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Theories explaining community assembly assume that biotic and abiotic filters sort species into communities based on the values of their traits and are thus based on between-species trait variability (BTV). Nevertheless, these filters act on individuals rather than on species. Consequently, the selection is also influenced by intraspecific trait variability (ITV) and its drivers. These drivers may be abiotic (e.g., water availability) or biotic (e.g., competition). Although closely related species should have similar traits, many of them coexist. We investigated the relative magnitudes of BTV and ITV in coexisting closely related species and how their individual traits differ under different drivers of ITV. We manipulated conditions in a greenhouse pot experiment with four common Carex species, where individuals of each species originated from four source localities. Individuals were grown in factorial combinations of two moisture levels, with and without a competitor (grass species Holcus lanatus, a frequent competitor). We analyzed the variability of six morphological traits on individuals in the greenhouse and three morphological traits in the source localities. Species identity was the main determinant of differences in most traits. Competition exerted a greater effect than water availability. For leaf dry matter content (LDMC) and vegetative height, competition's effect even exceeded the variability among species. On the contrary, for specific leaf area (SLA) and clonal spread, the interspecific differences exceeded ITV induced by experimental treatments. SLA measured in the greenhouse closely correlated with values measured in field populations, while LDMC did not. The variability caused by source locality of ramets in the greenhouse was small, although sometimes significant. Closely related species differ in their traits, but for some traits, ITV can exceed BTV. We can expect that ITV can modify the processes of community assembly, particularly among coexisting closely related species.

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The ability to recognize and differentiate between conspecifics and heterospecifics as well as their signals is critical for the coexistence of closely related species. In the genus Rattus, species are morphologically similar and multiple species often coexist. Here, we investigated the interspecific recognition and signal differentiation of two sympatric rat species, the brown rat (Rattus norvegicus, RN) and the Asian house rat (Rattus tanezumi, RT). In a two-way choice test, both RN and RT females showed a preference for conspecific male rats to heterospecific ones. RT females showed a significant preference for accessible urine of males of same species to those of other species, but not for the inaccessible urine. On the other hand, there were significant differences in the structural characteristics of the ultrasonic vocalization emitted by males of these two rat species. Sodium dodecyl sulphate‒polyacrylamide gel electrophoresis (SDS‒PAGE) and isoelectric focusing electrophoresis unveiled that major urinary proteins (MUPs) in voided urine were more highly expressed in RN males versus RT males. The interspecific differences of urinary volatile compounds were also discussed. In conclusion, female rats had the ability to distinguish between males of either species.

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BACKGROUND: Indian jointvetch (Aeschynomene indica) is a common and pernicious weed found in the upland direct-seeding rice fields in the lower reaches of the Yangtze River in China. However, there are few reports on the degree of harm, genetic characteristics, and management methods of this weed. The purpose of this study is to clarify the harm of Indian jointvetch to upland direct-seeding rice, analyze the genetic characteristics of this weed based on chloroplast genomics and identify its related species, and screen herbicides that are effective in managing this weed in upland direct-seeding rice fields. RESULTS: In a field investigation in upland direct-seeding rice paddies in Shanghai and Jiangsu, we determined that the plant height and maximum lateral distance of Indian jointvetch reached approximately 134.2 cm and 57.9 cm, respectively. With Indian jointvetch present at a density of 4/m2 and 8/m2, the yield of rice decreased by approximately 50% and 70%, respectively. We further obtained the first assembly of the complete chloroplast (cp.) genome sequence of Indian jointvetch (163,613 bp). There were 161 simple sequence repeats, 166 long repeats, and 83 protein-encoding genes. The phylogenetic tree and inverted repeat region expansion and contraction analysis based on cp. genomes demonstrated that species with closer affinity to A. indica included Glycine soja, Glycine max, and Sesbania cannabina. Moreover, a total of 3281, 3840, and 3838 single nucleotide polymorphisms were detected in the coding sequence regions of the cp. genomes of S. cannabina voucher IBSC, G. soja, and G. max compared with the A. indica sequence, respectively. A greenhouse pot experiment indicated that two pre-emergence herbicides, saflufenacil and oxyfluorfen, and two post-emergence herbicides, florpyrauxifen-benzyl and penoxsulam, can more effectively manage Indian jointvetch than other common herbicides in paddy fields. The combination of these two types of herbicides is recommended for managing Indian jointvetch throughout the entire growth period of upland direct-seeding rice. CONCLUSIONS: This study provides molecular resources for future research focusing on the identification of the infrageneric taxa, phylogenetic resolution, and biodiversity of Leguminosae plants, along with recommendations for reliable management methods to control Indian jointvetch.

