Crowhurst R N, Hawthorne B T, Rikkerink E H, Templeton M D
DSIR Plant Protection, Auckland, New Zealand.
Curr Genet. 1991 Nov;20(5):391-6. doi: 10.1007/BF00317067.
We have used a PCR-based technique, involving the random amplification of polymorphic DNA (RAPD), to assess genome variability between 21 isolates from F. solani f. sp. cucurbitae races 1 and 2. Based on RAPD marker patterns the isolates fell into two distinct groups corresponding to mating populations MPI and MPV. Four isolates that could not be assigned to one or other mating population by traditional means were distinguished by RAPD patterns. Seven polymorphic RAPD products were used to probe Southern blots of MPI and MPV genomic DNA. Six of the seven probes hybridized to single-copy sequences and five of the seven probes showed specificity for one or other mating population. We suggest that not only is the technique a rapid and reliable tool for isolate-typing of fungi but it also provides a rapid method for obtaining species- or race-specific hybridization probes.
我们使用了一种基于聚合酶链反应(PCR)的技术,即随机扩增多态性DNA(RAPD),来评估来自南瓜枯萎病菌1号和2号生理小种的21个分离株之间的基因组变异性。根据RAPD标记模式,这些分离株分为两个不同的组,分别对应交配群体MPI和MPV。通过传统方法无法归入任一交配群体的4个分离株,通过RAPD模式得以区分。7个多态性RAPD产物用于探测MPI和MPV基因组DNA的Southern杂交印迹。7个探针中的6个与单拷贝序列杂交,7个探针中的5个对一个或另一个交配群体具有特异性。我们认为,该技术不仅是一种快速可靠的真菌分离株分型工具,还提供了一种快速获得种特异性或生理小种特异性杂交探针的方法。