ITS2 Secondary Structure Research Articles

ITS secondary structure reconstruction to resolve taxonomy and phylogeny of the Betula L. genus.

The taxonomy and phylogeny of the Betula L. genus remain unresolved and are very difficult to assess due to several factors, especially because of frequent hybridization among different species. In the current study, we used nucleotide sequences of two internal transcribed spacer regions (ITS1 and ITS2), which are commonly used as phylogenetic markers. In addition to their nucleotide variation we reconstructed their secondary structure and used it to resolve phylogenetic relationships of some birch species. We explored whether consideration of secondary structure in phylogenetic analyses based on neighbor-joining, maximum parsimony, maximum likelihood, and Bayesian inference methods would help us obtain more solid support of the reconstructed phylogenetic trees. The results were not unambiguous. There were only a few clades with higher support when secondary structure was included into analysis. The phylogenetic trees generated using different methods were mostly in agreement with each other. However, the resolving power of these markers is still insufficient to reliably discriminate some closely related species. To achieve this aim more reliably there is a need for application of modern genomic approaches in combination with traditional ones.

Molecular phylogeny and morphology of Cephaleuros (Trentepohliales, Chlorophyta) from southern China

ABSTRACT Cephaleuros is a subaerial green alga widely distributed in tropical and subtropical regions. In this study, we investigated the taxonomy of Cephaleuros in southern China. Eight morphospecies (C. virescens, C. karstenii, C. expansus, C. drouetii, C. diffusus, C. tumidae-setae, C. lagerheimii and C. parasiticus) were identified according to previous descriptions, among which C. karstenii was the most frequently recorded. Subsequently, molecular phylogeny of Cephaleuros was inferred from 18S rDNA, ITS rDNA, 28S rDNA and rbcL sequences. Cephaleuros was subdivided into nine clades based on ITS analysis, which were recovered in phylogenies of two concatenated datasets (18S+ITS+28S and 18S+ITS+28S+rbcL). Molecular analyses revealed that Cephaleuros was composed of two main lineages, with one lineage comprising mostly samples of C. karstenii and C. virescens and the other comprising other species. The molecular phylogenies were not congruent with conventional morphological classifications, although some clades were distinguished by one or more diagnostic features. While the morphological features used to distinguish Cephaleuros spp were not reliable, the rbcL gene proved to be a good marker for identification of Cephaleuros. The ITS2 secondary structure prediction model showed that Cephaleuros spp have one ring structure with four helices, with Helix III being the longest without a branch in clades 3, 5 and 8. The present study represents the most detailed taxonomic assessment of Cephaleuros available to date. Further taxonomic research incorporating samples from the type locality is necessary for the reassessment of C. virescens.

Application of the ITS secondary structure in the identification of Nostoc species

The study of the species composition of Nostocales cyanobacteria in samples taken from different steppe ecosystems allowed us to analyze for the first time the diversity of the genus Nostoc in the Ukrainian steppe using molecular genetic methods. In the analysis, special attention was paid to the study of the predicted secondary structure of the ITS Box-B and V3 loops. During the analysis, it was shown that a number of isolated Nostoc strains, in addition to morphological ones, have significant genetic differences, which will allow us to propose new species for science based on further research.

Monilinema gen. nov., a homocytous genus (Cyanobacteria, Leptolyngbyaceae) from saline-alkaline lakes of Pantanal wetlands, Brazil.

Cyanobacteria morphotypes with simple morphology, mainly thin filamentous homocytous strains, comprise a taxonomically complex group and represent a challenge in systematic studies. However, the polyphasic approach applied nowadays to investigate the cyanobacterial diversity has emerged as a powerful tool to undercover cryptic taxa and to set up a more natural classification system. Yet, studies exploring the cultured diversity of cyanobacteria from extreme tropical environments have paved the way to the discovery of new cyanobacteria taxa, enabling the description of Pantanalinema, Alkalinema, and Cephalothrix as novel genera from saline-alkaline lakes in Pantanal wetlands (Brazil). The present study investigated a set of cyanobacterial strains resembling Leptolyngbya morphotypes by means of its morphology, 16S ribosomal RNA (16S rRNA) phylogeny, and ITS secondary structures. Based on the shape and structure of their trichomes, the low levels of 16S rRNA identity coupled with the distinct phylogenetic position with regard to well establish genera, and their divergent ecological feature, these strains must be accommodated into a novel genus, named as Monilinema gen. nov., described under the provisions of the International Code of Nomenclature for algae, fungi, and plants. Accordingly, based on the singularity of the D1-D1' and V3 helixes, these strains should be considered as a single species, Monilinema alkalinum.

