Druciarek Tobiasz, Lewandowski Mariusz, Tzanetakis Ioannis
Plant Pathology, University of Arkansas, Fayetteville, AR, 72701, USA.
Department of Applied Entomology, Faculty of Horticulture and Landscape Architecture, University of Life Sciences - SGGW, Nowoursynowska 159, 02-776, Warsaw, Poland.
Exp Appl Acarol. 2019 Jun;78(2):247-261. doi: 10.1007/s10493-019-00382-4. Epub 2019 May 25.
Eriophyoids affect crops around the globe directly or indirectly as virus vectors. Eriophyoid systematics initiated over a century ago, yet more than 90% of their fauna remain undescribed. Morphological identification is challenging because of a limited number of traits, cryptic speciation and complex life cycle reported for many species in the group. Nucleic acids extraction for mite identification is challenging due to their microscopic size with researchers using pooled samples leading to polymorphisms and inconclusive results. Identification of mite virus vectors is a tiresome task that could be simplified with a protocol that allows for the detection of viruses in the individual specimen. This communication describes an innovative, highly efficient extraction and detection pipeline. Direct Reverse Transcriptase - Polymerase Chain Reaction (Drt-PCR) assays were implemented in the molecular identification of eriophyoids and detection of viruses present in their bodies. The reverse transcription step allows for amplification from a single mite or egg, as in addition to the genomic DNA, it incorporates the abundant transcripts of targeted genes, whereas it also allows for the amplification of viruses. This communication provides an efficient, sensitive and cost-effective alternative that can be implemented in pest identification and detection as well as biological and ecological studies.
瘿螨作为病毒载体直接或间接地影响着全球的农作物。瘿螨分类学始于一个多世纪前，但仍有超过90%的瘿螨种类未被描述。由于该类群中许多物种的特征数量有限、存在隐存物种形成以及复杂的生命周期，形态学鉴定具有挑战性。由于瘿螨体型微小，提取核酸用于螨类鉴定具有挑战性，研究人员使用混合样本会导致多态性和不确定的结果。鉴定螨类病毒载体是一项繁琐的任务，若有一个能在单个样本中检测病毒的方案，这项任务便可简化。本通讯介绍了一种创新的、高效的提取和检测流程。直接逆转录-聚合酶链反应（Drt-PCR）检测法被用于瘿螨的分子鉴定及其体内病毒的检测。逆转录步骤允许从单个螨或卵进行扩增，因为除了基因组DNA外，它还包含靶向基因的丰富转录本，同时也允许对病毒进行扩增。本通讯提供了一种高效、灵敏且经济高效的替代方法，可用于害虫鉴定与检测以及生物学和生态学研究。
