A Comparison of Morphological and Molecular-Based Surveys to Estimate the Species Richness of Chaetoceros and Thalassiosira (Bacillariophyta), in the Bay of Fundy

The goal of this study was to compare the ability of morphology and molecular-based surveys to estimate species richness for two species-rich diatom genera, Chaetoceros Ehrenb. and Thalassiosira Cleve, in the Bay of Fundy. Phytoplankton tows were collected from two sites at intervals over two years and subsampled for morphology-based surveys (2010, 2011), a culture-based DNA reference library (DRL; 2010), and a molecular-based survey (2011). The DRL and molecular-based survey utilized the 3′ end of the RUBISCO large subunit (rbcL-3P) to identify genetic species groups (based on 0.1% divergence in rbcL-3P), which were subsequently identified morphologically to allow comparisons to the morphology-based survey. Comparisons were compiled for the year (2011) by site (n = 2) and by season (n = 3). Of the 34 taxa included in the comparisons, 50% of taxa were common to both methods, 35% were unique to the molecular-based survey, and 12% were unique to the morphology-based survey, while the remaining 3% of taxa were unidentified genetic species groups. The morphology-based survey excelled at identifying rare taxa in individual tow subsamples, which were occasionally missed with the molecular approach used here, while the molecular methods (the DRL and molecular-based survey), uncovered nine cryptic species pairs and four previously overlooked species. The last mentioned were typically difficult to identify and were generically assigned to Thalassiosira spp. during the morphology-based survey. Therefore, for now we suggest a combined approach encompassing routine morphology-based surveys accompanied by periodic molecular-based surveys to monitor for cryptic and difficult to identify taxa. As sequencing technologies improve, molecular-based surveys should become routine, leading to a more accurate representation of species composition and richness in monitoring programs.     

Application of molecular genetics and geometric morphometrics to taxonomy and conservation of cave beetles in central Italy

Integration of molecular genetic techniques and geometric morphometrics represent a valuable tool in the resolution of taxonomic uncertainty and the identification of significant units for conservation. We combined mitochondrial DNA cytochrome c oxidase subunit II gene sequence data and geometric morphometric analysis to examine taxonomic status and identify units for conservation in four species of the hypogean beetle Duvalius (Coleoptera, Trechinae) using mainly museum specimens collected in central Italy. Previous taxonomic studies based on morphological traits described several subspecies often inhabiting geographically distinct caves. Phylogenetic analysis identified two well supported monophyletic lineages and a number of different clades with relatively small genetic differences, suggesting a short divergence time in line with known geological history of the study area. Geometric morphometrics, on the other hand, recovered a high level of distinctiveness among specimens. Both genetic and morphometric analyses did not entirely corroborate former taxonomic nomenclature, suggesting possible rearrangements and the definition of evolutionary significant units. Beetles of the genus Duvalius are protected by regional laws and the majority of taxa considered in this study inhabit caves located outside protected areas. Our study advocates the importance of devoting protection efforts to networks of cave ecosystems rather than single locations or species.     

After 7 years and 1000 citations: comparative assessment of the DNA barcoding and the DNA taxonomy proposals for taxonomists and non-taxonomists

In 2003, two different approaches-DNA taxonomy and DNA barcoding-were simultaneously proposed to overcome some of the perceived intrinsic weaknesses of the traditional morphology-based taxonomical system, and to help non-taxonomists to resolve their crucial need for accurate and rapid species identification tools. After 7 years, it seems unlikely that a completely new taxonomical system based on molecular characters only (DNA taxonomy) will develop in the future. It is more likely that both morphological and molecular data will be simultaneously analyzed, developing what has been coined as "integrative taxonomy". Concerning DNA barcoding, it is now clear that it does not focus on building a tree-of-life nor to perform DNA taxonomy, but rather to produce a universal molecular identification key based on strong taxonomic knowledge that is collated in the barcode reference library. The indisputable success of the DNA barcoding project is chiefly due to the fact that DNA barcoding standards considerably enhance current practices in the molecular identification field, and standardization offers virtually endless applications for various users.     

