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Production of cecropin A antimicrobial peptide in rice seed endosperm

Mireia B., Montesinos L., Izquierdo E., Campo S., Mieulet D., Guiderdoni E., Rossignol M., Badosa E., Montesinos E., San Segundo B., Coca M.. 2014. BMC Plant Biology, 14 (102) : 29 p..

DOI: 10.1186/1471-2229-14-102

Background Cecropin A is a natural antimicrobial peptide that exhibits rapid, potent and long-lasting lytic activity against a broad spectrum of pathogens, thus having great biotechnological potential. Here, we report a system for producing bioactive cecropin A in rice seeds. Results Transgenic rice plants expressing a codon-optimized synthetic cecropin A gene drived by an endosperm-specific promoter, either the glutelin B1 or glutelin B4 promoter, were generated. The signal peptide sequence from either the glutelin B1 or the glutelin B4 were N-terminally fused to the coding sequence of the cecropin A. We also studied whether the presence of the KDEL endoplasmic reticulum retention signal at the C-terminal has an effect on cecropin A subcellular localization and accumulation. The transgenic rice plants showed stable transgene integration and inheritance. We show that cecropin A accumulates in protein storage bodies in the rice endosperm, particularly in type II protein bodies, supporting that the glutelin Nterminal signal peptides play a crucial role in directing the cecropin A to this organelle, independently of being tagged with the KDEL endoplasmic reticulum retention signal. The production of cecropin A in transgenic rice seeds did not affect seed viability or seedling growth. Furthermore, transgenic cecropin A seeds exhibited resistance to infection by fungal and bacterial pathogens (Fusarium verticillioides and Dickeya dadantii, respectively) indicating that the in planta-produced cecropin A is biologically active. Conclusions Rice seeds can sustain bioactive cecropin A production and accumulation in protein bodies. The system might benefit the production of this antimicrobial agent for subsequent applications in crop protection and food preservation.

Mots-clés : oryza sativa; grain; endosperme; bactéricide; biosynthèse; peptide; propriété antimicrobienne; résistance génétique; résistance aux organismes nuisibles; enterobacteriaceae; fusarium; plante transgénique; amélioration des plantes; physiologie végétale; protection des plantes; préservation des aliments; dickeya dadantii; fusarium verticilliodes

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