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果胶杆菌噬菌体 vB_PcaM_P7_Pc 是. 属的一个新成员

Pectobacterium carotovorum Phage vB_PcaM_P7_Pc Is a New Member of the Genus .

机构信息

Department of Microbiology, Faculty of Science, University of Kelaniyagrid.45202.31, Kelaniya, Sri Lanka.

出版信息

Microbiol Spectr. 2022 Dec 21;10(6):e0312622. doi: 10.1128/spectrum.03126-22. Epub 2022 Nov 8.

Abstract

Pectobacterium carotovorum is an economically important phytopathogen and has been identified as the major causative agent of bacterial soft rot in carrots. Control of this phytopathogen is vital to minimizing carrot harvest losses. As fully efficient control measures to successfully avoid the disease are unavailable, the phage-mediated biocontrol of the pathogen has recently gained scientific attention. In this study, we present a comprehensive characterization of the phage vB_PcaM_P7_Pc (abbreviated as P7_Pc) that was isolated from infected carrot samples with characteristic soft rot symptoms, which were obtained from storage facilities at market places in Gampaha District, Sri Lanka. P7_Pc is a myovirus, and it exhibits growth characteristics of an exclusively lytic life cycle. It showed visible lysis against four of the tested P. carotovorum strains and one Pectobacterium aroidearum strain. This phage also showed a longer latent period (125 min) than other related phages; however, this did not affect its high phage titter (>10 PFU/mL). The final assembled genome of P7_Pc is 147,299 bp in length with a G+C content of 50.34%. Of the 298 predicted open reading frames (ORFs) of the genome of P7_Pc, putative functions were assigned to 53 ORFs. Seven tRNA-coding genes were predicted in the genome, while the genome lacked any major genes coding for lysogeny-related products, confirming its virulent nature. The P7_Pc genome shares 96.12% and 95.74% average nucleotide identities with phages CR8 and PBES02, respectively. Phylogenetic and phylogenomic analyses of the genome revealed that P7_Pc clusters well within the clade with the members representing the genus . Currently, there are only 4 characterized phages ( phages phiTE and CB7 and phages DU_PP_I and DU_PP_IV) that are classified under the genus, making the phage P7_Pc the first reported member of the genus isolated using the host bacterium The results of this study provide a detailed characterization of the phage P7_Pc, enabling its careful classification into the genus . The knowledge gathered on the phage based on the shared biology of the genus will further aid in the future selection of phage P7_Pc as a biocontrol agent. Bacterial soft rot disease, caused by spp., can lead to significant losses in carrot yields. As current control measures involving the use of chemicals or antibiotics are not recommended in many countries, bacteriophage-mediated biocontrol strategies are being explored for the successful control of these phytopathogens. The successful implementation of such biocontrol strategies relies heavily upon the proper understanding of the growth characteristics and genomic properties of the phage. Further, the selection of taxonomically different phages for the formulation of phage cocktails in biocontrol applications is critical to combat potential bacterial resistance development. This study was conducted to carefully characterize and resolve the phylogenetic placement of the phage vB_PcaM_P7_Pc by using its biological and genomic properties. Phage P7_Pc has a myovirus morphotype with an exclusively lytic life cycle, and the absence of genes related to lysogeny, toxin production, and antibiotic resistance in its genome confirmed its suitability to be used in environmental applications. Furthermore, P7_Pc is classified under the genus , making it the first reported phage of the genus of the host species, .

摘要

胡萝卜欧文氏菌(Pectobacterium carotovorum)是一种具有重要经济意义的植物病原菌,已被确定为胡萝卜细菌性软腐病的主要病原体。控制这种植物病原菌对于最大限度地减少胡萝卜收获损失至关重要。由于目前还没有完全有效的控制措施来成功避免这种疾病,因此最近科学界开始关注噬菌体介导的病原菌生物防治。在本研究中,我们对从斯里兰卡甘帕哈区市场储存设施中具有典型软腐病症状的感染胡萝卜样本中分离出的噬菌体 vB_PcaM_P7_Pc(简称 P7_Pc)进行了全面表征。P7_Pc 是一种肌尾噬菌体,表现出专性裂解生命周期的生长特征。它对测试的 4 种胡萝卜欧文氏菌菌株和 1 种 Pectobacterium aroidearum 菌株表现出明显的裂解作用。该噬菌体的潜伏期(125 分钟)也比其他相关噬菌体长;然而,这并不影响其高噬菌体滴度(>10PFU/mL)。P7_Pc 的最终组装基因组长度为 147299bp,G+C 含量为 50.34%。在 P7_Pc 基因组的 298 个预测开放阅读框(ORFs)中,对 53 个 ORFs 赋予了推定功能。基因组中预测了 7 个 tRNA 编码基因,而基因组中缺乏任何编码溶原相关产物的主要基因,证实了其毒性特征。P7_Pc 基因组与噬菌体 CR8 和 PBES02 的平均核苷酸同一性分别为 96.12%和 95.74%。基因组的系统发育和系统基因组分析表明,P7_Pc 聚类很好,与代表属的成员聚类在一起。目前,仅有 4 种被表征的 噬菌体(噬菌体 phiTE 和 CB7 和 噬菌体 DU_PP_I 和 DU_PP_IV)被分类为该属,这使得噬菌体 P7_Pc 成为使用宿主菌 分离的第一个被报道的属成员。本研究对噬菌体 P7_Pc 进行了详细的特征描述,使其能够被小心地分类为属。基于属的共同生物学知识获得的噬菌体知识将进一步有助于未来选择噬菌体 P7_Pc 作为生物防治剂。由 spp.引起的细菌性软腐病可导致胡萝卜产量显著损失。由于目前在许多国家不建议使用化学物质或抗生素等化学物质进行控制,因此正在探索噬菌体介导的生物防治策略来成功控制这些植物病原菌。这种生物防治策略的成功实施在很大程度上依赖于对噬菌体生长特性和基因组特性的正确理解。此外,在生物防治应用中,为了对抗潜在的细菌耐药性发展,选择分类上不同的噬菌体来配制噬菌体鸡尾酒至关重要。本研究旨在通过使用噬菌体的生物学和基因组特性,仔细表征和确定 噬菌体 vB_PcaM_P7_Pc 的系统发育位置。噬菌体 P7_Pc 具有肌尾病毒形态,具有专性裂解生命周期,其基因组中缺乏与溶原、毒素产生和抗生素耐药性相关的基因,证实了其在环境应用中的适用性。此外,P7_Pc 被分类为属,使其成为第一个被报道的宿主物种的属成员。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/559b/9769974/fa64084e055f/spectrum.03126-22-f001.jpg

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