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Habitat partitioning among co-occurring, ecologically similar species is widespread in nature and thought to be an important mechanism for coexistence. The factors that cause habitat partitioning, however, are unknown for most species. We experimentally tested among three alternative hypotheses to explain habitat partitioning among two species of co-occurring burying beetle (Nicrophorus) that occupy forest (Nicrophorus orbicollis) and wetland (Nicrophorus hebes) habitats. Captive experiments revealed that the larger N. orbicollis (forest) was consistently dominant to N. hebes (wetland) in competitive interactions for carcasses that they require for reproduction. Transplant enclosure experiments in nature revealed that N. hebes had poor reproductive success whenever the dominant N. orbicollis was present. In the absence of N. orbicollis, N. hebes performed as well, or better, in forest versus its typical wetland habitat. In contrast, N. orbicollis performed poorly in wetlands regardless of the presence of N. hebes. These results support the competitive exclusion-tolerance rule where the competitively dominant N. orbicollis excludes the subordinate N. hebes from otherwise suitable or preferable forest habitat, while the subordinate N. hebes is uniquely able to tolerate the challenges of breeding in wetlands. Transplant experiments further showed that carcass burial depth-an important trait thought to enhance the competitive ability of the dominant N. orbicollis-is costly in wetland habitats. In the presence of N. hebes, N. orbicollis buried carcasses deeper; deeper burial is thought to provide a competitive advantage in forests but further compromised the reproductive success of N. orbicollis in wetlands. Overall, results provide evidence that the competitive exclusion-tolerance rule underlies habitat partitioning among ecologically similar species and that the traits important for competitive dominance in relatively benign environments are costly in more challenging environments, consistent with a trade-off.

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Objective To study the potential distribution area of a new species of Coptis huanjiangensis in China;To provide a basis for the rational development of new species resources protection policy,promoting the stable development of Coptidis Rhizoma species diversity and regional division of exploitation and utilization.Methods The distribution points of Coptis chinensis and Coptis chinensis var.brevisepala were identified and screened through resource census and online database.Based on the hypothesis of"the relatives in the same domain may have similar habitability characteristics",the maximum entropy model was used to obtain the distribution probability≥0.5 of Coptis chinensis and Coptis chinensis var.brevisepala respectively.ArcGIS software was used to obtain the areas with similar ecological environment characteristics to the sampling points of Coptis huanjiangensis.The spatial superposition was carried out with the distribution probability≥0.5 of Coptis chinensis and Coptis chinensis var.brevisepala.Results The potential distribution regions of Coptis chinensis and Coptis chinensis var.brevisepala with distribution probability≥0.5 were respectively normalized and then spatially superimposed,including the central of Sichuan Province,Hunan Province,Jiangxi Province,Guangxi Zhuang Autonomous Region,Fujian Province,Zhejiang Province,and the south of Anhui Province.Influenced by precipitation,temperature,sunshine duration,soil composition,topography,vegetation types and cold index,the potential distribution area of Coptis huanjiangensis was primarily located in the northern part of Guangxi Zhuang Autonomous Region and a small portion of southeastern Guizhou Province.After normalization and binarization treatment,the 25 continuous variables of Coptis huanjiangensis were overlaid with the distribution probability≥0.5 of the related species Coptis chinensis and Coptis chinensis var.brevisepala after respectively normalization treatment,primarily lied along the border between Guangxi Zhuang Autonomous Region and Guizhou Province.Conclusion The potential distribution area of Coptis huanjiangensis is located at the border of Guangxi Zhuang Autonomous Region and Guizhou Province,which can provide references for the study of Coptids Rhizoma species diversity.

