High Intraspecific Variability Research Articles

Evaluating the genetic diversity of Erythropalum scandens based on using inter-simple sequence repeat markers

Erythropalum scandens Blume, an emerging medicinal plant with great potential for drug development, also possesses high edible value. In this study, we investigated the genetic diversity of the germplasm of E. scandens obtained from different geographical locations using inter simple sequence repeat (ISSR) markers. For this purpose, 18 ISSR primer pairs with a distinct background and adequate polymorphism were selected. We established an optimal ISSR–PCR reaction system (20 µL) with the following parameters: 1 µL DNA template (60 ng µL−1), 1.2 µL primers (10 µmol µL−1), 10 µL MasterMix, and 7.8 µL H2O. A total of 183 loci were amplified using the 18 primer pairs, of which 121 (66.12%) indicated polymorphism. Moreover, 34 germplasms of E. scandens exhibited genetic similarity coefficients ranging from 0.7104 to 0.9563, genetic distances ranging from 0.0447 to 0.3420, Nei’s genetic diversity index of 0.1946, and Shannon’s information index of 0.2982, suggesting high intraspecific genetic diversity. UPGMA cluster and PCoA analyses distinguished the germplasm of E. scandens obtained from Guangxi from those collected from Guangdong, Hainan, Fujian, and Guizhou. However, the Mantel correlation analysis revealed that the genetic variation among the 34 germplasms of E. scandens was not significantly related to geographical distance. The analysis of the genetic background of wild and cultivated germplasms of E. scandens can help guide variety selection and breeding. Furthermore, the present study revealed the genetic background and affinities among 34 germplasms of E. scandens. Overall, our findings lay the foundation for the conservation and utilization of germplasm resources, identification and classification of varieties, and variety selection and improvement of E. scandens at the molecular level.

Comparative molecular and morphological species delineation of Halammohydra Remane, 1927 (Hydrozoa)—with the description of four new species

Whereas most cnidarians are macrofaunal, a few microscopic lineages have evolved, and some of them inhabit marine sediments. The meiofaunal genus with the most species is Halammohydra, comprising nine nominal species. Species are described with high intraspecific variability in, e.g., the number of tentacles and statocysts and the shape and length of tentacles and body, complicating morphological identification to species level. Additionally, there is not much molecular data available. This study aims to revise already described species with morphological and molecular methods, as well as, to delineate potential new species answering questions about their geographical distribution. For this, specimens were sampled at 16 locations in the Northwest Atlantic and two localities in the East Atlantic, documented with light microscopy, and fixed individually for sequencing (16S, 18S, and CO1). Herewith, morphological characters were linked to a specific sequence, enabling the testing of character variation within one molecular phylogenetic group. Phylogenetic analyses were conducted (Bayesian Interference and Maximum Likelihood) in combination with species delimitation tests (ABGD, GMYC, and bPTP). Four already described species were identified in the data sets, and all of these were found at multiple localities. Four new species are described. Overall, the combined molecular and morphological data acquisition revealed multiple new species and a high degree of sympatry in Halammohydra. This, together with the confirmed excessive intraspecific variation in morphological traits, underlines the necessity of molecular sequencing for the taxonomy and species identification of Halammohydra.

Choose Wisely: Great Variation among Genotypes of Promising Paludiculture Crop Phragmites australis.

Measures mitigating the climate crisis, such as paludiculture, which is the agriculture on rewetted peatlands, are urgently needed. The cosmopolitan species Phragmites australis has the potential to be used in paludiculture worldwide but is known for its high intraspecific variation. This raises the questions of whether (i) P. australis genotypes differ even at a regional scale, making them differently well suited for paludiculture and (ii) P. australis performance can be predicted by linking the variation in genotypes to strategies in the plant economics spectrum. Five P. australis genotypes from Mecklenburg-Western Pomerania were cultivated in two 10-month mesocosm experiments along gradients of water level and nutrient addition. We compared growth, morphology (height, growing density), above- and belowground biomass, functional and ecophysiological traits (SLA, LDMC, SRL, RDMC, root porosity, photosynthetic rate) as well as gene expression. Our results demonstrate a high variability of P. australis genotypes even at a regional scale, revealing genotype-specific productivity, morphology, and gene expression and implying that the selection of suitable genotypes will be crucial for the success of a paludiculture. However, trait covariation did not indicate distinct plant economic strategies to predict genotype performance. Instead, large-scale genotype trials are needed to select suitable genotypes for paludiculture.

