鉴定用于猕猴桃溃疡病植物病原体丁香假单胞菌猕猴桃致病变种生物防治的噬菌体。
Identification of bacteriophages for biocontrol of the kiwifruit canker phytopathogen Pseudomonas syringae pv. actinidiae.
作者信息
Frampton Rebekah A, Taylor Corinda, Holguín Moreno Angela V, Visnovsky Sandra B, Petty Nicola K, Pitman Andrew R, Fineran Peter C
机构信息
Department of Microbiology and Immunology, University of Otago, Dunedin, New Zealand.
出版信息
Appl Environ Microbiol. 2014 Apr;80(7):2216-28. doi: 10.1128/AEM.00062-14. Epub 2014 Jan 31.
Pseudomonas syringae pv. actinidiae is a reemerging pathogen which causes bacterial canker of kiwifruit (Actinidia sp.). Since 2008, a global outbreak of P. syringae pv. actinidiae has occurred, and in 2010 this pathogen was detected in New Zealand. The economic impact and the development of resistance in P. syringae pv. actinidiae and other pathovars against antibiotics and copper sprays have led to a search for alternative management strategies. We isolated 275 phages, 258 of which were active against P. syringae pv. actinidiae. Extensive host range testing on P. syringae pv. actinidiae, other pseudomonads, and bacteria isolated from kiwifruit orchards showed that most phages have a narrow host range. Twenty-four were analyzed by electron microscopy, pulse-field gel electrophoresis, and restriction digestion. Their suitability for biocontrol was tested by assessing stability and the absence of lysogeny and transduction. A detailed host range was performed, phage-resistant bacteria were isolated, and resistance to other phages was examined. The phages belonged to the Caudovirales and were analyzed based on morphology and genome size, which showed them to be representatives of Myoviridae, Podoviridae, and Siphoviridae. Twenty-one Myoviridae members have similar morphologies and genome sizes yet differ in restriction patterns, host range, and resistance, indicating a closely related group. Nine of these Myoviridae members were sequenced, and each was unique. The most closely related sequenced phages were a group infecting Pseudomonas aeruginosa and characterized by phages JG004 and PAK_P1. In summary, this study reports the isolation and characterization of P. syringae pv. actinidiae phages and provides a framework for the intelligent formulation of phage biocontrol agents against kiwifruit bacterial canker.
猕猴桃溃疡病菌是一种再度出现的病原菌,可引发猕猴桃细菌性溃疡病。自2008年以来,全球爆发了猕猴桃溃疡病菌疫情,2010年在新西兰检测到了这种病原菌。猕猴桃溃疡病菌及其他致病型对抗生素和铜制剂产生抗药性,带来了经济影响,促使人们寻找替代管理策略。我们分离出275株噬菌体,其中258株对猕猴桃溃疡病菌有活性。对猕猴桃溃疡病菌、其他假单胞菌以及从猕猴桃果园分离出的细菌进行了广泛的宿主范围测试,结果表明大多数噬菌体的宿主范围较窄。通过电子显微镜、脉冲场凝胶电泳和限制性酶切分析了24株噬菌体。通过评估稳定性、无溶原性和转导性来测试它们用于生物防治的适用性。进行了详细的宿主范围研究,分离出噬菌体抗性细菌,并检测了对其他噬菌体的抗性。这些噬菌体属于有尾噬菌体目,根据形态和基因组大小进行分析,结果表明它们分别代表肌尾噬菌体科、短尾噬菌体科和长尾噬菌体科。21株肌尾噬菌体科成员具有相似的形态和基因组大小,但限制性图谱、宿主范围和抗性不同,表明它们是一个密切相关的群体。对其中9株肌尾噬菌体科成员进行了测序,每株都独一无二。序列上最密切相关的噬菌体是一组感染铜绿假单胞菌的噬菌体,以噬菌体JG004和PAK_P1为特征。总之,本研究报告了猕猴桃溃疡病菌噬菌体的分离和特性,为智能配制防治猕猴桃细菌性溃疡病的噬菌体生物防治剂提供了框架。
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