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We developed maximum likelihood method for detecting positive selection or balancing selection using multilocus or genomic polymorphism and divergence data from two species. The method is especially useful for investigating natural selection in noncoding regions. Simulations demonstrate that the method outperforms existing methods in detecting both positive and balancing selection. We apply the method to population genomic data from human and chimpanzee. The list of genes identified under selection in the noncoding regions is prominently enriched in pathways related to the brain and nervous system. Therefore, our method will serve as a useful tool for comparative population genomic analysis.

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Four new species groups of the Tenuipalpus sensu lato group are proposed in the present study based on the total number of dorsal opisthosomal setae, namely, carolinensis with ten pairs of setae (214 species), dubinini with nine pairs of setae (33 species), granati with eight pairs of setae (29 species), and barticanus with seven pairs of setae (7 species) Additionally, diagnostic keys to species groups and 273 species of the Tenuipalpus sensu lato are provided. Three species, T. lustrabilis Chaudhri, T. guptai Sadana and Gupta, and T. solanensis Sadana and Gupta, are synonymized with T. punicae Pritchard and Baker. One species, T. rodionovi Chalilova, is suggested as a junior synonym of T. granati Sayed, and eight species, T. chiococcae De Leon, T. costarricensis Salas and Ochoa, T. ephedrae Livschitz and Mitrofanov, T. molinai Evans, T. santae Manson, T. simplychus Cromroy, T. tetrazygiae De Leon, and T. oxalis (Flechtmann), belonging to the carolinensis species group, are not included in the key. Furthermore, a new species of Tenuipalpus sensu lato, T. jazanensis sp. nov., is described and illustrated based on females collected from the Chamaerops spp. (Arecaceae).

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The use of natural immunostimulants is considered the most promising alternative to promote fish health, productive performance and quality, increasing the aquaculture profitability, sustainability and social acceptance. The purpose of this study was to evaluate the effect of the integration of a potential probiotic strain, Halobacterium salinarum, belonging to the Archaea domain, in the formulated diets of farmed gilthead seabream (Sparus aurata L.) in terms of growth performances and immunity responses. The experiment was set up to test two different levels of inclusion of the bacteria in the diet: 0.05% (D1) and 0.1% (D2). The effects on fish growth performances; humoral (peroxidase, protease, antiprotease and IgM levels) and cellular immunity parameters (phagocytosis, respiratory burst and myeloperoxidase), along with bactericidal activity, were evaluated after 15 and 30 days of experimental feeding. The obtained results showed that the inclusion of H. salinarum at the highest concentration (D2 0.1%) improved growth performances, bactericidal activity against Vibrio anguillarum and some parameters related both to the humoral and cellular immune response, suggesting exploring other aspects of welfare in view of future supplementations of this probiotic strain in the diet of S. aurata.

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Speciation is a central topic in evolutionary biology. However, how genomic divergence originates and accumulates in the face of gene flow during ecological adaptation remains poorly understood. Closely related species that have adapted to distinct environments but inhabit some overlapping ranges provide an ideal system to evaluate this issue. Here, we combine population genomics and species distribution models (SDMs) to examine genomic divergences between two sister plant species, Medicago ruthenica and M. archiducis-nicolai, that occur in northern China and the northeast Qinghai-Tibet Plateau, respectively, with overlapping distributions in the border of the two regions. M. ruthenica and M. archiducis-nicolai are well-delimited based on population genomic data, although hybrids exist in sympatric sampling locations. Coalescent simulations and SDMs suggest that the two species diverged from each other in the Quaternary but have been in continuous contact with gene flow occurring between the two species since then. We also discovered positive selection signatures associated with genes both outside and within genomic islands in both species that are probably involved in adaptations to arid and high-altitude environments. Our findings provide insights into how natural selection and climatic changes in the Quaternary initiated and maintained interspecific divergence of these two sister species.

