基于环境DNA宏条形码的洱海鱼类多样性研究

研究使用环境DNA宏条形码(eDNA metabarcoding)检测洱海鱼类多样性,探索适用于洱海鱼类多样性监测和保护的新方法。通过水样采集、过滤、eDNA提取、遗传标记扩增、测序与生物信息分析的环境DNA宏条形码标准化分析流程,从洱海16个采样点中获得可检测的9个采样点数据,共检测出17种鱼类,其中土著种5种、外来种12种;鲫(Carassius auratus)、鳙(Hypophthalmichthys nobilis)、麦穗鱼(Pseudorasbora parva)、泥鳅(Misgurnus anguillicaudatus)和食蚊鱼(Gambusia affinis)为优势种。研究结果表明虽然环境DNA宏条形码无法完全替代传统的鱼类监测方法,但作为一种新兴的生物多样性监测手段,其可用于快速检测洱海鱼类多样性及其空间分布。     

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...     

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。     

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...     

基于环境DNA宏条形码技术研究黄河三角洲典型潮沟系统鱼类多样性及其对水文连通性的响应

水文连通对于维护滨海湿地生物多样性至关重要,鱼类多样性作为生物多样性的重要组成部分,了解其对不同水文连通强度的响应具有重要意义。本研究基于环境DNA宏条形码技术(e DNA metabarcoding)检测黄河三角洲典型潮沟系统鱼类多样性,分析鱼类物种分布对不同水文连通强度潮沟生境差异的响应特征。利用12S r RNA经典鱼类引物对采集自三级潮沟系统的水样进行高通量测序,共检测出鱼类55种,其中本地鱼类27种、非本地鱼类28种,物种组成以鲈形目为主。各样点序列丰度均较高的鱼类有矛尾刺虾虎鱼(Acanthogobius hasta)、鮻(Planiliza haematocheilus)、长体刺虾虎鱼(Acanthogobius elongatus)等。鱼类多样性在不同水文连通性潮沟间差异明显,其中,二级潮沟群落多样性水平、丰富度指数、物种种类及各种鱼类类群中的个体均匀程度等都明显高于其他两级潮沟。RDA分析显示有6种环境因子与鱼类群落结构显著相关(P<0.05),分别为:硅酸盐(Si O₃^2––Si)、硝酸盐(NO₃     

外来种入侵与物种多样性

在入侵生态学研究方面 ,物种多样性与生物入侵之间的关系已成为当前研究和争论的焦点。自Elton的经典假说提出以来 ,物种丰富度高的群落比物种贫乏的群落更能抵抗外来种入侵的观点得到广泛接受。一些理论模型和多样性处理实验支持了该假说。但现在越来越多的野外观测和实验研究开始对这一经典假说提出异议 ,甚至反对。同时 ,在入侵生态学广泛受到关注的今天 ,大量的实验研究也提出了一些新的观点。本文对Elton经典假说提出以来全球有关物种多样性与生物入侵关系的主要研究及其观点进行了评述 ,以期为我国有关研究工作的开展提供参考。     

利用宏DNA条形码研究浮游动物多样性——以鸭绿江口为例

为评价宏DNA条形码技术在我国海洋生物多样性监测中的应用潜力,采集了22份鸭绿江口浮游动物样品,分别利用宏条形码分子鉴定和形态鉴定方法对优势类群桡足类进行多样性的比较研究。结果显示:(1)利用宏条形码分子鉴定方法共鉴定出4目23科32属229个操作分类单元(Operational Taxonomic Units, OTUs),形态方法共鉴定出3目5科5属6种;同时,利用形态鉴定得到的分类阶元多数(目:100%、科:80%、属:80%)能用宏条形码分子鉴定方法鉴定出来,而宏条形码分子鉴定方法鉴定得到的分类阶元多数(目:25%、科:83%、属:88%)却未能用形态鉴定出来,表明宏条形码分子鉴定方法在鉴定物种丰富度方面具有明显优势。(2)利用宏条形码分子鉴定与形态鉴定桡足类的多样性指数呈显著的一致性(r=0.524,P=0.024),表明宏条形码鉴定方法与形态方法在评价物种多样性方面具有较好的可比性。本研究表明宏条形码分子鉴定方法在我国海洋浮游动物业务化监测中具有较高的应用潜力。     

