane overexpressing CaneCPI-1 negatively affects the growth and development of the sugarcane weevil Sphenophorus levis

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Transgenic sugarcane expressing CaneCPI-1 exhibits resistance to Sphenophorus levis larvae.

Abstract

Transgenic plants have widely been used to improve resistance against insect attack. Sugarcane is an economically important crop; however, great losses are caused by insect attack. Sphenophorus levis is a sugarcane weevil that digs tunnels in the stem base, leading to the destruction of the crop. This insect is controlled inefficiently by chemical insecticides. Transgenic plants expressing peptidase inhibitors represent an important strategy for impairing insect growth and development. Knowledge of the major peptidase group present in the insect gut is critical when choosing the most effective inhibitor. S. levis larvae use cysteine peptidases as their major digestive enzymes, primarily cathepsin L-like activity. In this study, we developed transgenic sugarcane plants that overexpress sugarcane cysteine peptidase inhibitor 1 (CaneCPI-1) and assessed their potential through feeding bioassays with S. levis larvae. Cystatin overexpression in the transgenic plants was evaluated using semi-quantitative RT-PCR, RT-qPCR, and immunoblot assays. A 50% reduction of the average weight was observed in larvae that fed on transgenic plants in comparison to larvae that fed on non-transgenic plants. In addition, transgenic sugarcane exhibited less damage caused by larval attack than the controls. Our results suggest that the overexpression of CaneCPI-1 in sugarcane is a promising strategy for improving resistance against this insect.

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Acknowledgements

The research was supported by the São Paulo Research Foundation (FAPESP, CBME, CEPID Proc. 98/14138-2). The authors thank Centro de Tecnologia Canavieira, Piracicaba, São Paulo, Brazil, for essential help in sugarcane transformation. FHS is recipient of a productivity scholarship from National Council for Scientific and Technological Development (CNPq) (##311745/2013-0). V.K.S. and A.S.C.F. received a Grant from FAPESP (2013/05370-0 and 2005/59833-5, respectively), and C.W.R. received a Grant from National Council for Scientific and Technological Development (CNPq).

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    Authors and Affiliations

    1. Laboratory of Molecular Biology, Department of Genetics and Evolution, Federal University of São Carlos, Rodovia Washigton Luis, Km 235, São Carlos, SP, 13565-905, Brazil

      Vanessa Karine Schneider, Andrea Soares-Costa, Mohan Chakravarthi, Carolina Ribeiro & Flavio Henrique-Silva

    2. Centro de Tecnologia Canavieira, Fazenda Santo Antônio, s/n, cp. 162, Piracicaba, SP, CEP 13400-970, Brazil

      Sabrina Moutinho Chabregas & Maria Cristina Falco

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    1. Vanessa Karine Schneider
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    7. Flavio Henrique-Silva
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    Correspondence to Flavio Henrique-Silva.

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    Communicated by P. Lakshmanan.

    V. K. Schneider and A. Soares-Costa contributed equally to this work.

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    Schneider, V.K., Soares-Costa, A., Chakravarthi, M. et al. Transgenic sugarcane overexpressing CaneCPI-1 negatively affects the growth and development of the sugarcane weevil Sphenophorus levis . Plant Cell Rep 36, 193–201 (2017). https://doi.org/10.1007/s00299-016-2071-2

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