环境DNA宏条形码在生物多样性研究与监测中的应用

环境DNA宏条形码(eDNA metabarcoding)技术通过提取水体、土壤、空气中的环境DNA,使用引物PCR扩增与高通量测序,进行物种鉴定与生物多样性评估.作为一种新的监测技术,相比于传统监测技术更加快捷、准确以及对自然环境的破坏小,因此在一定程度上改变了我们调查地球生物多样性的方式.本文综述了环境DNA宏条形...     

环境DNA技术在鱼类生态学中的应用研究进展

全球渔业衰退是21世纪人类面临的重要挑战之一。为了有效地遏制鱼类资源的衰退,精确的鱼类生态调查是其首要任务。传统的鱼类监测以渔获物采集与形态学鉴定为主,往往耗时耗力且效果不佳,已无法满足现阶段大尺度上的精确调查。环境DNA （eDNA）技术作为一种近年来新兴的鱼类生态调查方法,其与传统方法相比具有灵敏度高、经济高效、采样受限小且对生态系统无干扰的优势,目前其已被广泛地应用于鱼类物种监测、多样性调查、生物量评估以及繁殖活动监测等方面的研究。然而,eDNA技术在鱼类生态学研究的具体应用中暴露出的一些问题将会影响其监测结果的精确性,诸如操作流程的不规范、基因数据库的不完善以及eDNA在环境中生态学过程的不明确等。鉴于上述原因,首先对eDNA技术的发展历程、分析流程以及eDNA技术在鱼类生态学研究领域中的研究进展进行了综述,而后着重分析了eDNA技术的发展当前所面临的困难与挑战,并提出了相应的解决方案,最后对eDNA技术未来在鱼类生态学研究领域中的发展趋势做出了展望。通过本研究,以期能够为eDNA技术在鱼类生态学领域中的准确应用提供理论基础。     

DNA宏条形码技术在海洋浮游动物多样性和生态学研究中的应用

浮游动物是海洋生态系统的关键类群,其覆盖门类广泛,多样性高。传统形态鉴定技术需要检测人员具备专业的形态鉴定知识,且费时费力。宏条形码技术无需分离生物个体,而是提取拖网采集到的浮游动物混合样本的总DNA,或者水体中的环境DNA （eDNA）,依托高通量测序平台测序,能够实现对大规模样本快速、准确、经济的分析,在海洋浮游动物生态学研究中得到越来越广泛的应用。分析了DNA宏条形码技术常用的核糖体和线粒体分子标记,在浮游动物多样性和数量研究中的可靠性和不足,并给出在海洋浮游动物群落监测,食物关系分析及生物入侵早期预警等研究中的应用。未来,开发多基因片段组合条形码,发展完备的参考数据库及实现准确的量化研究是DNA宏条形码技术发展的重要方向。     

基于环境DNA宏条形码技术的秦淮河生物多样性研究

秦淮河是南京的母亲河,其生物多样性受城市化进程影响面临严重威胁,而物种资源调研是生物多样性保护的基础。环境DNA宏条形码技术较形态学监测是一种简单高效、灵敏度高的新型监测技术。为探究秦淮河浮游生物、底栖动物及鱼类的生物多样性,于2019年7月,采用环境DNA宏条形码技术对其进行了探究,并分析了秦淮河上下游间的差异及环境因子对其群落结构的影响。结果表明:秦淮河共监测到浮游动物13属22种407个操作分类单元(Operational Taxonomic Units, OTUs),浮游植物85属60种4445个OTUs,底栖动物16属17种212个OTUs,鱼类53属44种1663个OTUs。其中浮游动物以游泳轮虫目(Ploima)和双甲目(Diplostraca)为主,共占浮游动物63.37%,浮游植物以隐藻门(Cryptomonas)和褐藻门(Ochrophyta)为主,共占浮游植物88.11%,底栖动物中节肢动物门(Arthropoda)占比最高,达91.67%,鱼类中鲤形目(Cypriniformes)占比最高,达69.99%。与秦淮河历史形态学监测数据相比,环境DNA宏条形码技术在...     