入侵种的DNA条形码鉴定

生物安全研究的范畴涉及任何由生物威胁所造成的风险。随着害虫优先考虑级别的不断变化,以及国家和部门间相互协作的不断加强,对DNA分子鉴定技术的标准化提出了更迫切的要求,而DNA条形码的出现为此类问题的解决提供了很好的机遇。我们以前人对毒蛾和果蝇的研究为例,比较了DNA条形码技术与PCR-RFLP等传统方法的鉴定效果,在此基础上提出了建立一个可对不同入侵种进行快速和准确鉴定的条形码技术平台的构想。     

基于DNA宏条形码技术的蓝彩带蜂粉源植物多样性研究

蓝彩带蜂(Nomia chalybeate Smith, 1875)隶属隧蜂科彩带蜂属,是一种典型的广访花性野生蜜蜂,广泛分布于我国东部地区及日本、韩国、印度、缅甸等国家。本研究基于我国东部9个省份18个不同样地20份蓝彩带蜂蜂粮样本,包括安徽省(AHGZ、AHJZ)、重庆市(CQSMS、CQWS)、福建省(FJSM)、广西省(GXHP、GXLZ)、湖南省(HNHY、HNQY)、海南省(HNLS、HNWC)、江西省(JXGX、JXJG6、JXJG8、JXQY6、JXQY8、JXYZ)、山东省(SDLL)、浙江省(ZJCS)。利用DNA宏条形码研究该蜂粉源植物的多样性,为全面掌握该蜂的粉源植物及保护策略提供理论依据。结果表明该蜂粉源植物共涉及47科,99属,124种,其中黄荆(Vitex negundo)是最优势种、其次是罗伞树(Ardisia quinquegona)和山莓(Rubus corchorifolius)。Alpha多样性指数分析显示：SDLL样本的物种丰富度和多样性最高,FJSM样本的丰富度和多样性最低。不同样地的粉源植物多样性具有差异,但该蜂同一样地不同时期的粉源植物多...     

基于环境DNA宏条码和底拖网的珠江河口鱼类多样性

文章采用环境DNA宏条码和底拖网对珠江河口鱼类多样性进行了研究, 并对两种方法进行了比较。利用环境DNA宏条码检测到了175种鱼类, 而利用底拖网采集到了47种鱼类, 结合两种方法共检测出179种鱼类, 隶属于15 目63科128属。其中两种方法共同识别了鱼类43种, 占总检测物种的24.02%, 基于底拖网的调查未能收集到基于环境DNA宏条码检测到的大多数物种。根据Shannon指数和Simpson指数显示, DNA宏条码所检测珠江河口鱼类群落α多样性显著高于底拖网方法(P<0.05)。两种方法的PCoA结果均显示珠江河口鱼类群落存在空间结构, 基于环境DNA宏条码的分析显示空间重叠更多。两种方法基于冗余分析均显示溶解氧和盐度是影响鱼类群落结构的主要环境因子。研究表明, 环境DNA 宏条形码是一种环保且可靠的评估方法, 将其搭载到现有调查可以更好地了解河口鱼类多样性。     

Quantification of mesocosm fish and amphibian species diversity via environmental DNA metabarcoding

Freshwater fauna are particularly sensitive to environmental change and disturbance. Management agencies frequently use fish and amphibian biodiversity as indicators of ecosystem health and a way to prioritize and assess management strategies. Traditional aquatic bioassessment that relies on capture of organisms via nets, traps and electrofishing gear typically has low detection probabilities for rare species and can injure individuals of protected species. Our objective was to determine whether environmental DNA (eDNA) sampling and metabarcoding analysis can be used to accurately measure species diversity in aquatic assemblages with differing structures. We manipulated the density and relative abundance of eight fish and one amphibian species in replicated 206‐L mesocosms. Environmental DNA was filtered from water samples, and six mitochondrial gene fragments were Illumina‐sequenced to measure species diversity in each mesocosm. Metabarcoding detected all nine species in all treatment replicates. Additionally, we found a modest, but positive relationship between species abundance and sequencing read abundance. Our results illustrate the potential for eDNA sampling and metabarcoding approaches to improve quantification of aquatic species diversity in natural environments and point the way towards using eDNA metabarcoding as an index of macrofaunal species abundance.     