Two New and Nontoxigenic Pseudo-nitzschia species (Bacillariophyceae) from Chinese Southeast Coastal Waters.

To explore the species diversity and toxin profile of Pseudo-nitzschia, monoclonal strains were established from Chinese southeast coastal waters. The morphology was examined under light and transmission electron microscopy. The internal transcribed spacer region of ribosomal DNA was sequenced for phylogenetic analyses, and the secondary structure of ITS2 was predicted and compared among allied taxa. A combination of morphological and molecular data showed the presence of two new species, Pseudo-nitzschia hainanensis sp. nov. and Pseudo-nitzschia taiwanensis sp. nov. Pseudo-nitzschiahainanensis was characterized by a dumpy-lanceolate valve with slightly blunt apicesand a central nodule, as well as striae comprising two rows of poroids. Pseudo-nitzschiataiwanensis was characterized by a slender-lanceolate valve, and striae comprising one row of split poroids. The poroid structure mainly comprised two sectors. Both taxa constituted their own monophyletic lineage in the phylogenetic analyses inferred from ITS2 rDNA and were well differentiated from other Pseudo-nitzschia species. Morphologically, P.hainanensis and P.taiwanensis could be assigned to the Pseudo-nitzschia delicatissima and the Pseudo-nitzschia pseudodelicatissima complex, respectively. Particulate domoic acid was measured using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS), but no detectable pDA was found. With the description of the two new species, the species diversity of genus Pseudo-nitzschia reaches 58 worldwide, among which 31 have been recorded from Chinese coastal waters.

The occurrence of Affixifilum gen. nov. and Neolyngbya (Oscillatoriaceae) in South Florida (USA), with the description of A. floridanum sp. nov. and N. biscaynensis sp. nov.

South Florida (USA) has a subtropical to tropical climate with an extensive and diverse coastline that supports the growth of benthic cyanobacterial mats (BCMs). These BCMs are widespread and potentially house numerous bioactive compounds; however, the extent of the cyanobacterial diversity within these mats remains largely unknown. To elucidate this diversity, BCMs from select locations in South Florida were sampled and isolated into unicyanobacterial cultures for morphological and molecular studies. Phylogenetic relationships of isolated taxa were assessed using the markers 16S rRNA and 16S-23S rRNA ITS by both maximum likelihood and Bayesian inference. We propose Affixifilum gen. nov. based on morphological characteristics and the 16S rRNA phylogeny. Two species are included: Affixifilum granulosum comb nov. (=Neolyngbya granulosa) found in Brazil and Florida (USA) and A. floridanum sp. nov. Several other features, including pair-wise distance of 16S rRNA and 16S-23S rRNA ITS, 16S-23S rRNA ITS secondary structure, morphology, and ecology, provide support for Affixifilum. We also propose the transfer of Lyngbya regalis to Neolyngbya as N. regalis comb. nov. and include the description of one novel species, N. biscaynensis sp. nov.

Purpureonostoc, a new name for a recently described genus of Nostoc-like cyanobacteria

A new cyanobacterial genus was recently named Purpurea, based on morphology, 16S rRNA gene sequence and 16-23S ITS secondary structure, which differentiated it from the other Nostoc-like genera (Cai et al. 2020). designation is invalid as it is contrary to Art. 20.2 (Art. 20.2. The name of a genus may not coincide with a Latin technical term in use in morphology at the time of publication unless it was published before 1 January 1912.). As this new genus is similar to Nostoc, Purpurea F. Cai et R. Li nom. inval. is here renamed as Purpureonostoc.