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Comparing the responses of closely related species to environmental changes is an efficient method to explore adaptive divergence, for a better understanding of the adaptive evolution of marine species under rapidly changing climates. Oysters are keystone species thrive in intertidal and estuarine areas where frequent environmental disturbance occurs including fluctuant salinity. The evolutionary divergence of two sister species of sympatric estuarine oysters, Crassostrea hongkongensis and Crassostrea ariakensis, in response to euryhaline habitats on phenotypes and gene expression, and the relative contribution of species effect, environment effect, and their interaction to the divergence were explored. After a 2-month outplanting at high- and low-salinity locations in the same estuary, the high growth rate, percent survival, and high tolerance indicated by physiological parameters suggested that the fitness of C. ariakensis was higher under high-salinity conditions and that of C. hongkongensis was higher under low-salinity conditions. Moreover, a transcriptomic analysis showed the two species exhibited differentiated transcriptional expression in high- and low-salinity habitats, largely caused by the species effect. Several of the important pathways enriched in divergent genes between species were also salinity-responsive pathways. Specifically, the pyruvate and taurine metabolism pathway and several solute carriers may contribute to the hyperosmotic adaptation of C. ariakensis, and some solute carriers may contribute to the hypoosmotic adaptation of C. hongkongensis. Our findings provide insights into the phenotypic and molecular mechanisms underlying salinity adaptation in marine mollusks, which will facilitate the assessment of the adaptive capacity of marine species in the context of climate change and will also provide practical information for marine resource conservation and aquaculture.

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In the process of species differentiation and adaption, the relative influence of natural selection on gene expression variation often remains unclear (especially its impact on phenotypic divergence). In this study, we used differentially expressed genes from brain, cochlea, and liver samples collected from two species of bats to determine the gene expression variation forced by natural selection when comparing at the interspecific (Rhinolophus siamensis and R. episcopus episcopus) and the intraspecific (R. e. episcopus and R. episcopus spp.) levels. In both cases, gene expression variation was extensively adaptive (>66.0%) and mainly governed by directional selection, followed by stabilizing selection, and finally balancing selection. The expression variation related to acoustic signals (resting frequency, RF) and body size (forearm length, FA) was also widely governed by natural selection (>69.1%). Different functional patterns of RF- or FA-related adaptive expression variation were found between the two comparisons, which manifested as abundant immune-related regulations between subspecies (indicating a relationship between immune response and phenotypic adaption). Our study verifies the extensive adaptive expression variation between both species and subspecies and provides insight into the effects of natural selection on species differentiation and adaptation as well as phenotypic divergence at the expression level.

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BACKGROUND: Improving crop resistance to insect herbivores is a major research objective in breeding programs. Although genomic technologies have increased the speed at which large populations can be genotyped, breeding programs still suffer from phenotyping constraints. The pollen beetle (Brassicogethes aeneus) is a major pest of oilseed rape for which no resistant cultivar is available to date, but previous studies have highlighted the potential of white mustard as a source of resistance and introgression of this resistance appears to be a promising strategy. Here we present a phenotyping protocol allowing mid-throughput (i.e., increased throughput compared to current methods) acquisition of resistance data, which could then be used for genetic mapping of QTLs. RESULTS: Contrasted white mustard genotypes were selected from an initial field screening and then evaluated for their resistance under controlled conditions using a standard phenotyping method on entire plants. We then upgraded this protocol for mid-throughput phenotyping, by testing two alternative methods. We found that phenotyping on detached buds did not provide the same resistance contrasts as observed with the standard protocol, in contrast to the phenotyping protocol with miniaturized plants. This protocol was then tested on a large panel composed of hundreds of plants. A significant variation in resistance among genotypes was observed, which validates the large-scale application of this new phenotyping protocol. CONCLUSION: The combination of this mid-throughput phenotyping protocol and white mustard as a source of resistance against the pollen beetle offers a promising avenue for breeding programs aiming to improve oilseed rape resistance. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

