Gene Cytochrome Research Articles

In Vitro vs. In Vivo Transcriptomic Approach Revealed Core Pathways of Nitrogen Deficiency Response in Tea Plant (Camellia sinensis (L.) Kuntze)

For the first time, we used an in vitro vs. in vivo experimental design to reveal core pathways under nitrogen deficiency (ND) in an evergreen tree crop. These pathways were related to lignin biosynthesis, cell redox homeostasis, the defense response to fungus, the response to Karrikin, amino acid transmembrane transport, the extracellular region, the cellular protein catabolic process, and aspartic-type endopeptidase activity. In addition, the mitogen-activated protein kinase pathway and ATP synthase (ATP)-binding cassette transporters were significantly upregulated under nitrogen deficiency in vitro and in vivo. Most of the MAPK downstream genes were related to calcium signaling (818 genes) rather than hormone signaling (157 genes). Moreover, the hormone signaling pathway predominantly contained auxin- and abscisic acid-related genes, indicating the crucial role of these hormones in ND response. Overall, 45 transcription factors were upregulated in both experiments, 5 WRKYs, 3 NACs, 2 MYBs, 2 ERFs, HD-Zip, RLP12, bHLH25, RADIALIS-like, and others, suggesting their ND regulation is independent from the presence of a root system. Gene network reconstruction displayed that these transcription factors participate in response to fungus/chitin, suggesting that nitrogen response and pathogen response have common regulation. The upregulation of lignin biosynthesis genes, cytochrome genes, and strigalactone response genes was much more pronounced under in vitro ND as compared to in vivo ND. Several cell wall-related genes were closely associated with cytochromes, indicating their important role in flavanols biosynthesis in tea plant. These results clarify the signaling mechanisms and regulation of the response to nitrogen deficiency in evergreen tree crops.

Phylogeography of the Red‐Headed Manakin Supports the River‐Refuge Hypothesis

ABSTRACTAimThe Amazon rainforest is one of the most biodiverse regions on earth, but our understanding of the processes that have shaped its patterns of diversity remains incomplete. One hypothesis for Amazonian speciation is the river‐refuge hypothesis, which suggests that retraction of forests away from the periphery of Amazonia, where rivers are narrow, facilitated divergence of populations separated by wide rivers. Later re‐expansion of forests would have allowed secondary contact between these populations, and co‐occurring hybrid zones may reveal the location where expanding forests first reconnected. Here, we test whether a widespread Amazonian songbird species shows evidence of population contact zones in the eastern headwaters of the Tapajós river, hypothesised to be an area of secondary contact under the river‐refuge hypothesis.LocationAmazon and Atlantic forests of South America.TaxonCeratopipra rubrocapilla (Pipridae, Passeriformes).MethodsWe sampled 147 C. rubrocapilla (Red‐headed Manakin) across its vast range, with 70 samples sequenced using reduced‐representation genomic sequencing and 139 sequenced at the mitochondrial gene cytochrome b. We use population genetic and phylogenetic analyses to investigate patterns of gene flow and population structure across the range, with a particular focus on samples from the headwaters of the Tapajós River to evaluate the potential role of the river‐refuge hypothesis.ResultsWe observe shallow population structure and evidence for widespread gene flow across the range of C. rubrocapilla. Patterns in the Tapajós headwaters match predictions of the river‐refuge hypothesis, with eastern headwater samples most similar to samples farther west on the opposite bank of the Tapajós River.Main ConclusionsThe close co‐occurrence between population contact zones within C. rubrocapilla and other previously studied hybrid zones reinforces the hypothesis that the eastern Tapajós headwaters is where rainforest populations reconnected in the past, following predictions of the river‐refuge hypothesis of Amazonian speciation.

