Institut National de la Recherche Agronomique, Station de Pathologie Végétale, 49071 Beaucouzé, and Laboratoire d'Ecologie Microbienne du Sol, UMR CNRS 557, Institut National de la Recherche Agronomique, Université Lyon 1, 69622 Villeurbanne Cedex, France.
Appl Environ Microbiol. 1998 Apr;64(4):1180-7. doi: 10.1128/AEM.64.4.1180-1187.1998.
Recently, DNA pairing analyses showed that Pseudomonas syringae pv. tomato and related pathovars, including P. syringae pv. maculicola, form a genomic species (Pseudomonas tomato) (L. Gardan, H. L. Shafik, and P. A. D. Grimont, p. 445-448, in K. Rudolph, T. J. Burr, J. W. Mansfield, D. Stead, A. Vivian, and J. von Kietzell, ed., Pseudomonas syringae Pathovars and Related Pathogens, 1997). The genetic diversity of 23 strains belonging to this genomic species and 4 outgroup strains was analyzed with randomly amplified polymorphic DNA (RAPD) and amplified fragment length polymorphic (AFLP) techniques. Simple boiling of P. syringae cells was suitable for subsequent DNA amplification to obtain reliable patterns in RAPD and AFLP analyses. In general, the grouping of P. syringae strains by both analysis techniques corresponded well with the classification obtained from an RFLP analysis of ribosomal DNA operons, DNA pairing studies, and an analysis of pathogenicity data. However, two strains of P. syringae pv. maculicola produced distinct DNA patterns compared to the DNA patterns of other P. syringae pv. maculicola strains; these patterns led us to assume that horizontal transfer of DNA could occur between bacterial populations. Both techniques used in this study have high discriminating power because strains of P. syringae pv. tomato and P. syringae pv. maculicola which were indistinguishable by other techniques, including pathogenicity tests on tomato, were separated into two groups by both RAPD and AFLP analyses. In addition, data analysis showed that the AFLP method was more efficient for assessing intrapathovar diversity than RAPD analysis and allowed clear delineation between intraspecific and interspecific genetic distances, suggesting that it could be an alternative to DNA pairing studies. However, it was not possible to distinguish the two races of P. syringae pv. tomato on the basis of an analysis of the data provided by either the AFLP or RAPD technique.
最近，DNA 配对分析表明，丁香假单胞菌 pv.番茄及其相关的致病变种，包括丁香假单胞菌 pv. 斑点病，形成了一个基因组种（假单胞菌番茄）（L. Gardan、H. L. Shafik 和 P. A. D. Grimont，第 445-448 页，载于 K. Rudolph、T. J. Burr、J. W. Mansfield、D. Stead、A. Vivian 和 J. von Kietzell 编著的《丁香假单胞菌致病变种及相关病原体》，1997 年）。利用随机扩增多态性 DNA（RAPD）和扩增片段长度多态性（AFLP）技术分析了属于该基因组种的 23 株和 4 株外群菌株的遗传多样性。简单煮沸丁香假单胞菌细胞适合随后的 DNA 扩增，以在 RAPD 和 AFLP 分析中获得可靠的图谱。一般来说，两种分析技术对丁香假单胞菌菌株的分组与核糖体 DNA 操纵子的 RFLP 分析、DNA 配对研究和致病性数据分析的分类结果非常吻合。然而，与其他丁香假单胞菌 pv. 斑点病菌株的 DNA 图谱相比，2 株丁香假单胞菌 pv. 斑点病菌株产生了不同的 DNA 图谱；这些图谱使我们假设细菌种群之间可能发生 DNA 水平转移。本研究中使用的两种技术都具有较高的分辨力，因为番茄假单胞菌 pv. 番茄和番茄假单胞菌 pv. 斑点病的菌株用其他技术（包括对番茄的致病性试验）无法区分，但通过 RAPD 和 AFLP 分析，它们被分为两组。此外，数据分析表明，AFLP 方法比 RAPD 分析更有效地评估种内多样性，并能清晰地区分种内和种间遗传距离，表明它可能是 DNA 配对研究的替代方法。然而，无论是 AFLP 技术还是 RAPD 技术，都无法根据数据区分番茄假单胞菌 pv. 番茄的两个品种。