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PREMISE: Although ecological differentiation driven by altitude and soil is hypothesized to promote coexistence of sympatric tree species of Damburneya (Lauraceae), the mechanistic role of leaf functional variation on ecological differentiation among co-occurring species remains unexplored. We aimed to determine whether the patterns of leaf trait variation reflect ecological differences among sympatric Damburneya species. We tested whether trait correlations underlying functional strategies and average species traits vary in response to local soil heterogeneity along an altitudinal gradient, potentially affecting species distributions. METHODS: At two contrasting altitudes (100, 1100 m a.s.l.) in a Mexican tropical rainforest, we characterized soil chemical and physical properties and sampled four Damburneya species to quantify five leaf functional traits. We used linear models to analyze paired and multivariate trait correlations, spatial and interspecific effects on trait variation, and trait response to local soil heterogeneity. Relative contributions of intra- and interspecific variation to local trait variability were quantified with an ANOVA. RESULTS: Soil nutrient availability was higher at low altitude, but all species had a high leaf N:P ratio across altitudes suggesting a limited P supply for plants. Species distribution differed altitudinally, with some species constrained to low or high altitude, potentially reflecting soil nutrient availability. Leaf traits responded to altitude and local soil properties, suggesting interspecific differences in functional strategies according to the leaf economics spectrum (conservative vs. acquisitive). CONCLUSIONS: The interspecific divergence in functional strategies in response to local environmental conditions suggests that trait variation could underlie ecological differentiation among Damburneya sympatric species.

PREMISA: Aunque se ha propuesto que la diferenciación ecológica impulsada por la variación del suelo y la altitud ha promovido la coexistencia de especies arbóreas simpátricas de Damburneya (Lauraceae), el papel de la variación de funcional foliar como mecanismo para explicar las diferencias ecológicas entre especies que coocurren sigue sin explorarse. Nuestro objetivo fue determinar si los patrones de variación de los rasgos funcionales foliares reflejan diferencias ecológicas entre especies simpátricas de Damburneya. Hipotetizamos que, tanto las correlaciones entre los rasgos que determinan las estrategias funcionales, como los valores promedio de los rasgos de las especies, varían en respuesta a la heterogeneidad local del suelo a lo largo de un gradiente altitudinal, afectando potencialmente la distribución de las especies. MÉTODO: En dos altitudes contrastantes (100, 1000 m s.n.m) en una selva húmeda tropical mexicana caracterizamos propiedades físicas y químicas del suelo y muestreamos cuatro especies de Damburneya para cuantificar cinco rasgos funcionales foliares. Usamos modelos lineales para analizar las correlaciones pareadas y multivariadas entre rasgos, los efectos espaciales e interespecíficos en la variación de rasgos, y la respuesta de los rasgos a la heterogeneidad local del suelo. Además, cuantificamos la contribución relativa de la variación intra- e interespecífica en la varianza local de los rasgos con un análisis de varianza. RESULTADOS: La disponibilidad de nutrientes en el suelo fue mayor en la zona baja, pero todas las especies tuvieron valores altos de N:P foliar en ambas altitudes, lo que sugiere un suministro limitado de P para las plantas. La distribución de las especies difirió altitudinalmente y algunas de ellas se restringieron a zonas altas o bajas, reflejando potencialmente la disponibilidad de nutrientes en el suelo. Los rasgos foliares respondieron a la altitud y a las propiedades edáficas locales, sugiriendo diferencias interespecíficas en las estrategias funcionales según el espectro de economía foliar (conservativa vs. adquisitiva). CONCLUSIONES: La divergencia interespecífica en las estrategias funcionales en respuesta a las condiciones ambientales locales sugiere que la variación de rasgos podría explicar algunas diferencias ecológicas entre las especies simpátricas de Damburneya.

