State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou, China.
BMC Microbiol. 2012 Aug 18;12:182. doi: 10.1186/1471-2180-12-182.
Two important plant pathogenic bacteria Acidovorax oryzae and Acidovorax citrulli are closely related and often not easy to be differentiated from each other, which often resulted in a false identification between them based on traditional methods such as carbon source utilization profile, fatty acid methyl esters, and ELISA detection tests. MALDI-TOF MS and Fourier transform infrared (FTIR) spectra have recently been successfully applied in bacterial identification and classification, which provide an alternate method for differentiating the two species.
Characterization and comparison of the 10 A. oryzae strains and 10 A. citrulli strains were performed based on traditional bacteriological methods, MALDI-TOF MS, and FTIR spectroscopy. Our results showed that the identity of the two closely related plant pathogenic bacteria A. oryzae and A. citrulli was able to be confirmed by both pathogenicity tests and species-specific PCR, but the two species were difficult to be differentiated based on Biolog and FAME profile as well as 16 S rRNA sequence analysis. However, there were significant differences in MALDI-TOF MS and FTIR spectra between the two species of Acidovorax. MALDI-TOF MS revealed that 22 and 18 peaks were specific to A. oryzae and A. citrulli, respectively, while FTIR spectra of the two species of Acidovorax have the specific peaks at 1738, 1311, 1128, 1078, 989 cm(-1) and at 1337, 968, 933, 916, 786 cm(-1), respectively.
This study indicated that MALDI-TOF MS and FTIR spectra may give a new strategy for rapid bacterial identification and differentiation of the two closely related species of Acidovorax.
两种重要的植物病原菌——稻黄寡养单胞菌和酸沼寡养单胞菌——密切相关,通常难以通过传统方法(如碳源利用谱、脂肪酸甲酯和 ELISA 检测试验)将它们彼此区分开来。基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)和傅里叶变换红外(FTIR)光谱最近已成功应用于细菌鉴定和分类,为区分这两个物种提供了一种替代方法。
基于传统细菌学方法、MALDI-TOF MS 和 FTIR 光谱对 10 株稻黄寡养单胞菌和 10 株酸沼寡养单胞菌进行了特征描述和比较。结果表明,通过致病性试验和种特异性 PCR 能够确认这两种密切相关的植物病原菌稻黄寡养单胞菌和酸沼寡养单胞菌的身份,但基于 Biolog 和 FAME 图谱以及 16S rRNA 序列分析,这两个物种难以区分。然而,在稻黄寡养单胞菌和酸沼寡养单胞菌的 MALDI-TOF MS 和 FTIR 光谱之间存在显著差异。MALDI-TOF MS 显示,分别有 22 个和 18 个峰是稻黄寡养单胞菌和酸沼寡养单胞菌所特有的,而这两种酸沼寡养单胞菌的 FTIR 光谱则分别在 1738、1311、1128、1078、989 cm-1 和 1337、968、933、916、786 cm-1 处有特征峰。
本研究表明,MALDI-TOF MS 和 FTIR 光谱可能为快速细菌鉴定和区分这两种密切相关的酸沼寡养单胞菌提供一种新策略。