Molecular systematics and conservation of the langurs and leaf monkeys of South Asia

Numerous morphology-based classification schemes have been proposed for langurs and leaf monkeys of South Asia but there is very little agreement between them. An incorrect classification scheme when used as a basis for biogeographic studies can support erroneous hypotheses. Further, lack of taxonomic resolution will also confound conservation efforts, given that conservation biologists use traditional morphology-based-classification schemes to prioritize species for conservation. Here, I have revisited recent molecular phylogenetic studies done on langurs and leaf monkeys of South Asia. Results from these studies are in turn used to derive a rational and scientific basis for prioritizing species for conservation. Molecular data support the classification of langurs of the Indian subcontinent—Hanuman, Nilgiri and purple-faced langurs—in the genus Semnopithecus, whereas Phayre’s leaf monkey along with other Southeast Asian leaf monkeys form another distinct clade (Trachypithecus). The phylogenetic position of capped and golden langurs remains unresolved. Molecular data suggest that they are closely related to each other but this group might have evolved through past hybridization between Semnopithecus and Trachypithecus. Additionally, genetic data also support the splitting of the so-called Hanuman langurs into at least three species. The scores for taxonomic uniqueness of langurs and leaf monkeys of South Asia were revised using this molecular phylogeny-based classification. According to the revised scores, Phayres leaf monkey and golden langur are priority species for conservation followed by capped and Nilgiri langurs.     

淡水龟科具闭壳结构龟类分类及系统进化研究进展

闭壳龟类是淡水龟科(Bataguridae)中一群具有"闭壳"结构的水栖或半水栖龟类的总称,即腹甲的腹盾和胸盾之间以可动的铰链韧带相连,使背甲和腹甲可以全部或部分闭合,它们是盒龟属(Cistoclemmys)、闭壳龟属(Cuo-ra)、锯缘龟属(Pyxidea)、齿缘龟属(Cyclemys)和果龟属(Notochelys),前4个属的龟类在中国均有分布。近几十年来,由于食用、传统中医药开发和宠物贸易的兴起,导致许多闭壳龟类野生物种数量急剧减少,并且当前许多亚洲闭壳龟的分类工作过多的依赖于市场购买的物种样本,这就使我们对闭壳龟属及种的归属划分变的较困难,因此对闭壳龟类物种种的界限的划分及属间系统发育关系的研究仍然是一个艰巨的挑战。通过对淡水龟科具有闭壳结构龟类的形态学研究,细胞遗传学研究以及分子系统学研究等进行了综述,以期为闭壳龟类的相关保护研究提供参考。     

The taxonomic feedback loop: symbiosis of morphology and molecules

Here, we relate the ongoing taxonomic story of a species complex of problematic, cryptic Australian freshwater shrimp (Atyidae; Caridina) to highlight the relative strength and utility of different taxonomic methods in assessing species boundaries. We used popular 'DNA barcode' gene fragments cytochrome c oxidase 1 and 16S ribosomal DNA. We then assessed the morphological attributes of these specimens and developed an identification key to complement the molecular results, and conclude that, despite occasionally strident arguments in favour of either molecular or morphological taxonomy, the two are inseparably linked and form parts of a greater whole.     

Is DNA Barcoding Actually Cheaper and Faster than Traditional Morphological Methods: Results from a Survey of Freshwater Bioassessment Efforts in the United States?

Taxonomic identification accounts for a substantial portion of cost associated with bioassessment programs across the United States. New analytical approaches, such as DNA barcoding have been promoted as a way to reduce monitoring costs and improve efficiency, yet this assumption has not been thoroughly evaluated. We address this question by comparing costs for traditional morphology-based bioassessment, the standard Sanger sequencing-based DNA barcoding approach, and emerging next-generation (NGS) molecular methods. Market demand for molecular approaches is also assessed through a survey of the level of freshwater bioassessment effort in the United States across multiple habitat types (lakes, streams, wetlands) and indicators (benthic invertebrates, fish, algae). All state and regional level programs administered by public agencies and reported via agency web sites were included in the survey. Costs were based on surveys of labs and programs willing to provide such information. More than 19,500 sites are sampled annually across the United States, with the majority of effort occurring in streams. Benthic invertebrates are the most commonly used indicator, but algae and fish comprise between 35% and 21% of total sampling effort, respectively. We estimate that between $104 and $193 million is spent annually on routine freshwater bioassessment in the United States. Approximately 30% of the bioassessment costs are comprised of the cost to conduct traditional morphology-based taxonomy. Current barcoding costs using Sanger sequencing are between 1.7 and 3.4 times as expensive as traditional taxonomic approaches, excluding the cost of field sampling (which is common to both approaches). However, the cost of NGS methods are comparable (or slightly less expensive) than traditional methods depending on the indicator. The promise of barcoding as a cheaper alternative to current practices is not yet realized, although molecular methods may provide other benefits, such as a faster sample processing and increased taxonomic resolution.     