Geographic Variation in Skull Morphology of the Japanese Crocodile Newt, Echinotriton andersoni (Amphibia: Urodela: Salamandridae), Inferred from Geometric Morphometrics

The Anderson's crocodile newt, Echinotriton andersoni, is an endangered species distributed in the islands of Central Ryukyus, Japan. This species is well known to show high intraspecific genetic variation among islands, but its morphological variation has not been explored. We examined the skull variation in this species including enigmatic specimens collected from Taiwan by using geometric morphometrics, and found sexual differences and geographic variation mainly between the Okinawa group (Okinawajima Island) and the Amami Group (Amamioshima and Tokunoshima Islands). We further found close resemblance between the Taiwanese specimens and the female specimens from Amamioshima Island.

Assessing Intraspecific Genetic Variability in Cannabis sativa Populations of Punjab Pakistan Using RAPD and DNA Barcode Markers

Cannabis sativa possess substantial amount of omega-3 fatty acids and several medicinal benefits with potential secondary metabolites. This study employed rbcL, matK, and trnH-psbA barcode markers and RAPD marker to explore genetic diversity in C. sativa populations in Punjab, Pakistan. In population genetic study using DNA barcode markers, cladogram showed investigated samples were genetically similar; however, demonstrated significant genetic diversity with the available DNA sequences of this plant from other regions and countries. The nucleotide sequence variation (% divergence) indicated high intraspecific variation in plants of different geographical zones. However, the RAPD marker analysis displayed genetic diversity among collected Cannabis sativa samples. The principal component analysis reinforced these findings by providing a spatial representation of genetic distances among the samples that revealed a considerable genetic diversity among the collected material and significant intraspecific variation was observed, indicating diverse genetic profiles across geographical regions. This research highlights the potential of molecular markers in germplasm selection for breeding programs.

Genetic differentiation between two subspecies of Emberiza schoeniclus and open forest bunting's evolution inferred from mitogenomes

The reed bunting, Emberiza schoeniclus (Linnaeus 1758), is the only member of the genus adapted to Mediterranean wetlands, where some subspecies are critically endangered. The first complete mitogenome of the eastern Iberian reed bunting (E. s. witherbyi) is presented here and compared with an unpublished mitogenome obtained in northeast Asia (most likely E. s. pyrrhulina). Genetic distance analyses are consistent with the new reed bunting data corresponding to two distinct lineages of E. schoeniclus. A new fossil‐calibrated phylogeny suggests that open forest buntings have suffered two rapid speciation events from Late Miocene to Pleistocene, that seem to be correlated with major climatic changes and habitat shifts. Adaptation to a new ecological niche (i.e. wetlands) could have favoured the reed bunting expansion across the Palearctic. The high intraspecific variation observed today could result from the establishment of resident populations within small areas, potentially acting as a climatic refuge.

Molecular assessment of the taxonomic status of pine vole species Microtus subterraneus and Microtus majori in southeastern Europe and Turkey

Intra– and inter–specific phylogenetic relationships and the taxonomic status of pine vole populations have been controversial for years, and cryptic species are thought to exist, especially concering the species' distribution area. To clarify the taxonomic status of Turkish populations we analysed mitochondrial (cytochrome–b and cytochrome oxidase–I) and nuclear (interphotoreceptor retinoid binding protein) gene markers, adding GenBank data from Europe and Caucasia. Considering the data obtained based on mean genetic distance, genetic diversity and fixation index values, Bayesian trees and Median–joining networks, we found that M. subterraneus and M. majori are valid species which have diverged since 1.28 Mya. Findings also suggested that although Anatolian, Thracian (Turkish Thrace and Greece) and European populations of M. subterraneus and Anatolian and Caucasian populations of M. majori have been in the process of divergency since 0.528–0.337 Mya correspond to the Pleistocene glacial periods, these intrapopulations do not appear to be different species.Besides, considering the high intraspecific variation in M. subterraneus, it remains likely that new species could be identified in future studies.

