Production of BP178, a derivative of the synthetic antibacterial peptide BP100, in the rice seed endosperm
Saturday, 2017/03/25 | 06:33:47
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Laura Montesinos, Mireia Bundó, Esther Badosa, Blanca San Segundo, María Coca and Emilio Montesinos BMC Plant Biology 14 March 2017; DOI: 10.1186/s12870-017-1011-9 AbstractBackgroundBP178 peptide is a synthetic BP100-magainin derivative possessing strong inhibitory activity against plant pathogenic bacteria, offering a great potential for future applications in plant protection and other fields. Here we report the production and recovery of a bioactive BP178 peptide using rice seeds as biofactories. ResultsA synthetic gene encoding the BP178 peptide was prepared and introduced in rice plants. The gene was efficiently expressed in transgenic rice under the control of an endosperm-specific promoter. Among the three endosperm-specific rice promoters (Glutelin B1, Glutelin B4 or Globulin 1), best results were obtained when using the Globulin 1 promoter. The BP178 peptide accumulated in the seed endosperm and was easily recovered from rice seeds using a simple procedure with a yield of 21 μg/g. The transgene was stably inherited for at least three generations, and peptide accumulation remained stable during long term storage of transgenic seeds. The purified peptide showed in vitro activity against the bacterial plant pathogen Dickeya sp., the causal agent of the dark brown sheath rot of rice. Seedlings of transgenic events showed enhanced resistance to the fungal pathogen Fusarium verticillioides, supporting that the in planta produced peptide was biologically active. ConclusionsThe strategy developed in this work for the sustainable production of BP178 peptide using rice seeds as biofactories represents a promising system for future production of peptides for plant protection and possibly in other fields.
See https://bmcplantbiol.biomedcentral.com/articles/10.1186/s12870-017-1011-9
Fig. 1 Schematic representation of the plant expression vectors for the expression of the BP178 gene in the rice endosperm. a Three different constructs in pCAMBIA vector (pC) in which the synthetic gene was cloned between the endosperm-specific promoter including the signal peptide coding sequence of the corresponding seed storage protein (SP) and the nopaline synthase terminator (nos). Relevant restriction enzyme sites for cloning purposes are indicated. The hptII gene encoding resistance to hygromycin under the control of the CaMV35S promoter and terminator was contained into the T-DNA region of pC. LB, left border; RB, right border of T-DNA. Arrows indicate the orientation of the sequence. b The chimeric BP178 peptide corresponding to the fusion of BP134 (KKLFKKILKYL-OH, a cecropin A (1–7)-melittin (2–9) hybrid, in green) linked through the hinge sequence (AGPA, in orange) to a magainin fragment peptide (GIGKFLHSAKKFGKAFVGEIMNS –OH, in blue); and extended with ER retention signal (KDEL, in pink). For details, see Additional file 1: Figure S1 |
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