Applying DNA barcoding to conservation practice: a case study of endangered birds and large mammals in China

Owning to advantages over traditional species identification methods, DNA barcoding is suggested to be a promising tool in conservation research. However, the use of DNA barcoding to accurately identify unknown samples in conservation practices has not been well documented in the literature. To illustrate this issue, we implemented a survey of endangered birds and mammals in China based on mitochondrial Cytochrome c Oxidase subunit I (COI) gene. We included mostly confiscated specimens and non-invasive samples while concealing species information to simulate real-world scenarios of identification. In total, 47 avian and 39 mammalian specimen were re-identified by sequential analyses of online species assignment, genetic distances, phylogenetic reconstruction, and diagnostic nucleotide method. With this multiple analyses approach, 82 individuals were accurately assigned to the species level and four individuals to the genus level. 78.72% of the avian specimen and 87.18% of mammalian specimen identifications were consistent with morphological classification. Among those inconsistent with morphological classification, we identified several potential errors including misidentification based on morphology and mislabelling that may have occurred while combining results from different analytical methods. Our case study not only enriches the barcode database, but also reports a successful application of DNA barcoding identification to conservation practices, which could effectively facilitate species identification of unknown samples in conservation practices in the future.     

Defining hotspots of characteristic species for multiple taxonomic groups in the Netherlands

Biogeographical zonation based on single taxa poses major limitations on planning for nature conservation. This paper identifies biogeographical patterns of multiple taxa in the Netherlands, where no endemics are present at species level, on the basis of characteristic species. We used occurrence data on five species groups in order to identify spatially coherent, ecologically important regions. TWINSPAN was used to cluster grid squares according to similarity in species composition for each taxonomic group. Species that are characteristic of each of the clusters were identified using a preference index, and corresponding clusters among the taxonomic groups were identified with Kappa statistics. Regions containing characteristic species for several taxonomic groups were defined as ‘hotspots’. Stepwise discriminant analysis was then used to characterize these hotspots according to differences in environmental conditions. The analysis yielded five regions that are clearly distinct in terms of species composition for individual taxonomic groups. Each region is characterized by a set of unique species that occur in the zonation of at least two of the taxonomic groups. Stepwise discriminant analysis revealed significant environmental differences among these regions. The concept of hotspots as operationalized in this study can make nature conservation planning more efficient. In combination, the hotspots defined here comprise the majority of the species occurring in the Netherlands for the studied groups. Therefore, this regionalization should be taken into account when prioritizing nature conservation efforts.     

Colour patterns do not diagnose species: quantitative evaluation of a DNA barcoded cryptic bumblebee complex

Cryptic diversity within bumblebees (Bombus) has the potential to undermine crucial conservation efforts designed to reverse the observed decline in many bumblebee species worldwide. Central to such efforts is the ability to correctly recognise and diagnose species. The B. lucorum complex (Bombus lucorum, B. cryptarum and B. magnus) comprises one of the most abundant and important group of wild plant and crop pollinators in northern Europe. Although the workers of these species are notoriously difficult to diagnose morphologically, it has been claimed that queens are readily diagnosable from morphological characters. Here we assess the value of colour-pattern characters in species identification of DNA-barcoded queens from the B. lucorum complex. Three distinct molecular operational taxonomic units were identified each representing one species. However, no uniquely diagnostic colour-pattern character state was found for any of these three molecular units and most colour-pattern characters showed continuous variation among the units. All characters previously deemed to be unique and diagnostic for one species were displayed by specimens molecularly identified as a different species. These results presented here raise questions on the reliability of species determinations in previous studies and highlights the benefits of implementing DNA barcoding prior to ecological, taxonomic and conservation studies of these important key pollinators.     

