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对光动力方法与铜制剂在控制猕猴桃植物中丁香假单胞菌猕猴桃致病变种方面的深入研究。

An insight into the photodynamic approach versus copper formulations in the control of Pseudomonas syringae pv. actinidiae in kiwi plants.

作者信息

Jesus Vânia, Martins Diana, Branco Tatiana, Valério Nádia, Neves Maria G P M S, Faustino Maria A F, Reis Luís, Barreal Esther, Gallego Pedro P, Almeida Adelaide

机构信息

Departamento de Biologia, Universidade de Aveiro, Aveiro, Portugal.

出版信息

Photochem Photobiol Sci. 2018 Feb 14;17(2):180-191. doi: 10.1039/c7pp00300e.

Abstract

In the last decade, the worldwide production of kiwi fruit has been highly affected by Pseudomonas syringae pv. actinidiae (Psa), a phytopathogenic bacterium; this has led to severe economic losses that are seriously affecting the kiwi fruit trade. The available treatments for this disease are still scarce, with the most common involving frequently spraying the orchards with copper derivatives, in particular cuprous oxide (CuO). However, these copper formulations should be avoided due to their high toxicity; therefore, it is essential to search for new approaches for controlling Psa. Antimicrobial photodynamic therapy (aPDT) may be an alternative approach to inactivate Psa. aPDT consists in the use of a photosensitizer molecule (PS) that absorbs light and by transference of the excess of energy or electrons to molecular oxygen forms highly reactive oxygen species (ROS) that can affect different molecular targets, thus being very unlikely to lead to the development of microbe resistance. The aim of the present study was to evaluate the effectiveness of aPDT to photoinactivate Psa, using the porphyrin Tetra-Py-Me and different light intensities. The degree of inactivation of Psa was assessed using the PS at 5.0 μM under low irradiance (4.0 mW cm). Afterward, ex vivo experiments, using artificially contaminated kiwi leaves, were conducted with a PS at 50 μM under 150 mW cm and sunlight irradiation. A reduction of 6 log in the in vitro assays after 90 min of irradiation was observed. In the ex vivo tests, the decrease was lower, approximately 1.8 log reduction at an irradiance of 150 mW cm, 1.2 log at 4.0 mW cm, and 1.5 log under solar radiation. However, after three successive cycles of treatment under 150 mW cm, a 4 log inactivation was achieved. No negative effects were observed on leaves after treatment. Assays using CuO were also performed at the recommended concentration by law (50 g h L) and at concentrations 10 times lower, in which at both concentrations, Psa was efficiently inactivated (5 log inactivation) after a few minutes of treatment, but negative effects were observed on the leaves after treatment.

摘要

在过去十年中,全球猕猴桃产量受到丁香假单胞菌猕猴桃致病变种(Psa)这一植物病原菌的严重影响;这导致了严重的经济损失,严重影响了猕猴桃贸易。针对这种疾病的现有治疗方法仍然稀缺,最常见的方法是频繁地向果园喷洒铜衍生物,特别是氧化亚铜(CuO)。然而,由于这些铜制剂毒性高,应避免使用;因此,寻找控制Psa的新方法至关重要。抗菌光动力疗法(aPDT)可能是使Psa失活的一种替代方法。aPDT包括使用一种光敏剂分子(PS),该分子吸收光并通过将多余的能量或电子转移给分子氧形成高活性氧物种(ROS),这些ROS可以影响不同的分子靶点,因此极不可能导致微生物产生抗性。本研究的目的是评估使用卟啉四吡甲基(Tetra-Py-Me)和不同光强度时,aPDT对Psa进行光灭活的有效性。在低辐照度(4.0 mW/cm²)下,使用5.0 μM的PS评估Psa的失活程度。之后,使用人工污染的猕猴桃叶片进行离体实验,在150 mW/cm²和阳光照射下使用50 μM的PS。照射90分钟后,在体外试验中观察到Psa减少了6个对数。在离体试验中,减少幅度较小,在150 mW/cm²辐照度下约减少1.8个对数,在4.0 mW/cm²下减少1.2个对数,在太阳辐射下减少1.5个对数。然而,在150 mW/cm²下连续进行三个循环的处理后,实现了4个对数的失活。处理后在叶片上未观察到负面影响。还按照法定推荐浓度(50 g/h·L)和低10倍的浓度进行了使用CuO的试验,在这两种浓度下,处理几分钟后Psa均被有效灭活(5个对数的失活),但处理后在叶片上观察到了负面影响。

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