Background: Populations of the earthworm, Lumbricus rubellus, are commonly found across highly contaminated former mine sites and are considered to have under-gone selection for mitigating metal toxicity. Comparison of adapted populations with those found on less contaminated soils can provide insights into ecological processes that demonstrate the long-term effects of soil contamination. Contemporary sequencing methods allow for portrayal of demographic inferences and highlight genetic variation indicative of selection at specific genes. Furthermore, the occurrence of L. rubellus lineages across the UK allows for inferences of mechanisms associated with drivers of speciation and local adaptation.Results: Using RADseq, we were able to define population structure between the two lineages through the use of draft genomes for each, demonstrating an absence of admixture between lineages and that populations over extensive geographic distances form discrete populations. Between the two British lineages, we were able to provide evidence for selection near to genes associated with epigenetic and morphological functions, as well as near a gene encoding a pheromone. Earthworms inhabiting highly contaminated soils bare close genomic resemblance to those from proximal control soils. We were able to define a number of SNPs that largely segregate populations and are indicative of genes that are likely under selection for managing metal toxicity. This includes calcium and phosphatehandling mechanisms linked to lead and arsenic contaminants, respectively, while we also observed evidence for glutathione-related mechanisms, including metallothionein, across multiple populations. Population genomic end points demonstrate no consistent reduction in nucleotide diversity, or increase in inbreeding coefficient, relative to history of exposure.Conclusions: Though we can clearly define lineage membership using genomic markers, as well as population structure between geographic localities, it is difficult to resolve markers that segregate entirely between populations in response to soil metal concentrations. This may represent a highly variable series of traits in response to the heterogenous nature of the soil environment, but ultimately demonstrates the maintenance of lineage-specific genetic variation among local populations. L. rubellus appears to provide an exemplary system for exploring drivers for speciation, with a continuum of lineages coexisting across continental Europe, while distinct lineages exist in isolation throughout the UK.

High-throughput techniques based on restriction site-associated DNA sequencing (RADseq) are enabling the low-cost discovery and genotyping of thousands of genetic markers for any species, including non-model organisms, which is revolutionizing ecological, evolutionary and conservation genetics. Technical differences among these methods lead to important considerations for all steps of genomics studies, from the specific scientific questions that can be addressed, and the costs of library preparation and sequencing, to the types of bias and error inherent in the resulting data. In this Review, we provide a comprehensive discussion of RADseq methods to aid researchers in choosing among the many different approaches and avoiding erroneous scientific conclusions from RADseq data, a problem that has plagued other genetic marker types in the past.

We have determined the complete nucleotide (nt) sequence of the mitochondrial genome of an oligochaete annelid, the earthworm Lumbricus terrestris. This genome contains the 37 genes typical of metazoan mitochondrial DNA (mtDNA), including ATPase8, which is missing from some invertebrate mtDNAs. ATPase8 is not immediately upstream of ATPase6, a condition found previously only in the mtDNA of snails. All genes are transcribed from the same DNA strand. The largest noncoding region is 384 nt and is characterized by several homopolymer runs, a tract of alternating TA pairs, and potential secondary structures. All protein-encoding genes either overlap the adjacent downstream gene or end at an abbreviated stop codon. In Lumbricus mitochondria, the variation of the genetic code that is typical of most invertebrate mitochondrial genomes is used. Only the codon ATG is used for translation initiation. Lumbricus mtDNA is A + T rich, which appears to affect the codon usage pattern. The DHU arm appears to be unpaired not only in tRNAser(AGN), as is typical for metazoans, but perhaps also in tRNAser(UCN), a condition found previously only in a chiton and among nematodes. Relating the Lumbricus gene organization to those of other major protostome groups requires numerous rearrangements.

A total of 579 Megascolecidae species have been reported in China through 2017. Most belong to the genera Amynthas and Metaphire. The family’s rich diversity merits further investigation into its evolutionary history. The arc of research indicates that analysis based on taxonomical and molecular methods reveals the evolutionary history of the family Megascolecidae in China. This paper summarizes updated findings regarding origin, speciation and dispersal of Megascolecidae in China, and discusses the evolution of major familial characteristics (e. g. spermathecal pores, caeca). Several analyses suggest that the ancestors of Megascolecidae in China may have come from the Indo-China Peninsula. Speciation radiation likely occurred after the Cretaceous-Palaeogene mass extinction, and species richness increased rapidly during the Cenozoic period. Ancestral range reconstruction analysis shows that species-groups are polyphyletic and that evolutionary reversal often resulted in sharp evolution of caeca, so we suggest that the current taxonomic system of Megascolecidae, which is based on a few morphological characters, should be reconstructed. We also note that the specific mechanism of evolution in Megascolecidae has not been studied. Hence, future research to reveal the specific evolutionary mechanism of Megascolecidae earthworms requires more systematic sampling of this family, combined with morphological research, phylogeny construction and analysis of geographical patterns, geological history and environmental factors.