How to build vegetation patches in hydraulic studies: a hydrodynamic-ecological perspective on a biological object

Vegetation in freshwater and coastal ecosystems modifies flows, retains sediment, protects banks and shorelines from erosion. Hydraulic laboratory studies with live vegetation or artificial plant mimics, or numerical models with abstracted patches, are often used to quantify the effects of vegetation on water flow and sedimentation. However, the choice of plant and patch characteristics is often not supported by field observations of patch dimensions, density or spacing between consecutive patches. The discrepancy between plants in natural conditions and in flume experiments or numerical studies may affect the relevance of these findings for natural ecosystems. In this study, we provide guidelines for building realistic vegetation patches in hydraulic studies. We collected data on four species of fully submerged freshwater aquatic macrophytes that can grow into well-defined patches. We considered three relevant levels: individual plants (inside patches), isolated patches and multiple neighbouring patches. At the plant level, we observed significant differences in biomechanical traits (Young’s modulus, flexural stiffness), resulting in stem Cauchy numbers ranging from 85.25 to 325.84, and leaf Cauchy numbers from 163.81 to 2003.97. At the patch level, we found significant relationships between patch length, width and height, showing covariation among different patch characteristics. The relationships among patch dimensions differed significantly among sampling sites for three of the four species, suggesting high intraspecific variability in patch sizes. By providing a first set of guidelines for choosing correct and ecologically relevant plant characteristics, this dataset aims to improve our understanding of the complex processes occurring inside and around submerged vegetated patches.

Systematic revision of the snorkel snail genus Rhiostoma Benson, 1860 (Gastropoda, Caenogastropoda, Cyclophoridae) with descriptions of new species.

The snorkel snail genus Rhiostoma Benson, 1860 is comprised of terrestrial cyclophorid snails with wide-ranging species diversity and radiation in Southeast Asia. The typical characters of the genus are a depressed shell, a detached and descending portion of the last whorl with a distinctive peristomal breathing device attached, and a calcareous cup-shaped operculum. Herein, we have revised the systematics of extant species based on shell morphology combined with COI barcoding. From these thirty recognised species, twelve are described as new to science: R. ? amarapuraensesp. nov., R.anceyisp. nov., R.breviocollarsp. nov., R.ebenozosterasp. nov., R.cheliopegmasp. nov., R.furfurosumsp. nov., R.gnomus, sp. nov., R.lannaensesp. nov., R.laoensesp. nov., R.platymorphasp. nov., R.rhothonotaphrosasp. nov., and R.tigrinasp. nov. All conchological characters are provided via illustrations of type specimens and living snails, and descriptions of the shells and radulae. Phylogenetic analysis based on partial COI gene sequences strongly supports the designated morphospecies and a monophyletic Rhiostoma, confirming that all pterocyclinid snails with a calcareous, cup-shaped operculum belong to the same clade. A high intra-specific divergence was observed in R.jalorensis and R.housei populations from locations in close proximity, suggesting a lower dispersal and higher level of isolation. The low inter-specific divergence found in R.hainesi, R.samuiense, R.asiphon, and R.rhothonotaphrosasp. nov. supports their recent diversification and local adaptation, and is congruent with their marked morphological differences. Finally, nine formerly Rhiostoma-placed species were reclassified into either the genus Cyclotus or the genus Opisthoporus.

