Khademi Mitra, Varasteh-Shams Marzieh, Nazarian-Firouzabadi Farhad, Ismaili Ahmad
Department of Agronomy and Plant Breeding, Faculty of Agriculture, Lorestan University, Khorramabad, Iran.
Front Plant Sci. 2020 Aug 13;11:1236. doi: 10.3389/fpls.2020.01236. eCollection 2020.
Antimicrobial peptides have been long known to confer resistance to plant pathogens. In this study, new recombinant peptides constructed from a dermaseptin B1 (DrsB1) peptide fused to a chitin-binding domain (CBD) from Avr4 protein, were used for -mediated transformation of tobacco plants. Polymerase chain reaction (PCR), semi-quantitative RT-PCR, and western blotting analysis demonstrated the incorporation and expression of transgenes in tobacco genome and transgenic plants, respectively. experiments with recombinant peptides extracted from transgenic plants demonstrated a significant (P<0.01) inhibitory effect on the growth and development of plant pathogens. The DrsB1-CBD recombinant peptide had the highest antifungal activity against fungal pathogens. The expression of the recombinant peptides greatly protected transgenic plants from , and fungi, in comparison to sp. and . Expression of new recombinant peptides resulted in a delay in the colonization of fungi and appearance of fungal disease symptoms from 6 days to more than 7 weeks. Scanning electron microscopy images revealed that the structure of the fungal mycelia appeared segmented, cling together, and crushed following the antimicrobial activity of the recombinant peptides. Greenhouse bioassay analysis showed that transgenic plants were more resistant to and infections as compared with the control plants. Due to the high antimicrobial activity of the recombinant peptides against plant pathogens and novelty of recombinant peptides, this report shows the feasibility of this approach to generate disease resistance transgenic plants.
长期以来,人们都知道抗菌肽能赋予植物对病原体的抗性。在本研究中,由与来自Avr4蛋白的几丁质结合结构域(CBD)融合的皮肤抗菌肽B1(DrsB1)构建的新型重组肽被用于介导烟草植物的转化。聚合酶链反应(PCR)、半定量逆转录PCR和蛋白质印迹分析分别证明了转基因在烟草基因组和转基因植物中的整合和表达。对从转基因植物中提取的重组肽进行的实验表明,其对植物病原体的生长和发育具有显著(P<0.01)抑制作用。DrsB1-CBD重组肽对真菌病原体具有最高的抗真菌活性。与[具体菌种1]和[具体菌种2]相比,重组肽的表达极大地保护了转基因植物免受[具体菌种3]、[具体菌种4]和[具体菌种5]真菌的侵害。新重组肽的表达导致真菌定殖延迟,真菌病害症状出现的时间从6天推迟到7周以上。扫描电子显微镜图像显示,重组肽的抗菌活性作用后,真菌菌丝体的结构出现分段、粘连在一起并被破坏的现象。温室生物测定分析表明,与对照植物相比,转基因植物对[具体病害1]和[具体病害2]感染的抗性更强。由于重组肽对植物病原体具有高抗菌活性且重组肽具有新颖性,本报告显示了这种方法用于培育抗病转基因植物的可行性。