环境DNA metabarcoding及其在生态学研究中的应用

环境DNA metabarcoding(eDNA metabarcoding)是指利用环境样本(如土壤、水、粪便等)中分离的DNA进行高通量的多个物种(或高级分类单元)鉴定的方法。近年来,该方法引起了学者的广泛关注,逐渐应用于生物多样性研究、水生生物监测、珍稀濒危物种和外来入侵物种检测等生态学领域。介绍环境DNA metabarcoding的含义和研究方法;重点介绍环境DNA metabarcoding在物种监测、生物多样性研究和食性分析等生态学领域中的应用;总结环境DNA metabarcoding应用于生态学研究领域面临的挑战并对该方法的发展进行展望。     

不同环境样本类型对蚌类环境DNA监测的差异研究

为了阐明在使用环境DNA宏条形码技术时不同环境样本类型如何影响蚌类物种的可检测性,于2021年冬季和春季在鄱阳湖分别采集表层水、底层水和沉积物进行环境DNA宏条形码分析,并结合传统方法采集验证。基于环境DNA宏条形码技术共检测到鄱阳湖蚌类33种,传统方法共采集蚌类18种,环境DNA宏条形码技术能检测出传统方法采集到的所有蚌类物种。表层水和底层水注释到的蚌类物种数均分别高于沉积物的,且表层水和底层水注释到的蚌类物种分别完全覆盖沉积物的。基于环境DNA宏条形码技术的蚌类α多样性水平季节差异不显著,但蚌类β多样性水平季节差异显著。表层水和底层水的蚌类多样性均显著高于沉积物样本的, Beta多样性分析也显示水体样本(表层水和底层水分别)和沉积物样本存在显著性差异。但表层水和底层水的蚌类多样性和群落结构均无显著差异。鄱阳湖蚌类群落结构与环境因子的关联分析表明水深(WD)、透明度(SD)、水温(WT)和总氮(TN)显著影响蚌类群落结构。环境DNA宏条形码技术在蚌类的多样性监测中可行,且采水样比采沉积物效果好,表层水和底层水无显著差异。     

两栖动物在水体污染生物监测中作为指示生物的研究概况

简单介绍了两栖动物作为指示生物在环境监测中的优越性及其研究历史和现状,同时也概述了我国学者在这方面的研究,总结了污染水体中部分蝌蚪的形态行为特征,并且提出了利用两栖动物的形态和行为模式建立水体污染生物监测仪器的可能性,为生物监测提供了科学依据。     

动物食性分析在生态学中的应用研究进展——基于DNA宏条形码技术

动物食性分析是动物营养生态学的重要研究手段,可用于解析动物与环境因素的关联性、捕食者与猎物之间的关系,以及动物物种多样性等科学问题。近年来,基于新一代测序技术的DNA宏条形码技术被广泛应用到生态学多个研究领域,极大地促进了生命科学交叉学科的发展。其中,DNA宏条形码技术在动物食性分析中具有高分辨、高效率、低样本量等优势,具有重要的应用前景。综述了基于DNA宏条形码技术的动物食性分析在生态学中的应用研究进展,并进一步总结了DNA宏条形码技术原理和食性分析方法,着重探讨了基于DNA宏条形码技术的动物食性分析在珍稀濒危动物保护、生物多样性监测、农业害虫防治等生态学研究领域中的应用,并对DNA宏条形码技术在动物食性分析中存在的问题及应用前景进行小结与展望。     

DNA条形码及其在生物多样性研究中的应用

DNA条形码是利用生物体内标准的、有足够变异的、易扩增且相对较短的DNA片段对物种进行快速准确鉴定的技术。自2003年DNA条形码相关概念提出以来广受关注,国内外相继开展了DNA条形码及信息系统建设研究,为DNA条形码技术的发展提供了坚实的研究基础和生物信息学分析平台。DNA条形码技术弥补了传统分类学的不足,为生物多样性研究提供了新的思路和方法。本文介绍了DNA条形码的产生与发展过程,国内外DNA条形码技术与信息系统建设研究进展,重点阐述了DNA条形码技术在物种鉴定、濒危物种保护、隐存种发现、生物多样性评估等研究领域中的应用。最后结合DNA条形码技术目前存在的问题,对其在相关研究领域的应用前景进行了展望。     