截至2017年中国已记录巨蚓科蚯蚓579种(亚种), 优势类群为远盲蚓属(Amynthas)与腔蚓属(Metaphire), 其丰富的物种多样性值得深入研究。近年来的研究指出联合形态分类与分子系统发育可较好地探讨中国巨蚓科蚯蚓的起源、分化与扩散。本文概述了中国巨蚓科蚯蚓物种的起源、分化时间以及扩散历程, 探讨了受精囊孔、盲肠形状等重要特征的演化, 分析了现行分类系统的缺陷。多项研究表明, 中国巨蚓科主要类群的祖先起源于中南半岛, 于白垩纪末期至新生代初期进入中国, 在新生代得以繁荣发展; 受精囊孔的对数或位置是多起源的, 盲肠形状演化中有祖征重现的现象, 故使用少数形态特征进行类群划分的现行分类系统有待改进。此外, 中国巨蚓科蚯蚓的具体演化机制及影响因素仍不明确。因此, 在未来研究中整合形态特征、分子数据与地理格局、地质历史及环境因子等信息, 定量分析类群演化与古地理、生物与非生物因素间的关系, 将有助于全面厘清中国巨蚓科蚯蚓演化的具体机制。

Earthworms have been perceived as benevolent soil engineers since the time of Charles Darwin, but several recent syntheses link earthworm activities to higher greenhouse gas emissions, less soil biodiversity, and inferior plant defense against pests. Our study provides new field-based evidence of the multiple direct and indirect impacts of earthworms on ecosystem functions within an ecological multifunctionality framework (i.e., aggregated measures of the ability of ecosystems to simultaneously provide multiple ecosystem functions). Data from a 13-year field experiment describing 21 ecosystem functions showed that earthworm presence generally enhanced multifunctionality by indirect rather than direct effects. Specifically, earthworms enhanced multifunctionality by shifting the functional composition toward a soil community favoring the bacterial energy channel and strengthening the biotic associations of soil microbial and microfaunal communities. However, earthworm-mediated changes in soil physical structure, pH, and taxonomic diversity were not related to multifunctionality. We conclude that the coordinated actions of earthworms and their associated soil biota were responsible for the maintenance of multifunctionality at high levels in this rice-wheat cropping system. Management of crop residue inputs and reduction of soil physicochemical disturbances should encourage beneficial earthworm effects and support multiple ecosystem services that are vital to sustainable agriculture.Copyright © 2019 Elsevier Ltd. All rights reserved.

Spatial and temporal aspects of the evolution of cryptic species complexes have received less attention than species delimitation within them. The phylogeography of the cryptic complex Hormogaster elisae (Oligochaeta, Hormogastridae) lacks knowledge on several aspects, including the small-scale distribution of its lineages or the palaeogeographic context of their diversification. To shed light on these topics, a dense specimen collection was performed in the center of the Iberian Peninsula - resulting in 28 new H. elisae collecting points, some of them as close as 760m from each other- for a higher resolution of the distribution of the cryptic lineages and the relationships between the populations. Seven molecular regions were amplified: mitochondrial subunit 1 of cytochrome c oxidase (COI), 16S rRNA and tRNA Leu, Ala, and Ser (16S t-RNAs), one nuclear ribosomal gene (a fragment of 28S rRNA) and one nuclear protein-encoding gene (histone H3) in order to infer their phylogenetic relationships. Different representation methods of the pairwise divergence in the cytochrome oxidase I sequence (heatmap and genetic landscape graphs) were used to visualize the genetic structure of H. elisae. A nested approach sensu Mairal et al. (2015) (connecting the evolutionary rates of two datasets of different taxonomic coverage) was used to obtain one approximation to a time-calibrated phylogenetic tree based on external Clitellata fossils and a wide molecular dataset. Our results indicate that limited active dispersal ability and ecological or biotic barriers could explain the isolation of the different cryptic lineages, which never co-occur. Rare events of long distance dispersal through hydrochory appear as one of the possible causes of range expansion.Copyright © 2017 Elsevier Inc. All rights reserved.