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The gut microbiota influences the phenotype and fitness of a host; however, limited information is currently available on the diversity and functions of the gut microbiota in wild animals. Therefore, we herein examined the diversity, composition, and potential functions of the gut microbiota in three Sceloporus lizards: Sceloporus aeneus, S. bicanthalis, and S. grammicus, inhabiting different habitats in a mountainous ecosystem. The gut bacterial community of S. bicanthalis from alpine grasslands at 4,150| |m a.s.l. exhibited greater taxonomic, phylogenetic, and functional alpha diversities than its sister species S. aeneus from cornfields and human-induced grasslands at 2,600| |m| |a.s.l. Bacteria of the genus Blautia and metabolic functions related to the degradation of aromatic compounds were more abundant in S. bicanthalis than in S. aeneus, whereas Oscillibacter and predicted functions related to amino acid metabolism and fermentation were more abundant in S. aeneus. The structure of the dominant and most prevalent bacteria, i.e., the core microbiota, was similar between the sister species from different habitats, but differed between S. grammicus and S. aeneus cohabiting at 2,600| |m| |a.s.l. and between S. grammicus and S. bicanthalis cohabiting at 4,150| |m a.s.l. These results suggest that phylogenetic relatedness defines the core microbiota, while the transient, i.e., non-core, microbiota is influenced by environmental differences in the habitats. Our comparisons between phylogenetically close species provide further evidence for the specialized and complex associations between hosts and the gut microbiota as well as insights into the roles of phylogeny and ecological factors as drivers of the gut microbiota in wild vertebrates.

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Sporothrix schenckii is a worldwide-distributed thermally dimorphic fungus, which usually causes a subacute to chronic infection through traumatic implantation or inoculation of its infectious propagules. The fungus encompasses a group of phylogenetically closely related species, thus named the S. schenckii complex, of which S. schenckiisensu stricto and S. brasiliensis are main causative species of sporotrichosis. Owing to a multifaceted molecular dynamic, the S. schenckii complex can switch between the mycelium and the yeast form. This characteristic along with a varying cell wall composition account for significant species-specific differences in the host range, virulence, and susceptibility to antifungal drugs. While culture remains the gold standard to diagnose sporotrichosis, polymerase chain reaction (PCR) or matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry-based methods have become an essential for accurate species identification in many clinical laboratories. If directly applied on tissue samples, molecular methods are helpful to improve both sensitivity of and time to the etiological diagnosis of sporotrichosis. This mini-review aims to put together the old and new knowledge on the S. schenckii complex biology and identification, with particular emphasis on the laboratory diagnosis-related aspects of disease.

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Noncoding regions of the chloroplast (cpDNA) and mitochondrial (mtDNA) genomes are commonly used in plant phylogenetic and population studies. Consensus primers, which are homologous to most coding regions, but amplify variable noncoding regions, are very useful for this purpose. However, high genetic diversity of plants poses a problem in developing molecular methods that require conserved DNA sequences between species.This chapter describes the protocol for designing PCR primers suitable for analysis of closely related plant species. As an example, we used PCR primer design for cpDNA noncoding regions of the rye (Secale).

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Sexual traits that promote species recognition are important drivers of reproductive isolation, especially among closely related species. Identifying neural processes that shape species differences in recognition is crucial for understanding the causal mechanisms of reproductive isolation. Temporal patterns are salient features of sexual signals that are widely used in species recognition by several taxa, including anurans. Recent advances in our understanding of temporal processing by the anuran auditory system provide an opportunity to investigate the neural basis of species-specific recognition. The anuran inferior colliculus consists of neurons that are selective for temporal features of calls. Of potential relevance are auditory neurons known as interval-counting neurons (ICNs) that are often selective for the pulse rate of conspecific advertisement calls. Here, we tested the hypothesis that ICNs mediate acoustic species recognition by exploiting the known differences in temporal selectivity in two cryptic species of gray treefrog (Hyla chrysoscelis and Hyla versicolor). We examined the extent to which the threshold number of pulses required to elicit behavioral responses from females and neural responses from ICNs was similar within each species but potentially different between the two species. In support of our hypothesis, we found that a species difference in behavioral pulse number thresholds closely matched the species difference in neural pulse number thresholds. However, this relationship held only for ICNs that exhibited band-pass tuning for conspecific pulse rates. Together, these findings suggest that differences in temporal processing of a subset of ICNs provide a mechanistic explanation for reproductive isolation between two cryptic treefrog species.