Metabarcoding Is Powerful yet Still Blind: A Comparative Analysis of Morphological and Molecular Surveys of Seagrass Communities

In the context of the sixth wave of extinction, reliable surveys of biodiversity are increasingly needed to infer the cause and consequences of species and community declines, identify early warning indicators of tipping points, and provide reliable impact assessments before engaging in activities with potential environmental hazards. DNA metabarcoding has emerged as having potential to provide speedy assessment of community structure from environmental samples. Here we tested the reliability of metabarcoding by comparing morphological and molecular inventories of invertebrate communities associated with seagrasses through estimates of alpha and beta diversity, as well as the identification of the most abundant taxa. Sediment samples were collected from six Zostera marina seagrass meadows across Brittany, France. Metabarcoding surveys were performed using both mitochondrial (Cytochrome Oxidase I) and nuclear (small subunit 18S ribosomal RNA) markers, and compared to morphological inventories compiled by a long-term benthic monitoring network. A sampling strategy was defined to enhance performance and accuracy of results by preventing the dominance of larger animals, boosting statistical support through replicates, and using two genes to compensate for taxonomic biases. Molecular barcodes proved powerful by revealing a remarkable level of diversity that vastly exceeded the morphological survey, while both surveys identified congruent differentiation of the meadows. However, despite the addition of individual barcodes of common species into taxonomic reference databases, the retrieval of only 36% of these species suggest that the remaining were either not present in the molecular samples or not detected by the molecular screening. This finding exemplifies the necessity of comprehensive and well-curated taxonomic reference libraries and multi-gene surveys. Overall, results offer methodological guidelines and support for metabarcoding as a powerful and repeatable method of characterizing communities, while also presenting suggestions for improvement, including implementation of pilot studies prior to performing full “blind” metabarcoding assessments to optimize sampling and amplification protocols.     

Barcoding the Collembola of Churchill: a molecular taxonomic reassessment of species diversity in a sub‐Arctic area

Although their functional importance in ecosystems is increasingly recognized, soil‐dwelling micro‐arthropods are usually poorly known in comparison with their above‐ground counterparts. Collembola constitute a significant and species‐rich component of the soil biodiversity, but it remains a woefully understudied group because of the taxonomic impediment. The ever‐increasing use of molecular taxonomic tools, such as DNA barcoding, provides a possible solution. Here, we test the use of this approach through a diversity survey of Collembola from the vicinity of Churchill, Manitoba, Canada, and compare the results with previous surveys in the same area and in other sub‐Arctic regions. The systematic barcoding campaign at Churchill revealed a diverse collembolan fauna consisting of 97 species‐level MOTUs in six types of habitats. If all these MOTUs are confirmed as species, this richness would be far higher than prior records for Arctic Canada and could lead to reconsider the actual diversity of the group in Arctic environments.     

Sampling Plant Diversity and Rarity at Landscape Scales: Importance of Sampling Time in Species Detectability

Documenting and estimating species richness at regional or landscape scales has been a major emphasis for conservation efforts, as well as for the development and testing of evolutionary and ecological theory. Rarely, however, are sampling efforts assessed on how they affect detection and estimates of species richness and rarity. In this study, vascular plant richness was sampled in 356 quarter hectare time-unlimited survey plots in the boreal region of northeast Alberta. These surveys consisted of 15,856 observations of 499 vascular plant species (97 considered to be regionally rare) collected by 12 observers over a 2 year period. Average survey time for each quarter-hectare plot was 82 minutes, ranging from 20 to 194 minutes, with a positive relationship between total survey time and total plant richness. When survey time was limited to a 20-minute search, as in other Alberta biodiversity methods, 61 species were missed. Extending the survey time to 60 minutes, reduced the number of missed species to 20, while a 90-minute cut-off time resulted in the loss of 8 species. When surveys were separated by habitat type, 60 minutes of search effort sampled nearly 90% of total observed richness for all habitats. Relative to rare species, time-unlimited surveys had ∼65% higher rare plant detections post-20 minutes than during the first 20 minutes of the survey. Although exhaustive sampling was attempted, observer bias was noted among observers when a subsample of plots was re-surveyed by different observers. Our findings suggest that sampling time, combined with sample size and observer effects, should be considered in landscape-scale plant biodiversity surveys.     