Environmental DNA detection of rare and invasive fish species in two Great Lakes tributaries

The extraction and characterization of DNA from aquatic environmental samples offers an alternative, noninvasive approach for the detection of rare species. Environmental DNA, coupled with PCR and next‐generation sequencing (“metabarcoding”), has proven to be very sensitive for the detection of rare aquatic species. Our study used a custom‐designed group‐specific primer set and next‐generation sequencing for the detection of three species at risk (Eastern Sand Darter, Ammocrypta pellucida; Northern Madtom, Noturus stigmosus; and Silver Shiner, Notropis photogenis), one invasive species (Round Goby, Neogobius melanostomus) and an additional 78 native species from two large Great Lakes tributary rivers in southern Ontario, Canada: the Grand River and the Sydenham River. Of 82 fish species detected in both rivers using capture‐based and eDNA methods, our eDNA method detected 86.2% and 72.0% of the fish species in the Grand River and the Sydenham River, respectively, which included our four target species. Our analyses also identified significant positive and negative species co‐occurrence patterns between our target species and other identified species. Our results demonstrate that eDNA metabarcoding that targets the fish community as well as individual species of interest provides a better understanding of factors affecting the target species spatial distribution in an ecosystem than possible with only target species data. Additionally, eDNA is easily implemented as an initial survey tool, or alongside capture‐based methods, for improved mapping of species distribution patterns.     

Marine water environmental DNA metabarcoding provides a comprehensive fish diversity assessment and reveals spatial patterns in a large oceanic area

Current methods for monitoring marine fish (including bony fishes and elasmobranchs) diversity mostly rely on trawling surveys, which are invasive, costly, and time‐consuming. Moreover, these methods are selective, targeting a subset of species at the time, and can be inaccessible to certain areas. Here, we used environmental DNA (eDNA), the DNA present in the water column as part of shed cells, tissues, or mucus, to provide comprehensive information about fish diversity in a large marine area. Further, eDNA results were compared to the fish diversity obtained in pelagic trawls. A total of 44 5 L‐water samples were collected onboard a wide‐scale oceanographic survey covering about 120,000 square kilometers in Northeast Atlantic Ocean. A short region of the 12S rRNA gene was amplified and sequenced through metabarcoding generating almost 3.5 million quality‐filtered reads. Trawl and eDNA samples resulted in the same most abundant species (European anchovy, European pilchard, Atlantic mackerel, and blue whiting), but eDNA metabarcoding resulted in more detected bony fish and elasmobranch species (116) than trawling (16). Although an overall correlation between fishes biomass and number of reads was observed, some species deviated from the common trend, which could be explained by inherent biases of each of the methods. Species distribution patterns inferred from eDNA metabarcoding data coincided with current ecological knowledge of the species, suggesting that eDNA has the potential to draw sound ecological conclusions that can contribute to fish surveillance programs. Our results support eDNA metabarcoding for broad‐scale marine fish diversity monitoring in the context of Directives such as the Common Fisheries Policy or the Marine Strategy Framework Directive.     