Insights into the phylogeny and chloroplast genome evolution of Eriocaulon (Eriocaulaceae)

BackgroundEriocaulon is a wetland plant genus with important ecological value, and one of the famous taxonomically challenging groups among angiosperms, mainly due to the high intraspecific diversity and low interspecific variation in the morphological characters of species within this genus. In this study, 22 samples representing 15 Eriocaulon species from China, were sequenced and combined with published samples of Eriocaulon to test the phylogenetic resolution using the complete chloroplast genome. Furthermore, comparative analyses of the chloroplast genomes were performed to investigate the chloroplast genome evolution of Eriocaulon.ResultsThe 22 Eriocaulon chloroplast genomes and the nine published samples were proved highly similar in genome size, gene content, and order. The Eriocaulon chloroplast genomes exhibited typical quadripartite structures with lengths from 150,222 bp to 151,584 bp. Comparative analyses revealed that four mutation hotspot regions (psbK-trnS, trnE-trnT, ndhF-rpl32, and ycf1) could serve as effective molecular markers for further phylogenetic analyses and species identification of Eriocaulon species. Phylogenetic results supported Eriocaulon as a monophyletic group. The identified relationships supported the taxonomic treatment of section Heterochiton and Leucantherae, and the section Heterochiton was the first divergent group. Phylogenetic tree supported Eriocaulon was divided into five clades. The divergence times indicated that all the sections diverged in the later Miocene and most of the extant Eriocaulon species diverged in the Quaternary. The phylogeny and divergence times supported rapid radiation occurred in the evolution history of Eriocaulon.ConclusionOur study mostly supported the taxonomic treatment at the section level for Eriocaulon species in China and demonstrated the power of phylogenetic resolution using whole chloroplast genome sequences. Comparative analyses of the Eriocaulon chloroplast genome developed molecular markers that can help us better identify and understand the evolutionary history of Eriocaulon species in the future.

High acorn diversity of the introduced Quercus rubra indicates its ability to spread efficiently in the new range

The morphological traits of seeds have a fundamental meaning for spread and regeneration of non-native zoochorous plants, as they determine the level of establishment success and the ability of native fauna to disperse the alien plant. Here we studied the size, mass and viability of acorns of the North-American northern red oak Quercus rubra in its introduced range (in central Poland). The study revealed that acorns sampled in 2018 and 2019, in Q. rubra stands which had just reached sexual maturity, achieved optimum age for seed crop production, or were close to the rotation age, and were planted in sites suitable to European coniferous, mixed, and deciduous forests, differed in their dimensions and weight. Acorn size, mass and viability were affected both by stand age and forest site type, but more strongly by the first factor, i.e. the older the stand, the larger the acorns. At the initial stage of reproduction larger acorns were produced in coniferous than in other forest sites, but this advantage decreased with the age of stand in favour of more fertile mixed and deciduous forest sites. Acorn dimensions and viability also differed in sequential years: larger acorns with a high proportion of viability were produced in 2018, while in 2019 the proportion of viable acorns was significantly lower. The moisture of (viable) acorns was well above the lethal level for most acorns in all stands studied, however, it was impacted by the autumn weather.The high intraspecific variability in size and weight makes Q. rubra acorns attractive for different species of native seed hoarders. It conditions effective acorn spread and regeneration of this introduced (invasive) oak in European forests. Therefore, leaving single oak trees over 90 years old – which produce the best quality seeds – on plots cleared during forestry operations can favour Q. rubra regeneration.

Integrative taxonomy of Serrasentis gibsoni n. sp. (Acanthocephala: Isthmosacanthidae) from flatfishes in the Gulf of Mexico