昆虫病原线虫资源概况和分类技术进展

昆虫病原线虫是具有重要潜在应用价值的害虫生物防治资源,主要包括斯氏线虫科（Steinernematidae）的斯氏线虫属Steinernema与新斯氏线虫属Neosteinernema线虫和异小杆线虫科（Heterorhabditidae）的异小杆线虫属Heterorhabditis线虫。近10年来,分子生物学方法与传统的形态学方法相结合应用到线虫的鉴定与分类,昆虫病原线虫的分类进入稳定与发展时期,越来越多的新种或品系被发现及应用于生物防治。目前已描述的昆虫病原线虫种类达65种,其中斯氏线虫属52种,新斯氏线虫属1种,异小杆线虫属12种。本文整理列出了迄今报道的昆虫病原线虫种类及其来源,并综述了昆虫病原线虫分类现状以及鉴定与分类方法上的研究进展,重点阐述了分子生物学技术在昆虫病原线虫鉴定与分类的应用状况。     

Measuring beta‐diversity from taxonomic similarity

Question: The utility of beta (β‐) diversity measures that incorporate information about the degree of taxonomic (dis)similarity between species plots is becoming increasingly recognized. In this framework, the question for this study is: can we define an ecologically meaningful index of β‐diversity that, besides indicating simple species turnover, is able to account for taxonomic similarity amongst species in plots? Methods: First, the properties of existing measures of taxonomic similarity measures are briefly reviewed. Next, a new measure of plot‐to‐plot taxonomic similarity is presented that is based on the maximal common subgraph of two taxonomic trees. The proposed measure is computed from species presences and absences and include information about the degree of higher‐level taxonomic similarity between species plots. The performance of the proposed measure with respect to existing coefficients of taxonomic similarity and the coefficient of Jaccard is discussed using a small data set of heath plant communities. Finally, a method to quantify β‐diversity from taxonomic dissimilarities is discussed. Results: The proposed measure of taxonomic β‐diversity incorporates not only species richness, but also information about the degree of higher‐order taxonomic structure between species plots. In this view, it comes closer to a modern notion of biological diversity than more traditional measures of β‐di‐versity. From regression analysis between the new coefficient and existing measures of taxonomic similarity it is shown that there is an evident nonlinearity between the coefficients. This nonlinearity demonstrates that the new coefficient measures similarity in a conceptually different way from previous indices. Also, in good agreement with the findings of previous authors, the regression between the new index and the Jaccard coefficient of similarity shows that more than 80% of the variance of the former is explained by the community structure at the species level, while only the residual variance is explained by differences in the higher‐order taxonomic structure of the species plots. This means that a genuine taxonomic approach to the quantification of plot‐to‐plot similarity is only needed if we are interested in the residual system's variation that is related to the higher‐order taxonomic structure of a pair of species plots.     

Should universal guidelines be applied to taxonomic research?

In an attempt to facilitate the integration of various methods of species delimitation, Dayrat (2005 ) recommends a set of nomenclatorial guidelines. He proposes to restrict the application of new specific epithets where a recent taxonomic revision has not dealt with the totality of names and variation in the group, where specimens are not well-represented in collections, where DNA extraction from type specimens is not possible, and where putative novel species are supported only by a single type of data (e.g. morphological). Dayrat further recommends that putative novel species for which only one type of data exists be described with the abbreviation 'sp.', so as to avoid the permanent establishment of a specific epithet, as required by the current codes of nomenclature. Contradicting himself, Dayrat implies that putative novel species supported only by DNA sequences should be named as valid species. If adopted, Dayrat's guidelines would impede taxonomic progress, diminish the utility of taxonomy to its users (e.g. conservation biologists and biogeographers), and prevent the integration of methods of species delimitation.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 90 , 761–764.     