Mitochondrial markers revealed genetic panmixia in the data-deficient yellowfin snapper, Lutjanus xanthopinnis (Actinopterygii: Eupercaria: Lutjanidae), from a hotspot of the southern region of the South China Sea

Understanding the genetic structure and diversity of marine fish is crucial for a sustainable management program. We examined the genetic diversity and historical demographics of the yellowfin snapper, Lutjanus xanthopinnis Iwatsuki, Tanaka et Allen, 2015, in the coastal waters of east Peninsular Malaysia which is bordered by the southern part of the South China Sea using the mitochondrial genes (mtDNA) D-loop and Cytochrome b (Cyt-b). A total of 99 (D-loop) and 78 (Cyt-b) specimens of L. xanthopinnis were successfully sequenced from six locations within the range of species distribution along the Malaysian South China Sea. In the presently reported study, the lack of genetic differentiation among populations can be attributed to historical demographic events, eggs and planktonic larvae’ ability to disperse, spawning patterns, and the absence of physical barriers in the geographical landscape. Maximum likelihood gene trees demonstrated that the populations under study had limited structuring and formed a panmictic population that lacks support for internal clades. The AMOVA (Analysis of Molecular Variance) and population pairwise ФST values indicated high genetic exchange between the study areas. A high level of haplotype diversity (D-loop: 0.948–1.000; Cyt-b: 0.542–0.928), low nucleotide diversity (D-loop: 0.0095–0.0159; Cyt-b: 0.0022–0.0049) and starlike haplotype network indicates a recent expansion of L. xanthopinnis populations in Malaysian South China Sea. However, neutrality and goodness of fit tests revealed non-significant values. Furthermore, the BSP (Bayesian skyline plot) analysis estimated population expansion events during the late Pleistocene. During this epoch, the fluctuation in sea level may have led to an increase in the abundance of resources and favorable habitats for the yellowfin snapper. The presently reported findings could initiate efficient management strategies for L. xanthopinnis along the coastal areas of the Malaysian South China Sea and other nearby nations that share the same waterways.

Tracing the origin of the alien pest Cydia pomonella in Algeria through a worldwide comparison of the species’ DNA barcodes

Abstract DNA barcodes (i.e., gene sequences used as identifiers for species identification) constitute a very useful tool in invasive pest research. Using them, we studied, for the first time, the origin and population structure of the alien Cydia pomonella in Africa, a major apple pest. We sequenced a fragment of the mitochondrial gene Cytochrome c oxidase subunit I (COI) from C. pomonella caterpillars collected in northern Algeria and compared them with DNA barcodes from the rest of the world available at public repositories. The phylogeny built upon that COI database supports the European origin of the species: haplotypes at the base of the phylogeny were registered in this continent. In Algeria, the results suggest that this pest could have been introduced from Europe, with two unique African mutations probably favoured by C. pomonella multivoltinism and large population sizes. Population genetic analyses at three Algerian localities showed neither structure nor founder effects. However, to confirm the underlying process of the observed population patterns, it is necessary to perform analyses using genes with higher mutation rates. Compared to higher resolution markers, mitochondrial DNA barcodes are still a cost‐effective tool for taxonomic determination of unknown specimens and, sometimes, may also allow a preliminary tracing of its origin. This is crucial to detect and stop the spread of invasive pests (i.e., borders). We encourage further studies including functional genes to assess whether any mutations are promoting C. pomonella adaptation to the Northern Africa environment.

Phylogeography of the ambrosia beetle Euwallacea interjectus (Coleoptera: Curculionidae: Scolytinae): an emerging poplar pest and its Fusarium mutualists from poplar plantations in China.

Native to Asia, Euwallacea interjectus (Blandford) (Coleoptera: Curculionidae: Scolytinae) is a destructive and invasive pest of live trees, and now it has been found in the United States and Argentina. In recent years, this pest appeared in high densities in poplar monocultures from Eastern China (Jiangsu and Shanghai) and Argentina and caused significant poplar mortality. However, the origin of the pests related to tree damage and the Fusarium mutualists from some poplar zones in China remained unclear. Here, we provided a broader phylogeographic analysis of E. interjectus based on the mitochondrial gene (cytochrome c oxidase I) to determine the global genetic structure of this species. Five mitochondrial lineages were found in the native area. Populations introduced to the United States were originated from 4 localities. The Argentine population was derived from Japan. The species was observed with strikingly high level of cytochrome c oxidase I intraspecific divergence that exceeded interspecific divergence, but the high intraspecific variation was correlated with geographical locations among the native populations. Two nuclear genes (arginine kinase and carbamoyl-phosphate synthetase 2-aspartate transcarbamylase-dihydroorotase) were more conservative, and intraspecific differences were lower than interspecific differences. The mitochondrial genetic variation was probably caused by evolution of lineages among geographically isolated populations. But it is immature to infer the existence of cryptic species based on cytochrome c oxidase I differences. All samples collected from poplar populations were indigenous and formed close relationship with a specimen from eastern and southern China. Surprisingly, pests from poplar populations in Jiangsu and Shanghai showed different haplotypes and mutualists. This suggested that the control strategies should consider the genetic and mutualistic diversity of beetles at different poplar localities.

