Rainey P B, Thompson I P, Palleroni N J
Molecular Microbial Ecology, Institute of Virology and Environmental Microbiology, Oxford, United Kingdom.
Int J Syst Bacteriol. 1994 Jan;44(1):54-61. doi: 10.1099/00207713-44-1-54.
A genome and fatty acid analysis of 16 Pseudomonas stutzeri reference strains having DNA compositions ranging from 62.2 to 65.5 mol% G+C was performed by pulsed-field gel electrophoresis of XbaI and SpeI macrorestriction fragments and gas chromatography of total cellular fatty acids. Macrorestriction fragment patterns were evaluated by using previously described algorithms (D. Grothues and B. Tümmler, Mol. Microbiol. 5:2763-2776, 1991), and the results allowed us to subdivide the species into two groups which correlated with G+C content. Two examples of recent strain divergence were observed among clinical isolates, but in general a marked degree of heterogeneity was observed in the macrorestriction fragment patterns, and even phenotypically similar strains produced divergent patterns. While the differences were not sufficiently great to exclude any strain from P. stutzeri, they suggest that recombination and niche-specific selection may be significant factors responsible for generating and maintaining the heterogeneity inherent in the species. Genome sizes were estimated from the sums of SpeI restriction fragment sizes and ranged from 3.4 to 4.3 Mbp; the genome sizes of the low-G+C-content strains (G+C contents, approximately 62 mol%) were confined to a narrow range between 3.9 and 4.1 Mbp. An examination of the distributions of macrorestriction fragments resulting from digestion with XbaI and SpeI showed that both distributions differed significantly from the expected (random) distribution, suggesting that there is a supragenic level of chromosomal organization. An analysis of fatty acid methyl ester data by using Microbial Identification System software revealed a similar correlation between phenotype and G+C content, indicating that division of the species is possible by the method used in this study.(ABSTRACT TRUNCATED AT 250 WORDS)
对16株施氏假单胞菌参考菌株进行了基因组和脂肪酸分析,这些菌株的DNA组成中鸟嘌呤与胞嘧啶的含量在62.2%至65.5%(摩尔比)之间。通过对XbaI和SpeI酶切大片段进行脉冲场凝胶电泳以及对总细胞脂肪酸进行气相色谱分析来完成。使用先前描述的算法(D. Grothues和B. Tümmler,《分子微生物学》5:2763 - 2776,1991)对酶切大片段模式进行评估,结果使我们能够将该物种细分为与鸟嘌呤与胞嘧啶含量相关的两组。在临床分离株中观察到了近期菌株分化的两个例子,但总体而言,在酶切大片段模式中观察到了显著程度的异质性,甚至表型相似的菌株也产生了不同的模式。虽然这些差异不足以将任何菌株排除在施氏假单胞菌之外,但它们表明重组和生态位特异性选择可能是导致并维持该物种固有异质性的重要因素。根据SpeI酶切片段大小总和估计基因组大小,范围在3.4至4.3兆碱基对之间;低鸟嘌呤与胞嘧啶含量菌株(鸟嘌呤与胞嘧啶含量约为62%)的基因组大小局限于3.9至4.1兆碱基对的狭窄范围内。对用XbaI和SpeI酶切产生的大片段分布进行检查表明,这两种分布均与预期(随机)分布有显著差异,表明存在超基因水平的染色体组织。使用微生物鉴定系统软件对脂肪酸甲酯数据进行分析,揭示了表型与鸟嘌呤与胞嘧啶含量之间的类似相关性,表明通过本研究中使用的方法对该物种进行划分是可行的。(摘要截选至250字)