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在经济重要性的 Tetranychidae 和 Phytoseiidae 物种中分离和特征分析多态微卫星标记和交叉扩增。

Isolation and characterization of polymorphic microsatellite markers in Tetranychus urticae and cross amplification in other Tetranychidae and Phytoseiidae species of economic importance.

机构信息

Unidad Asociada Entomología UJI-IVIA, Centro de Protección Vegetal y Biotecnología, Instituto Valenciano de Investigaciones Agrarias (IVIA), Ctra. Moncada-Náquera Km. 4.5, 46113 Moncada, Valencia, Spain.

出版信息

Exp Appl Acarol. 2012 May;57(1):37-51. doi: 10.1007/s10493-012-9529-x. Epub 2012 Feb 16.

Abstract

Tetranychus urticae Koch is a cosmopolitan phytophagous mite considered as the most polyphagous species among spider mites. Population genetic studies using molecular markers such as microsatellites have proven to be extremely informative to address questions about population structure, phylogeography and host preferences. The aim of this study was to increase the available molecular tools to gain insight into the genetic structure of T. urticae populations of citrus orchards, which might help in their management. Five microsatellite DNA libraries were developed using probes with the motifs CT, CTT, GT and CAC following the FIASCO protocol. Positive clones, those that included the insert with the microsatellite, were detected using the PIMA-PCR technique. Combinations of primers were designed on 22 out of 32 new microsatellites loci and their polymorphism was tested in four populations sampled along the eastern coast of Spain. Eleven successful amplifications were obtained. Cross amplification was tested in the tetranychids Aphlonobia histricina, Eutetranychus banksi, E. orientalis, Oligonychus perseae, Panonychus citri, Tetranychus evansi, T. okinawanus and T. turkestani, and the phytoseiids Amblyseius swirskii, A. cucumeris, A. andersoni, Euseius stipulatus, Neoseiulus barkeri, N. californicus, Phytoseiulus persimilis and Typhlodromus phialatus. Eight successful cross amplifications were obtained.

摘要

橘全爪螨(Tetranychus urticae Koch)是一种世界性的植食性螨类,被认为是所有叶螨中最具多食性的物种。使用微卫星等分子标记的种群遗传研究已被证明对于解决种群结构、系统地理学和宿主偏好等问题非常有帮助。本研究旨在增加可用的分子工具,深入了解柑橘园橘全爪螨种群的遗传结构,这可能有助于对其进行管理。本研究使用 CT、CTT、GT 和 CAC 基序的探针,根据 FIASCO 方案开发了 5 个微卫星 DNA 文库。使用 PIMA-PCR 技术检测到包含微卫星插入片段的阳性克隆。在 32 个新的微卫星位点中,有 22 个位点设计了引物组合,并在西班牙东海岸的 4 个种群中对其多态性进行了测试。获得了 11 次成功扩增。在四种植绥螨(Aphlonobia histricina、Eutetranychus banksi、E. orientalis、Oligonychus perseae、Panonychus citri、Tetranychus evansi、T. okinawanus 和 T. turkestani)和 5 种捕食螨(Amblyseius swirskii、A. cucumeris、A. andersoni、Euseius stipulatus、Neoseiulus barkeri、N. californicus、Phytoseiulus persimilis 和 Typhlodromus phialatus)中进行了交叉扩增测试,获得了 8 次成功的交叉扩增。

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