Assessment of the Genetic Diversity of the Monogenean Gill Parasite Lamellodiscus echeneis (Monogenea) Infecting Wild and Cage-Reared Populations of Sparus aurata (Teleostei) from the Mediterranean Sea.

The diplectanid monogenean Lamellodiscus echeneis (Wagener, 1857) is a specific and common gill parasite of the gilthead seabream Sparus aurata Linnaeus, 1758, in the Mediterranean Sea. Few isolated molecular studies of this monogenean have been conducted, and its population structure and genetic diversity are poorly understood. This study represents the first analysis of the population genetics of L. echeneis, isolated from wild and cage-reared gilthead seabream from fifteen localities in both the Southern (Tunisia) and Northern (Italy and Spain) regions of the Mediterranean Sea, using nuclear ITS rDNA markers and a partial fragment of the mitochondrial gene cytochrome oxidase subunit I (COI). The phylogenetic trees based on the newly obtained dataset and the previously published sequences of L. echeneis corroborated the spread of only a single species throughout the Mediterranean Sea. The star-like haplotypes network, inferred by COI sequences, suggested a recent population expansion of L. echeneis. This is supported by the observed high haplotype diversity (Hd = 0.918) and low nucleotide diversity (Pi = 0.01595). Population structure-based AMOVA for two groups (the Adriatic Sea and the rest of the Mediterranean Sea) attributed 35.39% of the total variation to differences within populations, 16.63% to differences among populations within groups, and 47.99% to differences among groups. Fixation indices were significant, with a high FST value (0.64612), likely related to the divergence of the parasite populations from the Adriatic Sea and other Mediterranean regions. Phylogenetic analyses grouped all samples into the main clade corresponding to L. echeneis from several localities. This study provides insight into the genetic variation between L. echeneis populations, and did not show a clear genetic structure between populations of L. echeneis throughout Tunisian, Italian, and Spanish localities, which can be attributed to the considerable gene flow between the populations favoured by the potential for host dispersion within the Mediterranean Sea. Finally, haplotypes shared between wild and cage-reared hosts provided evidence for the potential for cross-infection between wild and farmed hosts in the Mediterranean Sea.

Specific interaction of resorufin to outer-membrane cytochrome OmcE of Geobacter sulfurreducens: A new insight on artificial electron mediators in promoting extracellular electron transfer

Bioelectrochemical system (BES) is a unique biotechnology for wastewater treatment and energy recovery, and extracellular electron transfer (EET) between microbe and electrode is the key to optimize the performance of BESs. Resazurin is an effective artificial compound that can promote EET in BESs, but the way how it transports electrons is not fully understood. In this study differential pulse voltammetry revealed that the redox potential of resorufin (RR) (intermediate of resazurin reduction, actual electron mediator) within Geobacter sulfurreducens biofilm was positively shifted by 100 mV than that of free RR, and this shift was attenuated by the mutation of outer-membrane cytochrome gene omcE but not by omcS and omcZ mutation, indicating that RR specifically interacted with OmcE. By using heterologously expressed OmcE monomers in Escherichia coli, it was found that RR bonded with OmcE monomers with a moderate intensity (dissociation constant of 720 nM), and their interaction obviously increased the content of α helix in OmcE monomers. Biomolecular analysis indicated that heme II of OmcE monomer might be the binding site for RR (binding energy of -7.01 kJ/mol), which were favorable for electron transfer within OmcE-RR complex. Comparative transcriptomics showed that RZ addition significantly upregulated the expression of omcE, periplasmic cytochrome gene ppcB, and outer-membrane genes omaB, ombB and omcB, thus, it was hypothesized that OmcE-bound RR might serve as potential electron acceptor of OmbB-OmaB-OmcB porin complex which passes electrons across outer membrane. Our work demonstrated a new pathway of artificial electron mediators in facilitating EET in Geobacter species, which may guide the application of electron mediator in improving the performance of BESs.