Nucleotide variation and secondary structures of nuclear ribosomal ITS2 in phylogeny relationships of some wheat species

This research aimed to estimate the nucleotide polymorphism and point mutation/substitutions for ITS2 region in 12 different Triticum and Aegilops species, and to demonstrate their phylogenetic relationships. The aligned ITS2 dataset showed a total of 67 SNPs and transitions (75%) predominant over transversions (25%). The C/T transitions were observed at highest level in the studied species. The highest nucleotide substitutions occurred in ITS2 sequences of T. aestivum, T. turgidum, and A. speltoides that showed similar substitutions on the base sites 76, 193, 215, 230 and 241 (TCCAG changed to ATTTA). T. boeticum and T. urartu showed similar substitutions on the base sites 85, 201, 231, 234 and 252 (GTGCC changed to AGATA). Remarkably, the nucleotide of base site 146 was C in all Triticum species, but T in all Aegilops species. The substitution rate in A. neglecta, and A. umbellulata was zero, it was concluded that their ITS2 is conserved. The secondary structures of ITS2 in the studied species was similar to that of the other flowering plants with four helices and the variation in helices length. The helix III had the longest length compared to the other helices. The length ranges of helices I, II, IV varied from 9 (T. turgidum, and T. urartu) to 10, 11 (A. cylindrica) 13 and 6 (A. speltoides) and 10 (T. boeticum) paired bases, respectively. Using pairwise ITS2 nucleotide comparisons, the closeness of A. caudata with A. cylindrica, T. boeticum with T. urartu, A. neglecta with A. umbellulata was observed. Phylogeny trees classified 12 species into four main clades. The highest point mutations occurred in ITS2 sequences related to the species grouped in the first clade.

Occurrence of Pseudo-nitzschia species and associated domoic acid production along the Guangdong coast, South China Sea

The diatom genus Pseudo-nitzschia, which has been associated with amnesic shellfish poisoning events globally, is also one of the key harmful microalga groups in Guangdong coastal waters, off the north coast of the South China Sea. In order to explore the diversity and toxigenic characteristics, Pseudo-nitzschia isolates were established. Based on a combination of morphological and molecular features, in total 26 different Pseudo-nitzschia taxa were identified, including two new species, P. uniseriata H.C. Dong & Yang Li and P. yuensis H.C. Dong & Yang Li. Morphologically, P. uniseriata is unique by having striae mainly comprising one row of poroids, which are simple without divided hymen internally, and each poroid containing one, seldom two sectors. Pseudo-nitzschia yuensis is characterized by having striae comprising one to two rows of poroids. In biseriate striae, the poroids are polygonal and irregularly distributed, and a discontinuous row of poroids may be present in the middle. In uniseriate striae, the poroids usually contain 1–5 sectors. Both taxa are well differentiated from other Pseudo-nitzschia species in phylogenetic analyses inferred from ITS2 sequence-structure information. Pseudo-nitzschia uniseriata is sister to P. lineola, whereas P. yuensis forms a group together with P. micropora and P. delicatissima. When comparing ITS2 secondary structure, two hemi-compensatory base change (HCBCs) are found between P. uniseriata and P. lineola. One compensatory base change (CBC) and four HCBCs are found between P. yuensis and P. delicatissima, and there is one CBC and five HCBCs between P. yuensis and P. micropora. The ability of cultured strains to produce particulate DA (pDA) revealed production of pDA in twenty-nine strains belonging to seven species: P. bipertita, P. caciantha, P. cuspidata, P. fraudulenta, P. fukuyoi, P. lundholmiae and P. multiseries. This is the first report of P. bipertita being toxic, with pDA content of 15.6 ± 2.1 fg cell–1. The presence of brine shrimps significantly increased pDA content in P. cuspidata, P. fukuyoi, P. lundholmiae and P. multiseries 1.4 to 7 times, and induced pDA production in P. fraudulenta from below detection limit to 17.5 ± 1.6 fg cell–1. The highest pDA concentration, 4830.5 ± 120.3 fg cell–1, was detected in P. multiseries, a level much lower than previous reports on P. multiseries from North America and Europe. Overall, the cellular toxin levels in Pseudo-nitzschia spp. were low in Guangdong coastal isolates.

Fortiea necridiiformans sp. nov., a soil-dwelling cyanobacterium from Pachmarhi Biosphere Reserve, India.

Cyanobacterial strain PS4G was isolated from seepage soil sampled at Pachmarhi, Madhya Pradesh, India, and was characterized using a polyphasic approach. The results of morphological analysis showed that strain PS4G had unique morphological characteristics which were not observed in the other described species of the genus Fortiea. In the 16S rRNA gene phylogenetic analysis inferred using Bayesian inference, maximum-likelihood and neighbour-joining methods, strain PS4G clustered within the clade consisting of the members of the genus Fortiea. Furthermore, in the secondary structure analysis using the D1-D1' helix and BoxB regions of 16S-23S ITS, strain PS4G showed marked differences in comparison with other members of the genus Fortiea. Overall, the morphological, phylogenetic and folded 16S-23S ITS secondary structure examination indicated that strain PS4G represents a novel species of the genus Fortiea. In accordance with the International Code of Nomenclature of Algae, Fungi and Plants we describe a novel species of Fortiea with the name Fortiea necridiiformans sp. nov.