Environmental DNA reveals a mismatch between diversity facets of Amazonian fishes in response to contrasting geographical,environmental and anthropogenic effects

Freshwater ecosystems are among the most endangered ecosystem in the world. Understanding how human activities affect these ecosystems requires disentangling and quantifying the contribution of the factors driving community assembly. While it has been largely studied in temperate freshwaters, tropical ecosystems remain challenging to study due to the high species richness and the lack of knowledge on species distribution. Here, the use of eDNA-based fish inventories combined to a community-level modelling approach allowed depicting of assembly rules and quantifying the relative contribution of geographic, environmental and anthropic factors to fish assembly. We then used the model predictions to map spatial biodiversity and assess the representativity of sites surveyed in French Guiana within the EU Water Framework Directive (WFD) and highlighted areas that should host unique freshwater fish assemblages. We demonstrated a mismatch between the taxonomic and functional diversity. Taxonomic assemblages between but also within basins were mainly the results of dispersal limitation resulting from basin isolation and natural river barriers. Contrastingly, functional assemblages were ruled by environmental and anthropic factors. The regional mapping of fish diversity indicated that the sites surveyed within the EU WFD had a better representativity of the regional functional diversity than taxonomic diversity. Importantly, we also showed that the assemblages expected to be the most altered by anthropic factors were the most poorly represented in terms of functional diversity in the surveyed sites. The predictions of unique functional and taxonomic assemblages could, therefore, guide the establishment of new survey sites to increase fish diversity representativity and improve this monitoring program.     

Propagule pressure and the invasion risks of non-native freshwater fishes: a case study in England

European countries in general, and England in particular, have a long history of introducing non-native fish species, but there exist no detailed studies of the introduction pathways and propagules pressure for any European country. Using the nine regions of England as a preliminary case study, the potential relationship between the occurrence in the wild of non-native freshwater fishes (from a recent audit of non-native species) and the intensity ( i.e. propagule pressure) and diversity of fish imports was investigated. The main pathways of introduction were via imports of fishes for ornamental use ( e.g. aquaria and garden ponds) and sport fishing, with no reported or suspected cases of ballast water or hull fouling introductions. The recorded occurrence of non-native fishes in the wild was found to be related to the time (number of years) since the decade of introduction. A shift in the establishment rate, however, was observed in the 1970s after which the ratio of established-to-introduced species declined. The number of established non-native fish species observed in the wild was found to increase significantly ( P < 0·05) with increasing import intensity (log₁₀ x + 1 of the numbers of fish imported for the years 2000–2004) and with increasing consignment diversity (log₁₀ x + 1 of the numbers of consignment types imported for the years 2000–2004). The implications for policy and management are discussed.     

Effects of sampling seasons and locations on fish environmental DNA metabarcoding in dam reservoirs

Environmental DNA (eDNA) analysis has seen rapid development in the last decade, as a novel biodiversity monitoring method. Previous studies have evaluated optimal strategies, at several experimental steps of eDNA metabarcoding, for the simultaneous detection of fish species. However, optimal sampling strategies, especially the season and the location of water sampling, have not been evaluated thoroughly. To identify optimal sampling seasons and locations, we performed sampling monthly or at two‐monthly intervals throughout the year in three dam reservoirs. Water samples were collected from 15 and nine locations in the Miharu and Okawa dam reservoirs in Fukushima Prefecture, respectively, and five locations in the Sugo dam reservoir in Hyogo Prefecture, Japan. One liter of water was filtered with glass‐fiber filters, and eDNA was extracted. By performing MiFish metabarcoding, we successfully detected a total of 21, 24, and 22 fish species in Miharu, Okawa, and Sugo reservoirs, respectively. From these results, the eDNA metabarcoding method had a similar level of performance compared to conventional long‐term data. Furthermore, it was found to be effective in evaluating entire fish communities. The number of species detected by eDNA survey peaked in May in Miharu and Okawa reservoirs, and in March and June in Sugo reservoir, which corresponds with the breeding seasons of many of fish species inhabiting the reservoirs. In addition, the number of detected species was significantly higher in shore, compared to offshore samples in the Miharu reservoir, and a similar tendency was found in the other two reservoirs. Based on these results, we can conclude that the efficiency of species detection by eDNA metabarcoding could be maximized by collecting water from shore locations during the breeding seasons of the inhabiting fish. These results will contribute in the determination of sampling seasons and locations for fish fauna survey via eDNA metabarcoding, in the future.     

