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一种直接检测地衣中空气传播扩散的方法。

A method for the direct detection of airborne dispersal in lichens.

机构信息

Royal Botanic Garden Edinburgh, Edinburgh, UK.

College of Medical, Veterinary & Life Sciences (MVLS), University of Glasgow, Glasgow, UK.

出版信息

Mol Ecol Resour. 2018 Mar;18(2):240-250. doi: 10.1111/1755-0998.12731. Epub 2017 Nov 30.

Abstract

This study sets out a novel method to determine dispersal distances in lichens. Direct measurement of dispersal often remains difficult for lichens and other small inconspicuous species because of the need to track microscopic reproductive propagules, which even if they can be captured, cannot be identified using traditional morphological approaches. A low-cost device (<£200) was developed to trap the reproductive propagules of lichens, capable of sampling around 0.1 m of air per minute. In parallel, molecular techniques were developed to enable species-specific detection of propagules caught by the devices, with identification using novel species-specific primers and optimization of a standard DNA extraction and nested PCR protocol. The methods were tested for both their sensitivity and specificity against a suite of lichen epiphytes, differing in their reproductive mechanisms, dispersal structures and rarity. Sensitivity tests showed that the molecular techniques could detect a single asexual propagule (soredium or isidium), or as few as 10 sexual spores. As proof of concept, propagule traps were deployed into a wooded landscape where the target epiphytes were present. Extractions from deployed propagule traps were sequenced, showing that the method was able to detect the presence of the target species in the atmosphere. As far as we are aware, this is the first attempt to use mechanized propagule traps in combination with DNA diagnostics to detect dispersal of lichens. The tests carried out here point the way for future dispersal studies of lichen epiphytes and other passively dispersed microscopic organisms including fungi or bryophytes.

摘要

本研究提出了一种确定地衣扩散距离的新方法。由于需要追踪微观繁殖孢子,因此对于地衣和其他小型不显眼的物种来说,直接测量扩散通常仍然很困难,即使可以捕获这些孢子,也无法使用传统的形态学方法对其进行识别。开发了一种低成本设备(<200 英镑)来捕获地衣的繁殖孢子,每分钟能够采集约 0.1 米的空气。同时,开发了分子技术,使设备捕获的孢子能够进行种特异性检测,使用新的种特异性引物进行鉴定,并优化了标准 DNA 提取和巢式 PCR 方案。该方法针对一系列在繁殖机制、扩散结构和稀有程度上存在差异的地衣外生菌进行了敏感性和特异性测试。敏感性测试表明,分子技术能够检测到单个无性繁殖体(叶斑或叶状地衣)或少至 10 个有性孢子。作为概念验证,将繁殖体诱捕器部署到存在目标外生菌的林地景观中。从部署的繁殖体诱捕器中提取的样本进行测序,结果表明该方法能够检测到空气中目标物种的存在。据我们所知,这是首次尝试使用机械化繁殖体诱捕器结合 DNA 诊断来检测地衣的扩散。这里进行的测试为未来地衣外生菌和其他被动扩散的微生物(包括真菌或苔藓植物)的扩散研究指明了方向。

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