title: Production of cecropin A antimicrobial peptide in rice seed endosperm.
abstract: 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 N-terminal 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.
authors.forename: Mireia
authors.initials: M
authors.lastname: Bundó
authors.forename: Laura
authors.initials: L
authors.lastname: Montesinos
authors.forename: Esther
authors.initials: E
authors.lastname: Izquierdo
authors.forename: Sonia
authors.initials: S
authors.lastname: Campo
authors.forename: Delphine
authors.initials: D
authors.lastname: Mieulet
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Guiderdoni
authors.forename: Michel
authors.initials: M
authors.lastname: Rossignol
authors.forename: Esther
authors.initials: E
authors.lastname: Badosa
authors.forename: Emilio
authors.initials: E
authors.lastname: Montesinos
authors.forename: Blanca
authors.initials: B
authors.lastname: San Segundo
authors.affiliation: Centre for Research in Agricultural Genomics (CRAG), CSIC-IRTA-UAB-UB, Edifici CRAG, Campus de la UAB, 08193 Bellaterra, Barcelona, Spain. maria.coca@cragenomica.es.
authors.forename: María
authors.initials: M
authors.lastname: Coca
mesh_terms: D000595:Amino Acid Sequence; D023181:Antimicrobial Cationic Peptides / Q000096:biosynthesis* / Q000737:chemistry; D060467:Disease Resistance / Q000276:immunology; D056625:Endosperm / Q000378:metabolism*; D005670:Fusarium / Q000502:physiology; D018628:Gene Dosage; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009928:Organ Specificity / Q000235:genetics; D015388:Organelles / Q000378:metabolism; D012275:Oryza / Q000378:metabolism*; D010935:Plant Diseases / Q000276:immunology / Q000382:microbiology; D030821:Plants, Genetically Modified; D015203:Reproducibility of Results; D013347:Subcellular Fractions / Q000378:metabolism; D019076:Transgenes / Q000235:genetics
chemical_list: D023181:Antimicrobial Cationic Peptides; C031161:cecropin A
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