Phytopathology. 1999 Dec;89(12):1169-75. doi: 10.1094/PHYTO.1999.89.12.1169.
ABSTRACT Strains of the carnation wilt pathogen, Fusarium oxysporum f. sp. dianthi, can be distinguished by DNA fingerprint patterns, using the fungal transposable elements Fot1 and impala as probes for Southern hybridization. The DNA fingerprints correspond to three groups of F. oxysporum f. sp. dianthi strains: the first group includes isolates of races 1 and 8; the second group includes isolates of races 2, 5 and 6; and the third group includes isolates of race 4. Genomic DNAs flanking race-associated insertion sites of Fot1 (from races 1, 2, and 8) or impala (from race 4) were amplified by the inverse polymerase chain reaction (PCR) technique. These regions were cloned and sequenced, and three sets of primers overlapping the 3' or 5' end of the transposon and its genomic insertion were designed. Using fungal genomic DNA as template in PCR experiments, primer pairs generated amplification products of 295, 564 and 1,315 bp, corresponding to races 1 and 8; races 2, 5, and 6; and race 4, respectively. When multiplex PCR was performed with genomic DNA belonging to races 1 and 8, 2, or 4, single amplimers were generated, allowing clear race determination of the isolate tested. PCR was successfully performed on DNA extracted from susceptible carnation cv. Indios infected with isolates representative of races 1, 2, 4, and 8.
摘要 香石竹枯萎病菌（Fusarium oxysporum f. sp. dianthi）的菌株可以通过 DNA 指纹图谱模式来区分，使用真菌转座元件 Fot1 和 impala 作为 Southern 杂交的探针。DNA 指纹图谱与 F. oxysporum f. sp. dianthi 菌株的三组相对应：第一组包括 1 号和 8 号小种的分离物；第二组包括 2 号、5 号和 6 号小种的分离物；第三组包括 4 号小种的分离物。利用反向聚合酶链反应（PCR）技术扩增 Fot1（来自 1 号、2 号和 8 号小种）或 impala（来自 4 号小种）与种族相关的插入位点侧翼的基因组 DNA。这些区域被克隆和测序，并设计了三套引物，重叠转座子及其基因组插入的 3'或 5'末端。在 PCR 实验中，使用真菌基因组 DNA 作为模板，引物对产生了 295、564 和 1315bp 的扩增产物，分别对应于 1 号和 8 号小种；2 号、5 号和 6 号小种；和 4 号小种。当用属于 1 号和 8 号、2 号或 4 号小种的基因组 DNA 进行多重 PCR 时，会产生单一的扩增子，从而可以清楚地确定被测试分离物的小种。PCR 成功地应用于从感病香石竹品种 Indios 中提取的 DNA 上，该品种感染了代表 1、2、4 和 8 号小种的分离物。