Phylogeographical patterns of Campanula gr. arvatica, an endemic group of the Cantabrian mountains (NW Iberian Peninsula), based on plastid and nuclear DNA polymorphisms

Endemicity is a key consideration for conservation, and taxa that are endemic to a single country are vaulable conservation targets. In this study, we investigate endemic species of the genus Campanula in northern Iberian Peninsula based on sequences of nrDNA (internal transcribed spacer, ITS) and plastid regions (rbcL, trnL-F and trnS-G) and additionally the secondary structure of ITS2. The main aim of this work was to evaluate the evolutionary history of Campanula gr. arvatica from the Cantabrian Range (northern Iberian Peninsula) and establish phylogeographical patterns. Our results showed that molecular data from nuclear DNA and plastid regions should be applied for management and conservation strategies to monitor this endemic group from northern Spain. Furthermore, molecular evidence indicates significant differences among the three taxa analysed and shows the importance of the Cantabrian Mountains as a refuge for endemic Iberian flora in which processes of neo-speciation and/or crypto-speciation are under way.

<p><strong>Taxonomy and nomenclature of the recently published chlamydomonad genera <em>Ostravamonas</em> and <em>Paludistella</em> (Volvocales, Chlorophyceae)</strong></p>

A recently published chlamydomonad green algal genus, Paludistella H. Susanti, Mas. Yoshida, Tak. Nakayama, Nakada & M.M. Watanabe (2020: 72), was found to be superfluous as it originally included the type of Ostravamonas Barcytë & Hodač in Barcytë et al. (2020: 214). Here, the taxonomic and nomenclatural status under these generic names were examined based on morphological, phylogenetic, and ITS-2 secondary structure analyses, proposing two new combinations for P. asymmetrica H. Susanti, Mas. Yoshida, Tak. Nakayama, Nakada & M.M. Watanabe (2020: 76) and P. trianguloculus H. Susanti, Mas. Yoshida, Tak. Nakayama, Nakada & M.M. Watanabe (2020: 76) under Ostravamonas.

Description of novel species of Aliinostoc, Desikacharya and Desmonostoc using a polyphasic approach.

Five cyanobacterial strains with Nostoc-like morphology from different localities of the Mazandaran province of Iran were characterized using a polyphasic approach. Three strains clustered within the Aliinostoc clade whereas one each of the remaining two strains clustered within the genera Desmonostoc and Desikacharya. The phylogenetic positioning of all the strains by the bayesian inference, neighbour joining and maximum parsimony methods inferred using 16S rRNA gene indicated them to represent novel species of the genera Aliinostoc, Desmonostoc and Desikacharya. The 16S-23S ITS secondary structure analysis revealed that all five strains under study represented novel species unknown to science. In accordance with the International Code of Nomenclature for algae, fungi and plants we describe three novel species of the genus Aliinostoc and one species each of the genera Desmonostoc and Desikacharya.

Chlorocladiella gen. nov. (Pithophoraceae, Cladophorales, Chlorophyta), Including Four New Species From Various Freshwater Habitats in China.

Samples of Pithophoraceae, collected in diverse freshwater and damp terrestrial habitats from tropical and subtropical China, were characterized morphologically and ultrastructurally, and their phylogenetic position was determined based on nuclear ribosomal DNA sequences. Our phylogenetic analysis resolved a novel lineage of Pithophoraceae, sister to Aegagropilopsis. Based on our phylogenetic results, morphological observations and comparative rDNA ITS2 secondary structure analysis, we propose Chlorocladiella gen. nov., characterized by a well-developed system of prostrate filaments, and describe four new species, C.cochlea sp. nov., C. erecta sp. nov., C.medogensis sp. nov., and C. pisformis sp. nov. Two species were found growing on damp soil, which is an unusual habitat for cladophoralean green algae, indicating that the diversity of Cladophorales in terrestrial habitats may be greater than currently recognized.