Under the Sea: Investigation of Telson Morphology and Cryptic Diversity within Eucopia sculpticauda, a Deep-Sea Lophogastrid from the Gulf of Mexico (Peracarida: Lophogastrida).

The field of phylogenetics employs a variety of methods and techniques to study the evolution of life across the planet. Understanding evolutionary relationships is crucial to enriching our understanding of how genes and organisms have evolved throughout time and how they could possibly evolve in the future. Eucopia sculpticauda Faxon, 1893 is a deep-water peracarid in the order Lophogastrida Boas, 1883, which can often be found in high abundances in pelagic trawls. The species can be found along the Mariana Trench, in the Mid-Atlantic Ridge, west Atlantic and east Pacific Oceans, and in the Gulf of Mexico and as deep as 7526 m. Recent collections of E. sculpticauda in the Gulf of Mexico have revealed putative cryptic diversity within the species based on both molecular and morphological evidence. Previous studies have documented two different morphotypes of the telson: the terminal part of the pleon (abdomen) and part of the tail fan. In adults, the morphotypes can be distinguished by lateral constrictions in the telson. This evidence, combined with a previous barcoding study, led to the speculation that telson morphology may be a distinguishing character useful to define cryptic diversity within E. sculpticauda. This study presents additional molecular data from the mitochondrial genes cytochrome c oxidase subunit I, and the large ribosomal subunit (16S), and the nuclear histone 3 gene (H3) to investigate telson morphotypes in relation to evolutionary history within this species. Molecular data identified two strongly supported clades, lending support for potential cryptic diversification within the Gulf of Mexico. Investigations into telson morphology suggest that this character may be informative, but the morphotypes were sometimes ambiguous and additional characters could not be found that discriminate clades. At present, our data suggest early evidence for cryptic diversification within Gulf of Mexico populations, but additional morphological characters and geographic sampling are needed before a new species can be described.

High Expression of AhR and Environmental Pollution as AhR-Linked Ligands Impact on Oncogenic Signaling Pathways in Western Patients with Gastric Cancer-A Pilot Study.

The vast majority of gastric cancer (GC) cases are adenocarcinomas including intestinal and diffuse GC. The incidence of diffuse GC, often associated with poor overall survival, has constantly increased in Western countries. Epidemiological studies have reported increased mortality from GC after occupational exposure to pro-carcinogens that are metabolically activated by cytochrome P450 enzymes through aryl hydrocarbon receptor (AhR). However, little is known about the role of AhR and environmental AhR ligands in diffuse GC as compared to intestinal GC in Western patients. In a cohort of 29, we demonstrated a significant increase in AhR protein and mRNA expression levels in GCs independently of their subtypes and clinical parameters. AhR and RHOA mRNA expression were correlated in diffuse GC. Further, our study aimed to characterize in GC how AhR and the AhR-related genes cytochrome P450 1A1 (CYP1A1) and P450 1B1 (CYP1B1) affect the mRNA expression of a panel of genes involved in cancer development and progression. In diffuse GC, CYP1A1 expression correlated with genes involved in IGF signaling, epithelial-mesenchymal transition (Vimentin), and migration (MMP2). Using the poorly differentiated KATO III epithelial cell line, two well-known AhR pollutant ligands, namely 2-3-7-8 tetrachlorodibenzo-p-dioxin (TCDD) and benzo[a]pyrene (BaP), strongly increased the expression of CYP1A1 and Interleukin1β (IL1B), and to a lesser extend UGT1, NQO1, and AhR Repressor (AhRR). Moreover, the increased expression of CYP1B1 was seen in diffuse GC, and IHC staining indicated that CYP1B1 is mainly expressed in stromal cells. TCDD treatment increased CYP1B1 expression in KATO III cells, although at lower levels as compared to CYP1A1. In intestinal GC, CYP1B1 expression is inversely correlated with several cancer-related genes such as IDO1, a gene involved in the early steps of tryptophan metabolism that contributes to the endogenous AhR ligand kynurenine expression. Altogether, our data provide evidence for a major role of AhR in GC, as an environmental xenobiotic receptor, through different mechanisms and pathways in diffuse and intestinal GC. Our results support the continued efforts to clarify the identity of exogenous AhR ligands in diffuse GC in order to define new therapeutic strategies.