DNA条形码在生态学研究中的应用与展望

DNA条形码是利用标准的DNA片段对物种进行快速鉴定的技术,已在生物学各相关领域得到广泛应用。随着DNA条形码技术的不断发展和完善,已成功应用于生态学领域的相关研究中。本文综述了DNA条形码在物种快速鉴定和隐存种发现、群落系统发育重建和生态取证、群落内物种间相互关系研究等方面的应用,并介绍了DNAmetabarcoding技术和环境DNA条形码在生物多样性和生态学研究领域中的应用。最后,结合新的测序技术和未来大科学装置的发展,在相关数据库逐渐完善,新分析方法和计算模型不断开发使用的情景下,对DNA条形码在生态学相关领域的应用前景进行了展望。     

基于环境DNA-宏条形码技术的底栖动物监测及水质评价研究进展

底栖动物是淡水生态系统中物种多样性最高的类群,也是应用最广泛的水质监测指示生物之一。传统的底栖动物监测以形态学为基础,耗时费力,无法满足流域尺度大规模监测的需求。环境DNA-宏条形码技术是一种新兴的生物监测方法,其与传统方法相比优势在于采样方法简单、低成本、高灵敏度,不受生物样本和环境状况的影响,不依赖分类专家和鉴定资料,能够快速准确地对多个类群进行大规模、高通量的物种鉴定。然而,在实际应用中该方法的效果受诸多因素的影响,不同的方法、流程往往会产生差异较大的结果。鉴于此,着重分析总结了应用环境DNA-宏条形码技术监测底栖动物的关键影响因素,包括样品采集与处理流程、分子标记选择、引物设计、PCR偏好性、参考数据库的完整性及相应的优化。并基于此探讨了提高环境DNA-宏条形码技术在底栖动物监测效率和准确率的途径,以期为底栖动物环境DNA-宏条形码监测方案的制定提供可靠的参考。最后对该技术在底栖动物监测和水质评价中的最新发展方向进行了展望。     

Next‐generation monitoring of aquatic biodiversity using environmental DNA metabarcoding

Global biodiversity in freshwater and the oceans is declining at high rates. Reliable tools for assessing and monitoring aquatic biodiversity, especially for rare and secretive species, are important for efficient and timely management. Recent advances in DNA sequencing have provided a new tool for species detection from DNA present in the environment. In this study, we tested whether an environmental DNA (eDNA) metabarcoding approach, using water samples, can be used for addressing significant questions in ecology and conservation. Two key aquatic vertebrate groups were targeted: amphibians and bony fish. The reliability of this method was cautiously validated in silico, in vitro and in situ. When compared with traditional surveys or historical data, eDNA metabarcoding showed a much better detection probability overall. For amphibians, the detection probability with eDNA metabarcoding was 0.97 (CI = 0.90–0.99) vs. 0.58 (CI = 0.50–0.63) for traditional surveys. For fish, in 89% of the studied sites, the number of taxa detected using the eDNA metabarcoding approach was higher or identical to the number detected using traditional methods. We argue that the proposed DNA‐based approach has the potential to become the next‐generation tool for ecological studies and standardized biodiversity monitoring in a wide range of aquatic ecosystems.     

Occupancy in dynamic systems: accounting for multiple scales and false positives using environmental DNA to inform monitoring

Occupancy is an important metric to understand current and future trends in populations that have declined globally. In addition, occupancy can be an efficient tool for conducting landscape-scale and long-term monitoring. A challenge for occupancy monitoring programs is to determine the appropriate spatial scale of analysis and to obtain precise occupancy estimates for elusive species. We used a multi-scale occupancy model to assess occupancy of Columbia spotted frogs in the Great Basin, USA, based on environmental DNA (eDNA) detections. We collected three replicate eDNA samples at 220 sites across the Great Basin. We estimated and modeled ecological factors that described watershed and site occupancy at multiple spatial scales simultaneously while accounting for imperfect detection. Additionally, we conducted visual and dipnet surveys at all sites and used our paired detections to estimate the probability of a false positive detection for our eDNA sampling. We applied the estimated false positive rate to our multi-scale occupancy dataset and assessed changes in model selection. We had higher naïve occupancy estimates for eDNA (0.37) than for traditional survey methods (0.20). We estimated our false positive detection rate per qPCR replicate at 0.023 (95% CI: 0.016–0.033). When the false positive rate was applied to the multi-scale dataset, we did not observe substantial changes in model selection or parameter estimates. Conservation and resource managers have an increasing need to understand species occupancy in highly variable landscapes where the spatial distribution of habitat changes significantly over time due to climate change and human impact. A multi-scale occupancy approach can be used to obtain regional occupancy estimates that can account for spatially dynamic differences in availability over time, especially when assessing potential declines. Additionally, this study demonstrates how eDNA can be used as an effective tool for improved occupancy estimates across broad geographic scales for long-term monitoring.     