The Aporrectodea caliginosa species complex includes the most abundant earthworms in grasslands and agricultural ecosystems of the Paleartic region. Historically this complex consisted of the following taxa: A. caliginosa s.s.Savigny, 1826, A. trapezoides Dugés (1828), A. tuberculata (Eisen, 1874), and A. nocturna Evans (1946). These four taxa are morphologically very similar and difficult to differentiate because of their morphological variability. Consequently, their taxonomic status and their phylogenetic relationships have been a matter of discussion for more than a century. To study these questions, we sequenced the COII (686 bp), 12S (362 bp), 16S (1200 bp), ND1 (917 bp), and tRNAs(Asn-Asp-Val-Leu-Ala-Ser-Leu) (402 bp) mitochondrial and 28S (809 bp) nuclear gene regions for 85 European earthworms from 27 different localities belonging to the A. caliginosa species complex and four outgroup taxa. DNA sequences were analyzed using maximum parsimony, maximum likelihood, and Bayesian approaches of phylogenetic inference. The resulting trees were combined with morphological, ecological, and genomic evidence to test species boundaries (i.e., integrative approach). Our molecular analyses showed that A. caliginosa s.s. and A. tuberculata form a sister clade to A. trapezoides, A. longa, and A. nocturna, which indicates that A. longa is part of the A. caliginosa species complex. We confirm the species status of all these taxa and identify two hitherto unrecognized Aporrectodea species in Corsica (France). Moreover our analyses also showed the presence of highly divergent lineages within A. caliginosa, A. trapezoides, and A. longa, suggesting the existence of cryptic diversity within these taxa.

Soil organisms, including earthworms, are a key component of terrestrial ecosystems. However, little is known about their diversity, their distribution, and the threats affecting them. We compiled a global dataset of sampled earthworm communities from 6928 sites in 57 countries as a basis for predicting patterns in earthworm diversity, abundance, and biomass. We found that local species richness and abundance typically peaked at higher latitudes, displaying patterns opposite to those observed in aboveground organisms. However, high species dissimilarity across tropical locations may cause diversity across the entirety of the tropics to be higher than elsewhere. Climate variables were found to be more important in shaping earthworm communities than soil properties or habitat cover. These findings suggest that climate change may have serious implications for earthworm communities and for the functions they provide.Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.

There are many peregrine European earthworm species that are found in Siberia. In contrast, it is generally considered that the only Siberian species, E. n. nordenskioldi, was capable to disperse in the reverse direction, from Siberia into Europe. We studied genetic diversity of E. n. nordenskioldi in Southern Urals and Eastern Europe using the mitochondrial cox1 gene. We found that E. n. nordenskioldi from that region represents a new genetic lineage distinct from the previously known populations of this species from Siberia. Molecular clock estimates suggest that this newly found lineage separated from the rest of the species in Lower Pleistocene. Within the studied sample, we detected two geographically restricted groups, which also diverged long before the Holocene, one found in the East European Plain and the other restricted to the Urals. Those two groups were found in sympatry in only one population. Therefore, our results do not support the traditional viewpoint, suggesting that E. n. nordenskioldi is definitely not a recent invader in Europe.

The importance and beneficial effects of earthworms on soil structure and quality is well-established. In addition, earthworms have proved to be important model organisms for investigation of pollutant effects on soil ecosystems. In ecotoxicological investigations effects of various pollutants on earthworms were assessed. But some important issues regarding the effects of pollutants on earthworms still need to be comprehensively addressed. In this review several issues relevant to soil ecotoxicological investigations using earthworms are emphasized and guidelines that should be adopted in ecotoxicological investigations using earthworms are given. The inclusion of these guidelines in ecotoxicological studies will contribute to the better quantification of impacts of pollutants and will allow more accurate prediction of the real field effects of pollutants to earthworms.

Among oligochaetes, the Pheretima complex within the Megascolecidae is a major earthworm group. Recently, however, the systematics of the Pheretima complex based on morphology are challenged by molecular studies. Since little comparative analysis of earthworm complete mitochondrial genomes has been reported yet, we sequenced mitogenomes of four pheretimoid earthworm species to explore their phylogenetic relationships. The general earthworm genomic features are also found in four earthworms: all genes transcribed from the same strand, the same initiation codon ATG for each PCGs, and conserved structures of RNA genes. Interestingly we find an extra potential tRNA-leucine (CUN) in Amynthas longisiphonus. The earthworm mitochondrial ATP8 exhibits the highest evolutionary rate, while the gene CO1 evolves slowest. Phylogenetic analysis based on protein-coding genes (PCGs) strongly supports the monophyly of the Clitellata, Hirudinea, Oligochaeta, Megascolecidae and Pheretima complex. Our analysis, however, reveals non-monophyly within the genara Amynthas and Metaphire. Thus the generic divisions based on morphology in the Pheretima complex should be reconsidered.Copyright © 2015 Elsevier B.V. All rights reserved.