MORPHOLOGICAL AND MOLECULAR CHARACTERIZATION OF Donax faba (BIVALVIA: DONACIDAE) OBTAINED FROM KUTANG BEACH, LAMONGAN, INDONESIA

ARTICLE HIGLIGHTSThis study revealed 19 morphological variations of edible wedge clams, Donax faba (Bivalvia: Donacidae), from Kutang Beach, Lamongan, Indonesia, which has significant role as part of coastal ecosystem.This reseach also highlights the DNA barcoding of Donax faba based on COI gene (Cytochrome C Oxidase Subunit I).The average genetic distance of the research samples was 0.46%, while the value of this parameter between the research samples and the ingroup was found to be 1.51%. ABSTRACTDonax faba is a type of wedge clams with significant ecological and economic roles, as well as variations in color, pattern, and morphometric properties. Recently, a particular population of unidentified D. faba was reported from Kutang Beach, Lamongan, Indonesia. Therefore, this research aimed to assess the morphological variations and molecular characteristics of D. faba obtained from Kutang Beach based on COI gene. A total of 288 samples were collected during the lowest tide for morphological characterization of the color, pattern, and morphometrics of the shells. Additionally, molecular characterization was conducted based on the composition of nucleotide bases and amino acids of COI gene, genetic distance, as well as the relationships. The results showed that D. faba had 19 morphological variations, among which the most dominant type contained a whitish exterior with brown spots and a whitish purple interior. The average genetic distance of the samples was 0.46%, while the value was calculated as 1.51% between the samples and the ingroup. Automatic Barcode Gap Discovery (ABGD) analysis performed using a prior maximal distance of 0.001 showed the separation of these species into distinct categories.

One Genome, Multiple Phenotypes: Would Rhodnius milesi Carcavallo, Rocha, Galvão & Jurberg, 2001 (Hemiptera, Triatominae) Be a Valid Species or a Phenotypic Polymorphism of R. neglectus Lent, 1954?

Species of the Rhodnius genus have a complex taxonomy because the events of phenotypic plasticity and cryptic speciation make it difficult to correctly classify these vectors. During the taxonomic history of the genus, five synonymization events occurred. Additionally, some authors suggest that R. milesi possibly represent only phenotypic polymorphisms of R. neglectus. Thus, we analyzed the specific status of R. milesi in relation to R. neglectus using phylogenetic studies with the mitochondrial gene cytochrome B and the study of reproductive barriers. The phylogenetic reconstruction grouped R. milesi together with R. neglectus from different localities, demonstrating that these taxa represent the same species based on the phylogenetic species concept. Experimental crosses demonstrate the absence of pre- and postzygotic barriers under laboratory conditions. Additionally, when the hatch rates of crosses are compared to intraspecific crosses, it can be noted that they are high and very similar. Finally, the mortality rate of the hybrids does not indicate hybrid inviability, the absence of chromosome pairing errors does not indicate hybrid sterility, and the proportion between male and female hybrids demonstrates that Haldane’s rule was not acting. Therefore, we perform the formal synonymization of R. milesi with R. neglectus.

Genetic diversity of the Ruditapes decussatus and evidence of its hybridization with the alien R. philippinarum in the Western Mediterranean Sea

The introduction of alien species in marine ecosystems is often driven by the increasing demand of fishery resources. This is the case of the Manila clam (R. philippinarum), imported in Europe from Japan since the 1970s, to meet the growing demand for clams that the native species, the grooved carpet shell clam (Ruditapes decussatus), could not satisfy. Alien species introduction could threaten the genetic diversity and integrity of the native clam, also causing hybridization (i.e., gene flow from one species into the gene pool of another). Since R. philippinarum recently spread in a few important Mediterranean coastal areas, a combined approach based on morphological characteristics, length differences of two nuclear species-specific markers (ITS2, 5SrDNA) and the sequence of the mitochondrial gene cytochrome c oxidase subunit I (COI), was used to investigate the presence of hybrids in six Mediterranean wetlands (Sardinia, Italy). Eight individuals morphologically identified as R. decussatus were hybrids, having sequences specific to both R. decussatus and R. philippinarum in their nuclear DNA (ITS2 and 5SrDNA). Most of these individuals were found to be post-first generation (F1) hybrids indicating that F1-hybrids may be fertile. Secondly, to study the genetic diversity of R. decussatus in the Sardinian wetlands as well as in its whole distribution area, >380 new COI sequences from the eastern Atlantic Ocean and Mediterranean Sea were analysed along with those available from public databases. Mitochondrial COI data revealed variable haplotype and nucleotide diversities in different areas, which were not dependent on sample sizes. The aquaculture breeding activities and clam transplantation between different countries, along with the long pelagic larval dispersal and the commercial import of other bivalve species might have promoted gene exchange between different sites and thus higher diversity levels in a few wild populations. Our research, evaluating the genetic makeup of wild and hatchery stocks and clarifying the degree of hybridization, can contribute to develop further recommendations for conserving the genetic integrity of R. decussatus.

