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用于对大流行病原体进行严格分型的全基因组单核苷酸多态性分析。

Whole-genome single nucleotide polymorphism analysis for typing the pandemic pathogen sensu stricto.

作者信息

Kulik Tomasz, Molcan Tomasz, Fiedorowicz Grzegorz, van Diepeningen Anne, Stakheev Alexander, Treder Kinga, Olszewski Jacek, Bilska Katarzyna, Beyer Marco, Pasquali Matias, Stenglein Sebastian

机构信息

Department of Botany and Nature Protection, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland.

Department of Bioinformatics, Institute of Biochemistry and Biophysics, Polish Academy of Sciences (PAN), Warsaw, Poland.

出版信息

Front Microbiol. 2022 Jul 18;13:885978. doi: 10.3389/fmicb.2022.885978. eCollection 2022.

Abstract

Recent improvements in microbiology and molecular epidemiology were largely stimulated by whole- genome sequencing (WGS), which provides an unprecedented resolution in discriminating highly related genetic backgrounds. WGS is becoming the method of choice in epidemiology of fungal diseases, but its application is still in a pioneer stage, mainly due to the limited number of available genomes. Fungal pathogens often belong to complexes composed of numerous cryptic species. Detecting cryptic diversity is fundamental to understand the dynamics and the evolutionary relationships underlying disease outbreaks. In this study, we explore the value of whole-genome SNP analyses in identification of the pandemic pathogen (.). This species is responsible for cereal diseases and negatively impacts grain production worldwide. The fungus belongs to the monophyletic fungal complex referred to as species complex including at least sixteen cryptic species, a few among them may be involved in cereal diseases in certain agricultural areas. We analyzed WGS data from a collection of 99 strains and 33 strains representing all known cryptic species belonging to the FGSC complex. As a first step, we performed a phylogenomic analysis to reveal species-specific clustering. A RAxML maximum likelihood tree grouped all analyzed strains of into a single clade, supporting the clustering-based identification approach. Although, phylogenetic reconstructions are essential in detecting cryptic species, a phylogenomic tree does not fulfill the criteria for rapid and cost-effective approach for identification of fungi, due to the time-consuming nature of the analysis. As an alternative, analysis of WGS information by mapping sequence data from individual strains against reference genomes may provide useful markers for the rapid identification of fungi. We provide a robust framework for typing through the web-based PhaME workflow available at EDGE bioinformatics. The method was validated through multiple comparisons of assembly genomes to reference strain PH-1. We showed that the difference between intra- and interspecies variability was at least two times higher than intraspecific variation facilitating successful typing of . This is the first study which employs WGS data for typing plant pathogenic fusaria.

摘要

微生物学和分子流行病学的最新进展在很大程度上受到全基因组测序(WGS)的推动,全基因组测序在区分高度相关的遗传背景方面提供了前所未有的分辨率。全基因组测序正在成为真菌疾病流行病学中的首选方法,但其应用仍处于开拓阶段,主要原因是可用基因组数量有限。真菌病原体通常属于由众多隐性物种组成的复合体。检测隐性多样性对于理解疾病爆发背后的动态和进化关系至关重要。在本研究中,我们探索了全基因组单核苷酸多态性分析在鉴定大流行病原体(.)中的价值。该物种导致谷物疾病,并对全球粮食生产产生负面影响。这种真菌属于单系真菌复合体,称为物种复合体,包括至少16个隐性物种,其中一些可能在某些农业地区导致谷物疾病。我们分析了来自99个菌株和33个菌株的全基因组测序数据,这些菌株代表了属于FGSC复合体的所有已知隐性物种。第一步,我们进行了系统基因组分析以揭示物种特异性聚类。一个RAxML最大似然树将所有分析的菌株聚为一个单系分支,支持基于聚类的鉴定方法。虽然系统发育重建对于检测隐性物种至关重要,但由于分析耗时,系统基因组树不符合快速且经济高效的真菌鉴定方法标准。作为替代方案,通过将单个菌株的序列数据与参考基因组进行比对来分析全基因组测序信息,可能为真菌的快速鉴定提供有用的标记。我们通过EDGE生物信息学提供的基于网络的PhaME工作流程,为分型提供了一个强大的框架。该方法通过将组装基因组与参考菌株PH-1进行多次比较得到验证。我们表明,种内和种间变异性的差异至少比种内变异高两倍,这有助于成功对进行分型。这是第一项利用全基因组测序数据对植物致病镰刀菌进行分型的研究。

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