Unlocking biodiversity and conservation studies in high‐diversity environments using environmental DNA (eDNA): A test with Guianese freshwater fishes

Determining the species compositions of local assemblages is a prerequisite to understanding how anthropogenic disturbances affect biodiversity. However, biodiversity measurements often remain incomplete due to the limited efficiency of sampling methods. This is particularly true in freshwater tropical environments that host rich fish assemblages, for which assessments are uncertain and often rely on destructive methods. Developing an efficient and nondestructive method to assess biodiversity in tropical freshwaters is highly important. In this study, we tested the efficiency of environmental DNA (eDNA) metabarcoding to assess the fish diversity of 39 Guianese sites. We compared the diversity and composition of assemblages obtained using traditional and metabarcoding methods. More than 7,000 individual fish belonging to 203 Guianese fish species were collected by traditional sampling methods, and ~17 million reads were produced by metabarcoding, among which ~8 million reads were assigned to 148 fish taxonomic units, including 132 fish species. The two methods detected a similar number of species at each site, but the species identities partially matched. The assemblage compositions from the different drainage basins were better discriminated using metabarcoding, revealing that while traditional methods provide a more complete but spatially limited inventory of fish assemblages, metabarcoding provides a more partial but spatially extensive inventory. eDNA metabarcoding can therefore be used for rapid and large‐scale biodiversity assessments, while at a local scale, the two approaches are complementary and enable an understanding of realistic fish biodiversity.     

胶州湾近岸浅水区鱼类群落结构及多样性

根据2009年3月-2010年2月在胶州湾西北部近岸浅水区进行的鱼类逐月定置网调查数据,采用生态多样性指数和多元统计分析等方法研究了该海域的鱼类群落结构及多样性特征.结果表明:调查共鉴定出44种鱼类,其中海洋性27种,河口性鱼类17种.鱼类种类组成与底层水温和盐度有密切关系.胶州湾近岸浅水区鱼类群落Margalef种类丰富度指数(R)变化范围为0.33-3.50,Shannon-Wiener多样性指数(H’)为0.42-2.25,Pielou均匀度指数(J’)为0.23-0.93.聚类分析(CLUSTER)和多维标度排序(MDS)分析表明,不同鱼类种类组成的各月份样品可划分为3个组分:水温较低的冬季组G1(12月-翌年4月)、水温次高的春秋季组G2(5-6月、10-11月)和水温最高的夏季组G3(7-9月).其中G1的典型种主要为尖海龙(Syngnathus acus)、方氏云鳚(Enedrias fangi)和玉筋鱼(Ammodytes personatus)等冷温性常栖类群种类,G2的主要典型种为鳀鱼(Engraulis japonicus)和尖海龙,G3的主要典型种为鲐鱼(Scomber japonicus)、细条天竺鱼(Apogonichthys lineatus)和普氏缰虾虎鱼(Amoya pflaumi)等洄游性鱼类.单因子相似性分析(ANOSIM)表明,3个月份组间鱼类群落结构差异显著,不同月份组间两两差异均显著.生物-环境分析(BIOENV)表明,底层水温是影响胶州湾近岸浅水区鱼类群落结构时间变化的主要环境因子.在胶州湾内近岸浅水区发现40余种幼鱼,但占优势的主要为低值鱼类,表明胶州湾的产卵、肥育场功能有所下降,应加强胶州湾近岸水域生态环境和渔业资源的保护及修复.     

厦门市外来物种入侵现状及其风险评价指标体系

通过对厦门地区外来物种入侵状况的实地调查,对已有文献资料的查阅和专家咨询,整理了厦门市已入侵或值得警惕的外来植物名录。并从气候、土壤、植被生态群落结构、地貌特征、人为和自然干扰状况以及港口交通贸易等方面分析了厦门市外来生物入侵现状的原因。结合前人在外来生物入侵的风险评价方面的研究成果,针对厦门市的外来植物入侵现状,构建了适应厦门地区外来植物入侵风险评价指标体系框架,该指标体系由“移居与建群的可能性”、“危害与影响”和“预防与控制”3大部分,共计6个一级指标,17个二级指标构成。