Lack of adequate taxonomic knowledge may hinder endemic mammal conservation in the Brazilian Atlantic Forest

The Atlantic Forest is one of the most diverse and threatened ecosystems of the world, being thus classified as one of the most important biodiversity hotspots. However, habitat loss, overexploitation, alien species, disease and pollution are not the only threats faced by native fauna and flora. The lack of adequate taxonomic knowledge hinders conservation and management efforts of endemic species. This is true even for mammals, which is the most charismatic group of animals and traditionally receive a good deal of attention from scientists and the public in general. A few examples show how this gap in local fauna information can be demise for species conservation, even misguiding management strategies: molecular data reveal a hidden marsupial diversity; the lack of taxonomic studies at the species level seriously threatens rodent conservation; and the taxonomic rearrangement of the genusBrachyteles revealed a new species and had a great impact on management strategies. New species are discovered, described and taxonomically rearranged at an astounding rate. We can only be successful in biodiversity conservation if we have at least a minimum level of knowledge about what we are trying to preserve. That is true both for researchers and for the general public. Recent taxonomic revisions may represent the turning point in Neotropical fauna knowledge, which, coupled with a greater awareness of local people about the rich biodiversity that dot their backyards, can represent a better conservation prospect for the endemics of the Atlantic Forest.     

Land use change has stronger effects on functional diversity than taxonomic diversity in tropical Andean hummingbirds

Land use change modifies the environment at multiple spatial scales, and is a main driver of species declines and deterioration of ecosystem services. However, most of the research on the effects of land use change has focused on taxonomic diversity, while functional diversity, an important predictor of ecosystem services, is often neglected. We explored how local and landscape scale characteristics influence functional and taxonomic diversity of hummingbirds in the Andes Mountains in southern Ecuador. Data was collected in six landscapes along a land use gradient, from an almost intact landscape to one dominated by cattle pastures. We used point counts to sample hummingbirds from 2011 to 2012 to assessed how local factors (i.e., vegetation structure, flowering plants richness, nectar availability) and landscape factors (i.e., landscape heterogeneity, native vegetation cover) influenced taxonomic and functional diversity. Then, we analyzed environment – trait relationships (RLQ test) to explore how different hummingbird functional traits influenced species responses to these factors. Taxonomic and functional diversity of hummingbirds were positively associated with landscape heterogeneity but only functional diversity was positively related to native vegetation coverage. We found a weak response of taxonomic and functional diversity to land use change at the local scale. Environment‐trait associations showed that body mass of hummingbirds likely influenced species sensitivity to land use change. In conclusion, landscape heterogeneity created by land use change can positively influence hummingbird taxonomic and functional diversity; however, a reduction of native vegetation cover could decrease functional diversity. Given that functional diversity can mediate ecosystem services, the conservation of native vegetation cover could play a key role in the maintenance of hummingbird pollination services in the tropical Andes. Moreover, there are particular functional traits, such as body mass, that increase a species sensitivity to land use change.     

中国淡水胭脂藻属植物形态及分子系统发育分析

以云南省玉溪市和河南省焦作市新采集的2株淡水胭脂藻标本为材料, 对其进行形态特征观察和分子系统发育分析, 联合我国已报道的其他胭脂藻序列信息, 构建该属植物可靠的系统发育关系。根据其现代地理分布模式, 对胭脂藻属进行了祖先地理起源重建。基于rbcL、psbA和UPA序列, 利用贝叶斯法、最大似然法和邻接法构建的系统发育树高度一致, 研究中的标本与鸡公山胭脂藻H. jigongshanensis聚合为一个独立分支, 并得到了很高的支持率, 根据分子生物学证据, 将其鉴定为鸡公山胭脂藻。通过研究中所采集2株鸡公山胭脂藻的形态特征比较, 发现传统的形态分类特征包括藻体高度和细胞直径, 在不同的环境条件下是可变的, 不适合作为胭脂藻属植物种类鉴定的依据。对于胭脂藻属这一形态结构较为简单的类群, 必须借助分子生物学证据和系统发育分析手段对其进行种类鉴定和系统发育分析。分子系统发育分析结果表明, 中国特有种鸡公山胭脂藻H. jigongshanensis形成单系类群, 其与日本胭脂藻H. japananense亲缘关系较近, 所有淡水胭脂藻种类形成一个独立的聚类群, 与海洋种类遗传差异较大。祖先地理重建的分析结果表明, 淡水胭脂藻的祖先起源地位于北美洲, 然后逐渐向东加勒比海区域、拉丁美洲和欧亚大陆扩散, 形成现代地理分布模式。胭脂藻属独特的种类地理分布特点, 可能与其较为古老的起源时间和漫长的进化历史相关, 同时淡水胭脂藻在北美洲区域起源时间较早, 进化历史漫长, 遗传多样性大, 这与该地理区域分布的种类H. angolensis为多系类群是相一致的。