Abstract The Isthmosacanthidae acanthocephalan species of the genus Serrasentis are parasites of marine teleosts and an elasmobranch. In this study, Serrasentis gibsoni n. sp. is described from the intestines of four flatfish species (Paralichthyidae), namely Ancyclopsetta quadrocellata, Cyclopsetta chittendeni, Syacium gunteri, and S. papillosum from 10 oceanic sites in the Gulf of Mexico (GoM). Twenty sequences of the ‘barcoding’ region of cytochrome C oxidase subunit I gene were obtained from 20 adults of Serrasentis gibsoni n. sp. Additionally, five sequences of the barcoding region were obtained from five adults of rhadinorhynchid Gorgorhynchus lepidus from C. chittendeni, S. papillosum and one species of Haemulidae, Haemulom aurolineatum, from five oceanic sites from the GoM. Two phylogenetic approaches were followed: Bayesian inference and maximum likelihood. In both phylogenetic reconstructions, the sequences of Serrasentis gibsoni n. sp. were recovered as a monophyletic group within the genus Serrasentis and placed as a sister group to G. lepidus. However, due to the lack of molecular data for species of the Isthmosacanthidae and Rhadinorhynchidea, these phylogenetic inferences must be taken with caution. Serrasentis gibsoni n. sp. is the first species of Serrasentis described from Paralichthyidae flatfish species from marine waters of the Americas and from the GoM. Based on the barcoding data set analyzed, Serrasentis gibsoni n. sp. appears to have high intraspecific genetic variation; thus, it is necessary to continue exploring the genetic diversity of this species to infer its intraspecific evolutionary patterns.

Integrative taxonomy of the Lauraceae-feeding species of the genus Stephanitis (Hemiptera, Heteroptera, Tingidae) from Japan

Stephanitis Stål, 1873 (Hemiptera, Heteroptera, Tingidae, Tinginae, Tingini) is one of the most diverse lace bug genera in East Asia with over 50 species, many of them feeding on Ericaceae and Lauraceae. The Japanese species feeding on Lauraceae have high intraspecific and poor interspecific variation, so identification of some previous distribution records (mainly S. (Norba) aperta Horváth, 1912 and closely related taxa) is doubtful. In the present study, 5,080 specimens of Lauraceae-feeding species of Stephanitis from Japan were identified to species based on morphology, and DNA barcodes of the mitochondrial cytochrome c oxidase subunit (COI) region for 53 of those individuals were also obtained. The combined morphological and molecular evidence dataset showed that 10 species of Stephanitis feed on lauraceous trees in Japan: S. (Stephanitis) ambigua Horváth, 1912, S. (Norba) aperta, S. (N.) exigua Horváth, 1912, S. (N.) hayashiisp. nov., S. (N.) hiurai Takeya, 1963, S. (N.) ishikawaisp. nov., S. (N.) mendica Horváth, 1912, S. (S.) tabidula Horváth, 1912, S. (S.) takeyai Drake & Maa, 1955 and S. (S.) tomokuniisp. nov. Additionally, a new synonymy, S. (S.) fasciicarina Takeya, 1931, syn. nov. with S. (S.) tabidula, is proposed, a key for identifying the 10 species is provided and their distribution ranges and host plant relationships are presented.

First record of Mordellistena semiferruginea (Coleoptera: Mordellidae) in Italy and analysis of intraspecific variation in the COI gene

Mordellistena semiferruginea Reitter, 1911, despite its wide distributional range that spans Europe and Central Asia, is a rarely encountered member of the family Mordellidae. In this paper, we present the first record of this species in Italy based on 15 individuals captured in Malaise traps. We have compared these individuals with type specimens and provided a differential diagnosis of the species together with high-resolution photographs of the habitus and male genitalia. Additionally, we generated 14 sequences of the standard DNA barcoding fragment of the mitochondrial gene coding for cytochrome c oxidase subunit I (COI). The analyses of COI sequences showed a relatively high intraspecific divergence reaching 3.65% (p-distance), which is the highest within-species genetic variation documented so far in Mordellidae. Of the total number of 30 nucleotide polymorphisms, only one led to a divergence in the amino acid sequences. The predicted 3D structure of the encoded proteins showed that the variation occurred in Loop 1–2, far from the active site of the protein.

Stranding of Mesopelagic Fishes in the Canary Islands.