Integrating Genetic and Cytogenetic Data: A Diversity Study of Astyanax and Psalidodon (Characidae) Species from the Paraná River Basin.

Astyanax is one of the most specious fish groups in the Neotropical region, with many cryptic species, which represents a challenge for correct identification through traditional taxonomic methods. Psalidodon is a recently resurrected genus group of species previously belonging to Astyanax, specifically those with extensive chromosomal variation of the A. scabripinnis and fasciatus complexes. In the present study, the mitochondrial genes cytochrome c oxidase subunit 1 (COI), mitochondrial ATP synthase 6 and 8 (ATPase 6/8), and NADH dehydrogenase subunit 2 (ND2) were used in conjunction with chromosomal data to characterize molecularly and cytogenetically populations of Astyanax and Psalidodon from rivers and streams of the Ivaí River Basin (Paraná Basin). The results demonstrated the effectiveness of the integrative use of molecular and cytogenetic techniques, with the confirmation of at least three species for the sampled sites: A. lacustris, P. paranae, and P. fasciatus, which showed inter- and intrapopulation karyotype variations. In addition, extensive haplotypic variation can be observed for these species within the Ivaí River Basin and throughout the Paraná River Basin. The data demonstrate a hidden diversity among the species analyzed, enrich the ichthyofaunistic knowledge of small rivers and streams, and contribute to future conservation projects in these areas.

Applicability and perspectives for DNA barcoding of soil invertebrates

Belowground invertebrate communities are dominated by species-rich and very small microarthropods that require long handling times and high taxonomic expertise for species determination. Molecular based methods like metabarcoding circumvent the morphological determination process by assigning taxa bioinformatically based on sequence information. The potential to analyse diverse and cryptic communities in short time at high taxonomic resolution is promising. However, metabarcoding studies revealed that taxonomic assignment below family-level in Collembola (Hexapoda) and Oribatida (Acariformes) is difficult and often fails. These are the most abundant and species-rich soil-living microarthropods, and the application of molecular-based, automated species determination would be most beneficial in these taxa. In this study, we analysed the presence of a barcoding gap in the standard barcoding gene cytochrome oxidase I (COI) in Collembola and Oribatida. The barcoding gap describes a significant difference between intra- and interspecific genetic distances among taxa and is essential for bioinformatic taxa assignment. We collected COI sequences of Collembola and Oribatida from BOLD and NCBI and focused on species with a wide geographic sampling to capture the range of their intraspecific variance. Our results show that intra- and interspecific genetic distances in COI overlapped in most species, impeding accurate assignment. When a barcoding gap was present, it exceeded the standard threshold of 3% intraspecific distances and also differed between species. Automatic specimen assignments also showed that most species comprised of multiple genetic lineages that caused ambiguous taxon assignments in distance-based methods. Character-based taxonomic assignment using phylogenetic trees and monophyletic clades as criteria worked for some species of Oribatida but failed completely for Collembola. Notably, parthenogenetic species showed lower genetic variance in COI and more accurate species assignment than sexual species. The different patterns in genetic diversity among species suggest that the different degrees of genetic variance result from deep evolutionary distances. This indicates that a single genetic threshold, or a single standard gene, will probably not be sufficient for the molecular species identification of many Collembola and Oribatida taxa. Our results also show that haplotype diversity in some of the investigated taxa was not even nearly covered, but coverage was better for Collembola than for Oribatida. Additional use of secondary barcoding genes and long-read sequencing of marker genes can improve metabarcoding studies. We also recommend the construction of pan-genomes and pan-barcodes of species lacking a barcoding gap. This will allow both to identify species boundaries, and to cover the full range of variability in the marker genes, making molecular identification also possible for species with highly diverse barcode sequences.

