Machado Henrique, Gram Lone
Department of Systems Biology, Technical University of Denmark, Lyngby, Denmark Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Hørsholm, Denmark
Department of Systems Biology, Technical University of Denmark, Lyngby, Denmark.
Appl Environ Microbiol. 2015 Apr;81(8):2745-52. doi: 10.1128/AEM.00058-15. Epub 2015 Feb 6.
Microbial taxonomy is essential in all areas of microbial science. The 16S rRNA gene sequence is one of the main phylogenetic species markers; however, it does not provide discrimination in the family Vibrionaceae, where other molecular techniques allow better interspecies resolution. Although multilocus sequence analysis (MLSA) has been used successfully in the identification of Vibrio species, the technique has several limitations. They include the fact that several locus amplifications and sequencing have to be performed, which still sometimes lead to doubtful identifications. Using an in silico approach based on genomes from 103 Vibrionaceae strains, we demonstrate here the high resolution of the fur gene in the identification of Vibrionaceae species and its usefulness as a phylogenetic marker. The fur gene showed within-species similarity higher than 95%, and the relationships inferred from its use were in agreement with those observed for 16S rRNA analysis and MLSA. Furthermore, we developed a fur PCR sequencing-based method that allowed identification of Vibrio species. The discovery of the phylogenetic power of the fur gene and the development of a PCR method that can be used in amplification and sequencing of the gene are of general interest whether for use alone or together with the previously suggested loci in an MLSA.
微生物分类学在微生物科学的各个领域都至关重要。16S rRNA基因序列是主要的系统发育物种标记之一;然而,它在弧菌科中无法提供区分能力,而其他分子技术在该科中能实现更好的种间分辨率。尽管多位点序列分析(MLSA)已成功用于弧菌属物种的鉴定,但该技术存在一些局限性。这些局限性包括必须进行多个位点的扩增和测序,而这有时仍会导致鉴定结果存疑。基于103株弧菌科菌株的基因组,我们采用计算机模拟方法,在此证明了fur基因在弧菌科物种鉴定中的高分辨率及其作为系统发育标记的实用性。fur基因在种内的相似度高于95%,基于该基因推断的亲缘关系与16S rRNA分析和MLSA观察到的结果一致。此外,我们开发了一种基于fur基因PCR测序的方法,可用于鉴定弧菌属物种。fur基因系统发育能力的发现以及一种可用于该基因扩增和测序的PCR方法的开发,无论是单独使用还是与MLSA中先前建议的位点一起使用,都具有普遍意义。