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BACKGROUND: Although molecular analysis offers a wide range of options for species identification, a universal methodology for classifying and distinguishing closely related species remains elusive. This study validated the effectiveness of utilizing the entire chloroplast (cp) genome as a super-barcode to help identify and classify closely related species. METHODS: We here compared 26 complete cp genomes of ten Fritillaria species including 18 new sequences sequenced in this study. Each species had repeats and the cp genomes were used as a whole DNA barcode to test whether they can distinguish Fritillaria species. RESULTS: The cp genomes of Fritillaria medicinal plants were conserved in genome structure, gene type, and gene content. Comparison analysis of the Fritillaria cp genomes revealed that the intergenic spacer regions were highly divergent compared with other regions. By constructing the phylogenetic tree by the maximum likelihood and maximum parsimony methods, we found that the entire cp genome showed a high discrimination power for Fritillaria species with individuals of each species in a monophyletic clade. These results indicate that cp genome can be used to effectively differentiate medicinal plants from the genus Fritillaria at the species level. CONCLUSIONS: This study implies that cp genome can provide distinguishing differences to help identify closely related Fritillaria species, and has the potential to be served as a universal super-barcode for plant identification.

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Understanding the processes shaping population structure and reproductive isolation of marine organisms can improve their management and conservation. Using genomic markers combined with estimation of individual ancestries, assignment tests, spatial ecology, and demographic modeling, we (i) characterized the contemporary population structure, (ii) assessed the influence of space, fishing depth, and sampling years on contemporary distribution, and (iii) reconstructed the speciation history of two cryptic redfish species, Sebastes mentella and S. fasciatus. We genotyped 860 individuals in the Northwest Atlantic Ocean using 24,603 filtered single nucleotide polymorphisms (SNPs). Our results confirmed the clear genetic distinctiveness of the two species and identified three ecotypes within S. mentella and five populations in S. fasciatus. Multivariate analyses highlighted the influence of spatial distribution and depth on the overall genomic variation, while demographic modeling revealed that secondary contact models best explained inter- and intragenomic divergence. These species, ecotypes, and populations can be considered as a rare and wide continuum of genomic divergence in the marine environment. This acquired knowledge pertaining to the evolutionary processes driving population divergence and reproductive isolation will help optimizing the assessment of demographic units and possibly to refine fishery management units.

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Baseline separation and analysis of multicomponent mixtures of closely related pharmaceuticals using single column selectivity can often be challenging, requiring the combination of orthogonal stationary and mobile phase methods to monitor all the species and optimize reaction outcomes. In recent years, two-dimensional liquid chromatography (2D-LC) has become a valuable tool for improving peak capacity and selectivity. Though powerful, standard 2D-LC instrumentation and software can often lead to tedious method development and has a requirement for very specific expertise that is poorly suited for a fast-paced industrial environment. In this regard, the introduction of an automated online 2D-LC setup that could screen multiple columns in both dimensions without manual intervention will undeniably serve to streamline column/mobile phase selection and secure the viability of 2D-LC as a mainstay instrument for industrial applications. Herein, we introduce and investigate a multicolumn online 2D-LC approach that simplifies column screening and method development dramatically. This setup incorporates 6-position column selection valve technology whose functionality enables us to combine multiple columns in the first and second dimensions. This strategy in conjunction with diode array detection (DAD) in both dimensions and mass spectrometry (MS) acquisition in the second dimension serves to explore different columns and mobile phases as a framework for screening targeted compounds in multicomponent mixtures without having to perform chromatographic purification. Multiple online heart cutting achiral RPLC - achiral RPLC and achiral RPLC - chiral RPLC coupled to DAD and ESI-MS methods combining several stationary phase selectivity in an automated fashion are successfully applied to the separation and analysis of complex mixtures of drug substances, where in many instances, traditional 1D-ultra-high performance liquid chromatography (UHPLC) fails or delivers sub-optimal results. This automated online multicolumn 2D-LC workflow enables rapid and efficient identification of column/eluent combinations, as well as sample analysis across multiple columns in both dimensions overnight with a single click.

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