Most mesopelagic fishes perform large diel vertical migrations from the deep-sea zone to the surface. Although there is a trade-off between a higher food availability at the upper layers and an energy cost and predation risk, incursion towards the surface also implies a transport by currents, where the fish are exposed to a stranding risk on the coast. Here, we reported the first documented stranding of mesopelagic fishes along the southeast shore of Gran Canaria Island. Our study hypothesized that (1) the influence of the Canary Current, (2) the dominant incidence of the Trade Winds during summer, and (3) the presence of an upwelling filament coupled with an anticyclonic eddy south of Gran Canaria Island were the causative mechanisms of the strandings. Diaphus dumerilii (Myctophidae family) was the main species found as observed from an external morphological analysis using traditional taxonomy. The otolith contour analysis suggested the presence of other Diaphus spp. and Lobianchia dofleini. Nevertheless, the otolith morphological features described in the literature suggested that all the specimens were actually D. dumerelii. Errors in the identification were mainly due to the high intraspecific variability found in the otolith morphology. Even so, two patterns of oval and elliptic shapes were described with significant differences in its morphometry.

Integrative analysis on the hitchhiking differential grasshopper, Melanoplus differentialis (Thomas, 1865) (Acrididae, Orthoptera)

AbstractWe conducted the monitoring of hitchhiker insect pests around the port area in Korea and reported the settlement of Melanoplus differentialis (Thomas, 1865) in Onsan Port, Ulsan, Korea. The occurrence pattern of the invasive species was determined using a line census method. Species identification was first performed with DNA barcoding, and then the DNA barcoded samples were morphologically re‐examined. Monitoring of the occurrence pattern revealed that the invasive populations of M. differentialis settled in a narrow artificial green area, and it occurred in large numbers (over 100 individuals per hour). Species identification of the invasive population using DNA barcoding indicated that the population showing high intraspecific divergence from null to 3.9% was difficult to identify as a separate species. However, in morphological re‐examination, the population was clearly identified as a single species, M. differentialis. In a haplotype analysis using DNA barcodes, the population included 45 haplotypes with polymorphism at 46 nucleotide sites. On the basis of these results, we considered that M. differentialis may have invaded into Onsan Port, Ulsan, Korea. Therefore, we provided the biological traits of the grasshopper for risk assessment and suggested the necessity of monitoring and controlling the dispersal of M. differentialis into the inlands of Korea in the early stages.

Understanding the Taxonomic Complexes and Species Delimitation within Sambucus L. (Viburnaceae)

The genus Sambucus belongs to the family Viburnaceae, and has long been a debatable taxon regarding its systematics and taxonomic assignment. It is known for its morphological and genetic variations and thus the taxonomic delimitation remains uncertain. Phenotypic and genotypic variations within plant species causes difficulties in the detection of species boundaries. In the present review, we compiled a set of studies that assessed species discrimination within Sambucus genus by morphological and molecular markers as well as the taxonomic uncertainties in the re-circumscription of the segregated genera i.e., Viburnum L., Sambucus L., and Adoxa L., (syn. Sinadoxa C.Y. Wu, Z.L. Wu & R.F. Huang/Tetradoxa C.Y. Wu) from Caprifoliaceae and their family assignment. The data was obtained from published journal articles and various online databases. Morphological and molecular diagnostic characters were employed to distinguish Sambucus species. Our findings indicated that high intraspecific variations within narrow-ranged populations and their widespread counterparts contribute to its taxonomic complications. Additionally, intraspecific variations within Sambucus species leads to misidentifications and the misapplication of names, occasioning the publication of many putative synonyms and ambiguous names. Possible interspecific hybridizations experienced between Sambucus species increases complexity in character evolution. A morphological analysis showed that the endocarp seed morphological characters can reliably support the segregation of Sambucus taxa into discrete taxonomic entities. In molecular analysis, the NeighborNet (NN) phylogenetic split network revealed three distinct genera including Viburnum, Sambucus, and Adoxa, which clustered separately. The Maximum Likelihood (ML) analysis indicated that the Sambucus species formed a monophyletic group and clustered into two major clades, a small clade containing S. maderensis, S. peruviana, S. nigra, and S. canadensis, and a large clade encompassing the rest of the species with significantly supported clades (<52%). The preferred family assignment for Sambucus is Viburnaceae.

