Flores Oriana, Retamales Julio, Núñez Mauricio, León Marcela, Salinas Paula, Besoain Ximena, Yañez Carolina, Bastías Roberto
Laboratory of Microbiology, Institute of Biology, Pontificia Universidad Católica de Valparaíso, Av. Universidad # 330, Curauma, 2340000 Valparaíso, Chile.
Facultad de Ciencias, Escuela de Biotecnología, Universidad Santo Tomás, 8370003 Santiago, Chile.
Microorganisms. 2020 Jun 29;8(7):974. doi: 10.3390/microorganisms8070974.
pv. (Psa) is the causal agent of a bacterial canker in kiwifruit plants and has caused economic losses worldwide. Currently, the primary strategies to control this pathogen include the use of copper-based compounds and even antibiotics. However, the emergence of isolates of Psa that are resistant to these agrochemicals has raised the need for new alternatives to control this pathogen. Bacteriophages have been proposed as an alternative to control bacterial infections in agriculture, including Psa. Here, we show the isolation and characterization of 13 phages with the potential to control Psa infections in kiwifruit plants. The phages were characterized according to their host range and restriction fragment length polymorphism (RFLP) pattern. Four phages were selected according to their lytic effect on the bacteria and their tolerance to different environmental conditions of pH (4-7), temperature (4-37 °C), and solar radiation exposure (30 and 60 min). The selected phages (CHF1, CHF7, CHF19, and CHF21) were sequenced, revealing a high identity with the podophage of Psa phiPSA2. In vitro assays with kiwifruit leaf samples demonstrated that the mixture of phages reduced the Psa bacterial load within three hours post-application and was able to reduce the damage index in 50% of cases. Similarly, assays with kiwifruit plants maintained in greenhouse conditions showed that these phages were able to reduce the Psa bacterial load in more than 50% of cases and produced a significant decrease in the damage index of treated plants after 30 days. Finally, none of the selected phages were able to infect the other bacteria present in the natural microbiota of kiwifruit plants. These results show that bacteriophages are an attractive alternative to control Psa infections in kiwifruit plants.
丁香假单胞菌猕猴桃致病变种(Psa)是猕猴桃细菌性溃疡病的病原体,已在全球范围内造成经济损失。目前,控制这种病原体的主要策略包括使用铜基化合物甚至抗生素。然而,对这些农用化学品具有抗性的Psa分离株的出现,增加了寻找控制这种病原体新方法的需求。噬菌体已被提议作为控制农业中细菌感染的一种替代方法,包括控制Psa。在此,我们展示了13种具有控制猕猴桃植株中Psa感染潜力的噬菌体的分离和特性。根据噬菌体的宿主范围和限制性片段长度多态性(RFLP)模式对其进行了特性分析。根据它们对细菌的裂解作用以及对不同pH值(4 - 7)、温度(4 - 37°C)和太阳辐射暴露(30和60分钟)环境条件的耐受性,选择了四种噬菌体。对所选噬菌体(CHF1、CHF7、CHF19和CHF21)进行测序,发现它们与Psa噬菌体phiPSA2具有高度同源性。对猕猴桃叶片样本进行的体外试验表明,噬菌体混合物在施用后三小时内降低了Psa细菌载量,并且在50%的情况下能够降低损伤指数。同样,在温室条件下对猕猴桃植株进行的试验表明,这些噬菌体在50%以上的情况下能够降低Psa细菌载量,并且在30天后使处理植株的损伤指数显著降低。最后,所选噬菌体均不能感染猕猴桃植株天然微生物群中存在的其他细菌。这些结果表明,噬菌体是控制猕猴桃植株中Psa感染的一种有吸引力的替代方法。