Syromyatnikov M Y, Kokina A V, Belyakova N A, Kozlova E G, Popov V N
Voronezh State University, Universitetskaya pl., Voronezh, 394018, Russia..
Zootaxa. 2018 Mar 13;4394(2):270-278. doi: 10.11646/zootaxa.4394.2.9.
Predatory mites from the Amblyseius (Neoseiulus) genus (Family Phytoseiidae, Order Parasitiformes) are widely used for protecting plants against pests, especially thrips. The differentiation of Amblyseius and Neoseiulus species by their morphological features is problematic despite the fact that they are taxonomically different genera. The Phytoseiidae family includes a lot of species that are extremely difficult to distinguish from each other. The discovery of new molecular genetic markers might considerably facilitate the express identification of commercial mite species. Despite their high morphological similarity, the three common commercial Amblyseius and Neoseiulus mite species (Neoseiulus cucumeris, Amblyseius swirskii, and Neoseiulus barkeri) differed significantly in the nucleotide sequences of DNA fragments containing the ITS1 and ITS2 internal transcribed spacers. We found that when PCR products of the amplified DNA fragments from A. swirskii, N. cucumeris, and N. barkeri were treated with a combination of AccB1I, AspLEI, and SspI endonucleases and the resulting products were separated by electrophoresis in agarose gel, the obtained picture was sufficiently specific to provide accurate identification of the analyzed mite species. The lengths of the digestion products were different enough to allow their resolution in agarose gels. The PCR-RFLP method developed by us allows the rapid and accurate identification of commercially used Amblyseius and Neoseiulus species without DNA sequencing. The combination of the three endonucleases (AccB1I, AspLEI, and SspI) and FaeI can be used for the differentiation of all other but less frequently commercially used Phytoseiidae mites.
植绥螨科（寄螨目）钝绥螨属（新小绥螨属）的捕食螨被广泛用于保护植物免受害虫侵害，尤其是蓟马。尽管钝绥螨属和新小绥螨属在分类学上是不同的属，但通过形态特征区分这两个属的物种存在问题。植绥螨科包含许多彼此极难区分的物种。新分子遗传标记的发现可能会极大地促进商业用螨物种的快速鉴定。尽管三种常见的商业用钝绥螨属和新小绥螨属捕食螨（胡瓜新小绥螨、斯氏钝绥螨和巴氏新小绥螨）在形态上高度相似，但包含ITS1和ITS2内部转录间隔区的DNA片段的核苷酸序列却有显著差异。我们发现，当用AccB1I、AspLEI和SspI内切酶组合处理斯氏钝绥螨、胡瓜新小绥螨和巴氏新小绥螨扩增DNA片段的PCR产物，并将所得产物在琼脂糖凝胶中进行电泳分离时，得到的图谱具有足够的特异性，能够准确鉴定所分析的螨类物种。消化产物的长度差异足够大，能够在琼脂糖凝胶中分辨出来。我们开发的PCR-RFLP方法无需DNA测序就能快速准确地鉴定商业用钝绥螨属和新小绥螨属物种。这三种内切酶（AccB1I、AspLEI和SspI）与FaeI的组合可用于区分所有其他但较少用于商业的植绥螨科螨类。
