Yuanyuan Niu, Xiaobo Yang, Shang Wang, Yutong Yang, Hongrui Zhou, Chenyu Li, Bin Xue, Xi Zhang, Chen Zhao, Zhiqiang Shen, Jingfeng Wang, Yun Ling, Pingfeng Yu, Zhigang Qiu
College of Marine Ecology and Environment, Shanghai Ocean University, Shanghai, China.
Key Laboratory of Risk Assessment and Control for Environment and Food Safety, TianJin Institute of Environmental and Operational Medicine, Tianjin, China.
Front Microbiol. 2022 Jul 14;13:946251. doi: 10.3389/fmicb.2022.946251. eCollection 2022.
Bacteriophages (phages) are capable of infecting specific bacteria, and therefore can be used as a biological control agent to control bacteria-induced animal, plant, and human diseases. In this study, two homolog phages (named PPAY and PPAT) that infect PAO1 were isolated and characterized. The results of the phage plaque assay showed that PPAT plaques were transparent dots, while the PPAY plaques were translucent dots with a halo. Transmission electron microscopy results showed that PPAT (65 nm) and PPAY (60 nm) strains are similar in size and have an icosahedral head and a short tail. Therefore, these belong to the short-tailed phage family . One-step growth curves revealed the latent period of 20 min and burst time of 30 min for PPAT and PPAY. The burst size of PPAT (953 PFUs/infected cell) was higher than that of PPAY (457 PFUs/infected cell). Also, the adsorption rate constant of PPAT (5.97 × 10 ml/min) was higher than that of PPAY (1.32 × 10 ml/min) at 5 min. Whole-genome sequencing of phages was carried out using the Illumina HiSeq platform. The genomes of PPAT and PPAY have 54,888 and 50,154 bp, respectively. Only 17 of the 352 predicted ORFs of PPAT could be matched to homologous genes of known function. Likewise, among the 351 predicted ORFs of PPAY, only 18 ORFs could be matched to genes of established functions. Homology and evolutionary analysis indicated that PPAT and PPAY are closely related to PA11. The presence of tail fiber proteins in PPAY but not in PPAT may have contributed to the halo effect of its plaque spots. In all, PPAT and PPAY, newly discovered phages, showed growth inhibitory effects on bacteria and can be used for research and clinical purposes.
噬菌体能够感染特定细菌,因此可作为生物防治剂来控制由细菌引起的动物、植物和人类疾病。在本研究中,分离并鉴定了两种感染PAO1的同源噬菌体(命名为PPAY和PPAT)。噬菌体噬菌斑测定结果表明,PPAT噬菌斑为透明斑点,而PPAY噬菌斑为带晕圈的半透明斑点。透射电子显微镜结果显示,PPAT(65纳米)和PPAY(60纳米)菌株大小相似,具有二十面体头部和短尾。因此,它们属于短尾噬菌体家族。一步生长曲线显示,PPAT和PPAY的潜伏期为20分钟,爆发时间为30分钟。PPAT的爆发量(953个噬菌斑形成单位/感染细胞)高于PPAY(457个噬菌斑形成单位/感染细胞)。此外,在5分钟时,PPAT的吸附速率常数(5.97×10毫升/分钟)高于PPAY(1.32×10毫升/分钟)。使用Illumina HiSeq平台对噬菌体进行全基因组测序。PPAT和PPAY的基因组分别有54,888和50,154碱基对。PPAT预测的352个开放阅读框中只有17个能与已知功能的同源基因匹配。同样,在PPAY预测的351个开放阅读框中,只有18个开放阅读框能与已确定功能的基因匹配。同源性和进化分析表明,PPAT和PPAY与PA11密切相关。PPAY中存在尾纤维蛋白而PPAT中不存在,这可能导致了其噬菌斑斑点的晕圈效应。总之,新发现的噬菌体PPAT和PPAY对细菌具有生长抑制作用,可用于研究和临床目的。