A typical coastal plain taxon in central Brazil: Relationships and description of a new species of non-annual killifish (Cyprinodontiformes; Rivulidae) from the Paraná River basin

We describe a new species of the genus Atlantirivulus from a small first-order stream on the right margin of the Paraná River basin in the Mato Grosso do Sul, Brazil. The region of occurrence of the new species is known to be a hotspot for Melanorivulus species. This area, however, is approximately 700 km away from the record of occurrence of its closest congeners, which are distributed along the Brazilian coast in different hydrographic basins. Atlantirivulus enigmaticus, new species, is distinguished from all congeners by it s unique coloration, which consists of the flank being purplish grey with light green iridescence with brownish red dots scattered along of the body, by the neuromasts of the infraorbital series being aligned around eye, and by the presence of the short ventral process of the posttemporal. The phylogenetic position of the new species within the Atlantirivulus genus was based on molecular phylogenetic analysis using sequences of the mitochondrial gene cytochrome oxidase subunit 1. The results show that the new species is closely related to Atlantirivulus luelingi from the coastal drainages of the Brazilian state Santa Catarina. The phylogenetic position of the new species among congeners endemic to coastal river basins may be related to ancestral connections between coastal drainages and the Paraná River basin. The new species constitutes the westernmost record for the genus, and the first record of Atlantirivulus in the Paraná River basin. Insufficient collection efforts in areas of potential occurrence for the group and extinction processes in the broad region between the locality of the new species and the distribution areas of the remaining species may be a possible explanation for this disjunct distribution.

Sargassum horneri extract fermented by Lactiplantibacillus pentosus SH803 mediates adipocyte metabolism in 3T3-L1 preadipocytes by regulating oxidative damage and inflammation

Sargassum horneri (S. horneri), a brown seaweed excessively proliferating along Asian coastlines, are damaging marine ecosystems. Thus, this study aimed to enhance nutritional value of S. horneri through lactic acid bacteria fermentation to increase S. horneri utilization as a functional food supplement, and consequently resolve coastal S. horneri accumulation. S. horneri supplemented fermentation was most effective with Lactiplantibacillus pentosus SH803, thus this product (F-SHWE) was used for further in vitro studies. F-SHWE normalized expressions of oxidative stress related genes NF-κB, p53, BAX, cytochrome C, caspase 9, and caspase 3, while non-fermented S. horneri (SHWE) did not, in a H2O2-induced HT-29 cell model. Moreover, in an LPS-induced HT-29 cell model, F-SHWE repaired expressions of inflammation marker genes ZO1, IL1β, IFNγ more effectively than SHWE. For further functional assessment, F-SHWE was also treated in 3T3-L1 adipocytes. As a result, F-SHWE decreased lipid accumulation, along with gene expression of adipogenesis markers PPARγ, C/EBPα, C/EBPβ, aP2, and Lpl; lipogenesis markers Lep, Akt, SREBP1, Acc, Fas; inflammation markers IFN-γ and NF-κB. Notably, gene expression of C/EBPβ, IFN-γ and NF-κB were suppressed only by F-SHWE, suggesting the enhancing effect of fermentation on obesity-related properties. Compositional analysis attributed the protective effects of F-SHWE to acetate, an organic acid significantly higher in F-SHWE than SHWE. Therefore, F-SHWE is a novel potential anti-obesity agent, providing a strategy to reduce excess S. horneri populations along marine ecosystems.

Patterns of Genetic Diversity and Gene Flow Associated With an Aridity Gradient in Populations of Common Mole-rats, Cryptomys hottentotus hottentotus.

Genetic adaptation is the change of a population toward a phenotype that best fits the present ecological conditions of the environment it inhabits. As environmental conditions change, allele frequencies shift, resulting in different populations of the same species possessing genetic variation and divergent phenotypes. Cooperatively breeding common mole-rats (Cryptomys hottentotus hottentotus) inhabit environments along an aridity gradient in South Africa, which provides an opportunity for local genetic adaptations to occur. Using one mitochondrial gene (cytochrome b) and 3,540 SNP loci across the whole genome, we determined the phylogenetic relationship, population structure and genetic diversity of five populations of C. h. hottentotus located along an aridity gradient. Mitochondrial data identified population-specific clades that were less distinct in the two mesic populations, potentially indicating historical or recent gene flow, or the retention of ancestral haplotypes. Arid and semi-arid populations formed a distinct cluster from the non-arid populations. Genetic diversity and gene flow were higher in arid-dwelling individuals, suggesting greater connectivity and interactions between colonies in arid regions in comparison to mesic ones. Using an Aridity Index, we determined that isolation by environment, rather than isolation by geographical distance, best explains the genetic distance between the populations. Further analyses using target loci may determine if there are differing underlying genetic adaptations among populations of C. h. hottentotus. These analyses could help unravel population differences in response to environmental factors within a subspecies of bathyergid mole-rat and determine the adaptive capacity of this small nonmigratory subterranean rodent species in response to aridification in the face of climate change.

