Montesinos Laura, Bundó Mireia, Badosa Esther, San Segundo Blanca, Coca María, Montesinos Emilio
Institute of Food and Agricultural Technology-CIDSAV-XaRTA, University of Girona, Girona, 17071, Spain.
Centre for Research in Agricultural Genomics (CRAG), CSIC-IRTA-UAB-UB. Edifici CRAG, Campus de la UAB, 08193, Bellaterra, Barcelona, Spain.
BMC Plant Biol. 2017 Mar 14;17(1):63. doi: 10.1186/s12870-017-1011-9.
BP178 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.
A 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.
The 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.
BP178肽是一种合成的BP100-蛙皮素衍生物,对植物病原菌具有很强的抑制活性,在植物保护等领域具有巨大的应用潜力。在此,我们报道了利用水稻种子作为生物工厂生产和回收具有生物活性的BP178肽。
制备了编码BP178肽的合成基因,并将其导入水稻植株。该基因在胚乳特异性启动子的控制下在转基因水稻中高效表达。在三种胚乳特异性水稻启动子(谷蛋白B1、谷蛋白B4或球蛋白1)中,使用球蛋白1启动子时效果最佳。BP178肽在种子胚乳中积累,通过简单的程序即可从水稻种子中轻松回收,产量为21μg/g。转基因至少稳定遗传三代,并且在转基因种子的长期储存过程中肽的积累保持稳定。纯化后的肽在体外对细菌性植物病原菌Dickeya sp.(水稻深褐鞘腐病的病原菌)具有活性。转基因事件的幼苗对真菌病原菌轮枝镰孢菌表现出增强的抗性,这支持了在植物中产生的肽具有生物活性。
本研究中开发的利用水稻种子作为生物工厂可持续生产BP178肽的策略,代表了一种未来用于植物保护及可能其他领域肽生产的有前景的系统。