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Stacked insecticidal genes in potatoes exhibit enhanced toxicity against Colorado potato beetle, Leptinotarsa decemlineata (Coleoptera: Chrysomelidae)

Abstract

The present study was performed to express stacked insecticidal genes in potato cv. Lady Olympia and Agria to encode resistance against Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say). Bacillus thuringiensis (Bt) gene (cry3A), synthetic hybrid (SN-19) and plant proteinase inhibitor Oryza cystatin II (OCII) cloned in pCAMBIA1301 binary vector in two different combinations as of DS-1 (cry3A + SN-19 genes) and DS-2 (OCII + SN-19 genes) constructs and further transformed to two potato cultivars using Agrobacterium-mediated transformation. All molecular analyses confirmed gene integration and expression in a total of 27 primary transformants in both Agria and Lady Olympia. Insecticidal effects of T0 progeny transgenic potato plants were tested against CPB under laboratory conditions. Transgenic plants of Agria and Lady Olympia transformed with DS-1 and DS-2 constructs caused 100% mortality to all larval stages and adults of CPB. However, 100% mortality of tested insects took a longer time in the adult stage (10–14 days) compared to larval stages (2–6 days). Foliage consumption by L2-L4 larval stages and adults of CPB was significantly reduced in Agria and Lady Olympia plants transformed with DS-1 and DS-2 constructs, as compared to their control plants. Lower foliage consumption of transgenic plants by L1 larval stages was also observed, but the reduction was only statistically significant for some of the tested plants. These promising results indicate that the transgenic potato plants exhibit a high potential in controlling CPB population and are a useful tool in the management of imidacloprid-resistant CPB.

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Acknowledgements

The present study was supported by Nigde Omer Halisdemir Scientific Research Unit (BAP Project No. FEB 2017/18 BAGEP). The authors are highly thankful to TÜBITAK for providing PhD fellowship (code 2215) to Dr. Muhammad Salim to complete his doctoral studies. Authors are grateful to Prof. Dr. Mehmet Emin Çalışkan, Nigde Omer Halisdemir University and Mr. Cengiz Kurt, Trakya Agriculture Research Institute for providing seed tuber of potato and rice cultivar, respectively. Special thanks to Dr. Emre Aksoy for his technical help during experiments. We are also thankful to Prof. Dr. Sebahattin Özcan, University of Ankara and Prof. Dr. Josef Vlasak, Academy of Sciences of Czech Republic for providing recombinant plasmids of pTF101 and pET-22b( +), respectively.

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The research concept was conceived by AB. AG secured funding of the project as principal investigator. MS constructed recombinant plasmids, performed genetic transformation and recorded all bioassays data. AB supervised molecular part of the studies whereas AG supervised the overall research activities of the project. MS, AB and AG wrote the manuscript and presented it in its current form.

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Correspondence to Allah Bakhsh.

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Salim, M., Bakhsh, A. & Gökçe, A. Stacked insecticidal genes in potatoes exhibit enhanced toxicity against Colorado potato beetle, Leptinotarsa decemlineata (Coleoptera: Chrysomelidae). Plant Biotechnol Rep 15, 197–215 (2021). https://doi.org/10.1007/s11816-021-00668-3

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  • DOI: https://doi.org/10.1007/s11816-021-00668-3

Keywords

  • Gene stacking
  • Potato cultivars
  • Insect resistance
  • Colorado potato beetle