Evaluation of Different Formulations on the Viability of Phages for Use in Agriculture

Viruses Pub Date : 2024-09-07 DOI:10.3390/v16091430
Marcela León, Jorge Araya, Mauricio Nuñez, Manuel Arce, Fanny Guzmán, Carolina Yáñez, Ximena Besoain, Roberto Bastías
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Abstract

Bacteriophages have been proposed as biological controllers to protect plants against different bacterial pathogens. In this scenario, one of the main challenges is the low viability of phages in plants and under adverse environmental conditions. This work explores the use of 12 compounds and 14 different formulations to increase the viability of a phage mixture that demonstrated biocontrol capacity against Pseudomonas syringae pv. actinidiae (Psa) in kiwi plants. The results showed that the viability of the phage mixture decreases at 44 °C, at a pH lower than 4, and under UV radiation. However, using excipients such as skim milk, casein, and glutamic acid can prevent the viability loss of the phages under these conditions. Likewise, it was demonstrated that the use of these compounds prolongs the presence of phages in kiwi plants from 48 h to at least 96 h. In addition, it was observed that phages remained stable for seven weeks when stored in powder with skim milk, casein, or sucrose after lyophilization and at 4 °C. Finally, the phages with glutamic acid, sucrose, or skim milk maintained their antimicrobial activity against Psa on kiwi leaves and persisted within kiwi plants when added through roots. This study contributes to overcoming the challenges associated with the use of phages as biological controllers in agriculture.
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评估不同配方对农业用噬菌体活力的影响
噬菌体被认为是保护植物免受不同细菌病原体侵害的生物控制剂。在这种情况下,面临的主要挑战之一是噬菌体在植物体内和不利环境条件下的存活率较低。这项研究探讨了如何使用 12 种化合物和 14 种不同配方来提高噬菌体混合物的活力,该混合物对猕猴桃植物中的绿脓杆菌 pv. actinidiae(Psa)具有生物防治能力。结果表明,噬菌体混合物的活力在 44 ℃、pH 值低于 4 和紫外线辐射下会降低。然而,使用脱脂奶、酪蛋白和谷氨酸等辅料可以防止噬菌体在这些条件下的活力下降。同样,实验证明,使用这些化合物可使噬菌体在猕猴桃植株中的存活时间从 48 小时延长到至少 96 小时。此外,实验还观察到,噬菌体在冻干后与脱脂奶、酪蛋白或蔗糖一起制成粉末,并在 4 °C 下保存 7 周后仍保持稳定。最后,含有谷氨酸、蔗糖或脱脂奶的噬菌体对猕猴桃叶片上的 Psa 保持抗菌活性,通过根部添加到猕猴桃植株中时,噬菌体在猕猴桃植株中也能保持活性。这项研究有助于克服在农业中使用噬菌体作为生物控制剂所面临的挑战。
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