Structural characterization of ITS2 and CBC species concept applications in the tribe Coluteocarpeae (Brassicaceae)

The taxonomic utility of internal transcribed spacer 2 ITS2 secondary structures in different plant groups, as well as in Brassicaceae, has been addressed by many studies. Although characterization and applications of ITS2 secondary structures for the members of main Brassicaceae lineages Lineages I, II, III, and expanded Lineage II have been studied, the utility of compensatory base change CBC species concept has not been the subject of any studies thus far. In the current study, the ITS2 secondary structures of 49 Coluteocarpeae expanded Lineage II specimens were investigated to determine relationships among the species. In addition to the utility of the CBC species concept, the availability of hemi-CBC and nonstructural substitutions NSTs , which are also used for generic and species delimitation, were tested and discussed. A maximum likelihood tree, based on the sequence-structural alignment of the 49 specimens, was constructed to test the different generic assumptions reported by different researchers in the literature. The structural analysis showed that the ITS2 secondary structures of all of the Coluteocarpeae members exhibited a 4-fingered hand model, which was common in the majority of the family members. No CBCs were observed, whereas hemi-CBCs and NSTs were common among the tribe members. Although hemi-CBCs and NSTs were useful for distinguishing most of the Coluteocarpeae species Noccaea aptera Velen. F.K. Mey., Noccaea aghrica P.H. Davis & Kit Tan M. Firat & Ozudogru, Noccaea fendleri A.Gray Holub subsp. glauca A. Nelson Al-Shehbaz & M. Koch, Noccaea griffithiana Boiss. F.K. Mey., Noccaea rubescens Schott & Kotschy ex Boiss. F.K. Mey., etc. , they were not effective for delimitating some problematic species, such as members of Thlaspiceras F.K. Mey. A phylogenetic tree based on the sequence-structural dataset of the ITS2 showed that generic delimitation of Al-Shehbaz was more acceptable due to the fact that Noccaea sensu Al-Shehbaz is monophyletic.

Phylogenetic evaluation of the genus Nostoc and description of Nostoc neudorfense sp. nov., from the Czech Republic.

Cyanobacterial strain ARC8 was isolated from seepage coming into the river Dračice, Františkov, Czech Republic, and was characterized using a polyphasic approach. Strain ARC8 showed a typical Nostoc-like morphology and in-depth morphological characterization indicated that it is a member of the genus Nostoc. Furthermore, in the 16S rRNA gene phylogeny inferred using Bayesian inference, maximum likelihood and neighbour joining methods, strain ARC8 clustered within the Nostoc sensu stricto clade. The phylogenetic distance and the positioning of strain ARC8 also indicated that it is a member of the genus Nostoc. Furthermore, the rbcL gene phylogeny along with the 16S-23S ITS secondary structure analysis also supported the findings from the 16S rRNA gene tree. In accordance with the International Code of Nomenclature for Algae, Fungi and Plants we describe a novel species of Nostoc with the name Nostoc neudorfense sp. nov.

Widespread inspection and comparative analysis of ITS secondary structure conservation and covariation of Coccinellidae

ITS1 and ITS2 of ribosomal DNA (rDNA) of six subfamilies i.e. Epilachaninae, Chilocorinae, Coccinellinae, Coccidulinae, Scymininae Ortaliinae and Sticholotidinae were examined, aligned and conserved motifs in folding patterns of stable consensus secondary structures were assessed. RNAalifold and RNA fold algorithms applied by RNAz program to compute consensus secondary structures and MFE efficiently and derive a number of statistical scores i.e. SCI, Z-score and MPI. Assembled subsets in ITS1 (three sets) and ITS2 (ten sets) were significantly conserved. In this study, analysis of these consensus secondary structures shows that a motif sequence is remain conserved in all the species. Helices in subsets of ITS1 showed compensatory base involvement to maintain the stability and conservation of secondary structures during natural selection. Secondary structures of ITS2 found more conserved than ITS1. MP, BI and ML methods used to establish the evolutionary relationship, compared the topology significance on the basis of SCI and MPI in defined sets. Coccinellinae, Epilachaninae and Scymninae (all strains of species) were monophyletic whereas Coccidulinae, and Chilocorinae had inadequate sequence information. Of Ortaliinae and Sticholotidinae, yet no ITS sequences have been submitted. Thus, analysis of the structural folding patterns of ITS regions provided further prime information of species relatedness and inter-intra-specific variations.

