Souza Prado G. auteur aut IRD Bamogo P. K. A. auteur aut IRD Cordeiro de Abreu J. A. auteur aut IRD Gillet F. X. auteur aut IRD Olinto dos Santos V. auteur aut IRD Mattar Silva M. C. auteur aut IRD Brizard Jean-Paul auteur aut IRD Porto Bemquerer M. auteur aut IRD Bangratz Martine auteur aut IRD Brugidou Christophe auteur aut IRD Sérémé D. auteur aut IRD Grossi-de-Sa M. F. auteur aut IRD Lacombe Séverine auteur aut IRD text journalArticle eng text/pdf reformatted digital access BackgroundInsect resistance in crops represents a main challenge for agriculture. Transgenic approaches based on proteins displaying insect resistance properties are widely used as efficient breeding strategies. To extend the spectrum of targeted pathogens and overtake the development of resistance, molecular evolution strategies have been used on genes encoding these proteins to generate thousands of variants with new or improved functions. The cotton boll weevil (Anthonomus grandis) is one of the major pests of cotton in the Americas. An -amylase inhibitor (-AIC3) variant previously developed via molecular evolution strategy showed inhibitory activity against A. grandis -amylase (AGA).ResultsWe produced in a few days considerable amounts of -AIC3 using an optimised transient heterologous expression system in Nicotiana benthamiana. This high -AIC3 accumulation allowed its structural and functional characterizations. We demonstrated via MALDI-TOF MS/MS technique that the protein was processed as expected. It could inhibit up to 100% of AGA biological activity whereas it did not act on -amylase of two non-pathogenic insects. These data confirmed that N. benthamiana is a suitable and simple system for high-level production of biologically active -AIC3. Based on other benefits such as economic, health and environmental that need to be considerate, our data suggested that -AIC3 could be a very promising candidate for the production of transgenic crops resistant to cotton boll weevil without lethal effect on at least two non-pathogenic insects.ConclusionsWe propose this expression system can be complementary to molecular evolution strategies to identify the most promising variants before starting long-lasting stable transgenic programs. specialized Transient protein expression alpha-amylase inhibitors Gene silencing suppressors Cotton boll weevil 076 020 084 19 art. 15 [13 p.] 2019 1472-6750 https://www.documentation.ird.fr/hor/fdi:010075306 10.1186/s12896-019-0507-9 1472-6750 [F B010075306] https://www.documentation.ird.fr/hor/fdi:010075306 https://horizon.documentation.ird.fr/exl-doc/pleins_textes/divers19-03/010075306.pdf IRD - Base Horizon / Pleins textes 2019-04-03 2019-04-04 fdi:010075306 fre