Making environmental DNA count

The arc of reception for a new technology or method – like the reception of new information itself – can pass through predictable stages, with audiences’ responses evolving from ‘I don't believe it’, through ‘well, maybe’ to ‘yes, everyone knows that’ to, finally, ‘old news’. The idea that one can sample a volume of water, sequence DNA out of it, and report what species are living nearby has experienced roughly this series of responses among biologists, beginning with the microbial biologists who developed genetic techniques to reveal the unseen microbiome. ‘Macrobial’ biologists and ecologists – those accustomed to dealing with species they can see and count – have been slower to adopt such molecular survey techniques, in part because of the uncertain relationship between the number of recovered DNA sequences and the abundance of whole organisms in the sampled environment. In this issue of Molecular Ecology Resources, Evans et al. ( 2015 ) quantify this relationship for a suite of nine vertebrate species consisting of eight fish and one amphibian. Having detected all of the species present with a molecular toolbox of six primer sets, they consistently find DNA abundances are associated with species’ biomasses. The strength and slope of this association vary for each species and each primer set – further evidence that there is no universal parameter linking recovered DNA to species abundance – but Evans and colleagues take a significant step towards being able to answer the next question audiences tend to ask: ‘Yes, but how many are there?’     

Testing multiple substrates for terrestrial biodiversity monitoring using environmental DNA metabarcoding

Biological surveys based on visual identification of the biota are challenging, expensive and time consuming, yet crucial for effective biomonitoring. DNA metabarcoding is a rapidly developing technology that can also facilitate biological surveys. This method involves the use of next generation sequencing technology to determine the community composition of a sample. However, it is uncertain as to what biological substrate should be the primary focus of metabarcoding surveys. This study aims to test multiple sample substrates (soil, scat, plant material and bulk arthropods) to determine what organisms can be detected from each and where they overlap. Samples (n = 200) were collected in the Pilbara (hot desert climate) and Swan Coastal Plain (hot Mediterranean climate) regions of Western Australia. Soil samples yielded little plant or animal DNA, especially in the Pilbara, probably due to conditions not conducive to long‐term preservation. In contrast, scat samples contained the highest overall diversity with 131 plant, vertebrate and invertebrate families detected. Invertebrate and plant sequences were detected in the plant (86 families), pitfall (127 families) and vane trap (126 families) samples. In total, 278 families were recovered from the survey, 217 in the Swan Coastal Plain and 156 in the Pilbara. Aside from soil, 22%–43% of the families detected were unique to the particular substrate, and community composition varied significantly between substrates. These results demonstrate the importance of selecting appropriate metabarcoding substrates when undertaking terrestrial surveys. If the aim is to broadly capture all biota then multiple substrates will be required.     

DNA复合条形码在太白山土壤动物多样性研究中的应用

DNA复合条形码技术(metabarcoding)将DNA条形码与高通量测序技术相结合,快速便捷地鉴定群落混合样本中的物种,成为监测群落中物种组成和丰富度的可靠方法。采用这一方法分析了秦岭太白山5种不同生境的中小型土壤动物多样性,共得到土壤动物3门9纲28目199科。群落组成分析显示生境的变化对土壤动物群落组成有一定的影响。α多样性分析显示土壤动物群落丰富度指数最高的生境为针叶林,最低的为农田;土壤动物群落多样性指数最高的生境为针叶林,最低的为落叶小叶林。群落相似性分析显示高山草甸、针叶林和农田3种生境的土壤动物群落组成相似性较高,落叶小叶林和落叶阔叶林的土壤动物群落组成与这3种生境的差异较大,落叶小叶林与落叶阔叶林的土壤动物群落组成差异也较大。