Yli-Mattila T, Paavanen-Huhtala S, Fenton B, Tuovinen T
Department of Biology, University of Turku, Finland.
Exp Appl Acarol. 2000;24(10-11):863-80. doi: 10.1023/a:1006496423090.
The variation within and between Finnish Euseius finlandicus populations was investigated by RAPD-PCR and ITS sequence analyses. Resin DNA extraction was found to be a suitable method for samples of single mites used in PCR. The banding patterns from 24 RAPD primers and 10 primer pairs were very similar and reproducible in all specimens of the predatory mite studied. However, the E. finlandicus K-strain could be distinguished from organophosphate-resistant predatory mites (R-strain), since almost all of them produced a 1,400 bp RAPD-PCR product, which was missing or very rare in other strains studied. Another RAPD band of ca. 680 bp was in turn much more common in other mites of E. finlandicus than in the K-strain mites. Mite specific primers were designed and used to follow the survival of the R-strain released on apple trees. The 680 bp band obtained with specific primers was specific to the species E. finlandicus mites studied, including those that had been negative with RAPD primers. The 1,400 bp specific primers could be used as a marker for following the survival of R-strain mites on apple trees. At the species level it was possible to distinguish adults and eggs of E. finlandicus from Anthoseius rhenanus and Phytonemus pallidus by RAPD-PCR. In addition, a band at 480bp was found to correspond to DNA of the predatory mite Phytoseius macropilis, when both specific primer pairs were used together. It was not possible to amplify the ITS region of E. finlandicus rDNA using several primer pairs that work in other mites and aphids. However, a basidiomycete rDNA sequence was amplified with one of these ITS primer pairs in K-strain mites. Finally, it was found that fungal rDNA-specific primers amplified an ITS region of ca. 650 bp in several strains of E. finlandicus. Internal primers, designed to amplify the central part of the 650 bp product, successfully amplified this product from all the mites.
通过随机扩增多态性DNA聚合酶链反应（RAPD-PCR）和内转录间隔区（ITS）序列分析，对芬兰普通真绥螨种群内部及种群之间的变异进行了研究。结果发现，树脂DNA提取法适用于PCR中使用的单头螨样本。在所研究的捕食螨的所有样本中，24种RAPD引物和10对引物对的条带模式非常相似且可重复。然而，普通真绥螨K株可与抗有机磷捕食螨（R株）区分开来，因为几乎所有的K株都产生了一个1400 bp的RAPD-PCR产物，而在所研究的其他株系中该产物缺失或非常罕见。相反，另一条约680 bp的RAPD条带在普通真绥螨的其他螨类中比在K株螨类中更为常见。设计了螨特异性引物，并用于追踪释放到苹果树上的R株的存活情况。用特异性引物获得的680 bp条带对所研究的普通真绥螨物种具有特异性，包括那些用RAPD引物检测呈阴性的螨类。1400 bp特异性引物可作为追踪苹果树上R株螨类存活情况的标记。在物种水平上，通过RAPD-PCR可以将普通真绥螨的成虫和卵与莱茵钝绥螨和苍白食植螨区分开来。此外，当同时使用两对特异性引物时，发现一条480 bp的条带对应于大长植绥螨的DNA。使用在其他螨类和蚜虫中起作用的几对引物无法扩增普通真绥螨核糖体DNA的ITS区域。然而，在K株螨类中，其中一对ITS引物扩增出了一个担子菌核糖体DNA序列。最后，发现真菌核糖体DNA特异性引物在几个普通真绥螨株系中扩增出了一个约650 bp的ITS区域。设计用于扩增650 bp产物中部的内部引物成功地从所有螨类中扩增出了该产物。