A new species of Homalernis Meyrick, 1908 (Lepidoptera, Tortricidae, Tortricinae) represents the first record of the tribe Schoenotenini in Japan

Homalernis fluctuosa Suzuki & Jinbo, sp. nov., is described and illustrated from Honshu, Shikoku, Kyushu, Tsushima Island, Amamiooshima Island, and Okinawajima Island, Japan. This is not only the first record of the genus Homalernis but also of the tribe Schoenotenini from warm temperate zones in the Palaearctic region. The association of males and females of the new species was confirmed based on the mitochondrial gene cytochrome oxidase submit 1 (COI). We discuss the taxonomic positions of two alleged Homalernis species from Malaysia and the taxonomic position of Homalernis within Schoenotenini.

Inoculum source determines the stress resistance of electroactive functional taxa in biofilms: A metagenomic perspective

The inoculum has a crucial impact on bioreactor initialization and performance. However, there is currently a lack of guidance on selecting appropriate inocula for applications in environmental biotechnology. In this study, we applied microbial electrolysis cells (MECs) as models to investigate the differences in the functional potential of electroactive microorganisms (EAMs) within anodic biofilms developed from four different inocula (natural or artificial), using shotgun metagenomic techniques. We specifically focused on extracellular electron transfer (EET) function and stress resistance, which affect the performance and stability of MECs. Community profiling revealed that the family Geobacteraceae was the key EAM taxon in all biofilms, with Geobacter as the dominant genus. The c-type cytochrome gene imcH showed universal importance for Geobacteraceae EET and was utilized as a marker gene to evaluate the EET potential of EAMs. Additionally, stress response functional genes were used to assess the stress resistance potential of Geobacter species. Comparative analysis of imcH gene abundance revealed that EAMs with comparable overall EET potential could be enriched from artificial and natural inocula (P > 0.05). However, quantification of stress response gene copy numbers in the genomes demonstrated that EAMs originating from natural inocula possessed superior stress resistance potential (196 vs. 163). Overall, this study provides novel perspectives on the inoculum effect in bioreactors and offers theoretical guidance for selecting inoculum in environmental engineering applications.

The Complete Mitochondrial Genome Sequence of Eimeria kongi (Apicomplexa: Coccidia).

Rabbit coccidiosis is caused by infection with one or, more commonly, several Eimeria species that parasitize the hepatobiliary ducts or intestinal epithelium of rabbits. Currently, there are eleven internationally recognized species of rabbit coccidia, with the complete mitochondrial (mt) genomes of six species commonly infecting rabbits having been sequenced and annotated. Eimeria kongi was initially discovered in 2011 and prompted a preliminary study on this species. Through traditional morphological analysis, E. kongi was identified as a novel species of rabbit coccidia. To further validate this classification, we sequenced and annotated its mitochondrial genome. The complete mt genome of E. kongi spans 6258 bp and comprises three cytochrome genes (cytb, cox1, cox3), fourteen gene fragments for the large subunit (LSU) rRNA, and nine gene fragments for the small subunit (SSU) rRNA, lacking transfer RNA (tRNA) genes. Moreover, phylogenetic analysis of the mitochondrial genome sequence of E. kongi revealed its clustering with six other species of rabbit coccidia into a monophyletic group. Additionally, E. irresidua and E. flavescens were grouped within the lineage lacking oocyst residuum, consistent with their morphological characteristics. Consistent with multiple molecular phylogenies, in this investigation, E. kongi was further confirmed as a new species of rabbit coccidia. Our research findings are of great significance for the classification of coccidia and for coccidiosis prevention and control in rabbits.