Reptodigitus Chapmanii (Nostocales, Hapalosiphonaceae) Gen. Nov.: A Unique Nostocalean (Cyanobacteria) Genus Based on a Polyphasic Approach1.

The Nostocales is a monophyletic, heterocytous lineage of cyanobacteria capable of akinete production and division in multiple planes, depending upon family-level clade. While present in a variety of ecosystems, the diversity of the Nostocales has been poorly elucidated. Due to environmentally -induced phenotypic plasticity, morphology alone is often insufficient to determine the true phylogenetic placement of these taxa. In order to bridge this gap, taxonomists now employ the polyphasic approach, combining methods such as morphological analysis, phylogenetic analysis based on DNA sequence and genetic identity based on ribosomal genes, and secondary structure of the 16S-23S ITS and 16S rRNA gene sequences, as well as ecological characterization. Using this combined approach, a new genus and species (Reptodigitus chapmanii gen. et sp. nov.) isolated from the St. Johns River (Jacksonville, Florida, USA) within the Nostocales is herein described. Phylogenetic analyses place this taxon within the Hapalosiphonaceae, sister to the clade containing Fischerella, Hapalosiphon, and Westiellopsis. The 16S-23S ITS secondary folding structure analysis also supports the erection of this new genus.

Hierarchical Nanostructuring Array Enhances Mid-Hybridization for Accurate Herbal Identification via ITS2 DNA Barcode.

It remains a technical challenge to accurately identify close species of herbal medicines, especially from adulterants, because of their highly identical phenotypes and chemical compositions. Here, we report a direct, sequencing-free, high-curvature nanostructuring-based electrochemical herb sensor (nanoE-herb sensor) to identify herbal species quickly and accurately using ITS2 barcodes. We engineer a nano-roughened carbon-supported gold nanostructuring array by photolithograph-free, one-step electrodeposition. The 3D fractal nanostructures exhibit a high deflection angle that largely enhances DNA hybridization efficiency, particularly for the midcomplementary hybridization, as compared to the 2D planar surface. More importantly, such a trans-scale array biointerface (including macroscale carbon and nanoscale gold branches) can overcome the detection barrier of slow diffusion of a long genomic sequence and inaccessibility of the sequestered variations in ITS2 secondary structures through the out-protruded 3D functional nanostructures. Our nanoE-herb sensor achieves a detection limit of 0.18 fM for the 64-mer fragment of saffron ITS2 barcode with midhybridization and shows superior specificity against even single-base mismatch. The sensor also precisely differentiates saffron from six other adulterants by directly detecting unpurified asymmetric PCR amplicons (∼500 bp) with ITS2 sequences, suggesting its great potential in the field identification of herbal medicinal species and pathogenic bacteria with specific DNA barcodes.

Evolutionary Origin and Host Range of Plagiotoma lumbrici (Ciliophora, Hypotrichia), an Obligate Gut Symbiont of Lumbricid Earthworms.

Four common earthworm species, the anecic Lumbricus terrestris, the endogeic Octolasion tyrteum as well as the epigeic Eisenia fetida and Dendrobaena veneta, were examined for the presence of the microbial gut symbiont Plagiotoma lumbrici. The evolutionary origin of this endobiotic microbe was reconstructed, using the 18S rRNA gene, the ITS1-5.8S-ITS2 region, and the first two domains of the 28S rRNA gene. Plagiotoma lumbrici was exclusively detected in the anecic Lumbricus terrestris. Multigene analyses and the ITS2 secondary structure robustly determined the phylogenetic home of Plagiotoma lumbrici populations within the oxytrichid Dorsomarginalia (Spirotrichea: Hypotrichia) as a sister taxon of the free-living Hemiurosomoida longa. This indicates that earthworms obtained their gut endosymbiont by ingesting soil/leaf litter containing oxytrichine ciliates that became adapted to the intestinal tract of earthworms. Interestingly, according to the literature data, Plagiotoma lumbrici was detected in multiple anecic and some epigeic but never in endogeic earthworms. These observations suggest that Plagiotoma lumbrici might be adapted to certain gut conditions and the lifestyle of anecic Lumbricidae, such as Lumbricus, Aporrectodea, and Scherotheca, as well as of some co-occurring epigeic Lumbricus species.