Specific and Intraspecific Diversity of Symphypleona and Neelipleona (Hexapoda: Collembola) in Southern High Appalachia (USA)

Collembola, commonly known as springtails, are important detritivores, abundant in leaf litter and soil globally. Springtails are wingless hexapods with many North American species having wide distributions ranging from as far as Alaska to Mexico. Here, we analyze the occurrence and intraspecific diversity of springtails with a globular body shape (Symphypleona and Neelipleona), in southern high Appalachia, a significant biodiversity hotspot. The peaks of high Appalachia represent ‘sky islands’ due to their physical isolation, and they host numerous endemic species in other taxa. We surveyed globular Collembola through COI metabarcoding, assessing geographic and genetic diversity across localities and species. Intraspecific diversity in globular Collembola was extremely high, suggesting that considerable cryptic speciation has occurred. While we were able to associate morphospecies with described species in most of the major families in the region (Dicyrtomidae, Katiannidae, Sminthuridae, and Sminthurididae), other families (Neelidae, and Arrhopalitidae) are in more pressing need of taxonomic revision before species identities can be confirmed. Due to poor representation in databases, and high intraspecific variability, no identifications were accomplished through comparison with available DNA barcodes.

New Genetic Variants of Leptospira spp Characterized by MLST from Peruvian Isolates.

Leptospirosis is a zoonotic disease caused by the genus Leptospira, presenting complex and dynamic epidemiology. To determine the genetic variability and its phylogenetic relationship of Leptospira spp isolates from three sources in Iquitos (Peruvian Amazon) from 2002 to 2013, seven MLST genes were analyzed to obtain the Sequence Type (ST) and these sequences were concatenated for phylogenetic analysis. The genetic relationship between STs was determined with the goeBURST algorithm and genetic diversity was determined using DnaSP. Of 51 isolates, 48 were pathogenic belonging to five different species: Leptospira interrogans Nascimento 2004, Leptospira santarosai Feil 2004, Leptospira noguchii Haake 2021, Leptospira borgpetersenii Levett 2021, and Leptospira kirschneri Levett 2021. Of 20 STs identified, 60% corresponded to new genotypes circulating only in Peru. The genotypes ST17, ST37, and ST301 were recorded in rodents and humans. A high intraspecific genetic diversity was demonstrated in L. noguchi. The goeBURST analysis revealed three clonal complexes (CCs) and 16 singletons. The STs were found to show high genetic variability and phylogenetic and goeBURST analysis determined that the genotypes found did not form specific groups according to the source of infection or origin, which confirms the zoonotic potential of these STs in an area highly endemic for leptospirosis.

Assessing Morphological Species and Interspecific Relationships in North American Grapeferns (Sceptridium; Ophioglossaceae) Using ISSR Markers

Abstract— Relationships among North American Sceptridium (sensu Škoda) species are often difficult to assess because of few stable distinguishing characters and high intraspecific variability. We used ISSR-PCR to examine relationships among four samples of Holubiella, 24 samples representing four diploid North American Sceptridium species, and a Russian Sceptridium sample. Nine ISSR primers yielded 147 polymorphic loci. We generated three neighbor-joining dendrograms using a Jaccard, Nei & Li, or Dice coefficient. All three coefficients cluster based solely on the presence of bands and not the absence. ISSR data analyses across all three coefficients were consistent with published analyses of DNA sequence data in supporting Holubiella as sister to all Sceptridium species sampled. Sister group relationships among the four Sceptridium species were not consistent across all three neighbor-joining analyses, and bootstrap support was generally low. However, 26 of the 29 samples consistently clustered with other samples of similar morphology, and these clusters generally supported current species concepts. In taxa such as Sceptridium, with low DNA sequence variability among species, ISSR-PCR may provide an important tool for evaluating morphologically defined species, but additional data are necessary for establishing robust hypotheses of phylogenetic relationships.